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Research ArticleChemical Bioactive and Antioxidant Potential ofTwenty Wild Culinary Mushroom Species
S K Sharma1 and N Gautam2
1Department of Plant Pathology CSK Himachal Pradesh Agriculture University Palampur 176 062 India2Centre for Environmental Science and Technology School of Environment and Earth Sciences Central University of PunjabBathinda 151 001 India
Correspondence should be addressed to N Gautam ngautam86gmailcom
Received 8 May 2015 Accepted 11 June 2015
Academic Editor Miroslav Pohanka
Copyright copy 2015 S K Sharma and N GautamThis is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in anymedium provided the originalwork is properly cited
The chemical bioactive and antioxidant potential of twenty wild culinary mushroom species being consumed by the people ofnorthern Himalayan regions has been evaluated for the first time in the present study Nutrients analyzed include protein crudefat fibres carbohydrates and monosaccharides Besides preliminary study on the detection of toxic compounds was done onthese species Bioactive compounds evaluated are fatty acids amino acids tocopherol content carotenoids (120573-carotene lycopene)flavonoids ascorbic acid and anthocyanidins Fruitbodies extract of all the species was tested for different types of antioxidantassays Although differences were observed in the net values of individual species all the species were found to be rich in proteinand carbohydrates and low in fat Glucose was found to be the major monosaccharide Predominance of UFA (65ndash70) over SFA(30ndash35) was observed in all the species with considerable amounts of other bioactive compounds All the species showed highereffectiveness for antioxidant capacities
1 Introduction
Wild mushrooms have long been considered as highlynutritious tasty food items from ancient time [1ndash3] Besidesnutritional importance wild edible mushrooms are now wellknown for their pharmaceutical constituents [4 5] Presentlythere are several mushroom species which have establishedtherapeutic properties [6ndash11] In addition mushroom extractis considered as important remedies for the prevention andtreatment of many diseases for thousands of years in sev-eral parts of the world [12 13] Mushrooms are known tocontain immunomodulating compounds which help toimprove immune function in cancer patients during radioand chemotherapy and help to prolong survival times insome types of cancer [14] Another aspect of mushrooms islowering blood pressure and free cholesterol in plasma [15]Major bioactive compounds extracted from mushroomsare well known for their antioxidant [16] antitumor andantimicrobial properties [17] The nutritive nutraceuticalspresent in mushrooms are dietary fibres polyunsaturatedfatty acids (PUFA) proteins amino acids keto acids min-erals antioxidative vitamins and other antioxidants [18ndash20]
Currently 14000 mushroom species are known to existOut of these about 50 species are reported to possessvarying degrees of edibility and almost 3000 species spreadover 31 genera are regarded as prime edible mushroomsTo date only 200 of them are experimentally grown 100of them are economically cultivated approximately 60 arecommercially cultivated and about 10 have reached industrialscale production in many countries [21] Furthermore about2000 aremedicinalmushroomswith variety of health benefitsand 270 species are now considered as potential therapeuticor preventative agents that are ensured for human healthperspective The poisonous mushrooms are relatively smallin number (approximately 1) but there is an estimate thatabout 10may have poisonous attributes while 30 species areconsidered to be lethal [21]
The northern Himalayan regions of India include thestates of Himachal Pradesh (30∘221015840 to 33∘121015840N latitudeand 75∘451015840 to 79∘041015840E longitude) Uttarakhand (28∘431015840N to31∘281015840N latitude and 77∘341015840E to 81∘031015840E longitude) and someparts of Jammu and Kashmir (34∘81015840N and 77∘341015840E) Theregions in these states have extensive areas under forest
Hindawi Publishing CorporationBioMed Research InternationalVolume 2015 Article ID 346508 12 pageshttpdxdoiorg1011552015346508
2 BioMed Research International
cover (ie more than 50) The people of these areas usedifferent mushroom species for culinary purposes Theyhave very little idea about the medicinal importance ofthese mushroom species Their methods of identification areprimarily based upon sweet smell and color Further theknowledge about the edible species is restricted to the oldaged people Documentations studies of these edible specieshave been done by several workers [22] but the studies onchemical composition and medicinal importance are stilllacking besides the presence of toxic compounds in them Inview of this under present investigations twenty of such wildculinary mushroom species have been evaluated for theirchemical bioactive and antioxidant potential
2 Materials and Methods
21 Collection andProcessing of Samples All the sampleswerecollected during the frequent surveys to the different regionsof northern Himalayas (Table 1) After this the samples werevacuum-dried and preserved in air-tight cellophane bagswith a small amount of 1-4-paradichlorobenzene in porouspackets to keep them free of insects for further analysis
22 Chemical Evaluation Samples were powdered and eval-uated for protein fat carbohydrates ash and crude fibresCrude protein content was estimated using the Kjeldahlmethod by calculating total nitrogen (N) and protein contentwas expressed by N times 438 [23] Crude fat was estimatedusing a Soxhlet apparatus by extraction of known weightof powered samples with petroleum ether Ash content wascalculated by incineration in silica dishes at 525 plusmn 20∘C con-taining 5ndash10 gsample Fibres content was estimated on a fat-free sample using the acid-alkali method (125 each) Totalcarbohydrates percentage was calculated by the difference asthe totalweightminus (moisture content +protein content + crudefat + ash content + crude fibres)
For toxic metal detection diluted HCl (2) and copperfoil (1 times 12 cm strip purified or pretreated with concentratedHNO
3or diluted 3 HCl and rinsed in distilled water) were
taken After that samples (powered samples) were acidifiedwith 10ndash20mL of 5 diluted HCl (2) until colour changedfrom fairy pink to litmus Then strips of copper foil wereadded and boiled for 30 minutes with addition of waterfrom time to time to replace the losses by evaporation Theheavy metals got deposited on the copper foil and gavecharacteristics color to it The color deposited on the Cu foilwas noted after 30 minutes and results were interpreted forthe presence of heavy metals [24]
For monosaccharide composition samples (01 g) wereextracted with 25mL and 15mL and finally with 1mL of 70aqueous methanol After this the extract was centrifuged at4000 rpm (4∘C) for 10min Supernatant was collected andvolumewasmade up to 5mLwith 70methanolThe extractwas passed through Millipore filter (045 120583m) and injected tothe HPLC [24]
23 Bioactive Evaluation231 Fatty Acid Composition Powdered samples were dis-solved in 1mL of solution (prepared by sodium hydroxide
Table 1 List of wild culinary species collected from differentlocalities of twenty species from northern Himalayas
pellets (45 g) in 300mL of 50 methanol and vortexing for1 minute then the solution was left for 5 minutes at 100∘Cvortexed again for 1 minute and left at 100∘C in a waterbath for 25 minutes) Methylation was done by adding 2mLof solution (6N hydrogen chloride in methanol) and thenvortexed for 1 minute followed by heating (80∘C) in a waterbath For extraction of fatty acids 125mL of solution (25mLmethyl ter-butyl ether added to hexane) was added and thesolution was shaken for 10 minutes followed by removal ofupper layer and addition of 3mL of solution (10 sodiumhydroxide in water while stirring) Finally the top phase(23) was removed and transferred into a gas chromatographyvial and injected Unsaturation index (UI) was calculated as(mol of each (poly)unsaturated fatty acid times number doublebonds per each fatty acid)100
BioMed Research International 3
232 Amino Acids Powdered samples (01 g) were extractedwith 25mL followed by 15mL and 1mL of 70 aqueousmethanol After this centrifugation was done for 10 minutes(4000 rpm) at 4∘C Supernatants were dissolved in aque-ous methanol and the volume was made up to 5mL Itwas now passed through Millipore filter (045 120583m) Afterthis samples (10 120583L) were dried using vacuum oven andto these dried samples 20 120583L derivatising agent (preparedby ethanol triethylamine water phenylisothiocaynate) wasmixed with it and redried it Now the samples were left for 25minutes at room temperature Lastly 1mL ethanol was addedand injected into UPLC (Waters India Pvt Ltd)
233 Tocopherol Composition Tocopherol composition wasestimated following standard method [25] For this sam-ples were mixed with butylated hydroxytoluene (BHT) inhexane (10mgmL 100 120583120583L) and IS solution in hexane (120575tocopherol 16 120583gmL 250 120583L) Thereafter samples (500mg)were vortexed for 1min with methanol (4mL)Then sampleswere again vortexed with hexane (4mL) After this 2mL ofsaturated NaCl aqueous solution was added and the mixturewas vortexed (1min) followed by centrifugation at 4000 gfor 5min and the upper layer was separated The sampleswere again reextracted twice with hexane The extractswere then vacuum-dried and redissolved in hexane (1mL)followed by dehydration with anhydrous sodium sulphatethen filtered and transferred into a dark injection vial andanalysed byHPLC (Waters India Pvt Ltd) Chromatographiccomparisons were made by authentic standards Tocopherolcontents in mushroom samples were expressed in 120583g per g ofdry mushroom
234 Evaluation of Other Bioactive Compounds For 120573-carotene and lycopene estimation dried powdered samples(sim5 g) were extracted with 100mL of methanol at 25∘C(150 rpm) for 24 hours and filtered through Whatman Num-ber 4 filter paper The residue was again reextracted with2 additional 100mL portions of methanol These extractswere evaporated to dryness at 42∘C then redissolved inmethanol at a concentration of 50mgmL and stored at 4∘CThe driedmethanolic extract (100mg) was shaken vigorouslywith 10mL of acetonehexanemixture (4 6) for 1 minute andfiltered The absorbance of the filtrate was measured at 453505 and 663 nm [26] 120573-carotene and lycopene content wereestimated using the following equation
Lycopene (mg100mL)
= (00458times119860663) + (0372times119860505)
minus (00806times119860453) 120573-carotene (mg100mL)
= (0216times119860663) minus (0304times119860505)
+ (0452times119860453)
(1)
For phenolic compounds quantification powdered sam-ples (1mL) were mixed with Folin and Ciocalteursquos phenolreagent (1mL) After 3 minutes 1mL of saturated sodiumcarbonate solution was added to the mixture and the volume
was adjusted to 10mL with distilled water The reaction waskept in the dark for 90 minutes after which the absorbancewas read at 725 nm Gallic acid was used to calculate thestandard curve (001ndash04mM 1198772 = 09999) and the resultswere expressed as milligrams of gallic acid equivalents pergram of extract [27]
Total flavonoids of the sample extracts were measuredby AlCl
3method [28] For this an aqueous extract (15mL)
was mixed with deionized distilled water (5mL) and 03mLof 5 NaNO
2 After five minutes of incubation at room
temperature 15mL of 2 aluminium trichloride (AlCl3)
solution was added After the next 6 minutes 2mL of 1MNaOH was added The mixture was vigorously shaken onorbital shaker for 5min at 200 rpm and the absorbance wasread at 510 nm against a blank Quercetin with differentconcentrations was used as a standard
For ascorbic acid quantification standard ascorbic acidsolution (5mL L-ascorbic acid in 3 phosphoric acid) wasadded to 5mL of phosphoric acid A microburette was filledwith dye and the samples were titrated with the dye solutionto a pink color which persisted for 15 seconds The dyefactor (milligrams of ascorbic acid per milliliter of dye usingformula 05titrate) was determined A sample was preparedby taking 10 g of sample grounded in metaphosphoric acidand the volume was increased up to 100mL It was titratedafter filtration until a pink color appeared [24] The amountof ascorbic acid was calculated with the use of the followingequation
mg of ascorbic acid per100 g or mL
=Titrate times Dye factor times Vol madeAliquot of extract times wt of sample
times 100(2)
Anthocyanidins were quantified by using standardmethod [29] Briefly 05 g of samples was mixed withthe solvent (mixture of 85 15 (vv) of ethyl alcohol andhydrochloric acid 15M) followed by ultrasonication for 15minutes and filtration throughWhatman filter paper number1 Standard solution was prepared with cyaniding chloridewith a concentration of 5ndash15120583gmL in solvent which wasused The absorption was measured at 546 nm The totalquantity of anthocyanins (expressed in g of cyanidingchloride100 g extract) = (119860
119901times 119898st times 119891 times 100)(119860 st times 119898119901)
where 119860119901is absorption rate of the sample solution 119898
119901is
mass of the processed sample in g 119860 st is absorption rate ofthe standard solution 119898st is mass of the processed standardsolution in g and 119891 is dilution coefficient
24 Antioxidant Assays DPPH scavenging activity was mea-sured with adding DPPH (200120583L) solution at differentconcentrations (2ndash10mgmL) to 005mL of the samples(dissolved in ethanol) An equal amount of ethanol was addedto the control Ascorbic acid was used as the control [30]Theabsorbancewas read after 20min at 517 nm and the inhibitionwas calculated using the formula
where 1198600was the absorbance of the control and 119860
119875was the
absorbance in the presence of the sampleFor ABTS radical scavenging activity 10120583L of the sample
was added to 4mL of the diluted ABTS∙+ solution (preparedby adding 7mM of the ABTS stock solution to 245mMpotassium persulfate kept in the dark at room temperaturefor 12ndash16 h before use) The solution was then diluted with5mM phosphate-buffered saline (pH 74) and absorbancewas measured at 730 nm after 30min [31] The ABTS radicalscavenging activity was calculated as
119878 = (119860controlminus119860 sample
119860control)times 100 (4)
For reducing power estimation samples (200 120583L) weremixed with sodium phosphate buffer (pH 66) 1mM FeSO
4
and 1 potassium ferricyanide and incubated for 20minat 50∘C after that trichloroacetic acid was added and themixtures were centrifuged Supernatant (25mL) was mixedwith an equal volume of water and 05mL 01 FeCl
3 The
absorbance was measured at 700 nm [32]For Fe2+ chelating activity 1mL of the sample (2ndash10mg
mL) was mixed with 37mL of ultrapure water after thatthe mixture was reacted with ferrous chloride (2mmolL01mL) and ferrozine (5mmolL 02mL) for 20min and theabsorbance was read at 562 nm with using EDTA as controlThe chelating activity was calculated using the formula
where 119860119887is the absorbance of the blank and 119860
119904is the
absorbance in the presence of the extract [33]The scavenging activity of superoxide anion radicals
was measured following standard method [34] Samples (0ndash20mgmL 1mL) and Tris-HCl buffer (500mM pH 823mL) were incubated in a water bath at 25∘C for 20min andafter this pyrogallic acid (50mM 04mL) was added HClsolution (80M 01mL) was added to terminate the reactionafter 4min The absorbance of the mixture was measuredat 320 nm The scavenging ability was calculated using thefollowing formula
scavenging ability () = (1 minus119860 sample
119860control)times 100 (6)
where 119860control is the absorbance of control without thepolysaccharide sample and 119860 sample is the absorbance in thepresence of the polysaccharide sample
For ferric reducing antioxidant power (FRAP) assayfirstly FRAP reagent was prepared by mixing TPTZ (25mL10mM in 40mM HCl) 25mL of 300mM acetate buffer and25mL of FeCl
3sdot6H2O After this freshly prepared FRAP
reagent (18mL) was taken in a test tube and incubated at30∘C in water bath for 10 minutes Then absorbance wasread at 0min (119905
0) After this 100 120583L of sample extract or
standard and 100 120583L of distilled water were added to the testtube mixed and incubated at 30∘C for 30 minutes Thenthe absorbance was taken at 593 nm (119905
30) Ferrous sulphate
was used as standard [35 36] FRAP activity was determinedagainst a standard curve of ferrous sulphate and the valueswere expressed as 120583M Fe2+ equivalents per gram of extractand calculated using the following equation
FRAP value = Absorbance (sample+ FRAP reagent)
minusAbsorbance (FRAP reagent) (7)
25 Statistical Analysis All experiments were performed 3times and with 3 replicates The results were analyzed usingone-way analysis of variance (ANOVA) 119901 lt 005 was con-sidered significant and SPSS software (SPSS Inc ChicagoIL USA) was used to calculate differences Tukey-HSD at119901 lt 005 test was used to determine significant differences
3 Results
31 Chemical Evaluation Nutrient composition of the wildculinary mushrooms is shown in Table 2 Protein was foundin high levels and varied between 516 in Inocybe splendensand 2263 in Agaricus arvensis Protein percentage inPleurotus cystidiosus (2069) Amanita caesarea (1972)Agaricus campestris (1838) Cantharellus cibarius (1819)and Lentinus cladopus (1859) was also found to be highAll the twenty culinary species were found to be low in fatcontent Fat ranged from010 in Laccaria laccata to 038 inInocybe splendens In general these wild culinary mushroomsconsumed by local people were found to be higher in proteinand low in fat although differences were observed in netvalue of individual species Crude fibres ranged from 108in Hygrocybe coccinea to 242 in Lentinus cladopus Ashcontent varied between 011 in Inocybe splendens and 096in Agaricus arvensis Carbohydrates calculated by differencewere also found to be in abundant amounts and theirpercentage was ranged from 3119 in Inocybe splendens to5712 in Agaricus arvensis Nutrient contents of Inocybesplendens Hygrocybe nivea and Conocybe tenera were foundto be less as compared to other species
There are several reports about the toxicity reports due tomushrooms on humans in these areas and hence preliminarystudies were done to check the toxicity level of mushroomsFor all the twenty species being used by the people forculinary purposes the test was found to be negative Thatmeans these species are nontoxic and hence recommendedfor consumption
All the culinarymushroom species contained glucose andrhamnose as the principal carbohydrates (Table 3) Neverthe-less the present study also describes the presence of xyloseand mannose in all the studied species However galactoseand fructose were detected in very low percentage in someof the species Russula mairei contained lowest percentageof glucose (2160) and Agaricus arvensis contained highestpercentage of glucose (6412)
32 Bioactive Evaluation The results of fatty acid composi-tion (total saturated fatty acids SFA monounsaturated fattyacids MUFA and polyunsaturated fatty acids PUFA) of allthe species are shown in Table 4 In general the major fatty
BioMed Research International 5
Table 2 Percent chemical composition of twenty wild culinary mushroom species collected from northern Himalayan regions
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 3 Percent monosaccharide composition of twenty species collected from northern Himalayan regions showing richness in glucoserhamnose mannose xylose and galactose and fructose in lower percentage
Russula lepida 515 plusmn 01d 3261 plusmn 36f 1316 plusmn 12e 231 plusmn 01c ND NDR mairei 426 plusmn 01c 2160 plusmn 21a 1281 plusmn 16d 212 plusmn 00c ND NDND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
6 BioMed Research International
Table4Percentfattyacid
compo
sitionof
allthe
twentywild
culin
arymushroo
mspeciesc
ollected
from
different
region
sofn
orthernHim
alayas
Species
C900
C100
C120
C160
C1611
C171
C181
C182
C202
UI
Agaricu
sarvensis
006plusmn00a
030plusmn001
a028plusmn00a
186plusmn003
a006plusmn00a
009plusmn00a
018plusmn00a
324plusmn00a
221plusmn02a
057plusmn002
a
Acampestr
is003plusmn00a
026plusmn002
a021plusmn00a
119plusmn002
a009plusmn00a
002plusmn00a
035plusmn00a
204plusmn00a
218plusmn03a
046plusmn002
a
Acomtulus
006plusmn00a
022plusmn00a
ND
117plusmn001
a007plusmn00a
009plusmn00a
038plusmn00a
173plusmn00a
134plusmn01a
036plusmn001
a
Asilvicola
008plusmn00a
033plusmn00a
022plusmn00a
156plusmn002
a009plusmn00a
001plusmn00a
015plusmn00a
154plusmn00a
221plusmn03a
040plusmn001
a
Amanita
caesarea
004plusmn00a
032plusmn002
a019plusmn00a
196plusmn002
a012plusmn00a
005plusmn00a
031plusmn00a
219plusmn00a
128plusmn00a
039plusmn001
a
Acitrin
a006plusmn00a
028plusmn001
aND
091plusmn00a
011plusmn00a
008plusmn00a
031plusmn00a
159plusmn00a
168plusmn00a
037plusmn001
a
Afulva
004plusmn00a
021plusmn001
a025plusmn002
a010plusmn00a
008plusmn00a
004plusmn00a
015plusmn00a
164plusmn00a
123plusmn00a
031plusmn002
a
Cantharellu
scibarius
007plusmn00a
033plusmn002
a022plusmn001
a110plusmn002
a007plusmn00a
001plusmn00a
037plusmn00a
294plusmn00a
118plusmn01a
045plusmn002
a
Conocybe
tenera
006plusmn00a
020plusmn00a
ND
007plusmn00a
005plusmn00a
002plusmn00a
035plusmn00a
014plusmn00a
138plusmn00a
019plusmn00a
Gymnopilusjun
onius
005plusmn00a
031plusmn001
a014plusmn001
a18
7plusmn003
a008plusmn00a
002plusmn00a
088plusmn00a
049plusmn00a
121plusmn
01a
026plusmn001
a
Hygrocybe
coccinea
004plusmn00a
025plusmn002
a017plusmn00a
033plusmn00a
001plusmn00a
005plusmn00a
017plusmn00a
024plusmn00a
119plusmn00a
016plusmn00a
Hnivea
007plusmn00a
022plusmn001
aND
005plusmn00a
004plusmn00a
001plusmn00a
053plusmn00a
004plusmn00a
138plusmn02a
020plusmn00a
Inocybesplendens
002plusmn00a
019plusmn001
a023plusmn001
a089plusmn00a
002plusmn00a
003plusmn00a
018plusmn00a
010plusmn00a
020plusmn00a
005plusmn00a
Lactariusp
ubescens
001plusmn00a
023plusmn001
a016plusmn00a
075plusmn00a
007plusmn00a
008plusmn00a
026plusmn00a
344plusmn00a
083plusmn00a
046plusmn002
a
Laccarialaccata
002plusmn00a
022plusmn00a
ND
101plusmn
00a
006plusmn00a
009plusmn00a
092plusmn00a
284plusmn00a
029plusmn00a
042plusmn001
a
Lepista
nuda
003plusmn00a
024plusmn00a
ND
043plusmn00a
001plusmn00a
002plusmn00a
020plusmn00a
204plusmn00a
037plusmn00a
026plusmn00a
Lentinus
cladopu
s009plusmn00a
033plusmn002
a032plusmn00a
146plusmn00a
055plusmn00a
001plusmn00a
032plusmn00a
245plusmn00a
295plusmn00a
057plusmn002
a
Pleurotuscystid
iosus
006plusmn00a
031plusmn001
a028plusmn00a
135plusmn00a
009plusmn00a
005plusmn00a
098plusmn00a
204plusmn00a
221plusmn02a
052plusmn002
a
Russu
lalep
ida
006plusmn00a
019plusmn001
aND
033plusmn00a
005plusmn00a
005plusmn00a
049plusmn00a
204plusmn03a
165plusmn00a
042plusmn001
a
Rmairei
001plusmn00a
017plusmn001
aND
026plusmn00a
008plusmn00a
008plusmn00a
063plusmn00a
201plusmn02a
010plusmn00a
028plusmn001
a
ND=no
tdetected
Values
aree
xpressed
asmeanplusmnSE
andlette
rsin
thes
uperscrip
tsrepresentthe
significantd
ifference
ineach
columnwith119901le005accordingto
Tukeyrsquos
test
BioMed Research International 7
Table 5 Percent amino acids composition of all the wild culinary species collected from northern Himalayas
Values are expressed as mean plusmn SE and letters in superscript represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
acids found in the studied species were linoleic acid (C182)followed by oleic acid (C181) and palmitic acid (C160)Besides these three main fatty acids already described sixmore were identified and quantified PUFA were the maingroup of fatty acids documented in all the species Agaricusarvensis Amanita caesarea Cantharellus cibarius Lentinuscladopus and Pleurotus cystidiosus contained lower value ofMUFA but higher percentage of PUFA as compared to otherspecies due to the higher percentage of linoleic acidHoweverUFApredominated over SFA in all the studied species rangingfrom 65 to 70
Amino acid composition of all the species is shown inTable 5 In all the species aspartic acid (019ndash039) wasfound to be predominated amino acid followed by tyrosine(010ndash021) arginine (012ndash029) alanine (004ndash014)and proline (001ndash007) Amanita caesarea Agaricus arven-sis Cantharellus cibarius Lentinus cladopus and Pleurotuscystidiosus contained maximum amount of these aminoacids Tocopherol contents in all the studied mushroomspecies including three wild are detailed in Table 6 120572-tocopherol and 120573-tocopherol were found to be present in allthe species However 120574-tocopherol was documented fromfew species only Tocopherol content was ranged from 090to 433 120583gg in all the species Cantharellus cibarius (433 plusmn00 120583gg) contained all the three isomers in higher amount ascompared to other species 120573-tocopherol was found in higheramounts as compared to 120572-tocopherol 120574-tocopherol wasdetected only in nine species Cantharellus cibarius (433 plusmn00 120583gg) contained higher amounts of 120574 tocopherol
Results obtained for 120573-carotene lycopene flavonoidsascorbic acid and anthocyanidins composition of all the
twenty species are presented in Table 7 Phenols werethe major antioxidant component detected in significantamounts from all the species (1912ndash6336mgg) followedby anthocyanidins (614ndash1425mg cyanidin chloride100 gextract) flavonoids (114ndash417mgg) ascorbic acid whichwas found in small amounts (020ndash099mgg) 120573-carotene(021ndash079 120583g100 g) and lycopene (019ndash038 120583g100 g) Eachspecies differed with other species in net amounts of all thesecomponents Species like Agaricus arvensis Amanita cae-sarea Gymnopilus junonius Lentinus cladopus and Pleurotuscystidiosus contained higher values of these components ascompared to other species
33 Antioxidant Evaluation Antioxidant properties of allthe species were expressed as EC
radical scavenging activity in all the species showed differ-ences in effectiveness in antioxidant properties Among allthe species Cantharellus cibarius showed lowest EC
50values
(176 plusmn 02mgmL) followed by Amanita caesarea (202 plusmn02mgmL) and Agaricus arvensis (212 plusmn 04mgmL) Otherspecies showed slightly higher EC
50values and therefore
lesserDPPH radical scavenging activityCantharellus cibariusshowed higher DPPH radical scavenging activity and Inocybesplendens showed lower DPPH radical scavenging activitythan other species
For ABTS radical scavenging activities EC50ranged from
426 to 145mgmL Lowest EC50
values were obtained forAmanita caesarea (145 plusmn 06mgmL) showing high antiox-idant activities of this species Higher EC
50values were
8 BioMed Research International
Table 6 Tocopherol composition (120583gg) of twenty species collected from northern Himalayan regions
ND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 7 Other bioactive compounds evaluated from all the species
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
BioMed Research International 9
Table 8 EC50 values for different antioxidant assays on twenty wild culinary species collected from northern Himalayas
obtained for Hygrocybe nivea (426 plusmn 05mgmL) showinglowest ABTS radical scavenging activities of this speciesReducing power of Cantharellus cibarius (162 plusmn 05mgmL)was measured higher than other species Higher EC
50values
for reducing activity were measured in Inocybe splendens(519 plusmn 01mgmL)
Higher effectiveness in ferrous ion chelating activitywas detected in Agaricus arvensis (114 plusmn 01mgmL) andlow effectiveness was detected in Russula mairei (295 plusmn03mgmL) Nevertheless Cantharellus cibarius (127 plusmn02mgmL) Amanita caesarea (140 plusmn 04mgmL) Hygro-cybe coccinea (135 plusmn 02mgmL) and H nivea (145 plusmn06mgmL) showed lower EC
50values than remaining
species EC50
values of scavenging ability on superoxideradical were found to be maximum in Amanita caesarea(044 plusmn 03mgmL) and minimum in Lactarius pubescens(221 plusmn 06mgmL) Gymnopilus junonius showed maximumFRAP activity with least EC
Although there are previous reports on documentation ofculinary edible species from the regions native to northernHimalayas but there are no reports on the evaluation stud-ies as well as toxicity status of all these culinary speciesThe wild edible species Agaricus bisporus Boletus edulis
Morchella esculenta Cordyceps sinensis and Lentinula edodeswhich have been extensively worked out in India and otherparts of the world for their compositional and medicinalaspects have not been undertaken for investigations presently[37ndash40] Compositional studies showed that most of theculinary species were rich in protein carbohydrates andlow in fat There are several reports on richness of wildedible mushrooms with protein and carbohydrate contentsand low fat levels which directly make them nutritionallyrich [24 25 41] Nevertheless under present studies thedifferences between the nutrient concentrations of all thespecies differed but Agaricus arvensis Pleurotus cystidiosusAmanita caesarea Agaricus campestris Cantharellus cibariusand Lentinus cladopus showed higher nutrient percentagewhich is comparable to other wild and commercially cul-tivated species [25 42] The crude fat content detected inall the species was not found to be significantly differentCrude fibres were detected in appreciable percentage fromall the species which make them important in nutritionalpoint of viewThe results are in conformity with the previousreports on several wild edible Pleurotus and Lentinus speciesfrom northwest India [24 43] The species Inocybe splendensHygrocybe nivea and Conocybe tenera were not found tocontain higher percentage of nutrients Although previousreports showed that nutrients composition in wild species isless as compared to cultivated species [25] however Agaricusarvensis Pleurotus cystidiosus Amanita caesarea Agaricuscampestris Cantharellus cibarius and Lentinus cladopus were
10 BioMed Research International
found to be rich in protein and carbohydrates similar tocommercially grown species [25]
Fatty acid composition showed the dominance of UFAover SFA in all the studied mushrooms species which is inconformity with the other studies [41] The differences wereobserved in net amounts in all the species Unsaturationindex of Agaricus arvensis and Lentinus cladopus (057 plusmn002) was found to be significantly higher than otherspecies Whereas Inocybe splendens (005 plusmn 00) showedleast Unsaturation Index High UFA shows the medicinalimportance of these culinary mushrooms as these increasethe HDL cholesterol and decrease LDL cholesterol triacyl-glycerol lipid oxidation and LDL susceptibility to oxidation[44] Predominance of UFA over SFA in all the species showssimilar results as obtained for other wild and commerciallycultivated species [24 25] 120572- and 120573-tocopherol were detectedin higher amounts than third isomer in all the studied speciesSimilar findings were made in other wild and cultivatedspecies with higher 120572- and 120573-tocopherol than 120574-tocopherol[25] The high levels of these two compounds correspondwith a higher oxidative activity which is associated withcardiovascular protection [45] Phenolic compounds weredetected in higher amounts than other bioactive compoundsPresence of high phenolic compounds accounts for the highantioxidant properties of all the species [42] 120573-carotenelycopene and ascorbic acids were detected in low amountsAnthocyanidins were also detected from these wild speciesin appreciable amounts The presence of these functionalmedicinal compounds inmedicinal andor edible mushroomis due to habitat or substrates in which these grow to be highin the functionalmoleculesThe categories of thesemoleculesare anthocyanidins beta-glucans selenium ganoderic acidtriterpenes or cordycepinThe compounds identified in theseextracts show that at least a part of the functional compoundsin medicinal andor edible mushroom is due to growingmushrooms on substrates that are high in the functionalmolecules To these categories can be added anthocyanidinsbeta-glucans selenium ganoderic acid triterpenes or cordy-cepinThe amounts of these have been found to vary with thetype of extraction as ethanolic extract yields higher amountsof anthocyanidins as compared to methanolic hot water andcold water extracts [29]
All the studied species showed significant antioxidantproperties measured on the basis of EC
50values Never-
theless each species showed different antioxidant activitywith highly effective and less effective EC
50values Better
antioxidant properties of some species are due to presence ofhigher phenolic compounds 120573-carotene lycopene ascorbicacids anthocyanidins and tocopherol amounts in themHigh reducing power of some species is due to the presence ofhigher amounts of reducers (antioxidants) in them Presentlyinvestigated species are commonly used for culinary pur-poses in the regions native to northern Himalayas Many ofthe species in these regions are not evaluated previously fordetailed compositional analysisTheir knowledge is restrictedto old aged villagers of the regions and neglected for thecommercial exploitations There are no positive reportson toxicity of these mushrooms analyzed presently hencethese are safe for further experimental work related to
drug discovery All the culinary species contained impor-tant and useful nutraceuticals such as unsaturated fattyacids phenolics carotenoids ascorbic acid tocopherolsand anthocyanidins besides these some important aminoacids were detected in these mushrooms which could beused for the purpose of being used as functional ingredi-ents Since nutraceuticals are powerful in maintaining andpromoting health longevity and life quality the commer-cial exploitation of these species will certainly create animpact on nutritional therapy and also will be beneficialtodayrsquos food industry Direct use of these species for con-sumption and other culinary aspects is safe and healthpromoting with advantage of the additive effects of allthe bioactive and antioxidant compounds present in thesespecies
Abbreviations
AlCl3 Aluminum trichloride
ANOVA Analysis of varianceDW Distilled waterDPPH 22-Diphenyl-1-picrylhydrazylFeCl3 Ferric chloride
FRAP Ferric reducing antioxidant powerg GramsGAEs Gallic acid equivalentsGC Gas ChromatographyHCl Hydrochloric acidH2O Water
H3PO4 Phosphoric acid
HPLC High performance liquid chromatographyK2HPO4 Potassium hydrogen phosphate
The authors declare that there is no conflict of interestsregarding the publication of this paper
BioMed Research International 11
Acknowledgment
The authors wish to thank Science and Engineering BoardDepartment of Science and Technology New Delhi forresearch Grant (SBFTLS-042013) to carry out presentstudies
References
[1] W M Breene ldquoNutritional and medicinal value of specialtymushroomsrdquo Journal of Food Protection vol 53 pp 883ndash8941990
[2] S Chang and P Miles ldquoMushroom biologymdasha new disciplinerdquoMycologist vol 6 no 2 pp 64ndash65 1992
[3] P Manzi L Gambelli S Marconi V Vivanti and L Pizzoferra-to ldquoNutrients in edible mushrooms an inter-species compara-tive studyrdquo Food Chemistry vol 65 no 4 pp 477ndash482 1999
[4] P Bobek O Ozdın and M Mikus ldquoDietary oyster mushroom(Pleurotus ostreatus) accelerates plasma cholesterol turnover inhypercholesterolaemic ratsrdquo Physiological Research vol 44 no5 pp 287ndash291 1995
[5] P Bobek and S Galbavy ldquoHypocholesterolemic and antiathero-genic effect of oystermushroom (Pleurotus ostreatus) in rabbitsrdquoNahrung vol 43 no 5 pp 339ndash342 1999
[6] S Khatun A Islam U Cakilcioglu and N Chatterjee ldquoRe-search on mushroom as a potential source of nutraceuticals areview on Indian perspectiverdquo American Journal of Experimen-tal Agriculture vol 2 no 1 pp 47ndash73 2012
[7] J A Vaz L Barros A Martins J S Morais M H Vasconcelosand I C F R Ferreira ldquoPhenolic profile of seventeen por-tuguese wild mushroomsrdquo LWT Food Science and Technologyvol 44 no 1 pp 343ndash346 2011
[8] J A Vaz L Barros A Martins C Santos-Buelga M HVasconcelos and I C F R Ferreira ldquoChemical compositionof wild edible mushrooms and antioxidant properties of theirwater soluble polysaccharidic and ethanolic fractionsrdquo FoodChemistry vol 126 no 2 pp 610ndash616 2011
[9] T Ozen C Darcan O Aktop and I Turkekul ldquoScreening ofantioxidant antimicrobial activities and chemical contents ofedible mushrooms wildly grown in the Black Sea region ofTurkeyrdquo Combinatorial Chemistry amp High Throughput Screen-ing vol 14 no 2 pp 72ndash84 2011
[10] V E C Ooi and F Liu ldquoA Review of pharmacological activitiesof mushroom polysaccharidesrdquo International Journal of Medic-inal Mushrooms vol 1 no 3 pp 195ndash206 1999
[11] S PWasser and A LWeis ldquoMedicinal properties of substancesoccurring in higher Basidiomycete mushroom current per-spectiverdquo International Journal of Medicinal Mushrooms vol 1pp 31ndash62 1999
[12] P M Kidd ldquoThe use of mushroom glucans and proteoglycansin cancer treatmentrdquo Alternative Medicine Review vol 5 no 1pp 4ndash27 2000
[13] C Israilides and A Philippoussis ldquoBio-technologies of recy-cling agro-industrial wastes for the production of commerciallyimportant fungal polysaccharides andmushroomsrdquo Biotechnol-ogy and Genetic Engineering Reviews vol 20 pp 247ndash259 2003
[14] T Mizuno ldquoBioactive biomolecules of mushrooms food func-tion and medicinal effect of mushroom fungirdquo Food ReviewsInternational vol 11 no 1 pp 5ndash21 1995
[15] Y Kabir and S Kimura ldquoDietary mushrooms reduce bloodpressure in spontaneously hypertensive rats (SHR)rdquo Journal of
Nutritional Science and Vitaminology vol 35 no 1 pp 91ndash941989
[16] J-L Mau S-Y Tsai Y-H Tseng and S-J Huang ldquoAntioxidantproperties of hot water extracts from Ganoderma tsugae Mur-rillrdquo LWTmdashFood Science and Technology vol 38 no 6 pp 589ndash597 2005
[17] A Smania F D Monache E F A Smania M L Gil L CBenchetrit and F S Cruz ldquoAntibacterial activity of a substanceproduced by the fungus Pycnoporus sanguineus (Fr) MurrrdquoJournal of Ethnopharmacology vol 45 no 3 pp 177ndash181 1995
[18] W Andlauer and P Furst ldquoNutraceuticals a piece of historypresent status and outlookrdquo Food Research International vol 35no 2-3 pp 171ndash176 2002
[19] C L Kruger and S W Mann ldquoSafety evaluation of functionalingredientsrdquo Food and Chemical Toxicology vol 41 no 6 pp793ndash805 2003
[20] P H Mattila V I Piironen E J Uusi-Rauva and P E Koivis-toinen ldquoVitamin D contents in edible mushroomsrdquo Journal ofAgricultural and Food Chemistry vol 42 no 11 pp 2449ndash24531994
[21] R P Tewari ldquoMushroom their role in nature and societyrdquo inFrontiers inMushroomBiotechnology R D Rai R C Upadhyayand S R Sharma Eds pp 1ndash8 NRCM Chambaghat India2005
[22] K C Semwal S L Stephenson V K Bhatt and R P BhattldquoEdible mushrooms of the Northwestern Himalaya India astudy of indigenous knowledge distribution and diversityrdquoMycosphere vol 5 no 3 pp 440ndash461 2014
[23] E V Crisan and A Sands ldquoEdible mushrooms nutritionalvaluerdquo in The Biology and Cultivation of Edible Mushrooms ST Chang and W A Hayes Eds pp 137ndash165 Academic PressNew York NY USA 1978
[24] N S Atri S K Sharma R Joshi A Gulati and A Gula-ti ldquoNutritional and neutraceutical composition of five wildculinary-medicinal species of genus Pleurotus (Higher Basid-iomycetes) from Northwest Indiardquo International Journal ofMedicinal Mushrooms vol 15 no 1 pp 49ndash56 2013
[25] L Barros T Cruz P Baptista L M Estevinho and I C FR Ferreira ldquoWild and commercial mushrooms as source ofnutrients and nutraceuticalsrdquo Food and Chemical Toxicologyvol 46 no 8 pp 2742ndash2747 2008
[26] M Nagata and I Yamashita ldquoSimple method for simultaneousdetermination of chlorophyll and carotenoids in tomato fruitrdquoNippon Shokuhin Kogyo Gakkaishi vol 39 no 10 pp 925ndash9281992
[27] V L Singleton and J A Rossi ldquoColorimetric detection oftotal phenolics with phosphomolybdic-phosphotungstic acidreagentsrdquo American Journal of Enology and Viticulture vol 16pp 144ndash158 1965
[28] L K Jagadish V Venkata Krishnan R Shenbhagaramanand V Kaviyarasan ldquoComparitive study on the antioxidantanticancer and antimicrobial property of Agaricus bisporus (JE Lange) Imbach before and after boilingrdquo African Journal ofBiotechnology vol 8 no 4 pp 654ndash661 2009
[29] E Vamanu and S Nita ldquoAntioxidant capacity and the cor-relation with major phenolic compounds anthocyanin andtocopherol content in various extracts from the wild edibleBoletus edulis mushroomrdquo BioMed Research International vol2013 Article ID 313905 11 pages 2013
[30] E Vamanu ldquoBiological activities of the polysaccharides pro-duced in submerged culture of two edible Pleurotus ostreatus
12 BioMed Research International
mushroomsrdquo Journal of Biomedicine and Biotechnology vol2012 Article ID 565974 8 pages 2012
[31] P L Li L Huo W Su et al ldquoFree radical-scavenging capacityantioxidant activity and phenolic content of Pouzolzia zeylan-icardquo Journal of the Serbian Chemical Society vol 76 no 5 pp709ndash717 2011
[32] C Papuc M Crivineanu G Goran V Nicorescu and N Dur-dun ldquoFree radicals scavenging and antioxidant activity ofEuropean mistletoe (Viscum album) and European birthwort(Aristolochia clematitis)rdquo Revista de Chimie vol 61 no 7 pp619ndash622 2010
[33] VOOyetayo CDong andY Yao ldquoAntioxidant and antimicro-bial properties of aqueous extract from Dictyophora indusiatardquoThe Open Mycology Journal vol 3 no 1 pp 20ndash26 2009
[34] S Marklund and G Marklund ldquoInvolvement of the superoxideanion radical in the autoxidation of pyrogallol and a convenientassay for superoxide dismutaserdquo European Journal of Biochem-istry vol 47 no 3 pp 469ndash474 1974
[35] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996
[36] D J Huang b Ou and R L Prior ldquoThe chemistry behindantioxidant capacity assaysrdquo Journal of Agricultural and FoodChemistry vol 53 no 6 pp 1841ndash1856 2005
[37] P Manzi A Aguzzi and L Pizzoferrato ldquoNutritional value ofmushrooms widely consumed in Italyrdquo Food Chemistry vol 73no 3 pp 321ndash325 2001
[38] P Manzi S Marconi A Aguzzi and L Pizzoferrato ldquoCommer-cial mushrooms nutritional quality and effect of cookingrdquo FoodChemistry vol 84 no 2 pp 201ndash206 2004
[39] I P Krbavcic and I C Baric ldquoInfluence of deep fat fryingon some nutritional parameters of novel food based on mush-rooms and fresh soft cheeserdquo Food Chemistry vol 84 no 3 pp417ndash419 2004
[40] D Agrahar-Murugkar andG Subbulakshmi ldquoNutritional valueof edible wild mushrooms collected from the Khasi hills ofMeghalayardquo Food Chemistry vol 89 no 4 pp 599ndash603 2005
[41] V A Dıez and A Alvarez ldquoCompositional and nutritionalstudies on two wild edible mushrooms from northwest SpainrdquoFood Chemistry vol 75 no 4 pp 417ndash422 2001
[42] L Barros M-J Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007
[43] S K Sharma and N S Atri ldquoNutraceutical composition ofwild species of genus Lentinus Fr fromNothern Indiardquo CurrentResearch in Environmental ampAppliedMycology vol 4 pp 11ndash322014
[44] P J Kanu K Zhu J Baby Kanu H Zhou H Qian and K ZhuldquoBiologically active components and nutraceuticals in sesameand related products a review and prospectrdquo Trends in FoodScience and Technology vol 18 no 12 pp 599ndash608 2007
[45] F S Reis E Pereira L Barros M J Sousa A Martins and I CF R Ferreira ldquoBiomolecule profiles in inediblewildmushroomswith antioxidant valuerdquoMolecules vol 16 no 6 pp 4328ndash43382011
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
cover (ie more than 50) The people of these areas usedifferent mushroom species for culinary purposes Theyhave very little idea about the medicinal importance ofthese mushroom species Their methods of identification areprimarily based upon sweet smell and color Further theknowledge about the edible species is restricted to the oldaged people Documentations studies of these edible specieshave been done by several workers [22] but the studies onchemical composition and medicinal importance are stilllacking besides the presence of toxic compounds in them Inview of this under present investigations twenty of such wildculinary mushroom species have been evaluated for theirchemical bioactive and antioxidant potential
2 Materials and Methods
21 Collection andProcessing of Samples All the sampleswerecollected during the frequent surveys to the different regionsof northern Himalayas (Table 1) After this the samples werevacuum-dried and preserved in air-tight cellophane bagswith a small amount of 1-4-paradichlorobenzene in porouspackets to keep them free of insects for further analysis
22 Chemical Evaluation Samples were powdered and eval-uated for protein fat carbohydrates ash and crude fibresCrude protein content was estimated using the Kjeldahlmethod by calculating total nitrogen (N) and protein contentwas expressed by N times 438 [23] Crude fat was estimatedusing a Soxhlet apparatus by extraction of known weightof powered samples with petroleum ether Ash content wascalculated by incineration in silica dishes at 525 plusmn 20∘C con-taining 5ndash10 gsample Fibres content was estimated on a fat-free sample using the acid-alkali method (125 each) Totalcarbohydrates percentage was calculated by the difference asthe totalweightminus (moisture content +protein content + crudefat + ash content + crude fibres)
For toxic metal detection diluted HCl (2) and copperfoil (1 times 12 cm strip purified or pretreated with concentratedHNO
3or diluted 3 HCl and rinsed in distilled water) were
taken After that samples (powered samples) were acidifiedwith 10ndash20mL of 5 diluted HCl (2) until colour changedfrom fairy pink to litmus Then strips of copper foil wereadded and boiled for 30 minutes with addition of waterfrom time to time to replace the losses by evaporation Theheavy metals got deposited on the copper foil and gavecharacteristics color to it The color deposited on the Cu foilwas noted after 30 minutes and results were interpreted forthe presence of heavy metals [24]
For monosaccharide composition samples (01 g) wereextracted with 25mL and 15mL and finally with 1mL of 70aqueous methanol After this the extract was centrifuged at4000 rpm (4∘C) for 10min Supernatant was collected andvolumewasmade up to 5mLwith 70methanolThe extractwas passed through Millipore filter (045 120583m) and injected tothe HPLC [24]
23 Bioactive Evaluation231 Fatty Acid Composition Powdered samples were dis-solved in 1mL of solution (prepared by sodium hydroxide
Table 1 List of wild culinary species collected from differentlocalities of twenty species from northern Himalayas
pellets (45 g) in 300mL of 50 methanol and vortexing for1 minute then the solution was left for 5 minutes at 100∘Cvortexed again for 1 minute and left at 100∘C in a waterbath for 25 minutes) Methylation was done by adding 2mLof solution (6N hydrogen chloride in methanol) and thenvortexed for 1 minute followed by heating (80∘C) in a waterbath For extraction of fatty acids 125mL of solution (25mLmethyl ter-butyl ether added to hexane) was added and thesolution was shaken for 10 minutes followed by removal ofupper layer and addition of 3mL of solution (10 sodiumhydroxide in water while stirring) Finally the top phase(23) was removed and transferred into a gas chromatographyvial and injected Unsaturation index (UI) was calculated as(mol of each (poly)unsaturated fatty acid times number doublebonds per each fatty acid)100
BioMed Research International 3
232 Amino Acids Powdered samples (01 g) were extractedwith 25mL followed by 15mL and 1mL of 70 aqueousmethanol After this centrifugation was done for 10 minutes(4000 rpm) at 4∘C Supernatants were dissolved in aque-ous methanol and the volume was made up to 5mL Itwas now passed through Millipore filter (045 120583m) Afterthis samples (10 120583L) were dried using vacuum oven andto these dried samples 20 120583L derivatising agent (preparedby ethanol triethylamine water phenylisothiocaynate) wasmixed with it and redried it Now the samples were left for 25minutes at room temperature Lastly 1mL ethanol was addedand injected into UPLC (Waters India Pvt Ltd)
233 Tocopherol Composition Tocopherol composition wasestimated following standard method [25] For this sam-ples were mixed with butylated hydroxytoluene (BHT) inhexane (10mgmL 100 120583120583L) and IS solution in hexane (120575tocopherol 16 120583gmL 250 120583L) Thereafter samples (500mg)were vortexed for 1min with methanol (4mL)Then sampleswere again vortexed with hexane (4mL) After this 2mL ofsaturated NaCl aqueous solution was added and the mixturewas vortexed (1min) followed by centrifugation at 4000 gfor 5min and the upper layer was separated The sampleswere again reextracted twice with hexane The extractswere then vacuum-dried and redissolved in hexane (1mL)followed by dehydration with anhydrous sodium sulphatethen filtered and transferred into a dark injection vial andanalysed byHPLC (Waters India Pvt Ltd) Chromatographiccomparisons were made by authentic standards Tocopherolcontents in mushroom samples were expressed in 120583g per g ofdry mushroom
234 Evaluation of Other Bioactive Compounds For 120573-carotene and lycopene estimation dried powdered samples(sim5 g) were extracted with 100mL of methanol at 25∘C(150 rpm) for 24 hours and filtered through Whatman Num-ber 4 filter paper The residue was again reextracted with2 additional 100mL portions of methanol These extractswere evaporated to dryness at 42∘C then redissolved inmethanol at a concentration of 50mgmL and stored at 4∘CThe driedmethanolic extract (100mg) was shaken vigorouslywith 10mL of acetonehexanemixture (4 6) for 1 minute andfiltered The absorbance of the filtrate was measured at 453505 and 663 nm [26] 120573-carotene and lycopene content wereestimated using the following equation
Lycopene (mg100mL)
= (00458times119860663) + (0372times119860505)
minus (00806times119860453) 120573-carotene (mg100mL)
= (0216times119860663) minus (0304times119860505)
+ (0452times119860453)
(1)
For phenolic compounds quantification powdered sam-ples (1mL) were mixed with Folin and Ciocalteursquos phenolreagent (1mL) After 3 minutes 1mL of saturated sodiumcarbonate solution was added to the mixture and the volume
was adjusted to 10mL with distilled water The reaction waskept in the dark for 90 minutes after which the absorbancewas read at 725 nm Gallic acid was used to calculate thestandard curve (001ndash04mM 1198772 = 09999) and the resultswere expressed as milligrams of gallic acid equivalents pergram of extract [27]
Total flavonoids of the sample extracts were measuredby AlCl
3method [28] For this an aqueous extract (15mL)
was mixed with deionized distilled water (5mL) and 03mLof 5 NaNO
2 After five minutes of incubation at room
temperature 15mL of 2 aluminium trichloride (AlCl3)
solution was added After the next 6 minutes 2mL of 1MNaOH was added The mixture was vigorously shaken onorbital shaker for 5min at 200 rpm and the absorbance wasread at 510 nm against a blank Quercetin with differentconcentrations was used as a standard
For ascorbic acid quantification standard ascorbic acidsolution (5mL L-ascorbic acid in 3 phosphoric acid) wasadded to 5mL of phosphoric acid A microburette was filledwith dye and the samples were titrated with the dye solutionto a pink color which persisted for 15 seconds The dyefactor (milligrams of ascorbic acid per milliliter of dye usingformula 05titrate) was determined A sample was preparedby taking 10 g of sample grounded in metaphosphoric acidand the volume was increased up to 100mL It was titratedafter filtration until a pink color appeared [24] The amountof ascorbic acid was calculated with the use of the followingequation
mg of ascorbic acid per100 g or mL
=Titrate times Dye factor times Vol madeAliquot of extract times wt of sample
times 100(2)
Anthocyanidins were quantified by using standardmethod [29] Briefly 05 g of samples was mixed withthe solvent (mixture of 85 15 (vv) of ethyl alcohol andhydrochloric acid 15M) followed by ultrasonication for 15minutes and filtration throughWhatman filter paper number1 Standard solution was prepared with cyaniding chloridewith a concentration of 5ndash15120583gmL in solvent which wasused The absorption was measured at 546 nm The totalquantity of anthocyanins (expressed in g of cyanidingchloride100 g extract) = (119860
119901times 119898st times 119891 times 100)(119860 st times 119898119901)
where 119860119901is absorption rate of the sample solution 119898
119901is
mass of the processed sample in g 119860 st is absorption rate ofthe standard solution 119898st is mass of the processed standardsolution in g and 119891 is dilution coefficient
24 Antioxidant Assays DPPH scavenging activity was mea-sured with adding DPPH (200120583L) solution at differentconcentrations (2ndash10mgmL) to 005mL of the samples(dissolved in ethanol) An equal amount of ethanol was addedto the control Ascorbic acid was used as the control [30]Theabsorbancewas read after 20min at 517 nm and the inhibitionwas calculated using the formula
where 1198600was the absorbance of the control and 119860
119875was the
absorbance in the presence of the sampleFor ABTS radical scavenging activity 10120583L of the sample
was added to 4mL of the diluted ABTS∙+ solution (preparedby adding 7mM of the ABTS stock solution to 245mMpotassium persulfate kept in the dark at room temperaturefor 12ndash16 h before use) The solution was then diluted with5mM phosphate-buffered saline (pH 74) and absorbancewas measured at 730 nm after 30min [31] The ABTS radicalscavenging activity was calculated as
119878 = (119860controlminus119860 sample
119860control)times 100 (4)
For reducing power estimation samples (200 120583L) weremixed with sodium phosphate buffer (pH 66) 1mM FeSO
4
and 1 potassium ferricyanide and incubated for 20minat 50∘C after that trichloroacetic acid was added and themixtures were centrifuged Supernatant (25mL) was mixedwith an equal volume of water and 05mL 01 FeCl
3 The
absorbance was measured at 700 nm [32]For Fe2+ chelating activity 1mL of the sample (2ndash10mg
mL) was mixed with 37mL of ultrapure water after thatthe mixture was reacted with ferrous chloride (2mmolL01mL) and ferrozine (5mmolL 02mL) for 20min and theabsorbance was read at 562 nm with using EDTA as controlThe chelating activity was calculated using the formula
where 119860119887is the absorbance of the blank and 119860
119904is the
absorbance in the presence of the extract [33]The scavenging activity of superoxide anion radicals
was measured following standard method [34] Samples (0ndash20mgmL 1mL) and Tris-HCl buffer (500mM pH 823mL) were incubated in a water bath at 25∘C for 20min andafter this pyrogallic acid (50mM 04mL) was added HClsolution (80M 01mL) was added to terminate the reactionafter 4min The absorbance of the mixture was measuredat 320 nm The scavenging ability was calculated using thefollowing formula
scavenging ability () = (1 minus119860 sample
119860control)times 100 (6)
where 119860control is the absorbance of control without thepolysaccharide sample and 119860 sample is the absorbance in thepresence of the polysaccharide sample
For ferric reducing antioxidant power (FRAP) assayfirstly FRAP reagent was prepared by mixing TPTZ (25mL10mM in 40mM HCl) 25mL of 300mM acetate buffer and25mL of FeCl
3sdot6H2O After this freshly prepared FRAP
reagent (18mL) was taken in a test tube and incubated at30∘C in water bath for 10 minutes Then absorbance wasread at 0min (119905
0) After this 100 120583L of sample extract or
standard and 100 120583L of distilled water were added to the testtube mixed and incubated at 30∘C for 30 minutes Thenthe absorbance was taken at 593 nm (119905
30) Ferrous sulphate
was used as standard [35 36] FRAP activity was determinedagainst a standard curve of ferrous sulphate and the valueswere expressed as 120583M Fe2+ equivalents per gram of extractand calculated using the following equation
FRAP value = Absorbance (sample+ FRAP reagent)
minusAbsorbance (FRAP reagent) (7)
25 Statistical Analysis All experiments were performed 3times and with 3 replicates The results were analyzed usingone-way analysis of variance (ANOVA) 119901 lt 005 was con-sidered significant and SPSS software (SPSS Inc ChicagoIL USA) was used to calculate differences Tukey-HSD at119901 lt 005 test was used to determine significant differences
3 Results
31 Chemical Evaluation Nutrient composition of the wildculinary mushrooms is shown in Table 2 Protein was foundin high levels and varied between 516 in Inocybe splendensand 2263 in Agaricus arvensis Protein percentage inPleurotus cystidiosus (2069) Amanita caesarea (1972)Agaricus campestris (1838) Cantharellus cibarius (1819)and Lentinus cladopus (1859) was also found to be highAll the twenty culinary species were found to be low in fatcontent Fat ranged from010 in Laccaria laccata to 038 inInocybe splendens In general these wild culinary mushroomsconsumed by local people were found to be higher in proteinand low in fat although differences were observed in netvalue of individual species Crude fibres ranged from 108in Hygrocybe coccinea to 242 in Lentinus cladopus Ashcontent varied between 011 in Inocybe splendens and 096in Agaricus arvensis Carbohydrates calculated by differencewere also found to be in abundant amounts and theirpercentage was ranged from 3119 in Inocybe splendens to5712 in Agaricus arvensis Nutrient contents of Inocybesplendens Hygrocybe nivea and Conocybe tenera were foundto be less as compared to other species
There are several reports about the toxicity reports due tomushrooms on humans in these areas and hence preliminarystudies were done to check the toxicity level of mushroomsFor all the twenty species being used by the people forculinary purposes the test was found to be negative Thatmeans these species are nontoxic and hence recommendedfor consumption
All the culinarymushroom species contained glucose andrhamnose as the principal carbohydrates (Table 3) Neverthe-less the present study also describes the presence of xyloseand mannose in all the studied species However galactoseand fructose were detected in very low percentage in someof the species Russula mairei contained lowest percentageof glucose (2160) and Agaricus arvensis contained highestpercentage of glucose (6412)
32 Bioactive Evaluation The results of fatty acid composi-tion (total saturated fatty acids SFA monounsaturated fattyacids MUFA and polyunsaturated fatty acids PUFA) of allthe species are shown in Table 4 In general the major fatty
BioMed Research International 5
Table 2 Percent chemical composition of twenty wild culinary mushroom species collected from northern Himalayan regions
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 3 Percent monosaccharide composition of twenty species collected from northern Himalayan regions showing richness in glucoserhamnose mannose xylose and galactose and fructose in lower percentage
Russula lepida 515 plusmn 01d 3261 plusmn 36f 1316 plusmn 12e 231 plusmn 01c ND NDR mairei 426 plusmn 01c 2160 plusmn 21a 1281 plusmn 16d 212 plusmn 00c ND NDND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
6 BioMed Research International
Table4Percentfattyacid
compo
sitionof
allthe
twentywild
culin
arymushroo
mspeciesc
ollected
from
different
region
sofn
orthernHim
alayas
Species
C900
C100
C120
C160
C1611
C171
C181
C182
C202
UI
Agaricu
sarvensis
006plusmn00a
030plusmn001
a028plusmn00a
186plusmn003
a006plusmn00a
009plusmn00a
018plusmn00a
324plusmn00a
221plusmn02a
057plusmn002
a
Acampestr
is003plusmn00a
026plusmn002
a021plusmn00a
119plusmn002
a009plusmn00a
002plusmn00a
035plusmn00a
204plusmn00a
218plusmn03a
046plusmn002
a
Acomtulus
006plusmn00a
022plusmn00a
ND
117plusmn001
a007plusmn00a
009plusmn00a
038plusmn00a
173plusmn00a
134plusmn01a
036plusmn001
a
Asilvicola
008plusmn00a
033plusmn00a
022plusmn00a
156plusmn002
a009plusmn00a
001plusmn00a
015plusmn00a
154plusmn00a
221plusmn03a
040plusmn001
a
Amanita
caesarea
004plusmn00a
032plusmn002
a019plusmn00a
196plusmn002
a012plusmn00a
005plusmn00a
031plusmn00a
219plusmn00a
128plusmn00a
039plusmn001
a
Acitrin
a006plusmn00a
028plusmn001
aND
091plusmn00a
011plusmn00a
008plusmn00a
031plusmn00a
159plusmn00a
168plusmn00a
037plusmn001
a
Afulva
004plusmn00a
021plusmn001
a025plusmn002
a010plusmn00a
008plusmn00a
004plusmn00a
015plusmn00a
164plusmn00a
123plusmn00a
031plusmn002
a
Cantharellu
scibarius
007plusmn00a
033plusmn002
a022plusmn001
a110plusmn002
a007plusmn00a
001plusmn00a
037plusmn00a
294plusmn00a
118plusmn01a
045plusmn002
a
Conocybe
tenera
006plusmn00a
020plusmn00a
ND
007plusmn00a
005plusmn00a
002plusmn00a
035plusmn00a
014plusmn00a
138plusmn00a
019plusmn00a
Gymnopilusjun
onius
005plusmn00a
031plusmn001
a014plusmn001
a18
7plusmn003
a008plusmn00a
002plusmn00a
088plusmn00a
049plusmn00a
121plusmn
01a
026plusmn001
a
Hygrocybe
coccinea
004plusmn00a
025plusmn002
a017plusmn00a
033plusmn00a
001plusmn00a
005plusmn00a
017plusmn00a
024plusmn00a
119plusmn00a
016plusmn00a
Hnivea
007plusmn00a
022plusmn001
aND
005plusmn00a
004plusmn00a
001plusmn00a
053plusmn00a
004plusmn00a
138plusmn02a
020plusmn00a
Inocybesplendens
002plusmn00a
019plusmn001
a023plusmn001
a089plusmn00a
002plusmn00a
003plusmn00a
018plusmn00a
010plusmn00a
020plusmn00a
005plusmn00a
Lactariusp
ubescens
001plusmn00a
023plusmn001
a016plusmn00a
075plusmn00a
007plusmn00a
008plusmn00a
026plusmn00a
344plusmn00a
083plusmn00a
046plusmn002
a
Laccarialaccata
002plusmn00a
022plusmn00a
ND
101plusmn
00a
006plusmn00a
009plusmn00a
092plusmn00a
284plusmn00a
029plusmn00a
042plusmn001
a
Lepista
nuda
003plusmn00a
024plusmn00a
ND
043plusmn00a
001plusmn00a
002plusmn00a
020plusmn00a
204plusmn00a
037plusmn00a
026plusmn00a
Lentinus
cladopu
s009plusmn00a
033plusmn002
a032plusmn00a
146plusmn00a
055plusmn00a
001plusmn00a
032plusmn00a
245plusmn00a
295plusmn00a
057plusmn002
a
Pleurotuscystid
iosus
006plusmn00a
031plusmn001
a028plusmn00a
135plusmn00a
009plusmn00a
005plusmn00a
098plusmn00a
204plusmn00a
221plusmn02a
052plusmn002
a
Russu
lalep
ida
006plusmn00a
019plusmn001
aND
033plusmn00a
005plusmn00a
005plusmn00a
049plusmn00a
204plusmn03a
165plusmn00a
042plusmn001
a
Rmairei
001plusmn00a
017plusmn001
aND
026plusmn00a
008plusmn00a
008plusmn00a
063plusmn00a
201plusmn02a
010plusmn00a
028plusmn001
a
ND=no
tdetected
Values
aree
xpressed
asmeanplusmnSE
andlette
rsin
thes
uperscrip
tsrepresentthe
significantd
ifference
ineach
columnwith119901le005accordingto
Tukeyrsquos
test
BioMed Research International 7
Table 5 Percent amino acids composition of all the wild culinary species collected from northern Himalayas
Values are expressed as mean plusmn SE and letters in superscript represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
acids found in the studied species were linoleic acid (C182)followed by oleic acid (C181) and palmitic acid (C160)Besides these three main fatty acids already described sixmore were identified and quantified PUFA were the maingroup of fatty acids documented in all the species Agaricusarvensis Amanita caesarea Cantharellus cibarius Lentinuscladopus and Pleurotus cystidiosus contained lower value ofMUFA but higher percentage of PUFA as compared to otherspecies due to the higher percentage of linoleic acidHoweverUFApredominated over SFA in all the studied species rangingfrom 65 to 70
Amino acid composition of all the species is shown inTable 5 In all the species aspartic acid (019ndash039) wasfound to be predominated amino acid followed by tyrosine(010ndash021) arginine (012ndash029) alanine (004ndash014)and proline (001ndash007) Amanita caesarea Agaricus arven-sis Cantharellus cibarius Lentinus cladopus and Pleurotuscystidiosus contained maximum amount of these aminoacids Tocopherol contents in all the studied mushroomspecies including three wild are detailed in Table 6 120572-tocopherol and 120573-tocopherol were found to be present in allthe species However 120574-tocopherol was documented fromfew species only Tocopherol content was ranged from 090to 433 120583gg in all the species Cantharellus cibarius (433 plusmn00 120583gg) contained all the three isomers in higher amount ascompared to other species 120573-tocopherol was found in higheramounts as compared to 120572-tocopherol 120574-tocopherol wasdetected only in nine species Cantharellus cibarius (433 plusmn00 120583gg) contained higher amounts of 120574 tocopherol
Results obtained for 120573-carotene lycopene flavonoidsascorbic acid and anthocyanidins composition of all the
twenty species are presented in Table 7 Phenols werethe major antioxidant component detected in significantamounts from all the species (1912ndash6336mgg) followedby anthocyanidins (614ndash1425mg cyanidin chloride100 gextract) flavonoids (114ndash417mgg) ascorbic acid whichwas found in small amounts (020ndash099mgg) 120573-carotene(021ndash079 120583g100 g) and lycopene (019ndash038 120583g100 g) Eachspecies differed with other species in net amounts of all thesecomponents Species like Agaricus arvensis Amanita cae-sarea Gymnopilus junonius Lentinus cladopus and Pleurotuscystidiosus contained higher values of these components ascompared to other species
33 Antioxidant Evaluation Antioxidant properties of allthe species were expressed as EC
radical scavenging activity in all the species showed differ-ences in effectiveness in antioxidant properties Among allthe species Cantharellus cibarius showed lowest EC
50values
(176 plusmn 02mgmL) followed by Amanita caesarea (202 plusmn02mgmL) and Agaricus arvensis (212 plusmn 04mgmL) Otherspecies showed slightly higher EC
50values and therefore
lesserDPPH radical scavenging activityCantharellus cibariusshowed higher DPPH radical scavenging activity and Inocybesplendens showed lower DPPH radical scavenging activitythan other species
For ABTS radical scavenging activities EC50ranged from
426 to 145mgmL Lowest EC50
values were obtained forAmanita caesarea (145 plusmn 06mgmL) showing high antiox-idant activities of this species Higher EC
50values were
8 BioMed Research International
Table 6 Tocopherol composition (120583gg) of twenty species collected from northern Himalayan regions
ND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 7 Other bioactive compounds evaluated from all the species
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
BioMed Research International 9
Table 8 EC50 values for different antioxidant assays on twenty wild culinary species collected from northern Himalayas
obtained for Hygrocybe nivea (426 plusmn 05mgmL) showinglowest ABTS radical scavenging activities of this speciesReducing power of Cantharellus cibarius (162 plusmn 05mgmL)was measured higher than other species Higher EC
50values
for reducing activity were measured in Inocybe splendens(519 plusmn 01mgmL)
Higher effectiveness in ferrous ion chelating activitywas detected in Agaricus arvensis (114 plusmn 01mgmL) andlow effectiveness was detected in Russula mairei (295 plusmn03mgmL) Nevertheless Cantharellus cibarius (127 plusmn02mgmL) Amanita caesarea (140 plusmn 04mgmL) Hygro-cybe coccinea (135 plusmn 02mgmL) and H nivea (145 plusmn06mgmL) showed lower EC
50values than remaining
species EC50
values of scavenging ability on superoxideradical were found to be maximum in Amanita caesarea(044 plusmn 03mgmL) and minimum in Lactarius pubescens(221 plusmn 06mgmL) Gymnopilus junonius showed maximumFRAP activity with least EC
Although there are previous reports on documentation ofculinary edible species from the regions native to northernHimalayas but there are no reports on the evaluation stud-ies as well as toxicity status of all these culinary speciesThe wild edible species Agaricus bisporus Boletus edulis
Morchella esculenta Cordyceps sinensis and Lentinula edodeswhich have been extensively worked out in India and otherparts of the world for their compositional and medicinalaspects have not been undertaken for investigations presently[37ndash40] Compositional studies showed that most of theculinary species were rich in protein carbohydrates andlow in fat There are several reports on richness of wildedible mushrooms with protein and carbohydrate contentsand low fat levels which directly make them nutritionallyrich [24 25 41] Nevertheless under present studies thedifferences between the nutrient concentrations of all thespecies differed but Agaricus arvensis Pleurotus cystidiosusAmanita caesarea Agaricus campestris Cantharellus cibariusand Lentinus cladopus showed higher nutrient percentagewhich is comparable to other wild and commercially cul-tivated species [25 42] The crude fat content detected inall the species was not found to be significantly differentCrude fibres were detected in appreciable percentage fromall the species which make them important in nutritionalpoint of viewThe results are in conformity with the previousreports on several wild edible Pleurotus and Lentinus speciesfrom northwest India [24 43] The species Inocybe splendensHygrocybe nivea and Conocybe tenera were not found tocontain higher percentage of nutrients Although previousreports showed that nutrients composition in wild species isless as compared to cultivated species [25] however Agaricusarvensis Pleurotus cystidiosus Amanita caesarea Agaricuscampestris Cantharellus cibarius and Lentinus cladopus were
10 BioMed Research International
found to be rich in protein and carbohydrates similar tocommercially grown species [25]
Fatty acid composition showed the dominance of UFAover SFA in all the studied mushrooms species which is inconformity with the other studies [41] The differences wereobserved in net amounts in all the species Unsaturationindex of Agaricus arvensis and Lentinus cladopus (057 plusmn002) was found to be significantly higher than otherspecies Whereas Inocybe splendens (005 plusmn 00) showedleast Unsaturation Index High UFA shows the medicinalimportance of these culinary mushrooms as these increasethe HDL cholesterol and decrease LDL cholesterol triacyl-glycerol lipid oxidation and LDL susceptibility to oxidation[44] Predominance of UFA over SFA in all the species showssimilar results as obtained for other wild and commerciallycultivated species [24 25] 120572- and 120573-tocopherol were detectedin higher amounts than third isomer in all the studied speciesSimilar findings were made in other wild and cultivatedspecies with higher 120572- and 120573-tocopherol than 120574-tocopherol[25] The high levels of these two compounds correspondwith a higher oxidative activity which is associated withcardiovascular protection [45] Phenolic compounds weredetected in higher amounts than other bioactive compoundsPresence of high phenolic compounds accounts for the highantioxidant properties of all the species [42] 120573-carotenelycopene and ascorbic acids were detected in low amountsAnthocyanidins were also detected from these wild speciesin appreciable amounts The presence of these functionalmedicinal compounds inmedicinal andor edible mushroomis due to habitat or substrates in which these grow to be highin the functionalmoleculesThe categories of thesemoleculesare anthocyanidins beta-glucans selenium ganoderic acidtriterpenes or cordycepinThe compounds identified in theseextracts show that at least a part of the functional compoundsin medicinal andor edible mushroom is due to growingmushrooms on substrates that are high in the functionalmolecules To these categories can be added anthocyanidinsbeta-glucans selenium ganoderic acid triterpenes or cordy-cepinThe amounts of these have been found to vary with thetype of extraction as ethanolic extract yields higher amountsof anthocyanidins as compared to methanolic hot water andcold water extracts [29]
All the studied species showed significant antioxidantproperties measured on the basis of EC
50values Never-
theless each species showed different antioxidant activitywith highly effective and less effective EC
50values Better
antioxidant properties of some species are due to presence ofhigher phenolic compounds 120573-carotene lycopene ascorbicacids anthocyanidins and tocopherol amounts in themHigh reducing power of some species is due to the presence ofhigher amounts of reducers (antioxidants) in them Presentlyinvestigated species are commonly used for culinary pur-poses in the regions native to northern Himalayas Many ofthe species in these regions are not evaluated previously fordetailed compositional analysisTheir knowledge is restrictedto old aged villagers of the regions and neglected for thecommercial exploitations There are no positive reportson toxicity of these mushrooms analyzed presently hencethese are safe for further experimental work related to
drug discovery All the culinary species contained impor-tant and useful nutraceuticals such as unsaturated fattyacids phenolics carotenoids ascorbic acid tocopherolsand anthocyanidins besides these some important aminoacids were detected in these mushrooms which could beused for the purpose of being used as functional ingredi-ents Since nutraceuticals are powerful in maintaining andpromoting health longevity and life quality the commer-cial exploitation of these species will certainly create animpact on nutritional therapy and also will be beneficialtodayrsquos food industry Direct use of these species for con-sumption and other culinary aspects is safe and healthpromoting with advantage of the additive effects of allthe bioactive and antioxidant compounds present in thesespecies
Abbreviations
AlCl3 Aluminum trichloride
ANOVA Analysis of varianceDW Distilled waterDPPH 22-Diphenyl-1-picrylhydrazylFeCl3 Ferric chloride
FRAP Ferric reducing antioxidant powerg GramsGAEs Gallic acid equivalentsGC Gas ChromatographyHCl Hydrochloric acidH2O Water
H3PO4 Phosphoric acid
HPLC High performance liquid chromatographyK2HPO4 Potassium hydrogen phosphate
The authors declare that there is no conflict of interestsregarding the publication of this paper
BioMed Research International 11
Acknowledgment
The authors wish to thank Science and Engineering BoardDepartment of Science and Technology New Delhi forresearch Grant (SBFTLS-042013) to carry out presentstudies
References
[1] W M Breene ldquoNutritional and medicinal value of specialtymushroomsrdquo Journal of Food Protection vol 53 pp 883ndash8941990
[2] S Chang and P Miles ldquoMushroom biologymdasha new disciplinerdquoMycologist vol 6 no 2 pp 64ndash65 1992
[3] P Manzi L Gambelli S Marconi V Vivanti and L Pizzoferra-to ldquoNutrients in edible mushrooms an inter-species compara-tive studyrdquo Food Chemistry vol 65 no 4 pp 477ndash482 1999
[4] P Bobek O Ozdın and M Mikus ldquoDietary oyster mushroom(Pleurotus ostreatus) accelerates plasma cholesterol turnover inhypercholesterolaemic ratsrdquo Physiological Research vol 44 no5 pp 287ndash291 1995
[5] P Bobek and S Galbavy ldquoHypocholesterolemic and antiathero-genic effect of oystermushroom (Pleurotus ostreatus) in rabbitsrdquoNahrung vol 43 no 5 pp 339ndash342 1999
[6] S Khatun A Islam U Cakilcioglu and N Chatterjee ldquoRe-search on mushroom as a potential source of nutraceuticals areview on Indian perspectiverdquo American Journal of Experimen-tal Agriculture vol 2 no 1 pp 47ndash73 2012
[7] J A Vaz L Barros A Martins J S Morais M H Vasconcelosand I C F R Ferreira ldquoPhenolic profile of seventeen por-tuguese wild mushroomsrdquo LWT Food Science and Technologyvol 44 no 1 pp 343ndash346 2011
[8] J A Vaz L Barros A Martins C Santos-Buelga M HVasconcelos and I C F R Ferreira ldquoChemical compositionof wild edible mushrooms and antioxidant properties of theirwater soluble polysaccharidic and ethanolic fractionsrdquo FoodChemistry vol 126 no 2 pp 610ndash616 2011
[9] T Ozen C Darcan O Aktop and I Turkekul ldquoScreening ofantioxidant antimicrobial activities and chemical contents ofedible mushrooms wildly grown in the Black Sea region ofTurkeyrdquo Combinatorial Chemistry amp High Throughput Screen-ing vol 14 no 2 pp 72ndash84 2011
[10] V E C Ooi and F Liu ldquoA Review of pharmacological activitiesof mushroom polysaccharidesrdquo International Journal of Medic-inal Mushrooms vol 1 no 3 pp 195ndash206 1999
[11] S PWasser and A LWeis ldquoMedicinal properties of substancesoccurring in higher Basidiomycete mushroom current per-spectiverdquo International Journal of Medicinal Mushrooms vol 1pp 31ndash62 1999
[12] P M Kidd ldquoThe use of mushroom glucans and proteoglycansin cancer treatmentrdquo Alternative Medicine Review vol 5 no 1pp 4ndash27 2000
[13] C Israilides and A Philippoussis ldquoBio-technologies of recy-cling agro-industrial wastes for the production of commerciallyimportant fungal polysaccharides andmushroomsrdquo Biotechnol-ogy and Genetic Engineering Reviews vol 20 pp 247ndash259 2003
[14] T Mizuno ldquoBioactive biomolecules of mushrooms food func-tion and medicinal effect of mushroom fungirdquo Food ReviewsInternational vol 11 no 1 pp 5ndash21 1995
[15] Y Kabir and S Kimura ldquoDietary mushrooms reduce bloodpressure in spontaneously hypertensive rats (SHR)rdquo Journal of
Nutritional Science and Vitaminology vol 35 no 1 pp 91ndash941989
[16] J-L Mau S-Y Tsai Y-H Tseng and S-J Huang ldquoAntioxidantproperties of hot water extracts from Ganoderma tsugae Mur-rillrdquo LWTmdashFood Science and Technology vol 38 no 6 pp 589ndash597 2005
[17] A Smania F D Monache E F A Smania M L Gil L CBenchetrit and F S Cruz ldquoAntibacterial activity of a substanceproduced by the fungus Pycnoporus sanguineus (Fr) MurrrdquoJournal of Ethnopharmacology vol 45 no 3 pp 177ndash181 1995
[18] W Andlauer and P Furst ldquoNutraceuticals a piece of historypresent status and outlookrdquo Food Research International vol 35no 2-3 pp 171ndash176 2002
[19] C L Kruger and S W Mann ldquoSafety evaluation of functionalingredientsrdquo Food and Chemical Toxicology vol 41 no 6 pp793ndash805 2003
[20] P H Mattila V I Piironen E J Uusi-Rauva and P E Koivis-toinen ldquoVitamin D contents in edible mushroomsrdquo Journal ofAgricultural and Food Chemistry vol 42 no 11 pp 2449ndash24531994
[21] R P Tewari ldquoMushroom their role in nature and societyrdquo inFrontiers inMushroomBiotechnology R D Rai R C Upadhyayand S R Sharma Eds pp 1ndash8 NRCM Chambaghat India2005
[22] K C Semwal S L Stephenson V K Bhatt and R P BhattldquoEdible mushrooms of the Northwestern Himalaya India astudy of indigenous knowledge distribution and diversityrdquoMycosphere vol 5 no 3 pp 440ndash461 2014
[23] E V Crisan and A Sands ldquoEdible mushrooms nutritionalvaluerdquo in The Biology and Cultivation of Edible Mushrooms ST Chang and W A Hayes Eds pp 137ndash165 Academic PressNew York NY USA 1978
[24] N S Atri S K Sharma R Joshi A Gulati and A Gula-ti ldquoNutritional and neutraceutical composition of five wildculinary-medicinal species of genus Pleurotus (Higher Basid-iomycetes) from Northwest Indiardquo International Journal ofMedicinal Mushrooms vol 15 no 1 pp 49ndash56 2013
[25] L Barros T Cruz P Baptista L M Estevinho and I C FR Ferreira ldquoWild and commercial mushrooms as source ofnutrients and nutraceuticalsrdquo Food and Chemical Toxicologyvol 46 no 8 pp 2742ndash2747 2008
[26] M Nagata and I Yamashita ldquoSimple method for simultaneousdetermination of chlorophyll and carotenoids in tomato fruitrdquoNippon Shokuhin Kogyo Gakkaishi vol 39 no 10 pp 925ndash9281992
[27] V L Singleton and J A Rossi ldquoColorimetric detection oftotal phenolics with phosphomolybdic-phosphotungstic acidreagentsrdquo American Journal of Enology and Viticulture vol 16pp 144ndash158 1965
[28] L K Jagadish V Venkata Krishnan R Shenbhagaramanand V Kaviyarasan ldquoComparitive study on the antioxidantanticancer and antimicrobial property of Agaricus bisporus (JE Lange) Imbach before and after boilingrdquo African Journal ofBiotechnology vol 8 no 4 pp 654ndash661 2009
[29] E Vamanu and S Nita ldquoAntioxidant capacity and the cor-relation with major phenolic compounds anthocyanin andtocopherol content in various extracts from the wild edibleBoletus edulis mushroomrdquo BioMed Research International vol2013 Article ID 313905 11 pages 2013
[30] E Vamanu ldquoBiological activities of the polysaccharides pro-duced in submerged culture of two edible Pleurotus ostreatus
12 BioMed Research International
mushroomsrdquo Journal of Biomedicine and Biotechnology vol2012 Article ID 565974 8 pages 2012
[31] P L Li L Huo W Su et al ldquoFree radical-scavenging capacityantioxidant activity and phenolic content of Pouzolzia zeylan-icardquo Journal of the Serbian Chemical Society vol 76 no 5 pp709ndash717 2011
[32] C Papuc M Crivineanu G Goran V Nicorescu and N Dur-dun ldquoFree radicals scavenging and antioxidant activity ofEuropean mistletoe (Viscum album) and European birthwort(Aristolochia clematitis)rdquo Revista de Chimie vol 61 no 7 pp619ndash622 2010
[33] VOOyetayo CDong andY Yao ldquoAntioxidant and antimicro-bial properties of aqueous extract from Dictyophora indusiatardquoThe Open Mycology Journal vol 3 no 1 pp 20ndash26 2009
[34] S Marklund and G Marklund ldquoInvolvement of the superoxideanion radical in the autoxidation of pyrogallol and a convenientassay for superoxide dismutaserdquo European Journal of Biochem-istry vol 47 no 3 pp 469ndash474 1974
[35] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996
[36] D J Huang b Ou and R L Prior ldquoThe chemistry behindantioxidant capacity assaysrdquo Journal of Agricultural and FoodChemistry vol 53 no 6 pp 1841ndash1856 2005
[37] P Manzi A Aguzzi and L Pizzoferrato ldquoNutritional value ofmushrooms widely consumed in Italyrdquo Food Chemistry vol 73no 3 pp 321ndash325 2001
[38] P Manzi S Marconi A Aguzzi and L Pizzoferrato ldquoCommer-cial mushrooms nutritional quality and effect of cookingrdquo FoodChemistry vol 84 no 2 pp 201ndash206 2004
[39] I P Krbavcic and I C Baric ldquoInfluence of deep fat fryingon some nutritional parameters of novel food based on mush-rooms and fresh soft cheeserdquo Food Chemistry vol 84 no 3 pp417ndash419 2004
[40] D Agrahar-Murugkar andG Subbulakshmi ldquoNutritional valueof edible wild mushrooms collected from the Khasi hills ofMeghalayardquo Food Chemistry vol 89 no 4 pp 599ndash603 2005
[41] V A Dıez and A Alvarez ldquoCompositional and nutritionalstudies on two wild edible mushrooms from northwest SpainrdquoFood Chemistry vol 75 no 4 pp 417ndash422 2001
[42] L Barros M-J Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007
[43] S K Sharma and N S Atri ldquoNutraceutical composition ofwild species of genus Lentinus Fr fromNothern Indiardquo CurrentResearch in Environmental ampAppliedMycology vol 4 pp 11ndash322014
[44] P J Kanu K Zhu J Baby Kanu H Zhou H Qian and K ZhuldquoBiologically active components and nutraceuticals in sesameand related products a review and prospectrdquo Trends in FoodScience and Technology vol 18 no 12 pp 599ndash608 2007
[45] F S Reis E Pereira L Barros M J Sousa A Martins and I CF R Ferreira ldquoBiomolecule profiles in inediblewildmushroomswith antioxidant valuerdquoMolecules vol 16 no 6 pp 4328ndash43382011
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
232 Amino Acids Powdered samples (01 g) were extractedwith 25mL followed by 15mL and 1mL of 70 aqueousmethanol After this centrifugation was done for 10 minutes(4000 rpm) at 4∘C Supernatants were dissolved in aque-ous methanol and the volume was made up to 5mL Itwas now passed through Millipore filter (045 120583m) Afterthis samples (10 120583L) were dried using vacuum oven andto these dried samples 20 120583L derivatising agent (preparedby ethanol triethylamine water phenylisothiocaynate) wasmixed with it and redried it Now the samples were left for 25minutes at room temperature Lastly 1mL ethanol was addedand injected into UPLC (Waters India Pvt Ltd)
233 Tocopherol Composition Tocopherol composition wasestimated following standard method [25] For this sam-ples were mixed with butylated hydroxytoluene (BHT) inhexane (10mgmL 100 120583120583L) and IS solution in hexane (120575tocopherol 16 120583gmL 250 120583L) Thereafter samples (500mg)were vortexed for 1min with methanol (4mL)Then sampleswere again vortexed with hexane (4mL) After this 2mL ofsaturated NaCl aqueous solution was added and the mixturewas vortexed (1min) followed by centrifugation at 4000 gfor 5min and the upper layer was separated The sampleswere again reextracted twice with hexane The extractswere then vacuum-dried and redissolved in hexane (1mL)followed by dehydration with anhydrous sodium sulphatethen filtered and transferred into a dark injection vial andanalysed byHPLC (Waters India Pvt Ltd) Chromatographiccomparisons were made by authentic standards Tocopherolcontents in mushroom samples were expressed in 120583g per g ofdry mushroom
234 Evaluation of Other Bioactive Compounds For 120573-carotene and lycopene estimation dried powdered samples(sim5 g) were extracted with 100mL of methanol at 25∘C(150 rpm) for 24 hours and filtered through Whatman Num-ber 4 filter paper The residue was again reextracted with2 additional 100mL portions of methanol These extractswere evaporated to dryness at 42∘C then redissolved inmethanol at a concentration of 50mgmL and stored at 4∘CThe driedmethanolic extract (100mg) was shaken vigorouslywith 10mL of acetonehexanemixture (4 6) for 1 minute andfiltered The absorbance of the filtrate was measured at 453505 and 663 nm [26] 120573-carotene and lycopene content wereestimated using the following equation
Lycopene (mg100mL)
= (00458times119860663) + (0372times119860505)
minus (00806times119860453) 120573-carotene (mg100mL)
= (0216times119860663) minus (0304times119860505)
+ (0452times119860453)
(1)
For phenolic compounds quantification powdered sam-ples (1mL) were mixed with Folin and Ciocalteursquos phenolreagent (1mL) After 3 minutes 1mL of saturated sodiumcarbonate solution was added to the mixture and the volume
was adjusted to 10mL with distilled water The reaction waskept in the dark for 90 minutes after which the absorbancewas read at 725 nm Gallic acid was used to calculate thestandard curve (001ndash04mM 1198772 = 09999) and the resultswere expressed as milligrams of gallic acid equivalents pergram of extract [27]
Total flavonoids of the sample extracts were measuredby AlCl
3method [28] For this an aqueous extract (15mL)
was mixed with deionized distilled water (5mL) and 03mLof 5 NaNO
2 After five minutes of incubation at room
temperature 15mL of 2 aluminium trichloride (AlCl3)
solution was added After the next 6 minutes 2mL of 1MNaOH was added The mixture was vigorously shaken onorbital shaker for 5min at 200 rpm and the absorbance wasread at 510 nm against a blank Quercetin with differentconcentrations was used as a standard
For ascorbic acid quantification standard ascorbic acidsolution (5mL L-ascorbic acid in 3 phosphoric acid) wasadded to 5mL of phosphoric acid A microburette was filledwith dye and the samples were titrated with the dye solutionto a pink color which persisted for 15 seconds The dyefactor (milligrams of ascorbic acid per milliliter of dye usingformula 05titrate) was determined A sample was preparedby taking 10 g of sample grounded in metaphosphoric acidand the volume was increased up to 100mL It was titratedafter filtration until a pink color appeared [24] The amountof ascorbic acid was calculated with the use of the followingequation
mg of ascorbic acid per100 g or mL
=Titrate times Dye factor times Vol madeAliquot of extract times wt of sample
times 100(2)
Anthocyanidins were quantified by using standardmethod [29] Briefly 05 g of samples was mixed withthe solvent (mixture of 85 15 (vv) of ethyl alcohol andhydrochloric acid 15M) followed by ultrasonication for 15minutes and filtration throughWhatman filter paper number1 Standard solution was prepared with cyaniding chloridewith a concentration of 5ndash15120583gmL in solvent which wasused The absorption was measured at 546 nm The totalquantity of anthocyanins (expressed in g of cyanidingchloride100 g extract) = (119860
119901times 119898st times 119891 times 100)(119860 st times 119898119901)
where 119860119901is absorption rate of the sample solution 119898
119901is
mass of the processed sample in g 119860 st is absorption rate ofthe standard solution 119898st is mass of the processed standardsolution in g and 119891 is dilution coefficient
24 Antioxidant Assays DPPH scavenging activity was mea-sured with adding DPPH (200120583L) solution at differentconcentrations (2ndash10mgmL) to 005mL of the samples(dissolved in ethanol) An equal amount of ethanol was addedto the control Ascorbic acid was used as the control [30]Theabsorbancewas read after 20min at 517 nm and the inhibitionwas calculated using the formula
where 1198600was the absorbance of the control and 119860
119875was the
absorbance in the presence of the sampleFor ABTS radical scavenging activity 10120583L of the sample
was added to 4mL of the diluted ABTS∙+ solution (preparedby adding 7mM of the ABTS stock solution to 245mMpotassium persulfate kept in the dark at room temperaturefor 12ndash16 h before use) The solution was then diluted with5mM phosphate-buffered saline (pH 74) and absorbancewas measured at 730 nm after 30min [31] The ABTS radicalscavenging activity was calculated as
119878 = (119860controlminus119860 sample
119860control)times 100 (4)
For reducing power estimation samples (200 120583L) weremixed with sodium phosphate buffer (pH 66) 1mM FeSO
4
and 1 potassium ferricyanide and incubated for 20minat 50∘C after that trichloroacetic acid was added and themixtures were centrifuged Supernatant (25mL) was mixedwith an equal volume of water and 05mL 01 FeCl
3 The
absorbance was measured at 700 nm [32]For Fe2+ chelating activity 1mL of the sample (2ndash10mg
mL) was mixed with 37mL of ultrapure water after thatthe mixture was reacted with ferrous chloride (2mmolL01mL) and ferrozine (5mmolL 02mL) for 20min and theabsorbance was read at 562 nm with using EDTA as controlThe chelating activity was calculated using the formula
where 119860119887is the absorbance of the blank and 119860
119904is the
absorbance in the presence of the extract [33]The scavenging activity of superoxide anion radicals
was measured following standard method [34] Samples (0ndash20mgmL 1mL) and Tris-HCl buffer (500mM pH 823mL) were incubated in a water bath at 25∘C for 20min andafter this pyrogallic acid (50mM 04mL) was added HClsolution (80M 01mL) was added to terminate the reactionafter 4min The absorbance of the mixture was measuredat 320 nm The scavenging ability was calculated using thefollowing formula
scavenging ability () = (1 minus119860 sample
119860control)times 100 (6)
where 119860control is the absorbance of control without thepolysaccharide sample and 119860 sample is the absorbance in thepresence of the polysaccharide sample
For ferric reducing antioxidant power (FRAP) assayfirstly FRAP reagent was prepared by mixing TPTZ (25mL10mM in 40mM HCl) 25mL of 300mM acetate buffer and25mL of FeCl
3sdot6H2O After this freshly prepared FRAP
reagent (18mL) was taken in a test tube and incubated at30∘C in water bath for 10 minutes Then absorbance wasread at 0min (119905
0) After this 100 120583L of sample extract or
standard and 100 120583L of distilled water were added to the testtube mixed and incubated at 30∘C for 30 minutes Thenthe absorbance was taken at 593 nm (119905
30) Ferrous sulphate
was used as standard [35 36] FRAP activity was determinedagainst a standard curve of ferrous sulphate and the valueswere expressed as 120583M Fe2+ equivalents per gram of extractand calculated using the following equation
FRAP value = Absorbance (sample+ FRAP reagent)
minusAbsorbance (FRAP reagent) (7)
25 Statistical Analysis All experiments were performed 3times and with 3 replicates The results were analyzed usingone-way analysis of variance (ANOVA) 119901 lt 005 was con-sidered significant and SPSS software (SPSS Inc ChicagoIL USA) was used to calculate differences Tukey-HSD at119901 lt 005 test was used to determine significant differences
3 Results
31 Chemical Evaluation Nutrient composition of the wildculinary mushrooms is shown in Table 2 Protein was foundin high levels and varied between 516 in Inocybe splendensand 2263 in Agaricus arvensis Protein percentage inPleurotus cystidiosus (2069) Amanita caesarea (1972)Agaricus campestris (1838) Cantharellus cibarius (1819)and Lentinus cladopus (1859) was also found to be highAll the twenty culinary species were found to be low in fatcontent Fat ranged from010 in Laccaria laccata to 038 inInocybe splendens In general these wild culinary mushroomsconsumed by local people were found to be higher in proteinand low in fat although differences were observed in netvalue of individual species Crude fibres ranged from 108in Hygrocybe coccinea to 242 in Lentinus cladopus Ashcontent varied between 011 in Inocybe splendens and 096in Agaricus arvensis Carbohydrates calculated by differencewere also found to be in abundant amounts and theirpercentage was ranged from 3119 in Inocybe splendens to5712 in Agaricus arvensis Nutrient contents of Inocybesplendens Hygrocybe nivea and Conocybe tenera were foundto be less as compared to other species
There are several reports about the toxicity reports due tomushrooms on humans in these areas and hence preliminarystudies were done to check the toxicity level of mushroomsFor all the twenty species being used by the people forculinary purposes the test was found to be negative Thatmeans these species are nontoxic and hence recommendedfor consumption
All the culinarymushroom species contained glucose andrhamnose as the principal carbohydrates (Table 3) Neverthe-less the present study also describes the presence of xyloseand mannose in all the studied species However galactoseand fructose were detected in very low percentage in someof the species Russula mairei contained lowest percentageof glucose (2160) and Agaricus arvensis contained highestpercentage of glucose (6412)
32 Bioactive Evaluation The results of fatty acid composi-tion (total saturated fatty acids SFA monounsaturated fattyacids MUFA and polyunsaturated fatty acids PUFA) of allthe species are shown in Table 4 In general the major fatty
BioMed Research International 5
Table 2 Percent chemical composition of twenty wild culinary mushroom species collected from northern Himalayan regions
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 3 Percent monosaccharide composition of twenty species collected from northern Himalayan regions showing richness in glucoserhamnose mannose xylose and galactose and fructose in lower percentage
Russula lepida 515 plusmn 01d 3261 plusmn 36f 1316 plusmn 12e 231 plusmn 01c ND NDR mairei 426 plusmn 01c 2160 plusmn 21a 1281 plusmn 16d 212 plusmn 00c ND NDND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
6 BioMed Research International
Table4Percentfattyacid
compo
sitionof
allthe
twentywild
culin
arymushroo
mspeciesc
ollected
from
different
region
sofn
orthernHim
alayas
Species
C900
C100
C120
C160
C1611
C171
C181
C182
C202
UI
Agaricu
sarvensis
006plusmn00a
030plusmn001
a028plusmn00a
186plusmn003
a006plusmn00a
009plusmn00a
018plusmn00a
324plusmn00a
221plusmn02a
057plusmn002
a
Acampestr
is003plusmn00a
026plusmn002
a021plusmn00a
119plusmn002
a009plusmn00a
002plusmn00a
035plusmn00a
204plusmn00a
218plusmn03a
046plusmn002
a
Acomtulus
006plusmn00a
022plusmn00a
ND
117plusmn001
a007plusmn00a
009plusmn00a
038plusmn00a
173plusmn00a
134plusmn01a
036plusmn001
a
Asilvicola
008plusmn00a
033plusmn00a
022plusmn00a
156plusmn002
a009plusmn00a
001plusmn00a
015plusmn00a
154plusmn00a
221plusmn03a
040plusmn001
a
Amanita
caesarea
004plusmn00a
032plusmn002
a019plusmn00a
196plusmn002
a012plusmn00a
005plusmn00a
031plusmn00a
219plusmn00a
128plusmn00a
039plusmn001
a
Acitrin
a006plusmn00a
028plusmn001
aND
091plusmn00a
011plusmn00a
008plusmn00a
031plusmn00a
159plusmn00a
168plusmn00a
037plusmn001
a
Afulva
004plusmn00a
021plusmn001
a025plusmn002
a010plusmn00a
008plusmn00a
004plusmn00a
015plusmn00a
164plusmn00a
123plusmn00a
031plusmn002
a
Cantharellu
scibarius
007plusmn00a
033plusmn002
a022plusmn001
a110plusmn002
a007plusmn00a
001plusmn00a
037plusmn00a
294plusmn00a
118plusmn01a
045plusmn002
a
Conocybe
tenera
006plusmn00a
020plusmn00a
ND
007plusmn00a
005plusmn00a
002plusmn00a
035plusmn00a
014plusmn00a
138plusmn00a
019plusmn00a
Gymnopilusjun
onius
005plusmn00a
031plusmn001
a014plusmn001
a18
7plusmn003
a008plusmn00a
002plusmn00a
088plusmn00a
049plusmn00a
121plusmn
01a
026plusmn001
a
Hygrocybe
coccinea
004plusmn00a
025plusmn002
a017plusmn00a
033plusmn00a
001plusmn00a
005plusmn00a
017plusmn00a
024plusmn00a
119plusmn00a
016plusmn00a
Hnivea
007plusmn00a
022plusmn001
aND
005plusmn00a
004plusmn00a
001plusmn00a
053plusmn00a
004plusmn00a
138plusmn02a
020plusmn00a
Inocybesplendens
002plusmn00a
019plusmn001
a023plusmn001
a089plusmn00a
002plusmn00a
003plusmn00a
018plusmn00a
010plusmn00a
020plusmn00a
005plusmn00a
Lactariusp
ubescens
001plusmn00a
023plusmn001
a016plusmn00a
075plusmn00a
007plusmn00a
008plusmn00a
026plusmn00a
344plusmn00a
083plusmn00a
046plusmn002
a
Laccarialaccata
002plusmn00a
022plusmn00a
ND
101plusmn
00a
006plusmn00a
009plusmn00a
092plusmn00a
284plusmn00a
029plusmn00a
042plusmn001
a
Lepista
nuda
003plusmn00a
024plusmn00a
ND
043plusmn00a
001plusmn00a
002plusmn00a
020plusmn00a
204plusmn00a
037plusmn00a
026plusmn00a
Lentinus
cladopu
s009plusmn00a
033plusmn002
a032plusmn00a
146plusmn00a
055plusmn00a
001plusmn00a
032plusmn00a
245plusmn00a
295plusmn00a
057plusmn002
a
Pleurotuscystid
iosus
006plusmn00a
031plusmn001
a028plusmn00a
135plusmn00a
009plusmn00a
005plusmn00a
098plusmn00a
204plusmn00a
221plusmn02a
052plusmn002
a
Russu
lalep
ida
006plusmn00a
019plusmn001
aND
033plusmn00a
005plusmn00a
005plusmn00a
049plusmn00a
204plusmn03a
165plusmn00a
042plusmn001
a
Rmairei
001plusmn00a
017plusmn001
aND
026plusmn00a
008plusmn00a
008plusmn00a
063plusmn00a
201plusmn02a
010plusmn00a
028plusmn001
a
ND=no
tdetected
Values
aree
xpressed
asmeanplusmnSE
andlette
rsin
thes
uperscrip
tsrepresentthe
significantd
ifference
ineach
columnwith119901le005accordingto
Tukeyrsquos
test
BioMed Research International 7
Table 5 Percent amino acids composition of all the wild culinary species collected from northern Himalayas
Values are expressed as mean plusmn SE and letters in superscript represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
acids found in the studied species were linoleic acid (C182)followed by oleic acid (C181) and palmitic acid (C160)Besides these three main fatty acids already described sixmore were identified and quantified PUFA were the maingroup of fatty acids documented in all the species Agaricusarvensis Amanita caesarea Cantharellus cibarius Lentinuscladopus and Pleurotus cystidiosus contained lower value ofMUFA but higher percentage of PUFA as compared to otherspecies due to the higher percentage of linoleic acidHoweverUFApredominated over SFA in all the studied species rangingfrom 65 to 70
Amino acid composition of all the species is shown inTable 5 In all the species aspartic acid (019ndash039) wasfound to be predominated amino acid followed by tyrosine(010ndash021) arginine (012ndash029) alanine (004ndash014)and proline (001ndash007) Amanita caesarea Agaricus arven-sis Cantharellus cibarius Lentinus cladopus and Pleurotuscystidiosus contained maximum amount of these aminoacids Tocopherol contents in all the studied mushroomspecies including three wild are detailed in Table 6 120572-tocopherol and 120573-tocopherol were found to be present in allthe species However 120574-tocopherol was documented fromfew species only Tocopherol content was ranged from 090to 433 120583gg in all the species Cantharellus cibarius (433 plusmn00 120583gg) contained all the three isomers in higher amount ascompared to other species 120573-tocopherol was found in higheramounts as compared to 120572-tocopherol 120574-tocopherol wasdetected only in nine species Cantharellus cibarius (433 plusmn00 120583gg) contained higher amounts of 120574 tocopherol
Results obtained for 120573-carotene lycopene flavonoidsascorbic acid and anthocyanidins composition of all the
twenty species are presented in Table 7 Phenols werethe major antioxidant component detected in significantamounts from all the species (1912ndash6336mgg) followedby anthocyanidins (614ndash1425mg cyanidin chloride100 gextract) flavonoids (114ndash417mgg) ascorbic acid whichwas found in small amounts (020ndash099mgg) 120573-carotene(021ndash079 120583g100 g) and lycopene (019ndash038 120583g100 g) Eachspecies differed with other species in net amounts of all thesecomponents Species like Agaricus arvensis Amanita cae-sarea Gymnopilus junonius Lentinus cladopus and Pleurotuscystidiosus contained higher values of these components ascompared to other species
33 Antioxidant Evaluation Antioxidant properties of allthe species were expressed as EC
radical scavenging activity in all the species showed differ-ences in effectiveness in antioxidant properties Among allthe species Cantharellus cibarius showed lowest EC
50values
(176 plusmn 02mgmL) followed by Amanita caesarea (202 plusmn02mgmL) and Agaricus arvensis (212 plusmn 04mgmL) Otherspecies showed slightly higher EC
50values and therefore
lesserDPPH radical scavenging activityCantharellus cibariusshowed higher DPPH radical scavenging activity and Inocybesplendens showed lower DPPH radical scavenging activitythan other species
For ABTS radical scavenging activities EC50ranged from
426 to 145mgmL Lowest EC50
values were obtained forAmanita caesarea (145 plusmn 06mgmL) showing high antiox-idant activities of this species Higher EC
50values were
8 BioMed Research International
Table 6 Tocopherol composition (120583gg) of twenty species collected from northern Himalayan regions
ND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 7 Other bioactive compounds evaluated from all the species
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
BioMed Research International 9
Table 8 EC50 values for different antioxidant assays on twenty wild culinary species collected from northern Himalayas
obtained for Hygrocybe nivea (426 plusmn 05mgmL) showinglowest ABTS radical scavenging activities of this speciesReducing power of Cantharellus cibarius (162 plusmn 05mgmL)was measured higher than other species Higher EC
50values
for reducing activity were measured in Inocybe splendens(519 plusmn 01mgmL)
Higher effectiveness in ferrous ion chelating activitywas detected in Agaricus arvensis (114 plusmn 01mgmL) andlow effectiveness was detected in Russula mairei (295 plusmn03mgmL) Nevertheless Cantharellus cibarius (127 plusmn02mgmL) Amanita caesarea (140 plusmn 04mgmL) Hygro-cybe coccinea (135 plusmn 02mgmL) and H nivea (145 plusmn06mgmL) showed lower EC
50values than remaining
species EC50
values of scavenging ability on superoxideradical were found to be maximum in Amanita caesarea(044 plusmn 03mgmL) and minimum in Lactarius pubescens(221 plusmn 06mgmL) Gymnopilus junonius showed maximumFRAP activity with least EC
Although there are previous reports on documentation ofculinary edible species from the regions native to northernHimalayas but there are no reports on the evaluation stud-ies as well as toxicity status of all these culinary speciesThe wild edible species Agaricus bisporus Boletus edulis
Morchella esculenta Cordyceps sinensis and Lentinula edodeswhich have been extensively worked out in India and otherparts of the world for their compositional and medicinalaspects have not been undertaken for investigations presently[37ndash40] Compositional studies showed that most of theculinary species were rich in protein carbohydrates andlow in fat There are several reports on richness of wildedible mushrooms with protein and carbohydrate contentsand low fat levels which directly make them nutritionallyrich [24 25 41] Nevertheless under present studies thedifferences between the nutrient concentrations of all thespecies differed but Agaricus arvensis Pleurotus cystidiosusAmanita caesarea Agaricus campestris Cantharellus cibariusand Lentinus cladopus showed higher nutrient percentagewhich is comparable to other wild and commercially cul-tivated species [25 42] The crude fat content detected inall the species was not found to be significantly differentCrude fibres were detected in appreciable percentage fromall the species which make them important in nutritionalpoint of viewThe results are in conformity with the previousreports on several wild edible Pleurotus and Lentinus speciesfrom northwest India [24 43] The species Inocybe splendensHygrocybe nivea and Conocybe tenera were not found tocontain higher percentage of nutrients Although previousreports showed that nutrients composition in wild species isless as compared to cultivated species [25] however Agaricusarvensis Pleurotus cystidiosus Amanita caesarea Agaricuscampestris Cantharellus cibarius and Lentinus cladopus were
10 BioMed Research International
found to be rich in protein and carbohydrates similar tocommercially grown species [25]
Fatty acid composition showed the dominance of UFAover SFA in all the studied mushrooms species which is inconformity with the other studies [41] The differences wereobserved in net amounts in all the species Unsaturationindex of Agaricus arvensis and Lentinus cladopus (057 plusmn002) was found to be significantly higher than otherspecies Whereas Inocybe splendens (005 plusmn 00) showedleast Unsaturation Index High UFA shows the medicinalimportance of these culinary mushrooms as these increasethe HDL cholesterol and decrease LDL cholesterol triacyl-glycerol lipid oxidation and LDL susceptibility to oxidation[44] Predominance of UFA over SFA in all the species showssimilar results as obtained for other wild and commerciallycultivated species [24 25] 120572- and 120573-tocopherol were detectedin higher amounts than third isomer in all the studied speciesSimilar findings were made in other wild and cultivatedspecies with higher 120572- and 120573-tocopherol than 120574-tocopherol[25] The high levels of these two compounds correspondwith a higher oxidative activity which is associated withcardiovascular protection [45] Phenolic compounds weredetected in higher amounts than other bioactive compoundsPresence of high phenolic compounds accounts for the highantioxidant properties of all the species [42] 120573-carotenelycopene and ascorbic acids were detected in low amountsAnthocyanidins were also detected from these wild speciesin appreciable amounts The presence of these functionalmedicinal compounds inmedicinal andor edible mushroomis due to habitat or substrates in which these grow to be highin the functionalmoleculesThe categories of thesemoleculesare anthocyanidins beta-glucans selenium ganoderic acidtriterpenes or cordycepinThe compounds identified in theseextracts show that at least a part of the functional compoundsin medicinal andor edible mushroom is due to growingmushrooms on substrates that are high in the functionalmolecules To these categories can be added anthocyanidinsbeta-glucans selenium ganoderic acid triterpenes or cordy-cepinThe amounts of these have been found to vary with thetype of extraction as ethanolic extract yields higher amountsof anthocyanidins as compared to methanolic hot water andcold water extracts [29]
All the studied species showed significant antioxidantproperties measured on the basis of EC
50values Never-
theless each species showed different antioxidant activitywith highly effective and less effective EC
50values Better
antioxidant properties of some species are due to presence ofhigher phenolic compounds 120573-carotene lycopene ascorbicacids anthocyanidins and tocopherol amounts in themHigh reducing power of some species is due to the presence ofhigher amounts of reducers (antioxidants) in them Presentlyinvestigated species are commonly used for culinary pur-poses in the regions native to northern Himalayas Many ofthe species in these regions are not evaluated previously fordetailed compositional analysisTheir knowledge is restrictedto old aged villagers of the regions and neglected for thecommercial exploitations There are no positive reportson toxicity of these mushrooms analyzed presently hencethese are safe for further experimental work related to
drug discovery All the culinary species contained impor-tant and useful nutraceuticals such as unsaturated fattyacids phenolics carotenoids ascorbic acid tocopherolsand anthocyanidins besides these some important aminoacids were detected in these mushrooms which could beused for the purpose of being used as functional ingredi-ents Since nutraceuticals are powerful in maintaining andpromoting health longevity and life quality the commer-cial exploitation of these species will certainly create animpact on nutritional therapy and also will be beneficialtodayrsquos food industry Direct use of these species for con-sumption and other culinary aspects is safe and healthpromoting with advantage of the additive effects of allthe bioactive and antioxidant compounds present in thesespecies
Abbreviations
AlCl3 Aluminum trichloride
ANOVA Analysis of varianceDW Distilled waterDPPH 22-Diphenyl-1-picrylhydrazylFeCl3 Ferric chloride
FRAP Ferric reducing antioxidant powerg GramsGAEs Gallic acid equivalentsGC Gas ChromatographyHCl Hydrochloric acidH2O Water
H3PO4 Phosphoric acid
HPLC High performance liquid chromatographyK2HPO4 Potassium hydrogen phosphate
The authors declare that there is no conflict of interestsregarding the publication of this paper
BioMed Research International 11
Acknowledgment
The authors wish to thank Science and Engineering BoardDepartment of Science and Technology New Delhi forresearch Grant (SBFTLS-042013) to carry out presentstudies
References
[1] W M Breene ldquoNutritional and medicinal value of specialtymushroomsrdquo Journal of Food Protection vol 53 pp 883ndash8941990
[2] S Chang and P Miles ldquoMushroom biologymdasha new disciplinerdquoMycologist vol 6 no 2 pp 64ndash65 1992
[3] P Manzi L Gambelli S Marconi V Vivanti and L Pizzoferra-to ldquoNutrients in edible mushrooms an inter-species compara-tive studyrdquo Food Chemistry vol 65 no 4 pp 477ndash482 1999
[4] P Bobek O Ozdın and M Mikus ldquoDietary oyster mushroom(Pleurotus ostreatus) accelerates plasma cholesterol turnover inhypercholesterolaemic ratsrdquo Physiological Research vol 44 no5 pp 287ndash291 1995
[5] P Bobek and S Galbavy ldquoHypocholesterolemic and antiathero-genic effect of oystermushroom (Pleurotus ostreatus) in rabbitsrdquoNahrung vol 43 no 5 pp 339ndash342 1999
[6] S Khatun A Islam U Cakilcioglu and N Chatterjee ldquoRe-search on mushroom as a potential source of nutraceuticals areview on Indian perspectiverdquo American Journal of Experimen-tal Agriculture vol 2 no 1 pp 47ndash73 2012
[7] J A Vaz L Barros A Martins J S Morais M H Vasconcelosand I C F R Ferreira ldquoPhenolic profile of seventeen por-tuguese wild mushroomsrdquo LWT Food Science and Technologyvol 44 no 1 pp 343ndash346 2011
[8] J A Vaz L Barros A Martins C Santos-Buelga M HVasconcelos and I C F R Ferreira ldquoChemical compositionof wild edible mushrooms and antioxidant properties of theirwater soluble polysaccharidic and ethanolic fractionsrdquo FoodChemistry vol 126 no 2 pp 610ndash616 2011
[9] T Ozen C Darcan O Aktop and I Turkekul ldquoScreening ofantioxidant antimicrobial activities and chemical contents ofedible mushrooms wildly grown in the Black Sea region ofTurkeyrdquo Combinatorial Chemistry amp High Throughput Screen-ing vol 14 no 2 pp 72ndash84 2011
[10] V E C Ooi and F Liu ldquoA Review of pharmacological activitiesof mushroom polysaccharidesrdquo International Journal of Medic-inal Mushrooms vol 1 no 3 pp 195ndash206 1999
[11] S PWasser and A LWeis ldquoMedicinal properties of substancesoccurring in higher Basidiomycete mushroom current per-spectiverdquo International Journal of Medicinal Mushrooms vol 1pp 31ndash62 1999
[12] P M Kidd ldquoThe use of mushroom glucans and proteoglycansin cancer treatmentrdquo Alternative Medicine Review vol 5 no 1pp 4ndash27 2000
[13] C Israilides and A Philippoussis ldquoBio-technologies of recy-cling agro-industrial wastes for the production of commerciallyimportant fungal polysaccharides andmushroomsrdquo Biotechnol-ogy and Genetic Engineering Reviews vol 20 pp 247ndash259 2003
[14] T Mizuno ldquoBioactive biomolecules of mushrooms food func-tion and medicinal effect of mushroom fungirdquo Food ReviewsInternational vol 11 no 1 pp 5ndash21 1995
[15] Y Kabir and S Kimura ldquoDietary mushrooms reduce bloodpressure in spontaneously hypertensive rats (SHR)rdquo Journal of
Nutritional Science and Vitaminology vol 35 no 1 pp 91ndash941989
[16] J-L Mau S-Y Tsai Y-H Tseng and S-J Huang ldquoAntioxidantproperties of hot water extracts from Ganoderma tsugae Mur-rillrdquo LWTmdashFood Science and Technology vol 38 no 6 pp 589ndash597 2005
[17] A Smania F D Monache E F A Smania M L Gil L CBenchetrit and F S Cruz ldquoAntibacterial activity of a substanceproduced by the fungus Pycnoporus sanguineus (Fr) MurrrdquoJournal of Ethnopharmacology vol 45 no 3 pp 177ndash181 1995
[18] W Andlauer and P Furst ldquoNutraceuticals a piece of historypresent status and outlookrdquo Food Research International vol 35no 2-3 pp 171ndash176 2002
[19] C L Kruger and S W Mann ldquoSafety evaluation of functionalingredientsrdquo Food and Chemical Toxicology vol 41 no 6 pp793ndash805 2003
[20] P H Mattila V I Piironen E J Uusi-Rauva and P E Koivis-toinen ldquoVitamin D contents in edible mushroomsrdquo Journal ofAgricultural and Food Chemistry vol 42 no 11 pp 2449ndash24531994
[21] R P Tewari ldquoMushroom their role in nature and societyrdquo inFrontiers inMushroomBiotechnology R D Rai R C Upadhyayand S R Sharma Eds pp 1ndash8 NRCM Chambaghat India2005
[22] K C Semwal S L Stephenson V K Bhatt and R P BhattldquoEdible mushrooms of the Northwestern Himalaya India astudy of indigenous knowledge distribution and diversityrdquoMycosphere vol 5 no 3 pp 440ndash461 2014
[23] E V Crisan and A Sands ldquoEdible mushrooms nutritionalvaluerdquo in The Biology and Cultivation of Edible Mushrooms ST Chang and W A Hayes Eds pp 137ndash165 Academic PressNew York NY USA 1978
[24] N S Atri S K Sharma R Joshi A Gulati and A Gula-ti ldquoNutritional and neutraceutical composition of five wildculinary-medicinal species of genus Pleurotus (Higher Basid-iomycetes) from Northwest Indiardquo International Journal ofMedicinal Mushrooms vol 15 no 1 pp 49ndash56 2013
[25] L Barros T Cruz P Baptista L M Estevinho and I C FR Ferreira ldquoWild and commercial mushrooms as source ofnutrients and nutraceuticalsrdquo Food and Chemical Toxicologyvol 46 no 8 pp 2742ndash2747 2008
[26] M Nagata and I Yamashita ldquoSimple method for simultaneousdetermination of chlorophyll and carotenoids in tomato fruitrdquoNippon Shokuhin Kogyo Gakkaishi vol 39 no 10 pp 925ndash9281992
[27] V L Singleton and J A Rossi ldquoColorimetric detection oftotal phenolics with phosphomolybdic-phosphotungstic acidreagentsrdquo American Journal of Enology and Viticulture vol 16pp 144ndash158 1965
[28] L K Jagadish V Venkata Krishnan R Shenbhagaramanand V Kaviyarasan ldquoComparitive study on the antioxidantanticancer and antimicrobial property of Agaricus bisporus (JE Lange) Imbach before and after boilingrdquo African Journal ofBiotechnology vol 8 no 4 pp 654ndash661 2009
[29] E Vamanu and S Nita ldquoAntioxidant capacity and the cor-relation with major phenolic compounds anthocyanin andtocopherol content in various extracts from the wild edibleBoletus edulis mushroomrdquo BioMed Research International vol2013 Article ID 313905 11 pages 2013
[30] E Vamanu ldquoBiological activities of the polysaccharides pro-duced in submerged culture of two edible Pleurotus ostreatus
12 BioMed Research International
mushroomsrdquo Journal of Biomedicine and Biotechnology vol2012 Article ID 565974 8 pages 2012
[31] P L Li L Huo W Su et al ldquoFree radical-scavenging capacityantioxidant activity and phenolic content of Pouzolzia zeylan-icardquo Journal of the Serbian Chemical Society vol 76 no 5 pp709ndash717 2011
[32] C Papuc M Crivineanu G Goran V Nicorescu and N Dur-dun ldquoFree radicals scavenging and antioxidant activity ofEuropean mistletoe (Viscum album) and European birthwort(Aristolochia clematitis)rdquo Revista de Chimie vol 61 no 7 pp619ndash622 2010
[33] VOOyetayo CDong andY Yao ldquoAntioxidant and antimicro-bial properties of aqueous extract from Dictyophora indusiatardquoThe Open Mycology Journal vol 3 no 1 pp 20ndash26 2009
[34] S Marklund and G Marklund ldquoInvolvement of the superoxideanion radical in the autoxidation of pyrogallol and a convenientassay for superoxide dismutaserdquo European Journal of Biochem-istry vol 47 no 3 pp 469ndash474 1974
[35] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996
[36] D J Huang b Ou and R L Prior ldquoThe chemistry behindantioxidant capacity assaysrdquo Journal of Agricultural and FoodChemistry vol 53 no 6 pp 1841ndash1856 2005
[37] P Manzi A Aguzzi and L Pizzoferrato ldquoNutritional value ofmushrooms widely consumed in Italyrdquo Food Chemistry vol 73no 3 pp 321ndash325 2001
[38] P Manzi S Marconi A Aguzzi and L Pizzoferrato ldquoCommer-cial mushrooms nutritional quality and effect of cookingrdquo FoodChemistry vol 84 no 2 pp 201ndash206 2004
[39] I P Krbavcic and I C Baric ldquoInfluence of deep fat fryingon some nutritional parameters of novel food based on mush-rooms and fresh soft cheeserdquo Food Chemistry vol 84 no 3 pp417ndash419 2004
[40] D Agrahar-Murugkar andG Subbulakshmi ldquoNutritional valueof edible wild mushrooms collected from the Khasi hills ofMeghalayardquo Food Chemistry vol 89 no 4 pp 599ndash603 2005
[41] V A Dıez and A Alvarez ldquoCompositional and nutritionalstudies on two wild edible mushrooms from northwest SpainrdquoFood Chemistry vol 75 no 4 pp 417ndash422 2001
[42] L Barros M-J Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007
[43] S K Sharma and N S Atri ldquoNutraceutical composition ofwild species of genus Lentinus Fr fromNothern Indiardquo CurrentResearch in Environmental ampAppliedMycology vol 4 pp 11ndash322014
[44] P J Kanu K Zhu J Baby Kanu H Zhou H Qian and K ZhuldquoBiologically active components and nutraceuticals in sesameand related products a review and prospectrdquo Trends in FoodScience and Technology vol 18 no 12 pp 599ndash608 2007
[45] F S Reis E Pereira L Barros M J Sousa A Martins and I CF R Ferreira ldquoBiomolecule profiles in inediblewildmushroomswith antioxidant valuerdquoMolecules vol 16 no 6 pp 4328ndash43382011
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
where 1198600was the absorbance of the control and 119860
119875was the
absorbance in the presence of the sampleFor ABTS radical scavenging activity 10120583L of the sample
was added to 4mL of the diluted ABTS∙+ solution (preparedby adding 7mM of the ABTS stock solution to 245mMpotassium persulfate kept in the dark at room temperaturefor 12ndash16 h before use) The solution was then diluted with5mM phosphate-buffered saline (pH 74) and absorbancewas measured at 730 nm after 30min [31] The ABTS radicalscavenging activity was calculated as
119878 = (119860controlminus119860 sample
119860control)times 100 (4)
For reducing power estimation samples (200 120583L) weremixed with sodium phosphate buffer (pH 66) 1mM FeSO
4
and 1 potassium ferricyanide and incubated for 20minat 50∘C after that trichloroacetic acid was added and themixtures were centrifuged Supernatant (25mL) was mixedwith an equal volume of water and 05mL 01 FeCl
3 The
absorbance was measured at 700 nm [32]For Fe2+ chelating activity 1mL of the sample (2ndash10mg
mL) was mixed with 37mL of ultrapure water after thatthe mixture was reacted with ferrous chloride (2mmolL01mL) and ferrozine (5mmolL 02mL) for 20min and theabsorbance was read at 562 nm with using EDTA as controlThe chelating activity was calculated using the formula
where 119860119887is the absorbance of the blank and 119860
119904is the
absorbance in the presence of the extract [33]The scavenging activity of superoxide anion radicals
was measured following standard method [34] Samples (0ndash20mgmL 1mL) and Tris-HCl buffer (500mM pH 823mL) were incubated in a water bath at 25∘C for 20min andafter this pyrogallic acid (50mM 04mL) was added HClsolution (80M 01mL) was added to terminate the reactionafter 4min The absorbance of the mixture was measuredat 320 nm The scavenging ability was calculated using thefollowing formula
scavenging ability () = (1 minus119860 sample
119860control)times 100 (6)
where 119860control is the absorbance of control without thepolysaccharide sample and 119860 sample is the absorbance in thepresence of the polysaccharide sample
For ferric reducing antioxidant power (FRAP) assayfirstly FRAP reagent was prepared by mixing TPTZ (25mL10mM in 40mM HCl) 25mL of 300mM acetate buffer and25mL of FeCl
3sdot6H2O After this freshly prepared FRAP
reagent (18mL) was taken in a test tube and incubated at30∘C in water bath for 10 minutes Then absorbance wasread at 0min (119905
0) After this 100 120583L of sample extract or
standard and 100 120583L of distilled water were added to the testtube mixed and incubated at 30∘C for 30 minutes Thenthe absorbance was taken at 593 nm (119905
30) Ferrous sulphate
was used as standard [35 36] FRAP activity was determinedagainst a standard curve of ferrous sulphate and the valueswere expressed as 120583M Fe2+ equivalents per gram of extractand calculated using the following equation
FRAP value = Absorbance (sample+ FRAP reagent)
minusAbsorbance (FRAP reagent) (7)
25 Statistical Analysis All experiments were performed 3times and with 3 replicates The results were analyzed usingone-way analysis of variance (ANOVA) 119901 lt 005 was con-sidered significant and SPSS software (SPSS Inc ChicagoIL USA) was used to calculate differences Tukey-HSD at119901 lt 005 test was used to determine significant differences
3 Results
31 Chemical Evaluation Nutrient composition of the wildculinary mushrooms is shown in Table 2 Protein was foundin high levels and varied between 516 in Inocybe splendensand 2263 in Agaricus arvensis Protein percentage inPleurotus cystidiosus (2069) Amanita caesarea (1972)Agaricus campestris (1838) Cantharellus cibarius (1819)and Lentinus cladopus (1859) was also found to be highAll the twenty culinary species were found to be low in fatcontent Fat ranged from010 in Laccaria laccata to 038 inInocybe splendens In general these wild culinary mushroomsconsumed by local people were found to be higher in proteinand low in fat although differences were observed in netvalue of individual species Crude fibres ranged from 108in Hygrocybe coccinea to 242 in Lentinus cladopus Ashcontent varied between 011 in Inocybe splendens and 096in Agaricus arvensis Carbohydrates calculated by differencewere also found to be in abundant amounts and theirpercentage was ranged from 3119 in Inocybe splendens to5712 in Agaricus arvensis Nutrient contents of Inocybesplendens Hygrocybe nivea and Conocybe tenera were foundto be less as compared to other species
There are several reports about the toxicity reports due tomushrooms on humans in these areas and hence preliminarystudies were done to check the toxicity level of mushroomsFor all the twenty species being used by the people forculinary purposes the test was found to be negative Thatmeans these species are nontoxic and hence recommendedfor consumption
All the culinarymushroom species contained glucose andrhamnose as the principal carbohydrates (Table 3) Neverthe-less the present study also describes the presence of xyloseand mannose in all the studied species However galactoseand fructose were detected in very low percentage in someof the species Russula mairei contained lowest percentageof glucose (2160) and Agaricus arvensis contained highestpercentage of glucose (6412)
32 Bioactive Evaluation The results of fatty acid composi-tion (total saturated fatty acids SFA monounsaturated fattyacids MUFA and polyunsaturated fatty acids PUFA) of allthe species are shown in Table 4 In general the major fatty
BioMed Research International 5
Table 2 Percent chemical composition of twenty wild culinary mushroom species collected from northern Himalayan regions
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 3 Percent monosaccharide composition of twenty species collected from northern Himalayan regions showing richness in glucoserhamnose mannose xylose and galactose and fructose in lower percentage
Russula lepida 515 plusmn 01d 3261 plusmn 36f 1316 plusmn 12e 231 plusmn 01c ND NDR mairei 426 plusmn 01c 2160 plusmn 21a 1281 plusmn 16d 212 plusmn 00c ND NDND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
6 BioMed Research International
Table4Percentfattyacid
compo
sitionof
allthe
twentywild
culin
arymushroo
mspeciesc
ollected
from
different
region
sofn
orthernHim
alayas
Species
C900
C100
C120
C160
C1611
C171
C181
C182
C202
UI
Agaricu
sarvensis
006plusmn00a
030plusmn001
a028plusmn00a
186plusmn003
a006plusmn00a
009plusmn00a
018plusmn00a
324plusmn00a
221plusmn02a
057plusmn002
a
Acampestr
is003plusmn00a
026plusmn002
a021plusmn00a
119plusmn002
a009plusmn00a
002plusmn00a
035plusmn00a
204plusmn00a
218plusmn03a
046plusmn002
a
Acomtulus
006plusmn00a
022plusmn00a
ND
117plusmn001
a007plusmn00a
009plusmn00a
038plusmn00a
173plusmn00a
134plusmn01a
036plusmn001
a
Asilvicola
008plusmn00a
033plusmn00a
022plusmn00a
156plusmn002
a009plusmn00a
001plusmn00a
015plusmn00a
154plusmn00a
221plusmn03a
040plusmn001
a
Amanita
caesarea
004plusmn00a
032plusmn002
a019plusmn00a
196plusmn002
a012plusmn00a
005plusmn00a
031plusmn00a
219plusmn00a
128plusmn00a
039plusmn001
a
Acitrin
a006plusmn00a
028plusmn001
aND
091plusmn00a
011plusmn00a
008plusmn00a
031plusmn00a
159plusmn00a
168plusmn00a
037plusmn001
a
Afulva
004plusmn00a
021plusmn001
a025plusmn002
a010plusmn00a
008plusmn00a
004plusmn00a
015plusmn00a
164plusmn00a
123plusmn00a
031plusmn002
a
Cantharellu
scibarius
007plusmn00a
033plusmn002
a022plusmn001
a110plusmn002
a007plusmn00a
001plusmn00a
037plusmn00a
294plusmn00a
118plusmn01a
045plusmn002
a
Conocybe
tenera
006plusmn00a
020plusmn00a
ND
007plusmn00a
005plusmn00a
002plusmn00a
035plusmn00a
014plusmn00a
138plusmn00a
019plusmn00a
Gymnopilusjun
onius
005plusmn00a
031plusmn001
a014plusmn001
a18
7plusmn003
a008plusmn00a
002plusmn00a
088plusmn00a
049plusmn00a
121plusmn
01a
026plusmn001
a
Hygrocybe
coccinea
004plusmn00a
025plusmn002
a017plusmn00a
033plusmn00a
001plusmn00a
005plusmn00a
017plusmn00a
024plusmn00a
119plusmn00a
016plusmn00a
Hnivea
007plusmn00a
022plusmn001
aND
005plusmn00a
004plusmn00a
001plusmn00a
053plusmn00a
004plusmn00a
138plusmn02a
020plusmn00a
Inocybesplendens
002plusmn00a
019plusmn001
a023plusmn001
a089plusmn00a
002plusmn00a
003plusmn00a
018plusmn00a
010plusmn00a
020plusmn00a
005plusmn00a
Lactariusp
ubescens
001plusmn00a
023plusmn001
a016plusmn00a
075plusmn00a
007plusmn00a
008plusmn00a
026plusmn00a
344plusmn00a
083plusmn00a
046plusmn002
a
Laccarialaccata
002plusmn00a
022plusmn00a
ND
101plusmn
00a
006plusmn00a
009plusmn00a
092plusmn00a
284plusmn00a
029plusmn00a
042plusmn001
a
Lepista
nuda
003plusmn00a
024plusmn00a
ND
043plusmn00a
001plusmn00a
002plusmn00a
020plusmn00a
204plusmn00a
037plusmn00a
026plusmn00a
Lentinus
cladopu
s009plusmn00a
033plusmn002
a032plusmn00a
146plusmn00a
055plusmn00a
001plusmn00a
032plusmn00a
245plusmn00a
295plusmn00a
057plusmn002
a
Pleurotuscystid
iosus
006plusmn00a
031plusmn001
a028plusmn00a
135plusmn00a
009plusmn00a
005plusmn00a
098plusmn00a
204plusmn00a
221plusmn02a
052plusmn002
a
Russu
lalep
ida
006plusmn00a
019plusmn001
aND
033plusmn00a
005plusmn00a
005plusmn00a
049plusmn00a
204plusmn03a
165plusmn00a
042plusmn001
a
Rmairei
001plusmn00a
017plusmn001
aND
026plusmn00a
008plusmn00a
008plusmn00a
063plusmn00a
201plusmn02a
010plusmn00a
028plusmn001
a
ND=no
tdetected
Values
aree
xpressed
asmeanplusmnSE
andlette
rsin
thes
uperscrip
tsrepresentthe
significantd
ifference
ineach
columnwith119901le005accordingto
Tukeyrsquos
test
BioMed Research International 7
Table 5 Percent amino acids composition of all the wild culinary species collected from northern Himalayas
Values are expressed as mean plusmn SE and letters in superscript represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
acids found in the studied species were linoleic acid (C182)followed by oleic acid (C181) and palmitic acid (C160)Besides these three main fatty acids already described sixmore were identified and quantified PUFA were the maingroup of fatty acids documented in all the species Agaricusarvensis Amanita caesarea Cantharellus cibarius Lentinuscladopus and Pleurotus cystidiosus contained lower value ofMUFA but higher percentage of PUFA as compared to otherspecies due to the higher percentage of linoleic acidHoweverUFApredominated over SFA in all the studied species rangingfrom 65 to 70
Amino acid composition of all the species is shown inTable 5 In all the species aspartic acid (019ndash039) wasfound to be predominated amino acid followed by tyrosine(010ndash021) arginine (012ndash029) alanine (004ndash014)and proline (001ndash007) Amanita caesarea Agaricus arven-sis Cantharellus cibarius Lentinus cladopus and Pleurotuscystidiosus contained maximum amount of these aminoacids Tocopherol contents in all the studied mushroomspecies including three wild are detailed in Table 6 120572-tocopherol and 120573-tocopherol were found to be present in allthe species However 120574-tocopherol was documented fromfew species only Tocopherol content was ranged from 090to 433 120583gg in all the species Cantharellus cibarius (433 plusmn00 120583gg) contained all the three isomers in higher amount ascompared to other species 120573-tocopherol was found in higheramounts as compared to 120572-tocopherol 120574-tocopherol wasdetected only in nine species Cantharellus cibarius (433 plusmn00 120583gg) contained higher amounts of 120574 tocopherol
Results obtained for 120573-carotene lycopene flavonoidsascorbic acid and anthocyanidins composition of all the
twenty species are presented in Table 7 Phenols werethe major antioxidant component detected in significantamounts from all the species (1912ndash6336mgg) followedby anthocyanidins (614ndash1425mg cyanidin chloride100 gextract) flavonoids (114ndash417mgg) ascorbic acid whichwas found in small amounts (020ndash099mgg) 120573-carotene(021ndash079 120583g100 g) and lycopene (019ndash038 120583g100 g) Eachspecies differed with other species in net amounts of all thesecomponents Species like Agaricus arvensis Amanita cae-sarea Gymnopilus junonius Lentinus cladopus and Pleurotuscystidiosus contained higher values of these components ascompared to other species
33 Antioxidant Evaluation Antioxidant properties of allthe species were expressed as EC
radical scavenging activity in all the species showed differ-ences in effectiveness in antioxidant properties Among allthe species Cantharellus cibarius showed lowest EC
50values
(176 plusmn 02mgmL) followed by Amanita caesarea (202 plusmn02mgmL) and Agaricus arvensis (212 plusmn 04mgmL) Otherspecies showed slightly higher EC
50values and therefore
lesserDPPH radical scavenging activityCantharellus cibariusshowed higher DPPH radical scavenging activity and Inocybesplendens showed lower DPPH radical scavenging activitythan other species
For ABTS radical scavenging activities EC50ranged from
426 to 145mgmL Lowest EC50
values were obtained forAmanita caesarea (145 plusmn 06mgmL) showing high antiox-idant activities of this species Higher EC
50values were
8 BioMed Research International
Table 6 Tocopherol composition (120583gg) of twenty species collected from northern Himalayan regions
ND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 7 Other bioactive compounds evaluated from all the species
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
BioMed Research International 9
Table 8 EC50 values for different antioxidant assays on twenty wild culinary species collected from northern Himalayas
obtained for Hygrocybe nivea (426 plusmn 05mgmL) showinglowest ABTS radical scavenging activities of this speciesReducing power of Cantharellus cibarius (162 plusmn 05mgmL)was measured higher than other species Higher EC
50values
for reducing activity were measured in Inocybe splendens(519 plusmn 01mgmL)
Higher effectiveness in ferrous ion chelating activitywas detected in Agaricus arvensis (114 plusmn 01mgmL) andlow effectiveness was detected in Russula mairei (295 plusmn03mgmL) Nevertheless Cantharellus cibarius (127 plusmn02mgmL) Amanita caesarea (140 plusmn 04mgmL) Hygro-cybe coccinea (135 plusmn 02mgmL) and H nivea (145 plusmn06mgmL) showed lower EC
50values than remaining
species EC50
values of scavenging ability on superoxideradical were found to be maximum in Amanita caesarea(044 plusmn 03mgmL) and minimum in Lactarius pubescens(221 plusmn 06mgmL) Gymnopilus junonius showed maximumFRAP activity with least EC
Although there are previous reports on documentation ofculinary edible species from the regions native to northernHimalayas but there are no reports on the evaluation stud-ies as well as toxicity status of all these culinary speciesThe wild edible species Agaricus bisporus Boletus edulis
Morchella esculenta Cordyceps sinensis and Lentinula edodeswhich have been extensively worked out in India and otherparts of the world for their compositional and medicinalaspects have not been undertaken for investigations presently[37ndash40] Compositional studies showed that most of theculinary species were rich in protein carbohydrates andlow in fat There are several reports on richness of wildedible mushrooms with protein and carbohydrate contentsand low fat levels which directly make them nutritionallyrich [24 25 41] Nevertheless under present studies thedifferences between the nutrient concentrations of all thespecies differed but Agaricus arvensis Pleurotus cystidiosusAmanita caesarea Agaricus campestris Cantharellus cibariusand Lentinus cladopus showed higher nutrient percentagewhich is comparable to other wild and commercially cul-tivated species [25 42] The crude fat content detected inall the species was not found to be significantly differentCrude fibres were detected in appreciable percentage fromall the species which make them important in nutritionalpoint of viewThe results are in conformity with the previousreports on several wild edible Pleurotus and Lentinus speciesfrom northwest India [24 43] The species Inocybe splendensHygrocybe nivea and Conocybe tenera were not found tocontain higher percentage of nutrients Although previousreports showed that nutrients composition in wild species isless as compared to cultivated species [25] however Agaricusarvensis Pleurotus cystidiosus Amanita caesarea Agaricuscampestris Cantharellus cibarius and Lentinus cladopus were
10 BioMed Research International
found to be rich in protein and carbohydrates similar tocommercially grown species [25]
Fatty acid composition showed the dominance of UFAover SFA in all the studied mushrooms species which is inconformity with the other studies [41] The differences wereobserved in net amounts in all the species Unsaturationindex of Agaricus arvensis and Lentinus cladopus (057 plusmn002) was found to be significantly higher than otherspecies Whereas Inocybe splendens (005 plusmn 00) showedleast Unsaturation Index High UFA shows the medicinalimportance of these culinary mushrooms as these increasethe HDL cholesterol and decrease LDL cholesterol triacyl-glycerol lipid oxidation and LDL susceptibility to oxidation[44] Predominance of UFA over SFA in all the species showssimilar results as obtained for other wild and commerciallycultivated species [24 25] 120572- and 120573-tocopherol were detectedin higher amounts than third isomer in all the studied speciesSimilar findings were made in other wild and cultivatedspecies with higher 120572- and 120573-tocopherol than 120574-tocopherol[25] The high levels of these two compounds correspondwith a higher oxidative activity which is associated withcardiovascular protection [45] Phenolic compounds weredetected in higher amounts than other bioactive compoundsPresence of high phenolic compounds accounts for the highantioxidant properties of all the species [42] 120573-carotenelycopene and ascorbic acids were detected in low amountsAnthocyanidins were also detected from these wild speciesin appreciable amounts The presence of these functionalmedicinal compounds inmedicinal andor edible mushroomis due to habitat or substrates in which these grow to be highin the functionalmoleculesThe categories of thesemoleculesare anthocyanidins beta-glucans selenium ganoderic acidtriterpenes or cordycepinThe compounds identified in theseextracts show that at least a part of the functional compoundsin medicinal andor edible mushroom is due to growingmushrooms on substrates that are high in the functionalmolecules To these categories can be added anthocyanidinsbeta-glucans selenium ganoderic acid triterpenes or cordy-cepinThe amounts of these have been found to vary with thetype of extraction as ethanolic extract yields higher amountsof anthocyanidins as compared to methanolic hot water andcold water extracts [29]
All the studied species showed significant antioxidantproperties measured on the basis of EC
50values Never-
theless each species showed different antioxidant activitywith highly effective and less effective EC
50values Better
antioxidant properties of some species are due to presence ofhigher phenolic compounds 120573-carotene lycopene ascorbicacids anthocyanidins and tocopherol amounts in themHigh reducing power of some species is due to the presence ofhigher amounts of reducers (antioxidants) in them Presentlyinvestigated species are commonly used for culinary pur-poses in the regions native to northern Himalayas Many ofthe species in these regions are not evaluated previously fordetailed compositional analysisTheir knowledge is restrictedto old aged villagers of the regions and neglected for thecommercial exploitations There are no positive reportson toxicity of these mushrooms analyzed presently hencethese are safe for further experimental work related to
drug discovery All the culinary species contained impor-tant and useful nutraceuticals such as unsaturated fattyacids phenolics carotenoids ascorbic acid tocopherolsand anthocyanidins besides these some important aminoacids were detected in these mushrooms which could beused for the purpose of being used as functional ingredi-ents Since nutraceuticals are powerful in maintaining andpromoting health longevity and life quality the commer-cial exploitation of these species will certainly create animpact on nutritional therapy and also will be beneficialtodayrsquos food industry Direct use of these species for con-sumption and other culinary aspects is safe and healthpromoting with advantage of the additive effects of allthe bioactive and antioxidant compounds present in thesespecies
Abbreviations
AlCl3 Aluminum trichloride
ANOVA Analysis of varianceDW Distilled waterDPPH 22-Diphenyl-1-picrylhydrazylFeCl3 Ferric chloride
FRAP Ferric reducing antioxidant powerg GramsGAEs Gallic acid equivalentsGC Gas ChromatographyHCl Hydrochloric acidH2O Water
H3PO4 Phosphoric acid
HPLC High performance liquid chromatographyK2HPO4 Potassium hydrogen phosphate
The authors declare that there is no conflict of interestsregarding the publication of this paper
BioMed Research International 11
Acknowledgment
The authors wish to thank Science and Engineering BoardDepartment of Science and Technology New Delhi forresearch Grant (SBFTLS-042013) to carry out presentstudies
References
[1] W M Breene ldquoNutritional and medicinal value of specialtymushroomsrdquo Journal of Food Protection vol 53 pp 883ndash8941990
[2] S Chang and P Miles ldquoMushroom biologymdasha new disciplinerdquoMycologist vol 6 no 2 pp 64ndash65 1992
[3] P Manzi L Gambelli S Marconi V Vivanti and L Pizzoferra-to ldquoNutrients in edible mushrooms an inter-species compara-tive studyrdquo Food Chemistry vol 65 no 4 pp 477ndash482 1999
[4] P Bobek O Ozdın and M Mikus ldquoDietary oyster mushroom(Pleurotus ostreatus) accelerates plasma cholesterol turnover inhypercholesterolaemic ratsrdquo Physiological Research vol 44 no5 pp 287ndash291 1995
[5] P Bobek and S Galbavy ldquoHypocholesterolemic and antiathero-genic effect of oystermushroom (Pleurotus ostreatus) in rabbitsrdquoNahrung vol 43 no 5 pp 339ndash342 1999
[6] S Khatun A Islam U Cakilcioglu and N Chatterjee ldquoRe-search on mushroom as a potential source of nutraceuticals areview on Indian perspectiverdquo American Journal of Experimen-tal Agriculture vol 2 no 1 pp 47ndash73 2012
[7] J A Vaz L Barros A Martins J S Morais M H Vasconcelosand I C F R Ferreira ldquoPhenolic profile of seventeen por-tuguese wild mushroomsrdquo LWT Food Science and Technologyvol 44 no 1 pp 343ndash346 2011
[8] J A Vaz L Barros A Martins C Santos-Buelga M HVasconcelos and I C F R Ferreira ldquoChemical compositionof wild edible mushrooms and antioxidant properties of theirwater soluble polysaccharidic and ethanolic fractionsrdquo FoodChemistry vol 126 no 2 pp 610ndash616 2011
[9] T Ozen C Darcan O Aktop and I Turkekul ldquoScreening ofantioxidant antimicrobial activities and chemical contents ofedible mushrooms wildly grown in the Black Sea region ofTurkeyrdquo Combinatorial Chemistry amp High Throughput Screen-ing vol 14 no 2 pp 72ndash84 2011
[10] V E C Ooi and F Liu ldquoA Review of pharmacological activitiesof mushroom polysaccharidesrdquo International Journal of Medic-inal Mushrooms vol 1 no 3 pp 195ndash206 1999
[11] S PWasser and A LWeis ldquoMedicinal properties of substancesoccurring in higher Basidiomycete mushroom current per-spectiverdquo International Journal of Medicinal Mushrooms vol 1pp 31ndash62 1999
[12] P M Kidd ldquoThe use of mushroom glucans and proteoglycansin cancer treatmentrdquo Alternative Medicine Review vol 5 no 1pp 4ndash27 2000
[13] C Israilides and A Philippoussis ldquoBio-technologies of recy-cling agro-industrial wastes for the production of commerciallyimportant fungal polysaccharides andmushroomsrdquo Biotechnol-ogy and Genetic Engineering Reviews vol 20 pp 247ndash259 2003
[14] T Mizuno ldquoBioactive biomolecules of mushrooms food func-tion and medicinal effect of mushroom fungirdquo Food ReviewsInternational vol 11 no 1 pp 5ndash21 1995
[15] Y Kabir and S Kimura ldquoDietary mushrooms reduce bloodpressure in spontaneously hypertensive rats (SHR)rdquo Journal of
Nutritional Science and Vitaminology vol 35 no 1 pp 91ndash941989
[16] J-L Mau S-Y Tsai Y-H Tseng and S-J Huang ldquoAntioxidantproperties of hot water extracts from Ganoderma tsugae Mur-rillrdquo LWTmdashFood Science and Technology vol 38 no 6 pp 589ndash597 2005
[17] A Smania F D Monache E F A Smania M L Gil L CBenchetrit and F S Cruz ldquoAntibacterial activity of a substanceproduced by the fungus Pycnoporus sanguineus (Fr) MurrrdquoJournal of Ethnopharmacology vol 45 no 3 pp 177ndash181 1995
[18] W Andlauer and P Furst ldquoNutraceuticals a piece of historypresent status and outlookrdquo Food Research International vol 35no 2-3 pp 171ndash176 2002
[19] C L Kruger and S W Mann ldquoSafety evaluation of functionalingredientsrdquo Food and Chemical Toxicology vol 41 no 6 pp793ndash805 2003
[20] P H Mattila V I Piironen E J Uusi-Rauva and P E Koivis-toinen ldquoVitamin D contents in edible mushroomsrdquo Journal ofAgricultural and Food Chemistry vol 42 no 11 pp 2449ndash24531994
[21] R P Tewari ldquoMushroom their role in nature and societyrdquo inFrontiers inMushroomBiotechnology R D Rai R C Upadhyayand S R Sharma Eds pp 1ndash8 NRCM Chambaghat India2005
[22] K C Semwal S L Stephenson V K Bhatt and R P BhattldquoEdible mushrooms of the Northwestern Himalaya India astudy of indigenous knowledge distribution and diversityrdquoMycosphere vol 5 no 3 pp 440ndash461 2014
[23] E V Crisan and A Sands ldquoEdible mushrooms nutritionalvaluerdquo in The Biology and Cultivation of Edible Mushrooms ST Chang and W A Hayes Eds pp 137ndash165 Academic PressNew York NY USA 1978
[24] N S Atri S K Sharma R Joshi A Gulati and A Gula-ti ldquoNutritional and neutraceutical composition of five wildculinary-medicinal species of genus Pleurotus (Higher Basid-iomycetes) from Northwest Indiardquo International Journal ofMedicinal Mushrooms vol 15 no 1 pp 49ndash56 2013
[25] L Barros T Cruz P Baptista L M Estevinho and I C FR Ferreira ldquoWild and commercial mushrooms as source ofnutrients and nutraceuticalsrdquo Food and Chemical Toxicologyvol 46 no 8 pp 2742ndash2747 2008
[26] M Nagata and I Yamashita ldquoSimple method for simultaneousdetermination of chlorophyll and carotenoids in tomato fruitrdquoNippon Shokuhin Kogyo Gakkaishi vol 39 no 10 pp 925ndash9281992
[27] V L Singleton and J A Rossi ldquoColorimetric detection oftotal phenolics with phosphomolybdic-phosphotungstic acidreagentsrdquo American Journal of Enology and Viticulture vol 16pp 144ndash158 1965
[28] L K Jagadish V Venkata Krishnan R Shenbhagaramanand V Kaviyarasan ldquoComparitive study on the antioxidantanticancer and antimicrobial property of Agaricus bisporus (JE Lange) Imbach before and after boilingrdquo African Journal ofBiotechnology vol 8 no 4 pp 654ndash661 2009
[29] E Vamanu and S Nita ldquoAntioxidant capacity and the cor-relation with major phenolic compounds anthocyanin andtocopherol content in various extracts from the wild edibleBoletus edulis mushroomrdquo BioMed Research International vol2013 Article ID 313905 11 pages 2013
[30] E Vamanu ldquoBiological activities of the polysaccharides pro-duced in submerged culture of two edible Pleurotus ostreatus
12 BioMed Research International
mushroomsrdquo Journal of Biomedicine and Biotechnology vol2012 Article ID 565974 8 pages 2012
[31] P L Li L Huo W Su et al ldquoFree radical-scavenging capacityantioxidant activity and phenolic content of Pouzolzia zeylan-icardquo Journal of the Serbian Chemical Society vol 76 no 5 pp709ndash717 2011
[32] C Papuc M Crivineanu G Goran V Nicorescu and N Dur-dun ldquoFree radicals scavenging and antioxidant activity ofEuropean mistletoe (Viscum album) and European birthwort(Aristolochia clematitis)rdquo Revista de Chimie vol 61 no 7 pp619ndash622 2010
[33] VOOyetayo CDong andY Yao ldquoAntioxidant and antimicro-bial properties of aqueous extract from Dictyophora indusiatardquoThe Open Mycology Journal vol 3 no 1 pp 20ndash26 2009
[34] S Marklund and G Marklund ldquoInvolvement of the superoxideanion radical in the autoxidation of pyrogallol and a convenientassay for superoxide dismutaserdquo European Journal of Biochem-istry vol 47 no 3 pp 469ndash474 1974
[35] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996
[36] D J Huang b Ou and R L Prior ldquoThe chemistry behindantioxidant capacity assaysrdquo Journal of Agricultural and FoodChemistry vol 53 no 6 pp 1841ndash1856 2005
[37] P Manzi A Aguzzi and L Pizzoferrato ldquoNutritional value ofmushrooms widely consumed in Italyrdquo Food Chemistry vol 73no 3 pp 321ndash325 2001
[38] P Manzi S Marconi A Aguzzi and L Pizzoferrato ldquoCommer-cial mushrooms nutritional quality and effect of cookingrdquo FoodChemistry vol 84 no 2 pp 201ndash206 2004
[39] I P Krbavcic and I C Baric ldquoInfluence of deep fat fryingon some nutritional parameters of novel food based on mush-rooms and fresh soft cheeserdquo Food Chemistry vol 84 no 3 pp417ndash419 2004
[40] D Agrahar-Murugkar andG Subbulakshmi ldquoNutritional valueof edible wild mushrooms collected from the Khasi hills ofMeghalayardquo Food Chemistry vol 89 no 4 pp 599ndash603 2005
[41] V A Dıez and A Alvarez ldquoCompositional and nutritionalstudies on two wild edible mushrooms from northwest SpainrdquoFood Chemistry vol 75 no 4 pp 417ndash422 2001
[42] L Barros M-J Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007
[43] S K Sharma and N S Atri ldquoNutraceutical composition ofwild species of genus Lentinus Fr fromNothern Indiardquo CurrentResearch in Environmental ampAppliedMycology vol 4 pp 11ndash322014
[44] P J Kanu K Zhu J Baby Kanu H Zhou H Qian and K ZhuldquoBiologically active components and nutraceuticals in sesameand related products a review and prospectrdquo Trends in FoodScience and Technology vol 18 no 12 pp 599ndash608 2007
[45] F S Reis E Pereira L Barros M J Sousa A Martins and I CF R Ferreira ldquoBiomolecule profiles in inediblewildmushroomswith antioxidant valuerdquoMolecules vol 16 no 6 pp 4328ndash43382011
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 3 Percent monosaccharide composition of twenty species collected from northern Himalayan regions showing richness in glucoserhamnose mannose xylose and galactose and fructose in lower percentage
Russula lepida 515 plusmn 01d 3261 plusmn 36f 1316 plusmn 12e 231 plusmn 01c ND NDR mairei 426 plusmn 01c 2160 plusmn 21a 1281 plusmn 16d 212 plusmn 00c ND NDND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
6 BioMed Research International
Table4Percentfattyacid
compo
sitionof
allthe
twentywild
culin
arymushroo
mspeciesc
ollected
from
different
region
sofn
orthernHim
alayas
Species
C900
C100
C120
C160
C1611
C171
C181
C182
C202
UI
Agaricu
sarvensis
006plusmn00a
030plusmn001
a028plusmn00a
186plusmn003
a006plusmn00a
009plusmn00a
018plusmn00a
324plusmn00a
221plusmn02a
057plusmn002
a
Acampestr
is003plusmn00a
026plusmn002
a021plusmn00a
119plusmn002
a009plusmn00a
002plusmn00a
035plusmn00a
204plusmn00a
218plusmn03a
046plusmn002
a
Acomtulus
006plusmn00a
022plusmn00a
ND
117plusmn001
a007plusmn00a
009plusmn00a
038plusmn00a
173plusmn00a
134plusmn01a
036plusmn001
a
Asilvicola
008plusmn00a
033plusmn00a
022plusmn00a
156plusmn002
a009plusmn00a
001plusmn00a
015plusmn00a
154plusmn00a
221plusmn03a
040plusmn001
a
Amanita
caesarea
004plusmn00a
032plusmn002
a019plusmn00a
196plusmn002
a012plusmn00a
005plusmn00a
031plusmn00a
219plusmn00a
128plusmn00a
039plusmn001
a
Acitrin
a006plusmn00a
028plusmn001
aND
091plusmn00a
011plusmn00a
008plusmn00a
031plusmn00a
159plusmn00a
168plusmn00a
037plusmn001
a
Afulva
004plusmn00a
021plusmn001
a025plusmn002
a010plusmn00a
008plusmn00a
004plusmn00a
015plusmn00a
164plusmn00a
123plusmn00a
031plusmn002
a
Cantharellu
scibarius
007plusmn00a
033plusmn002
a022plusmn001
a110plusmn002
a007plusmn00a
001plusmn00a
037plusmn00a
294plusmn00a
118plusmn01a
045plusmn002
a
Conocybe
tenera
006plusmn00a
020plusmn00a
ND
007plusmn00a
005plusmn00a
002plusmn00a
035plusmn00a
014plusmn00a
138plusmn00a
019plusmn00a
Gymnopilusjun
onius
005plusmn00a
031plusmn001
a014plusmn001
a18
7plusmn003
a008plusmn00a
002plusmn00a
088plusmn00a
049plusmn00a
121plusmn
01a
026plusmn001
a
Hygrocybe
coccinea
004plusmn00a
025plusmn002
a017plusmn00a
033plusmn00a
001plusmn00a
005plusmn00a
017plusmn00a
024plusmn00a
119plusmn00a
016plusmn00a
Hnivea
007plusmn00a
022plusmn001
aND
005plusmn00a
004plusmn00a
001plusmn00a
053plusmn00a
004plusmn00a
138plusmn02a
020plusmn00a
Inocybesplendens
002plusmn00a
019plusmn001
a023plusmn001
a089plusmn00a
002plusmn00a
003plusmn00a
018plusmn00a
010plusmn00a
020plusmn00a
005plusmn00a
Lactariusp
ubescens
001plusmn00a
023plusmn001
a016plusmn00a
075plusmn00a
007plusmn00a
008plusmn00a
026plusmn00a
344plusmn00a
083plusmn00a
046plusmn002
a
Laccarialaccata
002plusmn00a
022plusmn00a
ND
101plusmn
00a
006plusmn00a
009plusmn00a
092plusmn00a
284plusmn00a
029plusmn00a
042plusmn001
a
Lepista
nuda
003plusmn00a
024plusmn00a
ND
043plusmn00a
001plusmn00a
002plusmn00a
020plusmn00a
204plusmn00a
037plusmn00a
026plusmn00a
Lentinus
cladopu
s009plusmn00a
033plusmn002
a032plusmn00a
146plusmn00a
055plusmn00a
001plusmn00a
032plusmn00a
245plusmn00a
295plusmn00a
057plusmn002
a
Pleurotuscystid
iosus
006plusmn00a
031plusmn001
a028plusmn00a
135plusmn00a
009plusmn00a
005plusmn00a
098plusmn00a
204plusmn00a
221plusmn02a
052plusmn002
a
Russu
lalep
ida
006plusmn00a
019plusmn001
aND
033plusmn00a
005plusmn00a
005plusmn00a
049plusmn00a
204plusmn03a
165plusmn00a
042plusmn001
a
Rmairei
001plusmn00a
017plusmn001
aND
026plusmn00a
008plusmn00a
008plusmn00a
063plusmn00a
201plusmn02a
010plusmn00a
028plusmn001
a
ND=no
tdetected
Values
aree
xpressed
asmeanplusmnSE
andlette
rsin
thes
uperscrip
tsrepresentthe
significantd
ifference
ineach
columnwith119901le005accordingto
Tukeyrsquos
test
BioMed Research International 7
Table 5 Percent amino acids composition of all the wild culinary species collected from northern Himalayas
Values are expressed as mean plusmn SE and letters in superscript represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
acids found in the studied species were linoleic acid (C182)followed by oleic acid (C181) and palmitic acid (C160)Besides these three main fatty acids already described sixmore were identified and quantified PUFA were the maingroup of fatty acids documented in all the species Agaricusarvensis Amanita caesarea Cantharellus cibarius Lentinuscladopus and Pleurotus cystidiosus contained lower value ofMUFA but higher percentage of PUFA as compared to otherspecies due to the higher percentage of linoleic acidHoweverUFApredominated over SFA in all the studied species rangingfrom 65 to 70
Amino acid composition of all the species is shown inTable 5 In all the species aspartic acid (019ndash039) wasfound to be predominated amino acid followed by tyrosine(010ndash021) arginine (012ndash029) alanine (004ndash014)and proline (001ndash007) Amanita caesarea Agaricus arven-sis Cantharellus cibarius Lentinus cladopus and Pleurotuscystidiosus contained maximum amount of these aminoacids Tocopherol contents in all the studied mushroomspecies including three wild are detailed in Table 6 120572-tocopherol and 120573-tocopherol were found to be present in allthe species However 120574-tocopherol was documented fromfew species only Tocopherol content was ranged from 090to 433 120583gg in all the species Cantharellus cibarius (433 plusmn00 120583gg) contained all the three isomers in higher amount ascompared to other species 120573-tocopherol was found in higheramounts as compared to 120572-tocopherol 120574-tocopherol wasdetected only in nine species Cantharellus cibarius (433 plusmn00 120583gg) contained higher amounts of 120574 tocopherol
Results obtained for 120573-carotene lycopene flavonoidsascorbic acid and anthocyanidins composition of all the
twenty species are presented in Table 7 Phenols werethe major antioxidant component detected in significantamounts from all the species (1912ndash6336mgg) followedby anthocyanidins (614ndash1425mg cyanidin chloride100 gextract) flavonoids (114ndash417mgg) ascorbic acid whichwas found in small amounts (020ndash099mgg) 120573-carotene(021ndash079 120583g100 g) and lycopene (019ndash038 120583g100 g) Eachspecies differed with other species in net amounts of all thesecomponents Species like Agaricus arvensis Amanita cae-sarea Gymnopilus junonius Lentinus cladopus and Pleurotuscystidiosus contained higher values of these components ascompared to other species
33 Antioxidant Evaluation Antioxidant properties of allthe species were expressed as EC
radical scavenging activity in all the species showed differ-ences in effectiveness in antioxidant properties Among allthe species Cantharellus cibarius showed lowest EC
50values
(176 plusmn 02mgmL) followed by Amanita caesarea (202 plusmn02mgmL) and Agaricus arvensis (212 plusmn 04mgmL) Otherspecies showed slightly higher EC
50values and therefore
lesserDPPH radical scavenging activityCantharellus cibariusshowed higher DPPH radical scavenging activity and Inocybesplendens showed lower DPPH radical scavenging activitythan other species
For ABTS radical scavenging activities EC50ranged from
426 to 145mgmL Lowest EC50
values were obtained forAmanita caesarea (145 plusmn 06mgmL) showing high antiox-idant activities of this species Higher EC
50values were
8 BioMed Research International
Table 6 Tocopherol composition (120583gg) of twenty species collected from northern Himalayan regions
ND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 7 Other bioactive compounds evaluated from all the species
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
BioMed Research International 9
Table 8 EC50 values for different antioxidant assays on twenty wild culinary species collected from northern Himalayas
obtained for Hygrocybe nivea (426 plusmn 05mgmL) showinglowest ABTS radical scavenging activities of this speciesReducing power of Cantharellus cibarius (162 plusmn 05mgmL)was measured higher than other species Higher EC
50values
for reducing activity were measured in Inocybe splendens(519 plusmn 01mgmL)
Higher effectiveness in ferrous ion chelating activitywas detected in Agaricus arvensis (114 plusmn 01mgmL) andlow effectiveness was detected in Russula mairei (295 plusmn03mgmL) Nevertheless Cantharellus cibarius (127 plusmn02mgmL) Amanita caesarea (140 plusmn 04mgmL) Hygro-cybe coccinea (135 plusmn 02mgmL) and H nivea (145 plusmn06mgmL) showed lower EC
50values than remaining
species EC50
values of scavenging ability on superoxideradical were found to be maximum in Amanita caesarea(044 plusmn 03mgmL) and minimum in Lactarius pubescens(221 plusmn 06mgmL) Gymnopilus junonius showed maximumFRAP activity with least EC
Although there are previous reports on documentation ofculinary edible species from the regions native to northernHimalayas but there are no reports on the evaluation stud-ies as well as toxicity status of all these culinary speciesThe wild edible species Agaricus bisporus Boletus edulis
Morchella esculenta Cordyceps sinensis and Lentinula edodeswhich have been extensively worked out in India and otherparts of the world for their compositional and medicinalaspects have not been undertaken for investigations presently[37ndash40] Compositional studies showed that most of theculinary species were rich in protein carbohydrates andlow in fat There are several reports on richness of wildedible mushrooms with protein and carbohydrate contentsand low fat levels which directly make them nutritionallyrich [24 25 41] Nevertheless under present studies thedifferences between the nutrient concentrations of all thespecies differed but Agaricus arvensis Pleurotus cystidiosusAmanita caesarea Agaricus campestris Cantharellus cibariusand Lentinus cladopus showed higher nutrient percentagewhich is comparable to other wild and commercially cul-tivated species [25 42] The crude fat content detected inall the species was not found to be significantly differentCrude fibres were detected in appreciable percentage fromall the species which make them important in nutritionalpoint of viewThe results are in conformity with the previousreports on several wild edible Pleurotus and Lentinus speciesfrom northwest India [24 43] The species Inocybe splendensHygrocybe nivea and Conocybe tenera were not found tocontain higher percentage of nutrients Although previousreports showed that nutrients composition in wild species isless as compared to cultivated species [25] however Agaricusarvensis Pleurotus cystidiosus Amanita caesarea Agaricuscampestris Cantharellus cibarius and Lentinus cladopus were
10 BioMed Research International
found to be rich in protein and carbohydrates similar tocommercially grown species [25]
Fatty acid composition showed the dominance of UFAover SFA in all the studied mushrooms species which is inconformity with the other studies [41] The differences wereobserved in net amounts in all the species Unsaturationindex of Agaricus arvensis and Lentinus cladopus (057 plusmn002) was found to be significantly higher than otherspecies Whereas Inocybe splendens (005 plusmn 00) showedleast Unsaturation Index High UFA shows the medicinalimportance of these culinary mushrooms as these increasethe HDL cholesterol and decrease LDL cholesterol triacyl-glycerol lipid oxidation and LDL susceptibility to oxidation[44] Predominance of UFA over SFA in all the species showssimilar results as obtained for other wild and commerciallycultivated species [24 25] 120572- and 120573-tocopherol were detectedin higher amounts than third isomer in all the studied speciesSimilar findings were made in other wild and cultivatedspecies with higher 120572- and 120573-tocopherol than 120574-tocopherol[25] The high levels of these two compounds correspondwith a higher oxidative activity which is associated withcardiovascular protection [45] Phenolic compounds weredetected in higher amounts than other bioactive compoundsPresence of high phenolic compounds accounts for the highantioxidant properties of all the species [42] 120573-carotenelycopene and ascorbic acids were detected in low amountsAnthocyanidins were also detected from these wild speciesin appreciable amounts The presence of these functionalmedicinal compounds inmedicinal andor edible mushroomis due to habitat or substrates in which these grow to be highin the functionalmoleculesThe categories of thesemoleculesare anthocyanidins beta-glucans selenium ganoderic acidtriterpenes or cordycepinThe compounds identified in theseextracts show that at least a part of the functional compoundsin medicinal andor edible mushroom is due to growingmushrooms on substrates that are high in the functionalmolecules To these categories can be added anthocyanidinsbeta-glucans selenium ganoderic acid triterpenes or cordy-cepinThe amounts of these have been found to vary with thetype of extraction as ethanolic extract yields higher amountsof anthocyanidins as compared to methanolic hot water andcold water extracts [29]
All the studied species showed significant antioxidantproperties measured on the basis of EC
50values Never-
theless each species showed different antioxidant activitywith highly effective and less effective EC
50values Better
antioxidant properties of some species are due to presence ofhigher phenolic compounds 120573-carotene lycopene ascorbicacids anthocyanidins and tocopherol amounts in themHigh reducing power of some species is due to the presence ofhigher amounts of reducers (antioxidants) in them Presentlyinvestigated species are commonly used for culinary pur-poses in the regions native to northern Himalayas Many ofthe species in these regions are not evaluated previously fordetailed compositional analysisTheir knowledge is restrictedto old aged villagers of the regions and neglected for thecommercial exploitations There are no positive reportson toxicity of these mushrooms analyzed presently hencethese are safe for further experimental work related to
drug discovery All the culinary species contained impor-tant and useful nutraceuticals such as unsaturated fattyacids phenolics carotenoids ascorbic acid tocopherolsand anthocyanidins besides these some important aminoacids were detected in these mushrooms which could beused for the purpose of being used as functional ingredi-ents Since nutraceuticals are powerful in maintaining andpromoting health longevity and life quality the commer-cial exploitation of these species will certainly create animpact on nutritional therapy and also will be beneficialtodayrsquos food industry Direct use of these species for con-sumption and other culinary aspects is safe and healthpromoting with advantage of the additive effects of allthe bioactive and antioxidant compounds present in thesespecies
Abbreviations
AlCl3 Aluminum trichloride
ANOVA Analysis of varianceDW Distilled waterDPPH 22-Diphenyl-1-picrylhydrazylFeCl3 Ferric chloride
FRAP Ferric reducing antioxidant powerg GramsGAEs Gallic acid equivalentsGC Gas ChromatographyHCl Hydrochloric acidH2O Water
H3PO4 Phosphoric acid
HPLC High performance liquid chromatographyK2HPO4 Potassium hydrogen phosphate
The authors declare that there is no conflict of interestsregarding the publication of this paper
BioMed Research International 11
Acknowledgment
The authors wish to thank Science and Engineering BoardDepartment of Science and Technology New Delhi forresearch Grant (SBFTLS-042013) to carry out presentstudies
References
[1] W M Breene ldquoNutritional and medicinal value of specialtymushroomsrdquo Journal of Food Protection vol 53 pp 883ndash8941990
[2] S Chang and P Miles ldquoMushroom biologymdasha new disciplinerdquoMycologist vol 6 no 2 pp 64ndash65 1992
[3] P Manzi L Gambelli S Marconi V Vivanti and L Pizzoferra-to ldquoNutrients in edible mushrooms an inter-species compara-tive studyrdquo Food Chemistry vol 65 no 4 pp 477ndash482 1999
[4] P Bobek O Ozdın and M Mikus ldquoDietary oyster mushroom(Pleurotus ostreatus) accelerates plasma cholesterol turnover inhypercholesterolaemic ratsrdquo Physiological Research vol 44 no5 pp 287ndash291 1995
[5] P Bobek and S Galbavy ldquoHypocholesterolemic and antiathero-genic effect of oystermushroom (Pleurotus ostreatus) in rabbitsrdquoNahrung vol 43 no 5 pp 339ndash342 1999
[6] S Khatun A Islam U Cakilcioglu and N Chatterjee ldquoRe-search on mushroom as a potential source of nutraceuticals areview on Indian perspectiverdquo American Journal of Experimen-tal Agriculture vol 2 no 1 pp 47ndash73 2012
[7] J A Vaz L Barros A Martins J S Morais M H Vasconcelosand I C F R Ferreira ldquoPhenolic profile of seventeen por-tuguese wild mushroomsrdquo LWT Food Science and Technologyvol 44 no 1 pp 343ndash346 2011
[8] J A Vaz L Barros A Martins C Santos-Buelga M HVasconcelos and I C F R Ferreira ldquoChemical compositionof wild edible mushrooms and antioxidant properties of theirwater soluble polysaccharidic and ethanolic fractionsrdquo FoodChemistry vol 126 no 2 pp 610ndash616 2011
[9] T Ozen C Darcan O Aktop and I Turkekul ldquoScreening ofantioxidant antimicrobial activities and chemical contents ofedible mushrooms wildly grown in the Black Sea region ofTurkeyrdquo Combinatorial Chemistry amp High Throughput Screen-ing vol 14 no 2 pp 72ndash84 2011
[10] V E C Ooi and F Liu ldquoA Review of pharmacological activitiesof mushroom polysaccharidesrdquo International Journal of Medic-inal Mushrooms vol 1 no 3 pp 195ndash206 1999
[11] S PWasser and A LWeis ldquoMedicinal properties of substancesoccurring in higher Basidiomycete mushroom current per-spectiverdquo International Journal of Medicinal Mushrooms vol 1pp 31ndash62 1999
[12] P M Kidd ldquoThe use of mushroom glucans and proteoglycansin cancer treatmentrdquo Alternative Medicine Review vol 5 no 1pp 4ndash27 2000
[13] C Israilides and A Philippoussis ldquoBio-technologies of recy-cling agro-industrial wastes for the production of commerciallyimportant fungal polysaccharides andmushroomsrdquo Biotechnol-ogy and Genetic Engineering Reviews vol 20 pp 247ndash259 2003
[14] T Mizuno ldquoBioactive biomolecules of mushrooms food func-tion and medicinal effect of mushroom fungirdquo Food ReviewsInternational vol 11 no 1 pp 5ndash21 1995
[15] Y Kabir and S Kimura ldquoDietary mushrooms reduce bloodpressure in spontaneously hypertensive rats (SHR)rdquo Journal of
Nutritional Science and Vitaminology vol 35 no 1 pp 91ndash941989
[16] J-L Mau S-Y Tsai Y-H Tseng and S-J Huang ldquoAntioxidantproperties of hot water extracts from Ganoderma tsugae Mur-rillrdquo LWTmdashFood Science and Technology vol 38 no 6 pp 589ndash597 2005
[17] A Smania F D Monache E F A Smania M L Gil L CBenchetrit and F S Cruz ldquoAntibacterial activity of a substanceproduced by the fungus Pycnoporus sanguineus (Fr) MurrrdquoJournal of Ethnopharmacology vol 45 no 3 pp 177ndash181 1995
[18] W Andlauer and P Furst ldquoNutraceuticals a piece of historypresent status and outlookrdquo Food Research International vol 35no 2-3 pp 171ndash176 2002
[19] C L Kruger and S W Mann ldquoSafety evaluation of functionalingredientsrdquo Food and Chemical Toxicology vol 41 no 6 pp793ndash805 2003
[20] P H Mattila V I Piironen E J Uusi-Rauva and P E Koivis-toinen ldquoVitamin D contents in edible mushroomsrdquo Journal ofAgricultural and Food Chemistry vol 42 no 11 pp 2449ndash24531994
[21] R P Tewari ldquoMushroom their role in nature and societyrdquo inFrontiers inMushroomBiotechnology R D Rai R C Upadhyayand S R Sharma Eds pp 1ndash8 NRCM Chambaghat India2005
[22] K C Semwal S L Stephenson V K Bhatt and R P BhattldquoEdible mushrooms of the Northwestern Himalaya India astudy of indigenous knowledge distribution and diversityrdquoMycosphere vol 5 no 3 pp 440ndash461 2014
[23] E V Crisan and A Sands ldquoEdible mushrooms nutritionalvaluerdquo in The Biology and Cultivation of Edible Mushrooms ST Chang and W A Hayes Eds pp 137ndash165 Academic PressNew York NY USA 1978
[24] N S Atri S K Sharma R Joshi A Gulati and A Gula-ti ldquoNutritional and neutraceutical composition of five wildculinary-medicinal species of genus Pleurotus (Higher Basid-iomycetes) from Northwest Indiardquo International Journal ofMedicinal Mushrooms vol 15 no 1 pp 49ndash56 2013
[25] L Barros T Cruz P Baptista L M Estevinho and I C FR Ferreira ldquoWild and commercial mushrooms as source ofnutrients and nutraceuticalsrdquo Food and Chemical Toxicologyvol 46 no 8 pp 2742ndash2747 2008
[26] M Nagata and I Yamashita ldquoSimple method for simultaneousdetermination of chlorophyll and carotenoids in tomato fruitrdquoNippon Shokuhin Kogyo Gakkaishi vol 39 no 10 pp 925ndash9281992
[27] V L Singleton and J A Rossi ldquoColorimetric detection oftotal phenolics with phosphomolybdic-phosphotungstic acidreagentsrdquo American Journal of Enology and Viticulture vol 16pp 144ndash158 1965
[28] L K Jagadish V Venkata Krishnan R Shenbhagaramanand V Kaviyarasan ldquoComparitive study on the antioxidantanticancer and antimicrobial property of Agaricus bisporus (JE Lange) Imbach before and after boilingrdquo African Journal ofBiotechnology vol 8 no 4 pp 654ndash661 2009
[29] E Vamanu and S Nita ldquoAntioxidant capacity and the cor-relation with major phenolic compounds anthocyanin andtocopherol content in various extracts from the wild edibleBoletus edulis mushroomrdquo BioMed Research International vol2013 Article ID 313905 11 pages 2013
[30] E Vamanu ldquoBiological activities of the polysaccharides pro-duced in submerged culture of two edible Pleurotus ostreatus
12 BioMed Research International
mushroomsrdquo Journal of Biomedicine and Biotechnology vol2012 Article ID 565974 8 pages 2012
[31] P L Li L Huo W Su et al ldquoFree radical-scavenging capacityantioxidant activity and phenolic content of Pouzolzia zeylan-icardquo Journal of the Serbian Chemical Society vol 76 no 5 pp709ndash717 2011
[32] C Papuc M Crivineanu G Goran V Nicorescu and N Dur-dun ldquoFree radicals scavenging and antioxidant activity ofEuropean mistletoe (Viscum album) and European birthwort(Aristolochia clematitis)rdquo Revista de Chimie vol 61 no 7 pp619ndash622 2010
[33] VOOyetayo CDong andY Yao ldquoAntioxidant and antimicro-bial properties of aqueous extract from Dictyophora indusiatardquoThe Open Mycology Journal vol 3 no 1 pp 20ndash26 2009
[34] S Marklund and G Marklund ldquoInvolvement of the superoxideanion radical in the autoxidation of pyrogallol and a convenientassay for superoxide dismutaserdquo European Journal of Biochem-istry vol 47 no 3 pp 469ndash474 1974
[35] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996
[36] D J Huang b Ou and R L Prior ldquoThe chemistry behindantioxidant capacity assaysrdquo Journal of Agricultural and FoodChemistry vol 53 no 6 pp 1841ndash1856 2005
[37] P Manzi A Aguzzi and L Pizzoferrato ldquoNutritional value ofmushrooms widely consumed in Italyrdquo Food Chemistry vol 73no 3 pp 321ndash325 2001
[38] P Manzi S Marconi A Aguzzi and L Pizzoferrato ldquoCommer-cial mushrooms nutritional quality and effect of cookingrdquo FoodChemistry vol 84 no 2 pp 201ndash206 2004
[39] I P Krbavcic and I C Baric ldquoInfluence of deep fat fryingon some nutritional parameters of novel food based on mush-rooms and fresh soft cheeserdquo Food Chemistry vol 84 no 3 pp417ndash419 2004
[40] D Agrahar-Murugkar andG Subbulakshmi ldquoNutritional valueof edible wild mushrooms collected from the Khasi hills ofMeghalayardquo Food Chemistry vol 89 no 4 pp 599ndash603 2005
[41] V A Dıez and A Alvarez ldquoCompositional and nutritionalstudies on two wild edible mushrooms from northwest SpainrdquoFood Chemistry vol 75 no 4 pp 417ndash422 2001
[42] L Barros M-J Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007
[43] S K Sharma and N S Atri ldquoNutraceutical composition ofwild species of genus Lentinus Fr fromNothern Indiardquo CurrentResearch in Environmental ampAppliedMycology vol 4 pp 11ndash322014
[44] P J Kanu K Zhu J Baby Kanu H Zhou H Qian and K ZhuldquoBiologically active components and nutraceuticals in sesameand related products a review and prospectrdquo Trends in FoodScience and Technology vol 18 no 12 pp 599ndash608 2007
[45] F S Reis E Pereira L Barros M J Sousa A Martins and I CF R Ferreira ldquoBiomolecule profiles in inediblewildmushroomswith antioxidant valuerdquoMolecules vol 16 no 6 pp 4328ndash43382011
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Values are expressed as mean plusmn SE and letters in superscript represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
acids found in the studied species were linoleic acid (C182)followed by oleic acid (C181) and palmitic acid (C160)Besides these three main fatty acids already described sixmore were identified and quantified PUFA were the maingroup of fatty acids documented in all the species Agaricusarvensis Amanita caesarea Cantharellus cibarius Lentinuscladopus and Pleurotus cystidiosus contained lower value ofMUFA but higher percentage of PUFA as compared to otherspecies due to the higher percentage of linoleic acidHoweverUFApredominated over SFA in all the studied species rangingfrom 65 to 70
Amino acid composition of all the species is shown inTable 5 In all the species aspartic acid (019ndash039) wasfound to be predominated amino acid followed by tyrosine(010ndash021) arginine (012ndash029) alanine (004ndash014)and proline (001ndash007) Amanita caesarea Agaricus arven-sis Cantharellus cibarius Lentinus cladopus and Pleurotuscystidiosus contained maximum amount of these aminoacids Tocopherol contents in all the studied mushroomspecies including three wild are detailed in Table 6 120572-tocopherol and 120573-tocopherol were found to be present in allthe species However 120574-tocopherol was documented fromfew species only Tocopherol content was ranged from 090to 433 120583gg in all the species Cantharellus cibarius (433 plusmn00 120583gg) contained all the three isomers in higher amount ascompared to other species 120573-tocopherol was found in higheramounts as compared to 120572-tocopherol 120574-tocopherol wasdetected only in nine species Cantharellus cibarius (433 plusmn00 120583gg) contained higher amounts of 120574 tocopherol
Results obtained for 120573-carotene lycopene flavonoidsascorbic acid and anthocyanidins composition of all the
twenty species are presented in Table 7 Phenols werethe major antioxidant component detected in significantamounts from all the species (1912ndash6336mgg) followedby anthocyanidins (614ndash1425mg cyanidin chloride100 gextract) flavonoids (114ndash417mgg) ascorbic acid whichwas found in small amounts (020ndash099mgg) 120573-carotene(021ndash079 120583g100 g) and lycopene (019ndash038 120583g100 g) Eachspecies differed with other species in net amounts of all thesecomponents Species like Agaricus arvensis Amanita cae-sarea Gymnopilus junonius Lentinus cladopus and Pleurotuscystidiosus contained higher values of these components ascompared to other species
33 Antioxidant Evaluation Antioxidant properties of allthe species were expressed as EC
radical scavenging activity in all the species showed differ-ences in effectiveness in antioxidant properties Among allthe species Cantharellus cibarius showed lowest EC
50values
(176 plusmn 02mgmL) followed by Amanita caesarea (202 plusmn02mgmL) and Agaricus arvensis (212 plusmn 04mgmL) Otherspecies showed slightly higher EC
50values and therefore
lesserDPPH radical scavenging activityCantharellus cibariusshowed higher DPPH radical scavenging activity and Inocybesplendens showed lower DPPH radical scavenging activitythan other species
For ABTS radical scavenging activities EC50ranged from
426 to 145mgmL Lowest EC50
values were obtained forAmanita caesarea (145 plusmn 06mgmL) showing high antiox-idant activities of this species Higher EC
50values were
8 BioMed Research International
Table 6 Tocopherol composition (120583gg) of twenty species collected from northern Himalayan regions
ND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 7 Other bioactive compounds evaluated from all the species
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
BioMed Research International 9
Table 8 EC50 values for different antioxidant assays on twenty wild culinary species collected from northern Himalayas
obtained for Hygrocybe nivea (426 plusmn 05mgmL) showinglowest ABTS radical scavenging activities of this speciesReducing power of Cantharellus cibarius (162 plusmn 05mgmL)was measured higher than other species Higher EC
50values
for reducing activity were measured in Inocybe splendens(519 plusmn 01mgmL)
Higher effectiveness in ferrous ion chelating activitywas detected in Agaricus arvensis (114 plusmn 01mgmL) andlow effectiveness was detected in Russula mairei (295 plusmn03mgmL) Nevertheless Cantharellus cibarius (127 plusmn02mgmL) Amanita caesarea (140 plusmn 04mgmL) Hygro-cybe coccinea (135 plusmn 02mgmL) and H nivea (145 plusmn06mgmL) showed lower EC
50values than remaining
species EC50
values of scavenging ability on superoxideradical were found to be maximum in Amanita caesarea(044 plusmn 03mgmL) and minimum in Lactarius pubescens(221 plusmn 06mgmL) Gymnopilus junonius showed maximumFRAP activity with least EC
Although there are previous reports on documentation ofculinary edible species from the regions native to northernHimalayas but there are no reports on the evaluation stud-ies as well as toxicity status of all these culinary speciesThe wild edible species Agaricus bisporus Boletus edulis
Morchella esculenta Cordyceps sinensis and Lentinula edodeswhich have been extensively worked out in India and otherparts of the world for their compositional and medicinalaspects have not been undertaken for investigations presently[37ndash40] Compositional studies showed that most of theculinary species were rich in protein carbohydrates andlow in fat There are several reports on richness of wildedible mushrooms with protein and carbohydrate contentsand low fat levels which directly make them nutritionallyrich [24 25 41] Nevertheless under present studies thedifferences between the nutrient concentrations of all thespecies differed but Agaricus arvensis Pleurotus cystidiosusAmanita caesarea Agaricus campestris Cantharellus cibariusand Lentinus cladopus showed higher nutrient percentagewhich is comparable to other wild and commercially cul-tivated species [25 42] The crude fat content detected inall the species was not found to be significantly differentCrude fibres were detected in appreciable percentage fromall the species which make them important in nutritionalpoint of viewThe results are in conformity with the previousreports on several wild edible Pleurotus and Lentinus speciesfrom northwest India [24 43] The species Inocybe splendensHygrocybe nivea and Conocybe tenera were not found tocontain higher percentage of nutrients Although previousreports showed that nutrients composition in wild species isless as compared to cultivated species [25] however Agaricusarvensis Pleurotus cystidiosus Amanita caesarea Agaricuscampestris Cantharellus cibarius and Lentinus cladopus were
10 BioMed Research International
found to be rich in protein and carbohydrates similar tocommercially grown species [25]
Fatty acid composition showed the dominance of UFAover SFA in all the studied mushrooms species which is inconformity with the other studies [41] The differences wereobserved in net amounts in all the species Unsaturationindex of Agaricus arvensis and Lentinus cladopus (057 plusmn002) was found to be significantly higher than otherspecies Whereas Inocybe splendens (005 plusmn 00) showedleast Unsaturation Index High UFA shows the medicinalimportance of these culinary mushrooms as these increasethe HDL cholesterol and decrease LDL cholesterol triacyl-glycerol lipid oxidation and LDL susceptibility to oxidation[44] Predominance of UFA over SFA in all the species showssimilar results as obtained for other wild and commerciallycultivated species [24 25] 120572- and 120573-tocopherol were detectedin higher amounts than third isomer in all the studied speciesSimilar findings were made in other wild and cultivatedspecies with higher 120572- and 120573-tocopherol than 120574-tocopherol[25] The high levels of these two compounds correspondwith a higher oxidative activity which is associated withcardiovascular protection [45] Phenolic compounds weredetected in higher amounts than other bioactive compoundsPresence of high phenolic compounds accounts for the highantioxidant properties of all the species [42] 120573-carotenelycopene and ascorbic acids were detected in low amountsAnthocyanidins were also detected from these wild speciesin appreciable amounts The presence of these functionalmedicinal compounds inmedicinal andor edible mushroomis due to habitat or substrates in which these grow to be highin the functionalmoleculesThe categories of thesemoleculesare anthocyanidins beta-glucans selenium ganoderic acidtriterpenes or cordycepinThe compounds identified in theseextracts show that at least a part of the functional compoundsin medicinal andor edible mushroom is due to growingmushrooms on substrates that are high in the functionalmolecules To these categories can be added anthocyanidinsbeta-glucans selenium ganoderic acid triterpenes or cordy-cepinThe amounts of these have been found to vary with thetype of extraction as ethanolic extract yields higher amountsof anthocyanidins as compared to methanolic hot water andcold water extracts [29]
All the studied species showed significant antioxidantproperties measured on the basis of EC
50values Never-
theless each species showed different antioxidant activitywith highly effective and less effective EC
50values Better
antioxidant properties of some species are due to presence ofhigher phenolic compounds 120573-carotene lycopene ascorbicacids anthocyanidins and tocopherol amounts in themHigh reducing power of some species is due to the presence ofhigher amounts of reducers (antioxidants) in them Presentlyinvestigated species are commonly used for culinary pur-poses in the regions native to northern Himalayas Many ofthe species in these regions are not evaluated previously fordetailed compositional analysisTheir knowledge is restrictedto old aged villagers of the regions and neglected for thecommercial exploitations There are no positive reportson toxicity of these mushrooms analyzed presently hencethese are safe for further experimental work related to
drug discovery All the culinary species contained impor-tant and useful nutraceuticals such as unsaturated fattyacids phenolics carotenoids ascorbic acid tocopherolsand anthocyanidins besides these some important aminoacids were detected in these mushrooms which could beused for the purpose of being used as functional ingredi-ents Since nutraceuticals are powerful in maintaining andpromoting health longevity and life quality the commer-cial exploitation of these species will certainly create animpact on nutritional therapy and also will be beneficialtodayrsquos food industry Direct use of these species for con-sumption and other culinary aspects is safe and healthpromoting with advantage of the additive effects of allthe bioactive and antioxidant compounds present in thesespecies
Abbreviations
AlCl3 Aluminum trichloride
ANOVA Analysis of varianceDW Distilled waterDPPH 22-Diphenyl-1-picrylhydrazylFeCl3 Ferric chloride
FRAP Ferric reducing antioxidant powerg GramsGAEs Gallic acid equivalentsGC Gas ChromatographyHCl Hydrochloric acidH2O Water
H3PO4 Phosphoric acid
HPLC High performance liquid chromatographyK2HPO4 Potassium hydrogen phosphate
The authors declare that there is no conflict of interestsregarding the publication of this paper
BioMed Research International 11
Acknowledgment
The authors wish to thank Science and Engineering BoardDepartment of Science and Technology New Delhi forresearch Grant (SBFTLS-042013) to carry out presentstudies
References
[1] W M Breene ldquoNutritional and medicinal value of specialtymushroomsrdquo Journal of Food Protection vol 53 pp 883ndash8941990
[2] S Chang and P Miles ldquoMushroom biologymdasha new disciplinerdquoMycologist vol 6 no 2 pp 64ndash65 1992
[3] P Manzi L Gambelli S Marconi V Vivanti and L Pizzoferra-to ldquoNutrients in edible mushrooms an inter-species compara-tive studyrdquo Food Chemistry vol 65 no 4 pp 477ndash482 1999
[4] P Bobek O Ozdın and M Mikus ldquoDietary oyster mushroom(Pleurotus ostreatus) accelerates plasma cholesterol turnover inhypercholesterolaemic ratsrdquo Physiological Research vol 44 no5 pp 287ndash291 1995
[5] P Bobek and S Galbavy ldquoHypocholesterolemic and antiathero-genic effect of oystermushroom (Pleurotus ostreatus) in rabbitsrdquoNahrung vol 43 no 5 pp 339ndash342 1999
[6] S Khatun A Islam U Cakilcioglu and N Chatterjee ldquoRe-search on mushroom as a potential source of nutraceuticals areview on Indian perspectiverdquo American Journal of Experimen-tal Agriculture vol 2 no 1 pp 47ndash73 2012
[7] J A Vaz L Barros A Martins J S Morais M H Vasconcelosand I C F R Ferreira ldquoPhenolic profile of seventeen por-tuguese wild mushroomsrdquo LWT Food Science and Technologyvol 44 no 1 pp 343ndash346 2011
[8] J A Vaz L Barros A Martins C Santos-Buelga M HVasconcelos and I C F R Ferreira ldquoChemical compositionof wild edible mushrooms and antioxidant properties of theirwater soluble polysaccharidic and ethanolic fractionsrdquo FoodChemistry vol 126 no 2 pp 610ndash616 2011
[9] T Ozen C Darcan O Aktop and I Turkekul ldquoScreening ofantioxidant antimicrobial activities and chemical contents ofedible mushrooms wildly grown in the Black Sea region ofTurkeyrdquo Combinatorial Chemistry amp High Throughput Screen-ing vol 14 no 2 pp 72ndash84 2011
[10] V E C Ooi and F Liu ldquoA Review of pharmacological activitiesof mushroom polysaccharidesrdquo International Journal of Medic-inal Mushrooms vol 1 no 3 pp 195ndash206 1999
[11] S PWasser and A LWeis ldquoMedicinal properties of substancesoccurring in higher Basidiomycete mushroom current per-spectiverdquo International Journal of Medicinal Mushrooms vol 1pp 31ndash62 1999
[12] P M Kidd ldquoThe use of mushroom glucans and proteoglycansin cancer treatmentrdquo Alternative Medicine Review vol 5 no 1pp 4ndash27 2000
[13] C Israilides and A Philippoussis ldquoBio-technologies of recy-cling agro-industrial wastes for the production of commerciallyimportant fungal polysaccharides andmushroomsrdquo Biotechnol-ogy and Genetic Engineering Reviews vol 20 pp 247ndash259 2003
[14] T Mizuno ldquoBioactive biomolecules of mushrooms food func-tion and medicinal effect of mushroom fungirdquo Food ReviewsInternational vol 11 no 1 pp 5ndash21 1995
[15] Y Kabir and S Kimura ldquoDietary mushrooms reduce bloodpressure in spontaneously hypertensive rats (SHR)rdquo Journal of
Nutritional Science and Vitaminology vol 35 no 1 pp 91ndash941989
[16] J-L Mau S-Y Tsai Y-H Tseng and S-J Huang ldquoAntioxidantproperties of hot water extracts from Ganoderma tsugae Mur-rillrdquo LWTmdashFood Science and Technology vol 38 no 6 pp 589ndash597 2005
[17] A Smania F D Monache E F A Smania M L Gil L CBenchetrit and F S Cruz ldquoAntibacterial activity of a substanceproduced by the fungus Pycnoporus sanguineus (Fr) MurrrdquoJournal of Ethnopharmacology vol 45 no 3 pp 177ndash181 1995
[18] W Andlauer and P Furst ldquoNutraceuticals a piece of historypresent status and outlookrdquo Food Research International vol 35no 2-3 pp 171ndash176 2002
[19] C L Kruger and S W Mann ldquoSafety evaluation of functionalingredientsrdquo Food and Chemical Toxicology vol 41 no 6 pp793ndash805 2003
[20] P H Mattila V I Piironen E J Uusi-Rauva and P E Koivis-toinen ldquoVitamin D contents in edible mushroomsrdquo Journal ofAgricultural and Food Chemistry vol 42 no 11 pp 2449ndash24531994
[21] R P Tewari ldquoMushroom their role in nature and societyrdquo inFrontiers inMushroomBiotechnology R D Rai R C Upadhyayand S R Sharma Eds pp 1ndash8 NRCM Chambaghat India2005
[22] K C Semwal S L Stephenson V K Bhatt and R P BhattldquoEdible mushrooms of the Northwestern Himalaya India astudy of indigenous knowledge distribution and diversityrdquoMycosphere vol 5 no 3 pp 440ndash461 2014
[23] E V Crisan and A Sands ldquoEdible mushrooms nutritionalvaluerdquo in The Biology and Cultivation of Edible Mushrooms ST Chang and W A Hayes Eds pp 137ndash165 Academic PressNew York NY USA 1978
[24] N S Atri S K Sharma R Joshi A Gulati and A Gula-ti ldquoNutritional and neutraceutical composition of five wildculinary-medicinal species of genus Pleurotus (Higher Basid-iomycetes) from Northwest Indiardquo International Journal ofMedicinal Mushrooms vol 15 no 1 pp 49ndash56 2013
[25] L Barros T Cruz P Baptista L M Estevinho and I C FR Ferreira ldquoWild and commercial mushrooms as source ofnutrients and nutraceuticalsrdquo Food and Chemical Toxicologyvol 46 no 8 pp 2742ndash2747 2008
[26] M Nagata and I Yamashita ldquoSimple method for simultaneousdetermination of chlorophyll and carotenoids in tomato fruitrdquoNippon Shokuhin Kogyo Gakkaishi vol 39 no 10 pp 925ndash9281992
[27] V L Singleton and J A Rossi ldquoColorimetric detection oftotal phenolics with phosphomolybdic-phosphotungstic acidreagentsrdquo American Journal of Enology and Viticulture vol 16pp 144ndash158 1965
[28] L K Jagadish V Venkata Krishnan R Shenbhagaramanand V Kaviyarasan ldquoComparitive study on the antioxidantanticancer and antimicrobial property of Agaricus bisporus (JE Lange) Imbach before and after boilingrdquo African Journal ofBiotechnology vol 8 no 4 pp 654ndash661 2009
[29] E Vamanu and S Nita ldquoAntioxidant capacity and the cor-relation with major phenolic compounds anthocyanin andtocopherol content in various extracts from the wild edibleBoletus edulis mushroomrdquo BioMed Research International vol2013 Article ID 313905 11 pages 2013
[30] E Vamanu ldquoBiological activities of the polysaccharides pro-duced in submerged culture of two edible Pleurotus ostreatus
12 BioMed Research International
mushroomsrdquo Journal of Biomedicine and Biotechnology vol2012 Article ID 565974 8 pages 2012
[31] P L Li L Huo W Su et al ldquoFree radical-scavenging capacityantioxidant activity and phenolic content of Pouzolzia zeylan-icardquo Journal of the Serbian Chemical Society vol 76 no 5 pp709ndash717 2011
[32] C Papuc M Crivineanu G Goran V Nicorescu and N Dur-dun ldquoFree radicals scavenging and antioxidant activity ofEuropean mistletoe (Viscum album) and European birthwort(Aristolochia clematitis)rdquo Revista de Chimie vol 61 no 7 pp619ndash622 2010
[33] VOOyetayo CDong andY Yao ldquoAntioxidant and antimicro-bial properties of aqueous extract from Dictyophora indusiatardquoThe Open Mycology Journal vol 3 no 1 pp 20ndash26 2009
[34] S Marklund and G Marklund ldquoInvolvement of the superoxideanion radical in the autoxidation of pyrogallol and a convenientassay for superoxide dismutaserdquo European Journal of Biochem-istry vol 47 no 3 pp 469ndash474 1974
[35] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996
[36] D J Huang b Ou and R L Prior ldquoThe chemistry behindantioxidant capacity assaysrdquo Journal of Agricultural and FoodChemistry vol 53 no 6 pp 1841ndash1856 2005
[37] P Manzi A Aguzzi and L Pizzoferrato ldquoNutritional value ofmushrooms widely consumed in Italyrdquo Food Chemistry vol 73no 3 pp 321ndash325 2001
[38] P Manzi S Marconi A Aguzzi and L Pizzoferrato ldquoCommer-cial mushrooms nutritional quality and effect of cookingrdquo FoodChemistry vol 84 no 2 pp 201ndash206 2004
[39] I P Krbavcic and I C Baric ldquoInfluence of deep fat fryingon some nutritional parameters of novel food based on mush-rooms and fresh soft cheeserdquo Food Chemistry vol 84 no 3 pp417ndash419 2004
[40] D Agrahar-Murugkar andG Subbulakshmi ldquoNutritional valueof edible wild mushrooms collected from the Khasi hills ofMeghalayardquo Food Chemistry vol 89 no 4 pp 599ndash603 2005
[41] V A Dıez and A Alvarez ldquoCompositional and nutritionalstudies on two wild edible mushrooms from northwest SpainrdquoFood Chemistry vol 75 no 4 pp 417ndash422 2001
[42] L Barros M-J Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007
[43] S K Sharma and N S Atri ldquoNutraceutical composition ofwild species of genus Lentinus Fr fromNothern Indiardquo CurrentResearch in Environmental ampAppliedMycology vol 4 pp 11ndash322014
[44] P J Kanu K Zhu J Baby Kanu H Zhou H Qian and K ZhuldquoBiologically active components and nutraceuticals in sesameand related products a review and prospectrdquo Trends in FoodScience and Technology vol 18 no 12 pp 599ndash608 2007
[45] F S Reis E Pereira L Barros M J Sousa A Martins and I CF R Ferreira ldquoBiomolecule profiles in inediblewildmushroomswith antioxidant valuerdquoMolecules vol 16 no 6 pp 4328ndash43382011
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Values are expressed as mean plusmn SE and letters in superscript represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
acids found in the studied species were linoleic acid (C182)followed by oleic acid (C181) and palmitic acid (C160)Besides these three main fatty acids already described sixmore were identified and quantified PUFA were the maingroup of fatty acids documented in all the species Agaricusarvensis Amanita caesarea Cantharellus cibarius Lentinuscladopus and Pleurotus cystidiosus contained lower value ofMUFA but higher percentage of PUFA as compared to otherspecies due to the higher percentage of linoleic acidHoweverUFApredominated over SFA in all the studied species rangingfrom 65 to 70
Amino acid composition of all the species is shown inTable 5 In all the species aspartic acid (019ndash039) wasfound to be predominated amino acid followed by tyrosine(010ndash021) arginine (012ndash029) alanine (004ndash014)and proline (001ndash007) Amanita caesarea Agaricus arven-sis Cantharellus cibarius Lentinus cladopus and Pleurotuscystidiosus contained maximum amount of these aminoacids Tocopherol contents in all the studied mushroomspecies including three wild are detailed in Table 6 120572-tocopherol and 120573-tocopherol were found to be present in allthe species However 120574-tocopherol was documented fromfew species only Tocopherol content was ranged from 090to 433 120583gg in all the species Cantharellus cibarius (433 plusmn00 120583gg) contained all the three isomers in higher amount ascompared to other species 120573-tocopherol was found in higheramounts as compared to 120572-tocopherol 120574-tocopherol wasdetected only in nine species Cantharellus cibarius (433 plusmn00 120583gg) contained higher amounts of 120574 tocopherol
Results obtained for 120573-carotene lycopene flavonoidsascorbic acid and anthocyanidins composition of all the
twenty species are presented in Table 7 Phenols werethe major antioxidant component detected in significantamounts from all the species (1912ndash6336mgg) followedby anthocyanidins (614ndash1425mg cyanidin chloride100 gextract) flavonoids (114ndash417mgg) ascorbic acid whichwas found in small amounts (020ndash099mgg) 120573-carotene(021ndash079 120583g100 g) and lycopene (019ndash038 120583g100 g) Eachspecies differed with other species in net amounts of all thesecomponents Species like Agaricus arvensis Amanita cae-sarea Gymnopilus junonius Lentinus cladopus and Pleurotuscystidiosus contained higher values of these components ascompared to other species
33 Antioxidant Evaluation Antioxidant properties of allthe species were expressed as EC
radical scavenging activity in all the species showed differ-ences in effectiveness in antioxidant properties Among allthe species Cantharellus cibarius showed lowest EC
50values
(176 plusmn 02mgmL) followed by Amanita caesarea (202 plusmn02mgmL) and Agaricus arvensis (212 plusmn 04mgmL) Otherspecies showed slightly higher EC
50values and therefore
lesserDPPH radical scavenging activityCantharellus cibariusshowed higher DPPH radical scavenging activity and Inocybesplendens showed lower DPPH radical scavenging activitythan other species
For ABTS radical scavenging activities EC50ranged from
426 to 145mgmL Lowest EC50
values were obtained forAmanita caesarea (145 plusmn 06mgmL) showing high antiox-idant activities of this species Higher EC
50values were
8 BioMed Research International
Table 6 Tocopherol composition (120583gg) of twenty species collected from northern Himalayan regions
ND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 7 Other bioactive compounds evaluated from all the species
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
BioMed Research International 9
Table 8 EC50 values for different antioxidant assays on twenty wild culinary species collected from northern Himalayas
obtained for Hygrocybe nivea (426 plusmn 05mgmL) showinglowest ABTS radical scavenging activities of this speciesReducing power of Cantharellus cibarius (162 plusmn 05mgmL)was measured higher than other species Higher EC
50values
for reducing activity were measured in Inocybe splendens(519 plusmn 01mgmL)
Higher effectiveness in ferrous ion chelating activitywas detected in Agaricus arvensis (114 plusmn 01mgmL) andlow effectiveness was detected in Russula mairei (295 plusmn03mgmL) Nevertheless Cantharellus cibarius (127 plusmn02mgmL) Amanita caesarea (140 plusmn 04mgmL) Hygro-cybe coccinea (135 plusmn 02mgmL) and H nivea (145 plusmn06mgmL) showed lower EC
50values than remaining
species EC50
values of scavenging ability on superoxideradical were found to be maximum in Amanita caesarea(044 plusmn 03mgmL) and minimum in Lactarius pubescens(221 plusmn 06mgmL) Gymnopilus junonius showed maximumFRAP activity with least EC
Although there are previous reports on documentation ofculinary edible species from the regions native to northernHimalayas but there are no reports on the evaluation stud-ies as well as toxicity status of all these culinary speciesThe wild edible species Agaricus bisporus Boletus edulis
Morchella esculenta Cordyceps sinensis and Lentinula edodeswhich have been extensively worked out in India and otherparts of the world for their compositional and medicinalaspects have not been undertaken for investigations presently[37ndash40] Compositional studies showed that most of theculinary species were rich in protein carbohydrates andlow in fat There are several reports on richness of wildedible mushrooms with protein and carbohydrate contentsand low fat levels which directly make them nutritionallyrich [24 25 41] Nevertheless under present studies thedifferences between the nutrient concentrations of all thespecies differed but Agaricus arvensis Pleurotus cystidiosusAmanita caesarea Agaricus campestris Cantharellus cibariusand Lentinus cladopus showed higher nutrient percentagewhich is comparable to other wild and commercially cul-tivated species [25 42] The crude fat content detected inall the species was not found to be significantly differentCrude fibres were detected in appreciable percentage fromall the species which make them important in nutritionalpoint of viewThe results are in conformity with the previousreports on several wild edible Pleurotus and Lentinus speciesfrom northwest India [24 43] The species Inocybe splendensHygrocybe nivea and Conocybe tenera were not found tocontain higher percentage of nutrients Although previousreports showed that nutrients composition in wild species isless as compared to cultivated species [25] however Agaricusarvensis Pleurotus cystidiosus Amanita caesarea Agaricuscampestris Cantharellus cibarius and Lentinus cladopus were
10 BioMed Research International
found to be rich in protein and carbohydrates similar tocommercially grown species [25]
Fatty acid composition showed the dominance of UFAover SFA in all the studied mushrooms species which is inconformity with the other studies [41] The differences wereobserved in net amounts in all the species Unsaturationindex of Agaricus arvensis and Lentinus cladopus (057 plusmn002) was found to be significantly higher than otherspecies Whereas Inocybe splendens (005 plusmn 00) showedleast Unsaturation Index High UFA shows the medicinalimportance of these culinary mushrooms as these increasethe HDL cholesterol and decrease LDL cholesterol triacyl-glycerol lipid oxidation and LDL susceptibility to oxidation[44] Predominance of UFA over SFA in all the species showssimilar results as obtained for other wild and commerciallycultivated species [24 25] 120572- and 120573-tocopherol were detectedin higher amounts than third isomer in all the studied speciesSimilar findings were made in other wild and cultivatedspecies with higher 120572- and 120573-tocopherol than 120574-tocopherol[25] The high levels of these two compounds correspondwith a higher oxidative activity which is associated withcardiovascular protection [45] Phenolic compounds weredetected in higher amounts than other bioactive compoundsPresence of high phenolic compounds accounts for the highantioxidant properties of all the species [42] 120573-carotenelycopene and ascorbic acids were detected in low amountsAnthocyanidins were also detected from these wild speciesin appreciable amounts The presence of these functionalmedicinal compounds inmedicinal andor edible mushroomis due to habitat or substrates in which these grow to be highin the functionalmoleculesThe categories of thesemoleculesare anthocyanidins beta-glucans selenium ganoderic acidtriterpenes or cordycepinThe compounds identified in theseextracts show that at least a part of the functional compoundsin medicinal andor edible mushroom is due to growingmushrooms on substrates that are high in the functionalmolecules To these categories can be added anthocyanidinsbeta-glucans selenium ganoderic acid triterpenes or cordy-cepinThe amounts of these have been found to vary with thetype of extraction as ethanolic extract yields higher amountsof anthocyanidins as compared to methanolic hot water andcold water extracts [29]
All the studied species showed significant antioxidantproperties measured on the basis of EC
50values Never-
theless each species showed different antioxidant activitywith highly effective and less effective EC
50values Better
antioxidant properties of some species are due to presence ofhigher phenolic compounds 120573-carotene lycopene ascorbicacids anthocyanidins and tocopherol amounts in themHigh reducing power of some species is due to the presence ofhigher amounts of reducers (antioxidants) in them Presentlyinvestigated species are commonly used for culinary pur-poses in the regions native to northern Himalayas Many ofthe species in these regions are not evaluated previously fordetailed compositional analysisTheir knowledge is restrictedto old aged villagers of the regions and neglected for thecommercial exploitations There are no positive reportson toxicity of these mushrooms analyzed presently hencethese are safe for further experimental work related to
drug discovery All the culinary species contained impor-tant and useful nutraceuticals such as unsaturated fattyacids phenolics carotenoids ascorbic acid tocopherolsand anthocyanidins besides these some important aminoacids were detected in these mushrooms which could beused for the purpose of being used as functional ingredi-ents Since nutraceuticals are powerful in maintaining andpromoting health longevity and life quality the commer-cial exploitation of these species will certainly create animpact on nutritional therapy and also will be beneficialtodayrsquos food industry Direct use of these species for con-sumption and other culinary aspects is safe and healthpromoting with advantage of the additive effects of allthe bioactive and antioxidant compounds present in thesespecies
Abbreviations
AlCl3 Aluminum trichloride
ANOVA Analysis of varianceDW Distilled waterDPPH 22-Diphenyl-1-picrylhydrazylFeCl3 Ferric chloride
FRAP Ferric reducing antioxidant powerg GramsGAEs Gallic acid equivalentsGC Gas ChromatographyHCl Hydrochloric acidH2O Water
H3PO4 Phosphoric acid
HPLC High performance liquid chromatographyK2HPO4 Potassium hydrogen phosphate
The authors declare that there is no conflict of interestsregarding the publication of this paper
BioMed Research International 11
Acknowledgment
The authors wish to thank Science and Engineering BoardDepartment of Science and Technology New Delhi forresearch Grant (SBFTLS-042013) to carry out presentstudies
References
[1] W M Breene ldquoNutritional and medicinal value of specialtymushroomsrdquo Journal of Food Protection vol 53 pp 883ndash8941990
[2] S Chang and P Miles ldquoMushroom biologymdasha new disciplinerdquoMycologist vol 6 no 2 pp 64ndash65 1992
[3] P Manzi L Gambelli S Marconi V Vivanti and L Pizzoferra-to ldquoNutrients in edible mushrooms an inter-species compara-tive studyrdquo Food Chemistry vol 65 no 4 pp 477ndash482 1999
[4] P Bobek O Ozdın and M Mikus ldquoDietary oyster mushroom(Pleurotus ostreatus) accelerates plasma cholesterol turnover inhypercholesterolaemic ratsrdquo Physiological Research vol 44 no5 pp 287ndash291 1995
[5] P Bobek and S Galbavy ldquoHypocholesterolemic and antiathero-genic effect of oystermushroom (Pleurotus ostreatus) in rabbitsrdquoNahrung vol 43 no 5 pp 339ndash342 1999
[6] S Khatun A Islam U Cakilcioglu and N Chatterjee ldquoRe-search on mushroom as a potential source of nutraceuticals areview on Indian perspectiverdquo American Journal of Experimen-tal Agriculture vol 2 no 1 pp 47ndash73 2012
[7] J A Vaz L Barros A Martins J S Morais M H Vasconcelosand I C F R Ferreira ldquoPhenolic profile of seventeen por-tuguese wild mushroomsrdquo LWT Food Science and Technologyvol 44 no 1 pp 343ndash346 2011
[8] J A Vaz L Barros A Martins C Santos-Buelga M HVasconcelos and I C F R Ferreira ldquoChemical compositionof wild edible mushrooms and antioxidant properties of theirwater soluble polysaccharidic and ethanolic fractionsrdquo FoodChemistry vol 126 no 2 pp 610ndash616 2011
[9] T Ozen C Darcan O Aktop and I Turkekul ldquoScreening ofantioxidant antimicrobial activities and chemical contents ofedible mushrooms wildly grown in the Black Sea region ofTurkeyrdquo Combinatorial Chemistry amp High Throughput Screen-ing vol 14 no 2 pp 72ndash84 2011
[10] V E C Ooi and F Liu ldquoA Review of pharmacological activitiesof mushroom polysaccharidesrdquo International Journal of Medic-inal Mushrooms vol 1 no 3 pp 195ndash206 1999
[11] S PWasser and A LWeis ldquoMedicinal properties of substancesoccurring in higher Basidiomycete mushroom current per-spectiverdquo International Journal of Medicinal Mushrooms vol 1pp 31ndash62 1999
[12] P M Kidd ldquoThe use of mushroom glucans and proteoglycansin cancer treatmentrdquo Alternative Medicine Review vol 5 no 1pp 4ndash27 2000
[13] C Israilides and A Philippoussis ldquoBio-technologies of recy-cling agro-industrial wastes for the production of commerciallyimportant fungal polysaccharides andmushroomsrdquo Biotechnol-ogy and Genetic Engineering Reviews vol 20 pp 247ndash259 2003
[14] T Mizuno ldquoBioactive biomolecules of mushrooms food func-tion and medicinal effect of mushroom fungirdquo Food ReviewsInternational vol 11 no 1 pp 5ndash21 1995
[15] Y Kabir and S Kimura ldquoDietary mushrooms reduce bloodpressure in spontaneously hypertensive rats (SHR)rdquo Journal of
Nutritional Science and Vitaminology vol 35 no 1 pp 91ndash941989
[16] J-L Mau S-Y Tsai Y-H Tseng and S-J Huang ldquoAntioxidantproperties of hot water extracts from Ganoderma tsugae Mur-rillrdquo LWTmdashFood Science and Technology vol 38 no 6 pp 589ndash597 2005
[17] A Smania F D Monache E F A Smania M L Gil L CBenchetrit and F S Cruz ldquoAntibacterial activity of a substanceproduced by the fungus Pycnoporus sanguineus (Fr) MurrrdquoJournal of Ethnopharmacology vol 45 no 3 pp 177ndash181 1995
[18] W Andlauer and P Furst ldquoNutraceuticals a piece of historypresent status and outlookrdquo Food Research International vol 35no 2-3 pp 171ndash176 2002
[19] C L Kruger and S W Mann ldquoSafety evaluation of functionalingredientsrdquo Food and Chemical Toxicology vol 41 no 6 pp793ndash805 2003
[20] P H Mattila V I Piironen E J Uusi-Rauva and P E Koivis-toinen ldquoVitamin D contents in edible mushroomsrdquo Journal ofAgricultural and Food Chemistry vol 42 no 11 pp 2449ndash24531994
[21] R P Tewari ldquoMushroom their role in nature and societyrdquo inFrontiers inMushroomBiotechnology R D Rai R C Upadhyayand S R Sharma Eds pp 1ndash8 NRCM Chambaghat India2005
[22] K C Semwal S L Stephenson V K Bhatt and R P BhattldquoEdible mushrooms of the Northwestern Himalaya India astudy of indigenous knowledge distribution and diversityrdquoMycosphere vol 5 no 3 pp 440ndash461 2014
[23] E V Crisan and A Sands ldquoEdible mushrooms nutritionalvaluerdquo in The Biology and Cultivation of Edible Mushrooms ST Chang and W A Hayes Eds pp 137ndash165 Academic PressNew York NY USA 1978
[24] N S Atri S K Sharma R Joshi A Gulati and A Gula-ti ldquoNutritional and neutraceutical composition of five wildculinary-medicinal species of genus Pleurotus (Higher Basid-iomycetes) from Northwest Indiardquo International Journal ofMedicinal Mushrooms vol 15 no 1 pp 49ndash56 2013
[25] L Barros T Cruz P Baptista L M Estevinho and I C FR Ferreira ldquoWild and commercial mushrooms as source ofnutrients and nutraceuticalsrdquo Food and Chemical Toxicologyvol 46 no 8 pp 2742ndash2747 2008
[26] M Nagata and I Yamashita ldquoSimple method for simultaneousdetermination of chlorophyll and carotenoids in tomato fruitrdquoNippon Shokuhin Kogyo Gakkaishi vol 39 no 10 pp 925ndash9281992
[27] V L Singleton and J A Rossi ldquoColorimetric detection oftotal phenolics with phosphomolybdic-phosphotungstic acidreagentsrdquo American Journal of Enology and Viticulture vol 16pp 144ndash158 1965
[28] L K Jagadish V Venkata Krishnan R Shenbhagaramanand V Kaviyarasan ldquoComparitive study on the antioxidantanticancer and antimicrobial property of Agaricus bisporus (JE Lange) Imbach before and after boilingrdquo African Journal ofBiotechnology vol 8 no 4 pp 654ndash661 2009
[29] E Vamanu and S Nita ldquoAntioxidant capacity and the cor-relation with major phenolic compounds anthocyanin andtocopherol content in various extracts from the wild edibleBoletus edulis mushroomrdquo BioMed Research International vol2013 Article ID 313905 11 pages 2013
[30] E Vamanu ldquoBiological activities of the polysaccharides pro-duced in submerged culture of two edible Pleurotus ostreatus
12 BioMed Research International
mushroomsrdquo Journal of Biomedicine and Biotechnology vol2012 Article ID 565974 8 pages 2012
[31] P L Li L Huo W Su et al ldquoFree radical-scavenging capacityantioxidant activity and phenolic content of Pouzolzia zeylan-icardquo Journal of the Serbian Chemical Society vol 76 no 5 pp709ndash717 2011
[32] C Papuc M Crivineanu G Goran V Nicorescu and N Dur-dun ldquoFree radicals scavenging and antioxidant activity ofEuropean mistletoe (Viscum album) and European birthwort(Aristolochia clematitis)rdquo Revista de Chimie vol 61 no 7 pp619ndash622 2010
[33] VOOyetayo CDong andY Yao ldquoAntioxidant and antimicro-bial properties of aqueous extract from Dictyophora indusiatardquoThe Open Mycology Journal vol 3 no 1 pp 20ndash26 2009
[34] S Marklund and G Marklund ldquoInvolvement of the superoxideanion radical in the autoxidation of pyrogallol and a convenientassay for superoxide dismutaserdquo European Journal of Biochem-istry vol 47 no 3 pp 469ndash474 1974
[35] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996
[36] D J Huang b Ou and R L Prior ldquoThe chemistry behindantioxidant capacity assaysrdquo Journal of Agricultural and FoodChemistry vol 53 no 6 pp 1841ndash1856 2005
[37] P Manzi A Aguzzi and L Pizzoferrato ldquoNutritional value ofmushrooms widely consumed in Italyrdquo Food Chemistry vol 73no 3 pp 321ndash325 2001
[38] P Manzi S Marconi A Aguzzi and L Pizzoferrato ldquoCommer-cial mushrooms nutritional quality and effect of cookingrdquo FoodChemistry vol 84 no 2 pp 201ndash206 2004
[39] I P Krbavcic and I C Baric ldquoInfluence of deep fat fryingon some nutritional parameters of novel food based on mush-rooms and fresh soft cheeserdquo Food Chemistry vol 84 no 3 pp417ndash419 2004
[40] D Agrahar-Murugkar andG Subbulakshmi ldquoNutritional valueof edible wild mushrooms collected from the Khasi hills ofMeghalayardquo Food Chemistry vol 89 no 4 pp 599ndash603 2005
[41] V A Dıez and A Alvarez ldquoCompositional and nutritionalstudies on two wild edible mushrooms from northwest SpainrdquoFood Chemistry vol 75 no 4 pp 417ndash422 2001
[42] L Barros M-J Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007
[43] S K Sharma and N S Atri ldquoNutraceutical composition ofwild species of genus Lentinus Fr fromNothern Indiardquo CurrentResearch in Environmental ampAppliedMycology vol 4 pp 11ndash322014
[44] P J Kanu K Zhu J Baby Kanu H Zhou H Qian and K ZhuldquoBiologically active components and nutraceuticals in sesameand related products a review and prospectrdquo Trends in FoodScience and Technology vol 18 no 12 pp 599ndash608 2007
[45] F S Reis E Pereira L Barros M J Sousa A Martins and I CF R Ferreira ldquoBiomolecule profiles in inediblewildmushroomswith antioxidant valuerdquoMolecules vol 16 no 6 pp 4328ndash43382011
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
ND = not detectedValues are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
Table 7 Other bioactive compounds evaluated from all the species
Values are expressed as mean plusmn SE and different letters represent the significant difference in each column with 119901 le 005 according to Tukeyrsquos test
BioMed Research International 9
Table 8 EC50 values for different antioxidant assays on twenty wild culinary species collected from northern Himalayas
obtained for Hygrocybe nivea (426 plusmn 05mgmL) showinglowest ABTS radical scavenging activities of this speciesReducing power of Cantharellus cibarius (162 plusmn 05mgmL)was measured higher than other species Higher EC
50values
for reducing activity were measured in Inocybe splendens(519 plusmn 01mgmL)
Higher effectiveness in ferrous ion chelating activitywas detected in Agaricus arvensis (114 plusmn 01mgmL) andlow effectiveness was detected in Russula mairei (295 plusmn03mgmL) Nevertheless Cantharellus cibarius (127 plusmn02mgmL) Amanita caesarea (140 plusmn 04mgmL) Hygro-cybe coccinea (135 plusmn 02mgmL) and H nivea (145 plusmn06mgmL) showed lower EC
50values than remaining
species EC50
values of scavenging ability on superoxideradical were found to be maximum in Amanita caesarea(044 plusmn 03mgmL) and minimum in Lactarius pubescens(221 plusmn 06mgmL) Gymnopilus junonius showed maximumFRAP activity with least EC
Although there are previous reports on documentation ofculinary edible species from the regions native to northernHimalayas but there are no reports on the evaluation stud-ies as well as toxicity status of all these culinary speciesThe wild edible species Agaricus bisporus Boletus edulis
Morchella esculenta Cordyceps sinensis and Lentinula edodeswhich have been extensively worked out in India and otherparts of the world for their compositional and medicinalaspects have not been undertaken for investigations presently[37ndash40] Compositional studies showed that most of theculinary species were rich in protein carbohydrates andlow in fat There are several reports on richness of wildedible mushrooms with protein and carbohydrate contentsand low fat levels which directly make them nutritionallyrich [24 25 41] Nevertheless under present studies thedifferences between the nutrient concentrations of all thespecies differed but Agaricus arvensis Pleurotus cystidiosusAmanita caesarea Agaricus campestris Cantharellus cibariusand Lentinus cladopus showed higher nutrient percentagewhich is comparable to other wild and commercially cul-tivated species [25 42] The crude fat content detected inall the species was not found to be significantly differentCrude fibres were detected in appreciable percentage fromall the species which make them important in nutritionalpoint of viewThe results are in conformity with the previousreports on several wild edible Pleurotus and Lentinus speciesfrom northwest India [24 43] The species Inocybe splendensHygrocybe nivea and Conocybe tenera were not found tocontain higher percentage of nutrients Although previousreports showed that nutrients composition in wild species isless as compared to cultivated species [25] however Agaricusarvensis Pleurotus cystidiosus Amanita caesarea Agaricuscampestris Cantharellus cibarius and Lentinus cladopus were
10 BioMed Research International
found to be rich in protein and carbohydrates similar tocommercially grown species [25]
Fatty acid composition showed the dominance of UFAover SFA in all the studied mushrooms species which is inconformity with the other studies [41] The differences wereobserved in net amounts in all the species Unsaturationindex of Agaricus arvensis and Lentinus cladopus (057 plusmn002) was found to be significantly higher than otherspecies Whereas Inocybe splendens (005 plusmn 00) showedleast Unsaturation Index High UFA shows the medicinalimportance of these culinary mushrooms as these increasethe HDL cholesterol and decrease LDL cholesterol triacyl-glycerol lipid oxidation and LDL susceptibility to oxidation[44] Predominance of UFA over SFA in all the species showssimilar results as obtained for other wild and commerciallycultivated species [24 25] 120572- and 120573-tocopherol were detectedin higher amounts than third isomer in all the studied speciesSimilar findings were made in other wild and cultivatedspecies with higher 120572- and 120573-tocopherol than 120574-tocopherol[25] The high levels of these two compounds correspondwith a higher oxidative activity which is associated withcardiovascular protection [45] Phenolic compounds weredetected in higher amounts than other bioactive compoundsPresence of high phenolic compounds accounts for the highantioxidant properties of all the species [42] 120573-carotenelycopene and ascorbic acids were detected in low amountsAnthocyanidins were also detected from these wild speciesin appreciable amounts The presence of these functionalmedicinal compounds inmedicinal andor edible mushroomis due to habitat or substrates in which these grow to be highin the functionalmoleculesThe categories of thesemoleculesare anthocyanidins beta-glucans selenium ganoderic acidtriterpenes or cordycepinThe compounds identified in theseextracts show that at least a part of the functional compoundsin medicinal andor edible mushroom is due to growingmushrooms on substrates that are high in the functionalmolecules To these categories can be added anthocyanidinsbeta-glucans selenium ganoderic acid triterpenes or cordy-cepinThe amounts of these have been found to vary with thetype of extraction as ethanolic extract yields higher amountsof anthocyanidins as compared to methanolic hot water andcold water extracts [29]
All the studied species showed significant antioxidantproperties measured on the basis of EC
50values Never-
theless each species showed different antioxidant activitywith highly effective and less effective EC
50values Better
antioxidant properties of some species are due to presence ofhigher phenolic compounds 120573-carotene lycopene ascorbicacids anthocyanidins and tocopherol amounts in themHigh reducing power of some species is due to the presence ofhigher amounts of reducers (antioxidants) in them Presentlyinvestigated species are commonly used for culinary pur-poses in the regions native to northern Himalayas Many ofthe species in these regions are not evaluated previously fordetailed compositional analysisTheir knowledge is restrictedto old aged villagers of the regions and neglected for thecommercial exploitations There are no positive reportson toxicity of these mushrooms analyzed presently hencethese are safe for further experimental work related to
drug discovery All the culinary species contained impor-tant and useful nutraceuticals such as unsaturated fattyacids phenolics carotenoids ascorbic acid tocopherolsand anthocyanidins besides these some important aminoacids were detected in these mushrooms which could beused for the purpose of being used as functional ingredi-ents Since nutraceuticals are powerful in maintaining andpromoting health longevity and life quality the commer-cial exploitation of these species will certainly create animpact on nutritional therapy and also will be beneficialtodayrsquos food industry Direct use of these species for con-sumption and other culinary aspects is safe and healthpromoting with advantage of the additive effects of allthe bioactive and antioxidant compounds present in thesespecies
Abbreviations
AlCl3 Aluminum trichloride
ANOVA Analysis of varianceDW Distilled waterDPPH 22-Diphenyl-1-picrylhydrazylFeCl3 Ferric chloride
FRAP Ferric reducing antioxidant powerg GramsGAEs Gallic acid equivalentsGC Gas ChromatographyHCl Hydrochloric acidH2O Water
H3PO4 Phosphoric acid
HPLC High performance liquid chromatographyK2HPO4 Potassium hydrogen phosphate
The authors declare that there is no conflict of interestsregarding the publication of this paper
BioMed Research International 11
Acknowledgment
The authors wish to thank Science and Engineering BoardDepartment of Science and Technology New Delhi forresearch Grant (SBFTLS-042013) to carry out presentstudies
References
[1] W M Breene ldquoNutritional and medicinal value of specialtymushroomsrdquo Journal of Food Protection vol 53 pp 883ndash8941990
[2] S Chang and P Miles ldquoMushroom biologymdasha new disciplinerdquoMycologist vol 6 no 2 pp 64ndash65 1992
[3] P Manzi L Gambelli S Marconi V Vivanti and L Pizzoferra-to ldquoNutrients in edible mushrooms an inter-species compara-tive studyrdquo Food Chemistry vol 65 no 4 pp 477ndash482 1999
[4] P Bobek O Ozdın and M Mikus ldquoDietary oyster mushroom(Pleurotus ostreatus) accelerates plasma cholesterol turnover inhypercholesterolaemic ratsrdquo Physiological Research vol 44 no5 pp 287ndash291 1995
[5] P Bobek and S Galbavy ldquoHypocholesterolemic and antiathero-genic effect of oystermushroom (Pleurotus ostreatus) in rabbitsrdquoNahrung vol 43 no 5 pp 339ndash342 1999
[6] S Khatun A Islam U Cakilcioglu and N Chatterjee ldquoRe-search on mushroom as a potential source of nutraceuticals areview on Indian perspectiverdquo American Journal of Experimen-tal Agriculture vol 2 no 1 pp 47ndash73 2012
[7] J A Vaz L Barros A Martins J S Morais M H Vasconcelosand I C F R Ferreira ldquoPhenolic profile of seventeen por-tuguese wild mushroomsrdquo LWT Food Science and Technologyvol 44 no 1 pp 343ndash346 2011
[8] J A Vaz L Barros A Martins C Santos-Buelga M HVasconcelos and I C F R Ferreira ldquoChemical compositionof wild edible mushrooms and antioxidant properties of theirwater soluble polysaccharidic and ethanolic fractionsrdquo FoodChemistry vol 126 no 2 pp 610ndash616 2011
[9] T Ozen C Darcan O Aktop and I Turkekul ldquoScreening ofantioxidant antimicrobial activities and chemical contents ofedible mushrooms wildly grown in the Black Sea region ofTurkeyrdquo Combinatorial Chemistry amp High Throughput Screen-ing vol 14 no 2 pp 72ndash84 2011
[10] V E C Ooi and F Liu ldquoA Review of pharmacological activitiesof mushroom polysaccharidesrdquo International Journal of Medic-inal Mushrooms vol 1 no 3 pp 195ndash206 1999
[11] S PWasser and A LWeis ldquoMedicinal properties of substancesoccurring in higher Basidiomycete mushroom current per-spectiverdquo International Journal of Medicinal Mushrooms vol 1pp 31ndash62 1999
[12] P M Kidd ldquoThe use of mushroom glucans and proteoglycansin cancer treatmentrdquo Alternative Medicine Review vol 5 no 1pp 4ndash27 2000
[13] C Israilides and A Philippoussis ldquoBio-technologies of recy-cling agro-industrial wastes for the production of commerciallyimportant fungal polysaccharides andmushroomsrdquo Biotechnol-ogy and Genetic Engineering Reviews vol 20 pp 247ndash259 2003
[14] T Mizuno ldquoBioactive biomolecules of mushrooms food func-tion and medicinal effect of mushroom fungirdquo Food ReviewsInternational vol 11 no 1 pp 5ndash21 1995
[15] Y Kabir and S Kimura ldquoDietary mushrooms reduce bloodpressure in spontaneously hypertensive rats (SHR)rdquo Journal of
Nutritional Science and Vitaminology vol 35 no 1 pp 91ndash941989
[16] J-L Mau S-Y Tsai Y-H Tseng and S-J Huang ldquoAntioxidantproperties of hot water extracts from Ganoderma tsugae Mur-rillrdquo LWTmdashFood Science and Technology vol 38 no 6 pp 589ndash597 2005
[17] A Smania F D Monache E F A Smania M L Gil L CBenchetrit and F S Cruz ldquoAntibacterial activity of a substanceproduced by the fungus Pycnoporus sanguineus (Fr) MurrrdquoJournal of Ethnopharmacology vol 45 no 3 pp 177ndash181 1995
[18] W Andlauer and P Furst ldquoNutraceuticals a piece of historypresent status and outlookrdquo Food Research International vol 35no 2-3 pp 171ndash176 2002
[19] C L Kruger and S W Mann ldquoSafety evaluation of functionalingredientsrdquo Food and Chemical Toxicology vol 41 no 6 pp793ndash805 2003
[20] P H Mattila V I Piironen E J Uusi-Rauva and P E Koivis-toinen ldquoVitamin D contents in edible mushroomsrdquo Journal ofAgricultural and Food Chemistry vol 42 no 11 pp 2449ndash24531994
[21] R P Tewari ldquoMushroom their role in nature and societyrdquo inFrontiers inMushroomBiotechnology R D Rai R C Upadhyayand S R Sharma Eds pp 1ndash8 NRCM Chambaghat India2005
[22] K C Semwal S L Stephenson V K Bhatt and R P BhattldquoEdible mushrooms of the Northwestern Himalaya India astudy of indigenous knowledge distribution and diversityrdquoMycosphere vol 5 no 3 pp 440ndash461 2014
[23] E V Crisan and A Sands ldquoEdible mushrooms nutritionalvaluerdquo in The Biology and Cultivation of Edible Mushrooms ST Chang and W A Hayes Eds pp 137ndash165 Academic PressNew York NY USA 1978
[24] N S Atri S K Sharma R Joshi A Gulati and A Gula-ti ldquoNutritional and neutraceutical composition of five wildculinary-medicinal species of genus Pleurotus (Higher Basid-iomycetes) from Northwest Indiardquo International Journal ofMedicinal Mushrooms vol 15 no 1 pp 49ndash56 2013
[25] L Barros T Cruz P Baptista L M Estevinho and I C FR Ferreira ldquoWild and commercial mushrooms as source ofnutrients and nutraceuticalsrdquo Food and Chemical Toxicologyvol 46 no 8 pp 2742ndash2747 2008
[26] M Nagata and I Yamashita ldquoSimple method for simultaneousdetermination of chlorophyll and carotenoids in tomato fruitrdquoNippon Shokuhin Kogyo Gakkaishi vol 39 no 10 pp 925ndash9281992
[27] V L Singleton and J A Rossi ldquoColorimetric detection oftotal phenolics with phosphomolybdic-phosphotungstic acidreagentsrdquo American Journal of Enology and Viticulture vol 16pp 144ndash158 1965
[28] L K Jagadish V Venkata Krishnan R Shenbhagaramanand V Kaviyarasan ldquoComparitive study on the antioxidantanticancer and antimicrobial property of Agaricus bisporus (JE Lange) Imbach before and after boilingrdquo African Journal ofBiotechnology vol 8 no 4 pp 654ndash661 2009
[29] E Vamanu and S Nita ldquoAntioxidant capacity and the cor-relation with major phenolic compounds anthocyanin andtocopherol content in various extracts from the wild edibleBoletus edulis mushroomrdquo BioMed Research International vol2013 Article ID 313905 11 pages 2013
[30] E Vamanu ldquoBiological activities of the polysaccharides pro-duced in submerged culture of two edible Pleurotus ostreatus
12 BioMed Research International
mushroomsrdquo Journal of Biomedicine and Biotechnology vol2012 Article ID 565974 8 pages 2012
[31] P L Li L Huo W Su et al ldquoFree radical-scavenging capacityantioxidant activity and phenolic content of Pouzolzia zeylan-icardquo Journal of the Serbian Chemical Society vol 76 no 5 pp709ndash717 2011
[32] C Papuc M Crivineanu G Goran V Nicorescu and N Dur-dun ldquoFree radicals scavenging and antioxidant activity ofEuropean mistletoe (Viscum album) and European birthwort(Aristolochia clematitis)rdquo Revista de Chimie vol 61 no 7 pp619ndash622 2010
[33] VOOyetayo CDong andY Yao ldquoAntioxidant and antimicro-bial properties of aqueous extract from Dictyophora indusiatardquoThe Open Mycology Journal vol 3 no 1 pp 20ndash26 2009
[34] S Marklund and G Marklund ldquoInvolvement of the superoxideanion radical in the autoxidation of pyrogallol and a convenientassay for superoxide dismutaserdquo European Journal of Biochem-istry vol 47 no 3 pp 469ndash474 1974
[35] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996
[36] D J Huang b Ou and R L Prior ldquoThe chemistry behindantioxidant capacity assaysrdquo Journal of Agricultural and FoodChemistry vol 53 no 6 pp 1841ndash1856 2005
[37] P Manzi A Aguzzi and L Pizzoferrato ldquoNutritional value ofmushrooms widely consumed in Italyrdquo Food Chemistry vol 73no 3 pp 321ndash325 2001
[38] P Manzi S Marconi A Aguzzi and L Pizzoferrato ldquoCommer-cial mushrooms nutritional quality and effect of cookingrdquo FoodChemistry vol 84 no 2 pp 201ndash206 2004
[39] I P Krbavcic and I C Baric ldquoInfluence of deep fat fryingon some nutritional parameters of novel food based on mush-rooms and fresh soft cheeserdquo Food Chemistry vol 84 no 3 pp417ndash419 2004
[40] D Agrahar-Murugkar andG Subbulakshmi ldquoNutritional valueof edible wild mushrooms collected from the Khasi hills ofMeghalayardquo Food Chemistry vol 89 no 4 pp 599ndash603 2005
[41] V A Dıez and A Alvarez ldquoCompositional and nutritionalstudies on two wild edible mushrooms from northwest SpainrdquoFood Chemistry vol 75 no 4 pp 417ndash422 2001
[42] L Barros M-J Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007
[43] S K Sharma and N S Atri ldquoNutraceutical composition ofwild species of genus Lentinus Fr fromNothern Indiardquo CurrentResearch in Environmental ampAppliedMycology vol 4 pp 11ndash322014
[44] P J Kanu K Zhu J Baby Kanu H Zhou H Qian and K ZhuldquoBiologically active components and nutraceuticals in sesameand related products a review and prospectrdquo Trends in FoodScience and Technology vol 18 no 12 pp 599ndash608 2007
[45] F S Reis E Pereira L Barros M J Sousa A Martins and I CF R Ferreira ldquoBiomolecule profiles in inediblewildmushroomswith antioxidant valuerdquoMolecules vol 16 no 6 pp 4328ndash43382011
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
obtained for Hygrocybe nivea (426 plusmn 05mgmL) showinglowest ABTS radical scavenging activities of this speciesReducing power of Cantharellus cibarius (162 plusmn 05mgmL)was measured higher than other species Higher EC
50values
for reducing activity were measured in Inocybe splendens(519 plusmn 01mgmL)
Higher effectiveness in ferrous ion chelating activitywas detected in Agaricus arvensis (114 plusmn 01mgmL) andlow effectiveness was detected in Russula mairei (295 plusmn03mgmL) Nevertheless Cantharellus cibarius (127 plusmn02mgmL) Amanita caesarea (140 plusmn 04mgmL) Hygro-cybe coccinea (135 plusmn 02mgmL) and H nivea (145 plusmn06mgmL) showed lower EC
50values than remaining
species EC50
values of scavenging ability on superoxideradical were found to be maximum in Amanita caesarea(044 plusmn 03mgmL) and minimum in Lactarius pubescens(221 plusmn 06mgmL) Gymnopilus junonius showed maximumFRAP activity with least EC
Although there are previous reports on documentation ofculinary edible species from the regions native to northernHimalayas but there are no reports on the evaluation stud-ies as well as toxicity status of all these culinary speciesThe wild edible species Agaricus bisporus Boletus edulis
Morchella esculenta Cordyceps sinensis and Lentinula edodeswhich have been extensively worked out in India and otherparts of the world for their compositional and medicinalaspects have not been undertaken for investigations presently[37ndash40] Compositional studies showed that most of theculinary species were rich in protein carbohydrates andlow in fat There are several reports on richness of wildedible mushrooms with protein and carbohydrate contentsand low fat levels which directly make them nutritionallyrich [24 25 41] Nevertheless under present studies thedifferences between the nutrient concentrations of all thespecies differed but Agaricus arvensis Pleurotus cystidiosusAmanita caesarea Agaricus campestris Cantharellus cibariusand Lentinus cladopus showed higher nutrient percentagewhich is comparable to other wild and commercially cul-tivated species [25 42] The crude fat content detected inall the species was not found to be significantly differentCrude fibres were detected in appreciable percentage fromall the species which make them important in nutritionalpoint of viewThe results are in conformity with the previousreports on several wild edible Pleurotus and Lentinus speciesfrom northwest India [24 43] The species Inocybe splendensHygrocybe nivea and Conocybe tenera were not found tocontain higher percentage of nutrients Although previousreports showed that nutrients composition in wild species isless as compared to cultivated species [25] however Agaricusarvensis Pleurotus cystidiosus Amanita caesarea Agaricuscampestris Cantharellus cibarius and Lentinus cladopus were
10 BioMed Research International
found to be rich in protein and carbohydrates similar tocommercially grown species [25]
Fatty acid composition showed the dominance of UFAover SFA in all the studied mushrooms species which is inconformity with the other studies [41] The differences wereobserved in net amounts in all the species Unsaturationindex of Agaricus arvensis and Lentinus cladopus (057 plusmn002) was found to be significantly higher than otherspecies Whereas Inocybe splendens (005 plusmn 00) showedleast Unsaturation Index High UFA shows the medicinalimportance of these culinary mushrooms as these increasethe HDL cholesterol and decrease LDL cholesterol triacyl-glycerol lipid oxidation and LDL susceptibility to oxidation[44] Predominance of UFA over SFA in all the species showssimilar results as obtained for other wild and commerciallycultivated species [24 25] 120572- and 120573-tocopherol were detectedin higher amounts than third isomer in all the studied speciesSimilar findings were made in other wild and cultivatedspecies with higher 120572- and 120573-tocopherol than 120574-tocopherol[25] The high levels of these two compounds correspondwith a higher oxidative activity which is associated withcardiovascular protection [45] Phenolic compounds weredetected in higher amounts than other bioactive compoundsPresence of high phenolic compounds accounts for the highantioxidant properties of all the species [42] 120573-carotenelycopene and ascorbic acids were detected in low amountsAnthocyanidins were also detected from these wild speciesin appreciable amounts The presence of these functionalmedicinal compounds inmedicinal andor edible mushroomis due to habitat or substrates in which these grow to be highin the functionalmoleculesThe categories of thesemoleculesare anthocyanidins beta-glucans selenium ganoderic acidtriterpenes or cordycepinThe compounds identified in theseextracts show that at least a part of the functional compoundsin medicinal andor edible mushroom is due to growingmushrooms on substrates that are high in the functionalmolecules To these categories can be added anthocyanidinsbeta-glucans selenium ganoderic acid triterpenes or cordy-cepinThe amounts of these have been found to vary with thetype of extraction as ethanolic extract yields higher amountsof anthocyanidins as compared to methanolic hot water andcold water extracts [29]
All the studied species showed significant antioxidantproperties measured on the basis of EC
50values Never-
theless each species showed different antioxidant activitywith highly effective and less effective EC
50values Better
antioxidant properties of some species are due to presence ofhigher phenolic compounds 120573-carotene lycopene ascorbicacids anthocyanidins and tocopherol amounts in themHigh reducing power of some species is due to the presence ofhigher amounts of reducers (antioxidants) in them Presentlyinvestigated species are commonly used for culinary pur-poses in the regions native to northern Himalayas Many ofthe species in these regions are not evaluated previously fordetailed compositional analysisTheir knowledge is restrictedto old aged villagers of the regions and neglected for thecommercial exploitations There are no positive reportson toxicity of these mushrooms analyzed presently hencethese are safe for further experimental work related to
drug discovery All the culinary species contained impor-tant and useful nutraceuticals such as unsaturated fattyacids phenolics carotenoids ascorbic acid tocopherolsand anthocyanidins besides these some important aminoacids were detected in these mushrooms which could beused for the purpose of being used as functional ingredi-ents Since nutraceuticals are powerful in maintaining andpromoting health longevity and life quality the commer-cial exploitation of these species will certainly create animpact on nutritional therapy and also will be beneficialtodayrsquos food industry Direct use of these species for con-sumption and other culinary aspects is safe and healthpromoting with advantage of the additive effects of allthe bioactive and antioxidant compounds present in thesespecies
Abbreviations
AlCl3 Aluminum trichloride
ANOVA Analysis of varianceDW Distilled waterDPPH 22-Diphenyl-1-picrylhydrazylFeCl3 Ferric chloride
FRAP Ferric reducing antioxidant powerg GramsGAEs Gallic acid equivalentsGC Gas ChromatographyHCl Hydrochloric acidH2O Water
H3PO4 Phosphoric acid
HPLC High performance liquid chromatographyK2HPO4 Potassium hydrogen phosphate
The authors declare that there is no conflict of interestsregarding the publication of this paper
BioMed Research International 11
Acknowledgment
The authors wish to thank Science and Engineering BoardDepartment of Science and Technology New Delhi forresearch Grant (SBFTLS-042013) to carry out presentstudies
References
[1] W M Breene ldquoNutritional and medicinal value of specialtymushroomsrdquo Journal of Food Protection vol 53 pp 883ndash8941990
[2] S Chang and P Miles ldquoMushroom biologymdasha new disciplinerdquoMycologist vol 6 no 2 pp 64ndash65 1992
[3] P Manzi L Gambelli S Marconi V Vivanti and L Pizzoferra-to ldquoNutrients in edible mushrooms an inter-species compara-tive studyrdquo Food Chemistry vol 65 no 4 pp 477ndash482 1999
[4] P Bobek O Ozdın and M Mikus ldquoDietary oyster mushroom(Pleurotus ostreatus) accelerates plasma cholesterol turnover inhypercholesterolaemic ratsrdquo Physiological Research vol 44 no5 pp 287ndash291 1995
[5] P Bobek and S Galbavy ldquoHypocholesterolemic and antiathero-genic effect of oystermushroom (Pleurotus ostreatus) in rabbitsrdquoNahrung vol 43 no 5 pp 339ndash342 1999
[6] S Khatun A Islam U Cakilcioglu and N Chatterjee ldquoRe-search on mushroom as a potential source of nutraceuticals areview on Indian perspectiverdquo American Journal of Experimen-tal Agriculture vol 2 no 1 pp 47ndash73 2012
[7] J A Vaz L Barros A Martins J S Morais M H Vasconcelosand I C F R Ferreira ldquoPhenolic profile of seventeen por-tuguese wild mushroomsrdquo LWT Food Science and Technologyvol 44 no 1 pp 343ndash346 2011
[8] J A Vaz L Barros A Martins C Santos-Buelga M HVasconcelos and I C F R Ferreira ldquoChemical compositionof wild edible mushrooms and antioxidant properties of theirwater soluble polysaccharidic and ethanolic fractionsrdquo FoodChemistry vol 126 no 2 pp 610ndash616 2011
[9] T Ozen C Darcan O Aktop and I Turkekul ldquoScreening ofantioxidant antimicrobial activities and chemical contents ofedible mushrooms wildly grown in the Black Sea region ofTurkeyrdquo Combinatorial Chemistry amp High Throughput Screen-ing vol 14 no 2 pp 72ndash84 2011
[10] V E C Ooi and F Liu ldquoA Review of pharmacological activitiesof mushroom polysaccharidesrdquo International Journal of Medic-inal Mushrooms vol 1 no 3 pp 195ndash206 1999
[11] S PWasser and A LWeis ldquoMedicinal properties of substancesoccurring in higher Basidiomycete mushroom current per-spectiverdquo International Journal of Medicinal Mushrooms vol 1pp 31ndash62 1999
[12] P M Kidd ldquoThe use of mushroom glucans and proteoglycansin cancer treatmentrdquo Alternative Medicine Review vol 5 no 1pp 4ndash27 2000
[13] C Israilides and A Philippoussis ldquoBio-technologies of recy-cling agro-industrial wastes for the production of commerciallyimportant fungal polysaccharides andmushroomsrdquo Biotechnol-ogy and Genetic Engineering Reviews vol 20 pp 247ndash259 2003
[14] T Mizuno ldquoBioactive biomolecules of mushrooms food func-tion and medicinal effect of mushroom fungirdquo Food ReviewsInternational vol 11 no 1 pp 5ndash21 1995
[15] Y Kabir and S Kimura ldquoDietary mushrooms reduce bloodpressure in spontaneously hypertensive rats (SHR)rdquo Journal of
Nutritional Science and Vitaminology vol 35 no 1 pp 91ndash941989
[16] J-L Mau S-Y Tsai Y-H Tseng and S-J Huang ldquoAntioxidantproperties of hot water extracts from Ganoderma tsugae Mur-rillrdquo LWTmdashFood Science and Technology vol 38 no 6 pp 589ndash597 2005
[17] A Smania F D Monache E F A Smania M L Gil L CBenchetrit and F S Cruz ldquoAntibacterial activity of a substanceproduced by the fungus Pycnoporus sanguineus (Fr) MurrrdquoJournal of Ethnopharmacology vol 45 no 3 pp 177ndash181 1995
[18] W Andlauer and P Furst ldquoNutraceuticals a piece of historypresent status and outlookrdquo Food Research International vol 35no 2-3 pp 171ndash176 2002
[19] C L Kruger and S W Mann ldquoSafety evaluation of functionalingredientsrdquo Food and Chemical Toxicology vol 41 no 6 pp793ndash805 2003
[20] P H Mattila V I Piironen E J Uusi-Rauva and P E Koivis-toinen ldquoVitamin D contents in edible mushroomsrdquo Journal ofAgricultural and Food Chemistry vol 42 no 11 pp 2449ndash24531994
[21] R P Tewari ldquoMushroom their role in nature and societyrdquo inFrontiers inMushroomBiotechnology R D Rai R C Upadhyayand S R Sharma Eds pp 1ndash8 NRCM Chambaghat India2005
[22] K C Semwal S L Stephenson V K Bhatt and R P BhattldquoEdible mushrooms of the Northwestern Himalaya India astudy of indigenous knowledge distribution and diversityrdquoMycosphere vol 5 no 3 pp 440ndash461 2014
[23] E V Crisan and A Sands ldquoEdible mushrooms nutritionalvaluerdquo in The Biology and Cultivation of Edible Mushrooms ST Chang and W A Hayes Eds pp 137ndash165 Academic PressNew York NY USA 1978
[24] N S Atri S K Sharma R Joshi A Gulati and A Gula-ti ldquoNutritional and neutraceutical composition of five wildculinary-medicinal species of genus Pleurotus (Higher Basid-iomycetes) from Northwest Indiardquo International Journal ofMedicinal Mushrooms vol 15 no 1 pp 49ndash56 2013
[25] L Barros T Cruz P Baptista L M Estevinho and I C FR Ferreira ldquoWild and commercial mushrooms as source ofnutrients and nutraceuticalsrdquo Food and Chemical Toxicologyvol 46 no 8 pp 2742ndash2747 2008
[26] M Nagata and I Yamashita ldquoSimple method for simultaneousdetermination of chlorophyll and carotenoids in tomato fruitrdquoNippon Shokuhin Kogyo Gakkaishi vol 39 no 10 pp 925ndash9281992
[27] V L Singleton and J A Rossi ldquoColorimetric detection oftotal phenolics with phosphomolybdic-phosphotungstic acidreagentsrdquo American Journal of Enology and Viticulture vol 16pp 144ndash158 1965
[28] L K Jagadish V Venkata Krishnan R Shenbhagaramanand V Kaviyarasan ldquoComparitive study on the antioxidantanticancer and antimicrobial property of Agaricus bisporus (JE Lange) Imbach before and after boilingrdquo African Journal ofBiotechnology vol 8 no 4 pp 654ndash661 2009
[29] E Vamanu and S Nita ldquoAntioxidant capacity and the cor-relation with major phenolic compounds anthocyanin andtocopherol content in various extracts from the wild edibleBoletus edulis mushroomrdquo BioMed Research International vol2013 Article ID 313905 11 pages 2013
[30] E Vamanu ldquoBiological activities of the polysaccharides pro-duced in submerged culture of two edible Pleurotus ostreatus
12 BioMed Research International
mushroomsrdquo Journal of Biomedicine and Biotechnology vol2012 Article ID 565974 8 pages 2012
[31] P L Li L Huo W Su et al ldquoFree radical-scavenging capacityantioxidant activity and phenolic content of Pouzolzia zeylan-icardquo Journal of the Serbian Chemical Society vol 76 no 5 pp709ndash717 2011
[32] C Papuc M Crivineanu G Goran V Nicorescu and N Dur-dun ldquoFree radicals scavenging and antioxidant activity ofEuropean mistletoe (Viscum album) and European birthwort(Aristolochia clematitis)rdquo Revista de Chimie vol 61 no 7 pp619ndash622 2010
[33] VOOyetayo CDong andY Yao ldquoAntioxidant and antimicro-bial properties of aqueous extract from Dictyophora indusiatardquoThe Open Mycology Journal vol 3 no 1 pp 20ndash26 2009
[34] S Marklund and G Marklund ldquoInvolvement of the superoxideanion radical in the autoxidation of pyrogallol and a convenientassay for superoxide dismutaserdquo European Journal of Biochem-istry vol 47 no 3 pp 469ndash474 1974
[35] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996
[36] D J Huang b Ou and R L Prior ldquoThe chemistry behindantioxidant capacity assaysrdquo Journal of Agricultural and FoodChemistry vol 53 no 6 pp 1841ndash1856 2005
[37] P Manzi A Aguzzi and L Pizzoferrato ldquoNutritional value ofmushrooms widely consumed in Italyrdquo Food Chemistry vol 73no 3 pp 321ndash325 2001
[38] P Manzi S Marconi A Aguzzi and L Pizzoferrato ldquoCommer-cial mushrooms nutritional quality and effect of cookingrdquo FoodChemistry vol 84 no 2 pp 201ndash206 2004
[39] I P Krbavcic and I C Baric ldquoInfluence of deep fat fryingon some nutritional parameters of novel food based on mush-rooms and fresh soft cheeserdquo Food Chemistry vol 84 no 3 pp417ndash419 2004
[40] D Agrahar-Murugkar andG Subbulakshmi ldquoNutritional valueof edible wild mushrooms collected from the Khasi hills ofMeghalayardquo Food Chemistry vol 89 no 4 pp 599ndash603 2005
[41] V A Dıez and A Alvarez ldquoCompositional and nutritionalstudies on two wild edible mushrooms from northwest SpainrdquoFood Chemistry vol 75 no 4 pp 417ndash422 2001
[42] L Barros M-J Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007
[43] S K Sharma and N S Atri ldquoNutraceutical composition ofwild species of genus Lentinus Fr fromNothern Indiardquo CurrentResearch in Environmental ampAppliedMycology vol 4 pp 11ndash322014
[44] P J Kanu K Zhu J Baby Kanu H Zhou H Qian and K ZhuldquoBiologically active components and nutraceuticals in sesameand related products a review and prospectrdquo Trends in FoodScience and Technology vol 18 no 12 pp 599ndash608 2007
[45] F S Reis E Pereira L Barros M J Sousa A Martins and I CF R Ferreira ldquoBiomolecule profiles in inediblewildmushroomswith antioxidant valuerdquoMolecules vol 16 no 6 pp 4328ndash43382011
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
found to be rich in protein and carbohydrates similar tocommercially grown species [25]
Fatty acid composition showed the dominance of UFAover SFA in all the studied mushrooms species which is inconformity with the other studies [41] The differences wereobserved in net amounts in all the species Unsaturationindex of Agaricus arvensis and Lentinus cladopus (057 plusmn002) was found to be significantly higher than otherspecies Whereas Inocybe splendens (005 plusmn 00) showedleast Unsaturation Index High UFA shows the medicinalimportance of these culinary mushrooms as these increasethe HDL cholesterol and decrease LDL cholesterol triacyl-glycerol lipid oxidation and LDL susceptibility to oxidation[44] Predominance of UFA over SFA in all the species showssimilar results as obtained for other wild and commerciallycultivated species [24 25] 120572- and 120573-tocopherol were detectedin higher amounts than third isomer in all the studied speciesSimilar findings were made in other wild and cultivatedspecies with higher 120572- and 120573-tocopherol than 120574-tocopherol[25] The high levels of these two compounds correspondwith a higher oxidative activity which is associated withcardiovascular protection [45] Phenolic compounds weredetected in higher amounts than other bioactive compoundsPresence of high phenolic compounds accounts for the highantioxidant properties of all the species [42] 120573-carotenelycopene and ascorbic acids were detected in low amountsAnthocyanidins were also detected from these wild speciesin appreciable amounts The presence of these functionalmedicinal compounds inmedicinal andor edible mushroomis due to habitat or substrates in which these grow to be highin the functionalmoleculesThe categories of thesemoleculesare anthocyanidins beta-glucans selenium ganoderic acidtriterpenes or cordycepinThe compounds identified in theseextracts show that at least a part of the functional compoundsin medicinal andor edible mushroom is due to growingmushrooms on substrates that are high in the functionalmolecules To these categories can be added anthocyanidinsbeta-glucans selenium ganoderic acid triterpenes or cordy-cepinThe amounts of these have been found to vary with thetype of extraction as ethanolic extract yields higher amountsof anthocyanidins as compared to methanolic hot water andcold water extracts [29]
All the studied species showed significant antioxidantproperties measured on the basis of EC
50values Never-
theless each species showed different antioxidant activitywith highly effective and less effective EC
50values Better
antioxidant properties of some species are due to presence ofhigher phenolic compounds 120573-carotene lycopene ascorbicacids anthocyanidins and tocopherol amounts in themHigh reducing power of some species is due to the presence ofhigher amounts of reducers (antioxidants) in them Presentlyinvestigated species are commonly used for culinary pur-poses in the regions native to northern Himalayas Many ofthe species in these regions are not evaluated previously fordetailed compositional analysisTheir knowledge is restrictedto old aged villagers of the regions and neglected for thecommercial exploitations There are no positive reportson toxicity of these mushrooms analyzed presently hencethese are safe for further experimental work related to
drug discovery All the culinary species contained impor-tant and useful nutraceuticals such as unsaturated fattyacids phenolics carotenoids ascorbic acid tocopherolsand anthocyanidins besides these some important aminoacids were detected in these mushrooms which could beused for the purpose of being used as functional ingredi-ents Since nutraceuticals are powerful in maintaining andpromoting health longevity and life quality the commer-cial exploitation of these species will certainly create animpact on nutritional therapy and also will be beneficialtodayrsquos food industry Direct use of these species for con-sumption and other culinary aspects is safe and healthpromoting with advantage of the additive effects of allthe bioactive and antioxidant compounds present in thesespecies
Abbreviations
AlCl3 Aluminum trichloride
ANOVA Analysis of varianceDW Distilled waterDPPH 22-Diphenyl-1-picrylhydrazylFeCl3 Ferric chloride
FRAP Ferric reducing antioxidant powerg GramsGAEs Gallic acid equivalentsGC Gas ChromatographyHCl Hydrochloric acidH2O Water
H3PO4 Phosphoric acid
HPLC High performance liquid chromatographyK2HPO4 Potassium hydrogen phosphate
The authors declare that there is no conflict of interestsregarding the publication of this paper
BioMed Research International 11
Acknowledgment
The authors wish to thank Science and Engineering BoardDepartment of Science and Technology New Delhi forresearch Grant (SBFTLS-042013) to carry out presentstudies
References
[1] W M Breene ldquoNutritional and medicinal value of specialtymushroomsrdquo Journal of Food Protection vol 53 pp 883ndash8941990
[2] S Chang and P Miles ldquoMushroom biologymdasha new disciplinerdquoMycologist vol 6 no 2 pp 64ndash65 1992
[3] P Manzi L Gambelli S Marconi V Vivanti and L Pizzoferra-to ldquoNutrients in edible mushrooms an inter-species compara-tive studyrdquo Food Chemistry vol 65 no 4 pp 477ndash482 1999
[4] P Bobek O Ozdın and M Mikus ldquoDietary oyster mushroom(Pleurotus ostreatus) accelerates plasma cholesterol turnover inhypercholesterolaemic ratsrdquo Physiological Research vol 44 no5 pp 287ndash291 1995
[5] P Bobek and S Galbavy ldquoHypocholesterolemic and antiathero-genic effect of oystermushroom (Pleurotus ostreatus) in rabbitsrdquoNahrung vol 43 no 5 pp 339ndash342 1999
[6] S Khatun A Islam U Cakilcioglu and N Chatterjee ldquoRe-search on mushroom as a potential source of nutraceuticals areview on Indian perspectiverdquo American Journal of Experimen-tal Agriculture vol 2 no 1 pp 47ndash73 2012
[7] J A Vaz L Barros A Martins J S Morais M H Vasconcelosand I C F R Ferreira ldquoPhenolic profile of seventeen por-tuguese wild mushroomsrdquo LWT Food Science and Technologyvol 44 no 1 pp 343ndash346 2011
[8] J A Vaz L Barros A Martins C Santos-Buelga M HVasconcelos and I C F R Ferreira ldquoChemical compositionof wild edible mushrooms and antioxidant properties of theirwater soluble polysaccharidic and ethanolic fractionsrdquo FoodChemistry vol 126 no 2 pp 610ndash616 2011
[9] T Ozen C Darcan O Aktop and I Turkekul ldquoScreening ofantioxidant antimicrobial activities and chemical contents ofedible mushrooms wildly grown in the Black Sea region ofTurkeyrdquo Combinatorial Chemistry amp High Throughput Screen-ing vol 14 no 2 pp 72ndash84 2011
[10] V E C Ooi and F Liu ldquoA Review of pharmacological activitiesof mushroom polysaccharidesrdquo International Journal of Medic-inal Mushrooms vol 1 no 3 pp 195ndash206 1999
[11] S PWasser and A LWeis ldquoMedicinal properties of substancesoccurring in higher Basidiomycete mushroom current per-spectiverdquo International Journal of Medicinal Mushrooms vol 1pp 31ndash62 1999
[12] P M Kidd ldquoThe use of mushroom glucans and proteoglycansin cancer treatmentrdquo Alternative Medicine Review vol 5 no 1pp 4ndash27 2000
[13] C Israilides and A Philippoussis ldquoBio-technologies of recy-cling agro-industrial wastes for the production of commerciallyimportant fungal polysaccharides andmushroomsrdquo Biotechnol-ogy and Genetic Engineering Reviews vol 20 pp 247ndash259 2003
[14] T Mizuno ldquoBioactive biomolecules of mushrooms food func-tion and medicinal effect of mushroom fungirdquo Food ReviewsInternational vol 11 no 1 pp 5ndash21 1995
[15] Y Kabir and S Kimura ldquoDietary mushrooms reduce bloodpressure in spontaneously hypertensive rats (SHR)rdquo Journal of
Nutritional Science and Vitaminology vol 35 no 1 pp 91ndash941989
[16] J-L Mau S-Y Tsai Y-H Tseng and S-J Huang ldquoAntioxidantproperties of hot water extracts from Ganoderma tsugae Mur-rillrdquo LWTmdashFood Science and Technology vol 38 no 6 pp 589ndash597 2005
[17] A Smania F D Monache E F A Smania M L Gil L CBenchetrit and F S Cruz ldquoAntibacterial activity of a substanceproduced by the fungus Pycnoporus sanguineus (Fr) MurrrdquoJournal of Ethnopharmacology vol 45 no 3 pp 177ndash181 1995
[18] W Andlauer and P Furst ldquoNutraceuticals a piece of historypresent status and outlookrdquo Food Research International vol 35no 2-3 pp 171ndash176 2002
[19] C L Kruger and S W Mann ldquoSafety evaluation of functionalingredientsrdquo Food and Chemical Toxicology vol 41 no 6 pp793ndash805 2003
[20] P H Mattila V I Piironen E J Uusi-Rauva and P E Koivis-toinen ldquoVitamin D contents in edible mushroomsrdquo Journal ofAgricultural and Food Chemistry vol 42 no 11 pp 2449ndash24531994
[21] R P Tewari ldquoMushroom their role in nature and societyrdquo inFrontiers inMushroomBiotechnology R D Rai R C Upadhyayand S R Sharma Eds pp 1ndash8 NRCM Chambaghat India2005
[22] K C Semwal S L Stephenson V K Bhatt and R P BhattldquoEdible mushrooms of the Northwestern Himalaya India astudy of indigenous knowledge distribution and diversityrdquoMycosphere vol 5 no 3 pp 440ndash461 2014
[23] E V Crisan and A Sands ldquoEdible mushrooms nutritionalvaluerdquo in The Biology and Cultivation of Edible Mushrooms ST Chang and W A Hayes Eds pp 137ndash165 Academic PressNew York NY USA 1978
[24] N S Atri S K Sharma R Joshi A Gulati and A Gula-ti ldquoNutritional and neutraceutical composition of five wildculinary-medicinal species of genus Pleurotus (Higher Basid-iomycetes) from Northwest Indiardquo International Journal ofMedicinal Mushrooms vol 15 no 1 pp 49ndash56 2013
[25] L Barros T Cruz P Baptista L M Estevinho and I C FR Ferreira ldquoWild and commercial mushrooms as source ofnutrients and nutraceuticalsrdquo Food and Chemical Toxicologyvol 46 no 8 pp 2742ndash2747 2008
[26] M Nagata and I Yamashita ldquoSimple method for simultaneousdetermination of chlorophyll and carotenoids in tomato fruitrdquoNippon Shokuhin Kogyo Gakkaishi vol 39 no 10 pp 925ndash9281992
[27] V L Singleton and J A Rossi ldquoColorimetric detection oftotal phenolics with phosphomolybdic-phosphotungstic acidreagentsrdquo American Journal of Enology and Viticulture vol 16pp 144ndash158 1965
[28] L K Jagadish V Venkata Krishnan R Shenbhagaramanand V Kaviyarasan ldquoComparitive study on the antioxidantanticancer and antimicrobial property of Agaricus bisporus (JE Lange) Imbach before and after boilingrdquo African Journal ofBiotechnology vol 8 no 4 pp 654ndash661 2009
[29] E Vamanu and S Nita ldquoAntioxidant capacity and the cor-relation with major phenolic compounds anthocyanin andtocopherol content in various extracts from the wild edibleBoletus edulis mushroomrdquo BioMed Research International vol2013 Article ID 313905 11 pages 2013
[30] E Vamanu ldquoBiological activities of the polysaccharides pro-duced in submerged culture of two edible Pleurotus ostreatus
12 BioMed Research International
mushroomsrdquo Journal of Biomedicine and Biotechnology vol2012 Article ID 565974 8 pages 2012
[31] P L Li L Huo W Su et al ldquoFree radical-scavenging capacityantioxidant activity and phenolic content of Pouzolzia zeylan-icardquo Journal of the Serbian Chemical Society vol 76 no 5 pp709ndash717 2011
[32] C Papuc M Crivineanu G Goran V Nicorescu and N Dur-dun ldquoFree radicals scavenging and antioxidant activity ofEuropean mistletoe (Viscum album) and European birthwort(Aristolochia clematitis)rdquo Revista de Chimie vol 61 no 7 pp619ndash622 2010
[33] VOOyetayo CDong andY Yao ldquoAntioxidant and antimicro-bial properties of aqueous extract from Dictyophora indusiatardquoThe Open Mycology Journal vol 3 no 1 pp 20ndash26 2009
[34] S Marklund and G Marklund ldquoInvolvement of the superoxideanion radical in the autoxidation of pyrogallol and a convenientassay for superoxide dismutaserdquo European Journal of Biochem-istry vol 47 no 3 pp 469ndash474 1974
[35] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996
[36] D J Huang b Ou and R L Prior ldquoThe chemistry behindantioxidant capacity assaysrdquo Journal of Agricultural and FoodChemistry vol 53 no 6 pp 1841ndash1856 2005
[37] P Manzi A Aguzzi and L Pizzoferrato ldquoNutritional value ofmushrooms widely consumed in Italyrdquo Food Chemistry vol 73no 3 pp 321ndash325 2001
[38] P Manzi S Marconi A Aguzzi and L Pizzoferrato ldquoCommer-cial mushrooms nutritional quality and effect of cookingrdquo FoodChemistry vol 84 no 2 pp 201ndash206 2004
[39] I P Krbavcic and I C Baric ldquoInfluence of deep fat fryingon some nutritional parameters of novel food based on mush-rooms and fresh soft cheeserdquo Food Chemistry vol 84 no 3 pp417ndash419 2004
[40] D Agrahar-Murugkar andG Subbulakshmi ldquoNutritional valueof edible wild mushrooms collected from the Khasi hills ofMeghalayardquo Food Chemistry vol 89 no 4 pp 599ndash603 2005
[41] V A Dıez and A Alvarez ldquoCompositional and nutritionalstudies on two wild edible mushrooms from northwest SpainrdquoFood Chemistry vol 75 no 4 pp 417ndash422 2001
[42] L Barros M-J Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007
[43] S K Sharma and N S Atri ldquoNutraceutical composition ofwild species of genus Lentinus Fr fromNothern Indiardquo CurrentResearch in Environmental ampAppliedMycology vol 4 pp 11ndash322014
[44] P J Kanu K Zhu J Baby Kanu H Zhou H Qian and K ZhuldquoBiologically active components and nutraceuticals in sesameand related products a review and prospectrdquo Trends in FoodScience and Technology vol 18 no 12 pp 599ndash608 2007
[45] F S Reis E Pereira L Barros M J Sousa A Martins and I CF R Ferreira ldquoBiomolecule profiles in inediblewildmushroomswith antioxidant valuerdquoMolecules vol 16 no 6 pp 4328ndash43382011
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
The authors wish to thank Science and Engineering BoardDepartment of Science and Technology New Delhi forresearch Grant (SBFTLS-042013) to carry out presentstudies
References
[1] W M Breene ldquoNutritional and medicinal value of specialtymushroomsrdquo Journal of Food Protection vol 53 pp 883ndash8941990
[2] S Chang and P Miles ldquoMushroom biologymdasha new disciplinerdquoMycologist vol 6 no 2 pp 64ndash65 1992
[3] P Manzi L Gambelli S Marconi V Vivanti and L Pizzoferra-to ldquoNutrients in edible mushrooms an inter-species compara-tive studyrdquo Food Chemistry vol 65 no 4 pp 477ndash482 1999
[4] P Bobek O Ozdın and M Mikus ldquoDietary oyster mushroom(Pleurotus ostreatus) accelerates plasma cholesterol turnover inhypercholesterolaemic ratsrdquo Physiological Research vol 44 no5 pp 287ndash291 1995
[5] P Bobek and S Galbavy ldquoHypocholesterolemic and antiathero-genic effect of oystermushroom (Pleurotus ostreatus) in rabbitsrdquoNahrung vol 43 no 5 pp 339ndash342 1999
[6] S Khatun A Islam U Cakilcioglu and N Chatterjee ldquoRe-search on mushroom as a potential source of nutraceuticals areview on Indian perspectiverdquo American Journal of Experimen-tal Agriculture vol 2 no 1 pp 47ndash73 2012
[7] J A Vaz L Barros A Martins J S Morais M H Vasconcelosand I C F R Ferreira ldquoPhenolic profile of seventeen por-tuguese wild mushroomsrdquo LWT Food Science and Technologyvol 44 no 1 pp 343ndash346 2011
[8] J A Vaz L Barros A Martins C Santos-Buelga M HVasconcelos and I C F R Ferreira ldquoChemical compositionof wild edible mushrooms and antioxidant properties of theirwater soluble polysaccharidic and ethanolic fractionsrdquo FoodChemistry vol 126 no 2 pp 610ndash616 2011
[9] T Ozen C Darcan O Aktop and I Turkekul ldquoScreening ofantioxidant antimicrobial activities and chemical contents ofedible mushrooms wildly grown in the Black Sea region ofTurkeyrdquo Combinatorial Chemistry amp High Throughput Screen-ing vol 14 no 2 pp 72ndash84 2011
[10] V E C Ooi and F Liu ldquoA Review of pharmacological activitiesof mushroom polysaccharidesrdquo International Journal of Medic-inal Mushrooms vol 1 no 3 pp 195ndash206 1999
[11] S PWasser and A LWeis ldquoMedicinal properties of substancesoccurring in higher Basidiomycete mushroom current per-spectiverdquo International Journal of Medicinal Mushrooms vol 1pp 31ndash62 1999
[12] P M Kidd ldquoThe use of mushroom glucans and proteoglycansin cancer treatmentrdquo Alternative Medicine Review vol 5 no 1pp 4ndash27 2000
[13] C Israilides and A Philippoussis ldquoBio-technologies of recy-cling agro-industrial wastes for the production of commerciallyimportant fungal polysaccharides andmushroomsrdquo Biotechnol-ogy and Genetic Engineering Reviews vol 20 pp 247ndash259 2003
[14] T Mizuno ldquoBioactive biomolecules of mushrooms food func-tion and medicinal effect of mushroom fungirdquo Food ReviewsInternational vol 11 no 1 pp 5ndash21 1995
[15] Y Kabir and S Kimura ldquoDietary mushrooms reduce bloodpressure in spontaneously hypertensive rats (SHR)rdquo Journal of
Nutritional Science and Vitaminology vol 35 no 1 pp 91ndash941989
[16] J-L Mau S-Y Tsai Y-H Tseng and S-J Huang ldquoAntioxidantproperties of hot water extracts from Ganoderma tsugae Mur-rillrdquo LWTmdashFood Science and Technology vol 38 no 6 pp 589ndash597 2005
[17] A Smania F D Monache E F A Smania M L Gil L CBenchetrit and F S Cruz ldquoAntibacterial activity of a substanceproduced by the fungus Pycnoporus sanguineus (Fr) MurrrdquoJournal of Ethnopharmacology vol 45 no 3 pp 177ndash181 1995
[18] W Andlauer and P Furst ldquoNutraceuticals a piece of historypresent status and outlookrdquo Food Research International vol 35no 2-3 pp 171ndash176 2002
[19] C L Kruger and S W Mann ldquoSafety evaluation of functionalingredientsrdquo Food and Chemical Toxicology vol 41 no 6 pp793ndash805 2003
[20] P H Mattila V I Piironen E J Uusi-Rauva and P E Koivis-toinen ldquoVitamin D contents in edible mushroomsrdquo Journal ofAgricultural and Food Chemistry vol 42 no 11 pp 2449ndash24531994
[21] R P Tewari ldquoMushroom their role in nature and societyrdquo inFrontiers inMushroomBiotechnology R D Rai R C Upadhyayand S R Sharma Eds pp 1ndash8 NRCM Chambaghat India2005
[22] K C Semwal S L Stephenson V K Bhatt and R P BhattldquoEdible mushrooms of the Northwestern Himalaya India astudy of indigenous knowledge distribution and diversityrdquoMycosphere vol 5 no 3 pp 440ndash461 2014
[23] E V Crisan and A Sands ldquoEdible mushrooms nutritionalvaluerdquo in The Biology and Cultivation of Edible Mushrooms ST Chang and W A Hayes Eds pp 137ndash165 Academic PressNew York NY USA 1978
[24] N S Atri S K Sharma R Joshi A Gulati and A Gula-ti ldquoNutritional and neutraceutical composition of five wildculinary-medicinal species of genus Pleurotus (Higher Basid-iomycetes) from Northwest Indiardquo International Journal ofMedicinal Mushrooms vol 15 no 1 pp 49ndash56 2013
[25] L Barros T Cruz P Baptista L M Estevinho and I C FR Ferreira ldquoWild and commercial mushrooms as source ofnutrients and nutraceuticalsrdquo Food and Chemical Toxicologyvol 46 no 8 pp 2742ndash2747 2008
[26] M Nagata and I Yamashita ldquoSimple method for simultaneousdetermination of chlorophyll and carotenoids in tomato fruitrdquoNippon Shokuhin Kogyo Gakkaishi vol 39 no 10 pp 925ndash9281992
[27] V L Singleton and J A Rossi ldquoColorimetric detection oftotal phenolics with phosphomolybdic-phosphotungstic acidreagentsrdquo American Journal of Enology and Viticulture vol 16pp 144ndash158 1965
[28] L K Jagadish V Venkata Krishnan R Shenbhagaramanand V Kaviyarasan ldquoComparitive study on the antioxidantanticancer and antimicrobial property of Agaricus bisporus (JE Lange) Imbach before and after boilingrdquo African Journal ofBiotechnology vol 8 no 4 pp 654ndash661 2009
[29] E Vamanu and S Nita ldquoAntioxidant capacity and the cor-relation with major phenolic compounds anthocyanin andtocopherol content in various extracts from the wild edibleBoletus edulis mushroomrdquo BioMed Research International vol2013 Article ID 313905 11 pages 2013
[30] E Vamanu ldquoBiological activities of the polysaccharides pro-duced in submerged culture of two edible Pleurotus ostreatus
12 BioMed Research International
mushroomsrdquo Journal of Biomedicine and Biotechnology vol2012 Article ID 565974 8 pages 2012
[31] P L Li L Huo W Su et al ldquoFree radical-scavenging capacityantioxidant activity and phenolic content of Pouzolzia zeylan-icardquo Journal of the Serbian Chemical Society vol 76 no 5 pp709ndash717 2011
[32] C Papuc M Crivineanu G Goran V Nicorescu and N Dur-dun ldquoFree radicals scavenging and antioxidant activity ofEuropean mistletoe (Viscum album) and European birthwort(Aristolochia clematitis)rdquo Revista de Chimie vol 61 no 7 pp619ndash622 2010
[33] VOOyetayo CDong andY Yao ldquoAntioxidant and antimicro-bial properties of aqueous extract from Dictyophora indusiatardquoThe Open Mycology Journal vol 3 no 1 pp 20ndash26 2009
[34] S Marklund and G Marklund ldquoInvolvement of the superoxideanion radical in the autoxidation of pyrogallol and a convenientassay for superoxide dismutaserdquo European Journal of Biochem-istry vol 47 no 3 pp 469ndash474 1974
[35] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996
[36] D J Huang b Ou and R L Prior ldquoThe chemistry behindantioxidant capacity assaysrdquo Journal of Agricultural and FoodChemistry vol 53 no 6 pp 1841ndash1856 2005
[37] P Manzi A Aguzzi and L Pizzoferrato ldquoNutritional value ofmushrooms widely consumed in Italyrdquo Food Chemistry vol 73no 3 pp 321ndash325 2001
[38] P Manzi S Marconi A Aguzzi and L Pizzoferrato ldquoCommer-cial mushrooms nutritional quality and effect of cookingrdquo FoodChemistry vol 84 no 2 pp 201ndash206 2004
[39] I P Krbavcic and I C Baric ldquoInfluence of deep fat fryingon some nutritional parameters of novel food based on mush-rooms and fresh soft cheeserdquo Food Chemistry vol 84 no 3 pp417ndash419 2004
[40] D Agrahar-Murugkar andG Subbulakshmi ldquoNutritional valueof edible wild mushrooms collected from the Khasi hills ofMeghalayardquo Food Chemistry vol 89 no 4 pp 599ndash603 2005
[41] V A Dıez and A Alvarez ldquoCompositional and nutritionalstudies on two wild edible mushrooms from northwest SpainrdquoFood Chemistry vol 75 no 4 pp 417ndash422 2001
[42] L Barros M-J Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007
[43] S K Sharma and N S Atri ldquoNutraceutical composition ofwild species of genus Lentinus Fr fromNothern Indiardquo CurrentResearch in Environmental ampAppliedMycology vol 4 pp 11ndash322014
[44] P J Kanu K Zhu J Baby Kanu H Zhou H Qian and K ZhuldquoBiologically active components and nutraceuticals in sesameand related products a review and prospectrdquo Trends in FoodScience and Technology vol 18 no 12 pp 599ndash608 2007
[45] F S Reis E Pereira L Barros M J Sousa A Martins and I CF R Ferreira ldquoBiomolecule profiles in inediblewildmushroomswith antioxidant valuerdquoMolecules vol 16 no 6 pp 4328ndash43382011
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
mushroomsrdquo Journal of Biomedicine and Biotechnology vol2012 Article ID 565974 8 pages 2012
[31] P L Li L Huo W Su et al ldquoFree radical-scavenging capacityantioxidant activity and phenolic content of Pouzolzia zeylan-icardquo Journal of the Serbian Chemical Society vol 76 no 5 pp709ndash717 2011
[32] C Papuc M Crivineanu G Goran V Nicorescu and N Dur-dun ldquoFree radicals scavenging and antioxidant activity ofEuropean mistletoe (Viscum album) and European birthwort(Aristolochia clematitis)rdquo Revista de Chimie vol 61 no 7 pp619ndash622 2010
[33] VOOyetayo CDong andY Yao ldquoAntioxidant and antimicro-bial properties of aqueous extract from Dictyophora indusiatardquoThe Open Mycology Journal vol 3 no 1 pp 20ndash26 2009
[34] S Marklund and G Marklund ldquoInvolvement of the superoxideanion radical in the autoxidation of pyrogallol and a convenientassay for superoxide dismutaserdquo European Journal of Biochem-istry vol 47 no 3 pp 469ndash474 1974
[35] I F F Benzie and J J Strain ldquoThe ferric reducing ability ofplasma (FRAP) as a measure of lsquoantioxidant powerrsquo the FRAPassayrdquo Analytical Biochemistry vol 239 no 1 pp 70ndash76 1996
[36] D J Huang b Ou and R L Prior ldquoThe chemistry behindantioxidant capacity assaysrdquo Journal of Agricultural and FoodChemistry vol 53 no 6 pp 1841ndash1856 2005
[37] P Manzi A Aguzzi and L Pizzoferrato ldquoNutritional value ofmushrooms widely consumed in Italyrdquo Food Chemistry vol 73no 3 pp 321ndash325 2001
[38] P Manzi S Marconi A Aguzzi and L Pizzoferrato ldquoCommer-cial mushrooms nutritional quality and effect of cookingrdquo FoodChemistry vol 84 no 2 pp 201ndash206 2004
[39] I P Krbavcic and I C Baric ldquoInfluence of deep fat fryingon some nutritional parameters of novel food based on mush-rooms and fresh soft cheeserdquo Food Chemistry vol 84 no 3 pp417ndash419 2004
[40] D Agrahar-Murugkar andG Subbulakshmi ldquoNutritional valueof edible wild mushrooms collected from the Khasi hills ofMeghalayardquo Food Chemistry vol 89 no 4 pp 599ndash603 2005
[41] V A Dıez and A Alvarez ldquoCompositional and nutritionalstudies on two wild edible mushrooms from northwest SpainrdquoFood Chemistry vol 75 no 4 pp 417ndash422 2001
[42] L Barros M-J Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007
[43] S K Sharma and N S Atri ldquoNutraceutical composition ofwild species of genus Lentinus Fr fromNothern Indiardquo CurrentResearch in Environmental ampAppliedMycology vol 4 pp 11ndash322014
[44] P J Kanu K Zhu J Baby Kanu H Zhou H Qian and K ZhuldquoBiologically active components and nutraceuticals in sesameand related products a review and prospectrdquo Trends in FoodScience and Technology vol 18 no 12 pp 599ndash608 2007
[45] F S Reis E Pereira L Barros M J Sousa A Martins and I CF R Ferreira ldquoBiomolecule profiles in inediblewildmushroomswith antioxidant valuerdquoMolecules vol 16 no 6 pp 4328ndash43382011
Submit your manuscripts athttpwwwhindawicom
PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014