GJRMI - Volume 2, Issue 4, April 2013

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INDEX – GJRMI, Vol.2, Iss. 4, April 2013

MEDICINAL PLANTS RESEARCH

Laboratory Sciences & Ethno-Botany SCREENING FOR ANTIMICROBIAL ACTIVITY OF SOME PLANTS FROM SAUDI FOLK

MEDICINE

Abdallah Emad M, El-Ghazali Gamal E 189–197

Botany DIVERSITY AND AVAILABILITY STATUS OF ETHNOMEDICINAL PLANTS IN THE LOHBA

RANGE OF KEDARNATH FOREST DIVISION (KFD), GARHWAL HIMALAYA

Ballabha Radha, Singh Dinesh, Tiwari J K, Tiwari P 198–212

Ophthalmology & Pharmacology

ANTICATARACT ACTIVITY OF ERVATAMIA CORONARIA LEAF EXTRACT ON

CHEMICALLY INDUCED CATARACTOGENESIS IN RATS

Rathnakumar K, Jaikumar S, Duraisami R, Sengottuvelu S 213–218

Ethno-Botany STUDIES ON TRADITIONAL HERBAL PEDIATRICS PRACTICES IN JAISINGHPUR, DISTRICT

KANGRA (HIMACHAL PRADESH, INDIA)

Rawat Dhiraj S, Kharwal Anjna D 219–230

Bio-Chemistry

LIVER ENZYMES AND ITS ASSOCIATION WITH AGE AND SEX IN SICKLE CELL ANAEMIA

PATIENTS AND HAEMOGLOBIN S TRAIT CARRIES.

Chuku L C, Chinaka N C 231–237

Life Sciences CHELIDONIUM MAJUS L. - A REVIEW ON PHARMACOLOGICAL ACTIVITIES AND

CLINICAL EFFECTS

Biswas Surjyo Jyoti 238–245

INDIGENOUS MEDICINE

Ayurveda – Dravya Guna A DETAILED PHARMACOGNOSTICAL EVALUATION ON LEAF OF OLAX SCANDENS ROXB.

Naik Raghavendra, Borkar Sneha D, Harisha C R, Acharya R N 246–253

Ayurveda – Rachana Sharira

A CLINICAL EVALUATION ON RUJAKARA MARMA WITH SPECIAL REFERENCE TO PAIN

THRESHOLD

Benjwal Shobha 254–258

Ayurveda – Dravya Guna

PHARMACOGNOSTICAL AND PRELIMINARY PHYTOCHEMICAL INVESTIGATIONS ON

DIFFERENT PARTS OF BULBOPHYLLUM NEILGHERRENSE WIGHT. -AN ORCHID USED IN

FOLK MEDICINE.

Kumari Harshitha, Nishteswar K, Harisha C R 259–269

Ayurveda – Dravya Guna

A COMPARATIVE ACUTE TOXICITY EVALUATION OF ASHOKA KSHEERAPAKA PREPARED

FROM TWO DIFFERENT SPECIES OF SARACA (S. ASOCA & S. THAIPINGENSIS)

Chavan S S, Gamit R V, Ashok B K, Shukla V J, Das P, Ravishankar B 270–277

COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – BRANCH OF KEBUKA [CHEILOCOSTUS SPECIOSUS (J.

KONIG) C. SPECHT], OF THE FAMILY COSTACEAE PLACE – KOPPA, CHIKKAMAGALUR DISTRICT, KARNATAKA, INDIA

Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 4 | April 2013 | 189–197

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal

SCREENING FOR ANTIMICROBIAL ACTIVITY OF SOME PLANTS FROM

SAUDI FOLK MEDICINE

Abdallah Emad M1*, El-Ghazali Gamal E

2

1, 2

Department of Laboratory Sciences, College of Science and Arts, Al-Rass, P.O. Box 53, Qassim University,

Saudi Arabia.

*Corresponding author: [email protected]

Received: 24/02/2013; Revised: 28/03/2013; Accepted: 31/03/ 2013

ABSTRACT

Methanolic extract of nine medicinal plants from Saudi folk medicine (Tamarix aphylla,

Dactyloctenium aegyptium, Francoeuria crispa, Rhazya stricta, Trichodesma africanum, Haloxylon

salicornicum, Echinops spinosissimus, Zygophyllum simplex and Blepharis ciliaris) were examined

for their phytochemical compounds and antimicrobial potential against seven standard bacteria

(Proteus vulgaris NCTC 8196, Escherichia coli ATCC 25922, Bacillus cereus NCTC 8236,

Salmonella typhi NCTC 0650, Klebsiella pneumonia ATCC 53651, Pseudomonas aeruginosa ATCC

27853 and Staphylococcus aureus ATCC 25923) and one standard fungus (Candida albicans ATCC

7596). The phytochemical analysis showed presence of some active principles which correlates with

the antimicrobial activity of some plant extracts. Most plants showed some degree of antimicrobial

activity. However, the methanol extracts of Rhazya stricta, Francoeuria crispa and Blepharis ciliaris

respectively, recorded the maximum antimicrobial activities compared to Chloramphenicol as

antibacterial and Clotrimazole as antifungal antibiotic. The results of this investigation support the

use of these plants in Saudi folk medicine for treatment of ailments caused by microorganisms.

KEYWORDS: Medicinal plants, folk medicine, phytochemical, antimicrobial

Research article

Cite this article:

Abdallah Emad M, El-Ghazali Gamal E (2013), SCREENING FOR ANTIMICROBIAL ACTIVITY OF

SOME PLANTS FROM SAUDI FOLK MEDICINE, Global J Res. Med. Plants & Indigen. Med., Volume

2(4): 189–197

mailto:[email protected]



INTRODUCTION

No doubt that what we called it folk,

traditional or alternative medicine was the main

source for remedies since antiquity. On the

other side, what we called modern medicine is

now facing challenge due to its failure to

convoy the development of diseases.

Accordingly, there is a serious need to go back

to the Mother Nature and its rich sources of

natural remedies. It is well known that, man has

been employing natural products as remedies

since times immemorial, this knowledge

accumulated and passed from generation to

generation. Thus, the ethno-botanical

knowledge about drugs considered as a basic

source for new therapeutics.

Recently, the misuse of synthetic antibiotics

has promoted the emergence of antibiotic-

resistant pathogens, including multidrug

resistant strains (Kumar and Schweizer, 2005).

This predicament, impose to search for other

alternatives that are not centered upon standard

antibiotics therapy, or we risk the possibility of

eventually having no defense against these

antibiotic-resistant pathogens (Treadway,

1998). Medicinal plants, particularly those

employed in folk medicine could be a

promising alternative, as it could gives a new

source of antimicrobial agents with possibly

novel mechanisms of action (Runyoro et al.,

2006). The first step towards discovering novel

antimicrobials is the screening of such plants.

Saudi Arabia has a hot desert climate and

rainfall is scarce in most parts of the country.

The flora of Saudi Arabia as well as the other

countries in the peninsula has been neglected

for a long time due to its arid climate. The first

attempt to cover the flora of Saudi Arabia was

in 1974 (Alfarhan et al., 1998). However, folk

medicine, including medicinal herbs, occupies

a significant part of Saudi Arabia’s heritage and

it is widely practiced until now (Al-Essa et al.,

1998). Although, the growing development in

healthcare in Saudi Arabia which based on the

western modern medicine decreases the public

interest in the traditional medicine, particularly

in towns.

Tamarix aphylla, Dactyloctenium

aegyptium, Francoeuria crispa, Rhazya stricta,

Trichodesma africanum, Haloxylon

salicornicum, Echinops spinosissimus,

Zygophyllum simplex and Blepharis ciliaris are

chosen to study because they are already being

utilized in folk medicine as antimicrobials (in

treating wounds, sores, inflammations, cough

and cold, etc.) by native inhabitants and

Bedouin at Al-Rass province, Qassim district,

Saudi Arabia (El-Ghazali et al., 2010.) In the

present study, the antimicrobial activities

beside the preliminary phytochemical

investigation of the above mentioned plants

were evaluated.

MATERIALS AND METHODS

Plant material

Plants were collected based on information

of previous ethnobotanical survey on medicinal

plants used by native nomadic people (El-

Ghazali et al., 2010). In this study, nine

selected plants claimed to be used as

antimicrobials in folk medicine (Antidiarrheal,

antiseptic, anti-inflammatory, anti-cold and

cough, and in wound treatment) were collected

from Al-Rass province, Qassim district, Saudi

Arabia. All plants were identified by Gamal E.

El-Ghazali (Taxonomist). Information

regarding these plants is shown in Table 1.

Microbial strains

All microbial strains, 7 reference bacterial

strains representing the gram negatives

(Escherichia coli ATCC 25922, Klebsiella

pneumonia ATCC 53651, Pseudomonas

aeruginosa ATCC 27853, Proteus vulgaris

NCTC 8196, Salmonella typhi NCTC 0650)

and gram positives (Bacillus cereus NCTC

8236, Staphylococcus aureus ATCC 25923),

and one reference fungal strain (Candida

albicans ATCC 7596) were obtained from the

stock culture of the microbiology laboratory,

Medicinal and Aromatic Plants Research

Institute, Khartoum, Sudan.



Inoculum preparation

Active cultures were prepared by

inoculating the stock culture into sterile bottles

containing Nutrient broth (MAST Laboratories

LTD, UK) for bacteria and Malt extract broth

(OXOID, UK) for fungus and incubated for 24

h at 37°C and 72 h at 25°C respectively. The

turbidity of actively growing bacterial

suspension was adjusted to match the turbidity

standard of 0.5 McFarland. This turbidity is

equivalent to approximately 1–2 × 108 CFU/ml

for bacteria and about 2.0 × 105

spore/ml for

fungi. This suspension was used for the

antimicrobial examination.

Plant extracts

Plant parts (leaves or whole plant) were

washed with distilled water and air dried in

shade inside the laboratory for about 2–3 weeks

until totally dried and grounded with a grinder

machine. 50 g of each ground material was

soaked in 500 ml of methanol, for at least 72 h

with frequent shakings. The samples were

filtered using Whattman No.1 filter paper

(Whatman limited, UK). The filtrate was

evaporated to dryness under reduced pressure

at 40 o

C. All the extracts (in powder form) were

kept in refrigerator in dark bottles until used

(Samie et al., 2005).

Phytochemical screening

Screening for some active phytochemical

principles claimed having antimicrobial activity

were undertaken as described by Edeoga et al.,

(2005), Krishnaiah et al., (2009) and Abdallah

et al., (2009) for detection of tannins, saponins,

flavonoids, terpenoids, phenolic compounds,

alkaloids and anthraquinones.

Antimicrobial assay

The antimicrobial activity of the methanol

extract of the nominated plant extracts were

evaluated using agar-well diffusion method as

mentioned by Abdallah et al., (2012), with

minor modifications. The dry methanolic

extracts of plants under study were re-

constructed with 70% methanol to make a final

concentration 100 mg/ml and filtered using

0.22 μm pore-size black polycarbonate filters

(Millipore). To a sterile Petri-dish (Size

100 mm), 25 ml of molten Nutrient Agar

(MAST Laboratories Ltd, UK) or Potato

Dextrose agar (Oxoid Ltd, UK) was poured and

left to solidify. Fresh working cell suspensions

(Bacteria or fungus) were prepared and

adjusted to 0.5 McFarland's standard. Then

from each microorganism, 100 μl was spread

onto the surface of the plates of Nutrient Agar

for bacteria or Potatoes Dextrose Agar for

fungi. After about 15 min., 6 mm wells were

punched into the agar using a sterile cork borer.

Afterwards, 100 μl (10000 μg/wells) from each

concentration was loaded into the wells of the

previously prepared plates and incubated for 24

h at 37°C for bacterial strains and 72 h at 25°C

for fungus. Chloramphenicol 5 mg/ml

(50 μg/wells) (Riyadh Pharma. Co. Ltd, SA)

and Clotrimazole 10 mg/ml (100 μg/wells)

(Pharco Pharmaceuticals, Egypt) were

employed as antibacterial and antifungal

positive controls, respectively. 70% methanol

was employed as a negative control. Each test

was repeated twice and the mean zone of

inhibition was recorded.

RESULTS AND DISCUSSION

Nowadays, the natural products and

medicinal plants are a subject of great global

interest for the discovery of new antimicrobial

agents (Sashikala et al., 2009). This could be

related to the recent failure of antibiotics

against the dramatic emerging of the multidrug

resistant pathogens in addition to the rapid

spread of the new infections (Abdallah, 2011).

The ethnobotanical information of the studied

plants and their usage in the folk medicine at

Al-Rass province, Saudi Arabia are

summarized in Table 1. The ethnobotanical

approach has advantages over the random

screening, as this approach depends on the

human experience with diseases through

generations.

The results of the phytochemical screening

of the nominated plant extracts (Methanol

extracts) showed that, all the methanolic



extracts of the plants exhibited presence of at

least two or more of these phytochemical

compounds (Table 2). The phytochemical

compounds studied in this investigation having

antimicrobial potency, as reported by many

researchers such as Watt and Pretorius (2001)

for tannin and phenolic compounds, Deeni and

Sadiq (2002) and Rohit et al. (2012) for

anthraquinones and saponins, Rίos and Recio

(2005) for alkaloids, flavonoids and terpenoids.

It is known that the phytochemical compounds,

which are secondary metabolic products in

plants, produce some biological activities in

human and animal and responsible for their use

as a drug (Sofowora, 1984).

Table 1: Ethnobotanical information of some selected medicinal plants from Al- Rass

province

*NA: Not available.

Scientific name Family Common

names

Part tested Traditional uses

Tamarix aphylla

(L.) Karsten.

Tamaricaceae Taramisk,

Atheltrep, Salt

cedar, Athel,

Athel pine

Leaves Anti-inflammatory and

in treating wounds

Dactyloctenium

aegyptium

(L.) Willd.

Poaceae

(Graminae)

Crowfoot

grass, Star

grass, Beach

wire grass,

Button grass,

Buck grass

Whole plant In treating wounds

Francoeuria crispa

(Forssk.) Cass.

Asteraceae

(Compositae)

Francoeuria Whole plant Its smoke breathed to

treat the upper thorax

inflammations

Rhazya stricta

Decne.

Apocynaceae Harmal,

Senhwar,

Dogbane.

Leaves Dried leaves used in

healing wounds

Trichodesma

africanum

(L.) Lehm.

Boraginaceae NA* Whole plant Anti-cold

Haloxylon

salicornicum

(Moq.) Boiss.

Chenopodiaceae NA* Whole plant Its smoke breathed to

treat cold

Echinops

spinosissimus

Turra.

Asteraceae NA* Leaves In treating the upper

thorax infections.

Zygophyllum simplex

L.

Zygophyllaceae NA* Whole plant In treating ophthalmia

Blepharis ciliaris

(L.)B.L. Burtt.

Acanthaceae NA* Whole

plant

In treating wounds and

in renal disorder.



Table 2: Qualitative analysis of the phytochemicals of the medicinal plants

Plant Phytochemical compounds*

Tan Sap Fla Ter Phen Alk Anth

Tamarix aphylla ‒ + ‒ + + ± ‒

Dactyloctenium

aegyptium

‒ ± ‒ ‒ + + ‒

Francoeuria crispa + ‒ + + + + +

Rhazya stricta ‒ ‒ ‒ + + + +

Trichodesma africanum + + ± + ± ‒ ‒

Haloxylon salicornicum ‒ + ‒ + + + ‒

Echinops spinosissimus ± ‒ + + + ± ‒

Zygophyllum simplex + + + + + ± ±

Blepharis ciliaris + + + + + ‒ +

Negative control (D.W.) ‒ ‒ ‒ ‒ ‒ ‒ ‒

* Tan= Tannins, Sap= Saponins, Fla= Flavonoids, Ter= Terpenoids, Phen= Phenolic compounds,

Alk= Alkaloids, Anth= Anthraquinones.

+ = Presence, − = Absence, ± = Weak reaction, D.W. = Distilled water.

Table 3: Screening of the methanol extracts of some plant species for antimicrobial activity of

Gram-negative bacteria

Tested*

Mean zone of inhibition (mm) of microorganisms

(Mean±SEM)**

Pr Ec Kp Ps Sa

Tamarix aphylla 11.0± 2.0 9.0 ± 1.0 8.5 ± 0.5 10.5 ± 1.5 15.5 ± 0.5

Dactyloctenium aegyptium 12.5 ± 0.5 8.0 ± 0.0 12.0 ± 1.0 11.5 ± 1.5 13.0 ± 0.0

Francoeuria crispa 18.0 ± 1.0 19.0 ± 1.0 15.0 ± 0.0 15.5 ± 0.5 16.0 ± 0.0

Rhazya stricta 20.5 ± 0.5 19.5 ± 0.5 20.0 ± 2.0 24.0 ± 1.0 9.5 ± 0.5

Trichodesma africanum 9.5 ± 0.5 10.5 ± 0.5 12.5 ± 0.5 10.5 ± 0.5 14.0 ± 0.0

Haloxylon salicornicum 11.5 ± 1.5 13.0 ± 0.0 12.0 ± 2.0 12.0 ± 0.0 15.5 ± 0.5

Echinps spinosissimus 12.5 ± 1.5 12.0 ± 0.0 12.0 ± 1.5 10.5 ± 0.5 13.0 ± 0.0

Zygophyllum simplex 14.5 ± 0.5 10.0 ± 1.0 8.5 ± 0.5 9.5 ± 0.5 13.0 ± 0.0

Blepharis ciliaris 18.5 ± 0.5 14.5 ± 0.5 16.0 ± 0.0 15.0 ± 2.0 14.5 ± 1.5

Chloramphenicol 5 mg/ml 26.0 ± 1.0 13.7 ± 1.05 22.0 ± 0.2 24.0 ± 1.0 16.5 ± 1.5

* Plants tested are as methanol extracts at 100mg/ml, Chloramphenicol as antibacterial at 5 mg/ml.

**Mean ± Standard error of means (SEM ), mm=millimeter;

Microorganisms: Pr = Proteus vulgaris NCTC 8196, Ec = Escherichia coli ATCC 25922, Kp = Klebsiella

pneumonia ATCC 53651, Ps = Pseudomonas aeruginosa ATCC 27853, Sa = Staphylococcus aureus ATCC

25923.



Table 4: Screening of the methanol extracts of some plant species for antimicrobial activity of

Gram-positive bacteria and fungus

Tested*

Mean zone of inhibition (mm) of microorganisms

(Mean±SEM)**

Bs Sal Cand

Tamarix aphylla 12.0 ± 2.0 11.0 ±0.0 6.5 ± 0.5

Dactyloctenium aegyptium 11.0 ± 2.0 9.5 ± 0.5 11.5 ± 0.5

Francoeuria crispa 11.5 ±1.5 10.5 ± 0.5 18.0 ± 1.0

Rhazya stricta 10.0 ± 1.0 19.0 ± 0.5 14.0 ± 1.0

Trichodesma africanum 13.5 ± 1.5 11.5 ± 1.5 17.5 ± 0.5

Haloxylon salicornicum 12.5 ± 1.5 12.5 ± 2.5 14.5 ± 0.5

Echinps spinosissimus 13.0 ± 0.0 13.5 ± 0.5 15.0 ± 0.0

Zygophyllum simplex 9.5 ± 0.5 13.0 ± 2.0 13.5 ± 0.5

Blepharis ciliaris 11.5 ± 0.5 14.5 ± 0.5 18.0 ± 1.0

Chloramphenicol 5 mg/ml 21.0 ± 1.0 13.0 ± 1.0 ‒

Clotrimazole 10 mg/ml ‒ ‒ 13.0 ± 1.0

* Plants tested are as methanol extracts at 100 mg/ml, Chloramphenicol as antibacterial at 5 mg/ml and

Clotrimazole as antifungal at 10 mg/ml, ‒ = Not tested.

**Mean ± Standard error of means (SEM), mm=millimeter;

Microorganisms: Bs = Bacillus cereus NCTC 8236, Sal = Salmonella typhi NCTC 0650, Cand = Candida

albicans ATCC 7596.

The antibacterial and antifungal activities of

the methanolic plant’s extracts are shown in

Tables 3 and 4. Methanol as a solvent is the

most commonly used solvents for preliminary

studies of antimicrobial activities in plants (Das

et al., 2010). In general, most plants showed

some degree of antimicrobial activity against

tested microorganisms. However, Rhazya

stricta, Francoeuria crispa and Blepharis

ciliaris respectively, recorded the highest

antimicrobial activity among other plants, even

higher than that was recorded by the

commercial antibiotic itself (Tables 3 and 4). E.

coli was much susceptible to Rhazya stricta

(19.5 ± 0.5), Francoeuria crispa

(19.5 ± 0.5 mm) and Blepharis ciliaris

(14.5 ± 0.5 mm) when compared to

Chloramphenicol (13.7 ± 1.05 mm). Similarly,

Salmonella typhi was highly susceptible to

Rhazya stricta (19.0 ± 0.5 mm) and Blepharis

ciliaris (14.5 ± 0.5 mm), compared to

Chloramphenicol (13.0 ± 1.0 mm). Also, the

susceptibility of Pseudomonas aeruginosa to

the methanol extract of Rhazya stricta was

similar to that of Chloramphenicol

(24.0 ± 1.0 mm). Our results regarding the

antibacterial potential of the methanol extract

of Rhazya stricta are in harmony with Ahmad

et al., (2004) who mentioned that the crude

ethanolic extract of Rhazya stricta exhibited a

considerable antibacterial activity against a

wide range of gram-positive and gram negative

bacteria, the susceptibility of some of these

bacteria to this plant extract were higher than

that of the antibiotic Co-trimoxazole. Same

result recorded by Staphylococcus aureus

where the inhibition zones with the methanol

extract of Francoeuria crispa almost equal to

its inhibition zone with Chloramphenicol

(Tables 3 and 4). Similarly, the results of this

study regarding the methanol extract of

Francoeuria crispa (Tables 3 and 4) are in

agreement with the findings of El-Kamali and

Mahjoub (2009) there were found that the most

susceptible bacteria were E. coli and P.

vulgaris and Staphylococcus aureus and the



least susceptible were Salmonella para typhi

and B. subtillis. Also, Blepharis ciliaris was

reported in literature as a plant of antimicrobial

properties (El-Shanawany et al., 2012) agrees

with the findings of this investigation. The

antibiotic Chloramphenicol became available

commercially in 1948 and it was active against

all gram-positive and many gram-negative

bacteria at that time. However, within a few

years bacterial resistance to Chloramphenicol

was recorded (Shaw, 1984). Since Rhazya

stricta, Francoeuria crispa and Blepharis

ciliaris exhibited maximum antibacterial

activities higher than the Chloramphenicol

against some bacteria, it is believed that these

plants when extracted and purified may lead to

new effective antibacterial drugs.

As shown in Tables 3 and 4, the results of

the antifungal activities showed that, most plant

extracts exhibited antifungal activity higher

than the antifungal agent (Clotrimazole

10 mg/ml). The methanol extract of

Francoeuria crispa and Blepharis ciliaris

(18.0 ± 1.0 mm), besides Tricodesma

africanum (17.5 ± 0.5) revealed the maximum

antifungal activity against Candida albicans

compared to Clotrimazole (13.0 ± 1.0 mm).

Data regarding the antifungal activity of the

above mentioned plants are scanty. However,

these results are promising in order to introduce

new antifungal agents. Undoubtedly, the

antimicrobial agents derived from such plants

may eradicate bacteria or fungi by a mechanism

different than that occurred by the synthetic or

semi-synthetic antibiotics (Eloff, 1997), which

could be much effective with less side effects.

CONCLUSION

The results of the present study provide

support to the claim on these plants in Saudi

folk medicine against some of the mentioned

disorders and diseases. Plants of interest are

those which exhibited antimicrobial activity

higher than the antibiotics tested. These plants

(Rhazya stricta, Francoeuria crispa and

Blepharis ciliaris) should be subjected to

intensive studies such as fractionation, isolation

of the active constituents, toxicity against

animal and human cells and so forth.

ACKNOWLEDGEMENTS

This study was supported by the Deanship

of scientific research, Qassim University, Saudi

Arabia, grant No. SR-D-1105. Authors are

greatly thankful for this support.

REFERENCES

Abdallah EM, Khalid AS, Ibrahim N (2009).

Antibacterial activity of oleo-gum

resins of Commiphora molmol and

Boswellia papyrifera against methicillin

resistant Staphylococcus aureus

(MRSA). Sci. Res. Essay. 4 (4): 351–

356.

Abdallah EM (2011). Plants: An alternative

source for antimicrobials. J. Applied

Pharma. Sci., 1(6): 16–20.

Abdallah EM, Hsouna AB, Al-Khalifa KS

(2012). Antimicrobial, antioxidant and

phytochemical investigation of

Balanites aegyptiaca (L.) Del. edible

fruit from Sudan. Afri. J. Biotech.,

11(52): 11535–11542.

Ahmad S, Fatima K, Atiq-ur-Rahman (2004).

Antibacterial activity of Pakistani

Rhazya stricta. Pakistan J. Sci. Indust.

Res., 47 (1): 29–33.



Al-Essa MA, Al-Mehaidib A, Al-Gain S

(1998). Parental awareness of liver

disease among children in Saudi Arabia.

Ann. Saudi Med., 18 (1): 79–81.

Alfarhan A H, Chaudhary SA, Thomas J

(1998). Notes on the flora of Saudi

Arabia (Third edition). J. King Saud

Univ., 10(1): 31–40.

Das K, Tiwari R KS, Shrivastava D K (2010).

Techniques for evaluation of medicinal

plant products as antimicrobial agent:

Current methods and future trends. J.

Med. Plants Res., 4(2): 104–111.

Deeni YY, Sadiq NM (2002). Antimicrobial

properties and phytochemical

constituents of the leaves of African

mistletoe (Tapinanthus dodoneifolius

(DC) Danser) (Loranthaceae): an

ethnomedicinal plant of Hausaland,

North Nigeria J. of Ethnopharma., 83:

235–240.

Edeoga H O, Okwu D E, Mbaebie B O (2005).

Phytochemical constituents of some

Nigerian medicinal Plants. African J.

Biotech., 4 (7): 685–688.

El-Ghazali GE, Al-Khalifa KS, Saleem GA,

Abdallah EM (2010). Traditional

medicinal plants indigenous to Al-Rass

province, Saudi Arabia. J. Med. Plants

Res., 4(24): 2680–2683.

El-Kamali H H, Mahjoub SA (2009).

Antibacterial activity of Francoeuria

crispa, Pulicaria undulates, Ziziphus

spina-christi and Cucurbita pepo

anainst seven standard pathogenic

bacteria. Ethnobotanical Leaflets, 13:

722–733.

Eloff JN (1997). Which extract should be used

for the screening and isolation of

antimicrobial components from plants?.

J. Ethnopharma., 60: 1–8.

El-Shanawany MA, Sayed HM, Ibrahim SRM,

Fayed MMA, Radwan MM, Ross SA

(2012). A new isoflavone from

Blepharis ciliaris of Egyptian origin.

Med. Chem. Res.DOI 10.1007/S00044-

012-0228-2.

Krishnaiha D, Devi T, Bono A, Sarbatly R

(2009). Studies on phytochemical

constituents of six Malaysian medicinal

plants. J. Med. Plants Res., 3(2): 67–

72.

Kumar A, Schweizer HP (2005). Bacterial

resistance to antibiotics: Active efflux

and reduced uptake. Advanced Drug

Delivery Rev., 57: 1486–1513.

Rίos J L, Recio M C (2005). Medicinal plants

and antimicrobial activity. J.

Ethnopharma., 100: 80–84.

Rohit S, Thakur G S, Sanodiya B S,

Mukeshwar P, Prakash B (2012).

Saponin: a wonder drug from

Chlorophytum species. Global J Res.

Med. Plants & Indigen. Med.,1(10):

503–515.

Runyoro D, Matee M, Olipa N, Joseph C,

Mbwambo H (2006), Screening of

Tanzanian medicinal plants for anti-

Candida activity. BMC Complement

Altern. Med., 6(11). doi:10.1186/1472-

6882-6-11.

Samie A, Obi C L, Bessong PO, Namrita L

(2005). Activity profiles of fourteen

selected medicinal plants from Rural

Venda communities in South Africa

against fifteen clinical bacterial species.

African J. Biotech., 4 (12): 1443–1451.

Sashikala GD, Kottai AM, Satheesh DK, Rekha

S, Indhumathy, Nandhini R. (2009).

Studies on the antibacterial and

antifungal activities of the ethanolic

extracts of Luffa cylindrica (Linn) fruit.

Int. J. Drug Dev. Res., 1(1):105–109.



Shaw WV (1984). Bacterial resistance to

Chloramphenicol, British Med. Bull.,

40 (1): 36–41.

Sofowora A (1984). Medical Plants and

Traditional Medicine in Africa. (eds.)

John Willey and Sons Ltd., Ibadan,

Nigeria.

Treadway S (1998). Exploring the Universe of

Ayurvedic botanicals to manage

bacterial infections, Clinical Nutrition

Insights, 6 (17): 1–5.

Watt E, Pretorius J C (2001). Purification and

identification of active antibacterial

components in Carpobotus edulis L., J.

Ethnopharma., 76: 87–91.

Source of Support: Qassim University,

Saudi Arabia

Conflict of Interest: None Declared




DIVERSITY AND AVAILABILITY STATUS OF ETHNO-MEDICINAL

PLANTS IN THE LOHBA RANGE OF KEDARNATH FOREST DIVISION

(KFD), GARHWAL HIMALAYA

Ballabha Radha1*, Singh Dinesh

2, Tiwari J K

3, Tiwari P

4

1, 2, 3, 4Department of Botany and Microbiology, HNB Garhwal University, Srinagar Garhwal-246 174,

Uttarakhand, India

*Corresponding Author: E-mail: [email protected]

Received: 20/02/2013; Revised: 25/02/2013; Accepted: 27/03/2013

ABSTRACT

The present study has been carried out in the Lohba range of the Kedarnath Forest Division,

Garhwal Himalaya to document the diversity, ethno-medicinal uses and availability status of

medicinal plants. The inhabitants of the region are dependent up to a large extent on wild resources

for their therapeutic needs. The region is rich in ethnomedicinal plant diversity. A total of 140

species belonging to 126 genera and 64 families were recorded from the study area. Out of the

documented species 69 were herbs, 37 shrubs, 23 trees and the rest 11 were climbers. Out of the

recorded plant species, 17 were abundant, 83 common and 40 uncommon to this area. Plant parts are

used to cure cold, cough, fever, stomach disorders, joints pain, eye diseases, healing of cuts and

wounds, toothache, etc. This study will be helpful in developing a comprehensive data base on the

medicinal plant resources to strengthen the health care system in the area and in conserving the

traditional knowledge for the prosperity of the remote village areas.

KEYWORDS: Ethnomedicinal plants, availability status, Kedarnath Forest Division, Garhwal

Himalaya.

Research article

Cite this article:

Ballabha R, Singh D, Tiwari J K, Tiwari P (2013), DIVERSITY AND AVAILABILITY STATUS OF

ETHNO-MEDICINAL PLANTS IN THE LOHBA RANGE OF KEDARNATH FOREST DIVISION

(KFD), GARHWAL HIMALAYA, Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 198–212



INTRODUCTION

Ethnobotanical studies typically focus on

recording the knowledge of traditional societies

in remote places (Hodges and Bennett, 2006).

In the remote areas traditional customs and

beliefs are still maintained and modern trends

are yet to reach, which provide interesting

scope of ethnobotanical studies (Tiwari et al.,

2010a). Indigenous people of different parts of

the world have a vast knowledge of, and

capacity for, developing innovative practices

and products from their environment.

Indigenous knowledge grows from close

interdependence between knowledge, land,

environment and other aspects of culture in

indigenous societies, and the oral transmission

of knowledge in accordance with well

understood cultural principles and rules

regarding secrecy and sacredness that govern

the management of knowledge (Tripathi et al.,

2000). According to WHO approximately 80%

of world population in developing countries

depends on traditional medicines for primary

healthcare (WHO, 2002) and in modern

medicine too, nearly 25% are based on plant

derived drugs (Tripathi, 2002).

Garhwal Himalaya occupies an important

place in Indian subcontinent and has its

peculiar topography, vegetation, people and

traditions. About 80% of the total population is

rural and the inhabitants are called the

Garhwalis or Paharis. They have their own

cultures, medicines, foods, etc. and are well

versed with valuable knowledge accumulated

through a long period of experience. Even now

they are dependent on the natural resources

from the forests for their sustenance and for the

treatment of various ailments (Tiwari et al.,

2010a).

The plants are still serving as remedies for

various ailments in crude form, as modern

medicine has not adequately armed the

therapeutic arsenal of the natives of remote

areas. The literature abounds in investigations

on folk medicines in different parts of Garhwal

Himalaya (Gaur et al., 1984, 1985, 1987;

Tiwari, 1986; Negi et al., 1993; Maikhuri et al.,

1998; Gaur, 1999; Badoni and Badoni, 2001;

Negi et al., 2002; Semwal et al., 2010; Tiwari

et al., 2010b) but little attention has been paid

on plants used in ethonomedicine from the

Lohba range of the Kedarnath Forest Division

(KFD). Documentation of such practices is

required in view of gradual disappearance of

this knowledge in new generations. Therefore,

an attempt has been made to record the

diversity and indigenous uses of plants in

ethnomedicine in the Lohba range of KFD.


Study Area

KFD is situated in the north-west part of the

Himalaya and well known for its rich

biodiversity. The inhabitants of the area largely

depend on plants for food, fodder, medicine,

timber, fuel-wood, dye, beverage, and various

religious and cultural needs. Geographically,

the area stretches between 29° 57' 33" to 30°

06' 05" N latitudes and 79°11' 33" to 79° 20'

33" E longitudes with the altitude ranging from

1268 m to 3067 m asl (Fig. 1). The total

geographical area of region is about 16387.40

ha which represents 26.76 % of the Division.

Western Ramganga is the main river of this

area, which originates from the lesser

Himalayan mountain range (Dhudhatauli) and

enters into Corbett National Park after flowing

100 km with its tributaries. Besides providing

perennial water source it provides habitat to

many plant and animal communities.

The mountainous tract of the whole region

is varying in altitude which contributes

variation in the climatic conditions to play an

important role in the distribution of the

vegetation in the area. Summer, rainy and

winter seasons are well marked due to

fluctuating precipitation, temperature, light,

wind, humidity and even day- length. The

maximum temperature was recorded during the

months of May and June (25–30°C), whereas

minimum in the months of December and

January (10–20°C). Rainfall, snowfall,

hailstorm, dew, frost, etc, are the main form of

precipitation and the average annual rainfall



0

10

20

30

40

50

60

Number of species

was 110 mm in the region. The average annual

relative humidity was 42% which decreases

with increase in temperature and altitude in

study area. The maximum humidity reaches

near to 80–92% during July and August,

whereas the minimum humidity (25–38%) was

recorded in the months of January and

December.

The vegetation of the Lohba Range is

characterized by sub montane and montane

zone types. The area is represented by Pine-

mixed forest (1200–1500 m), Oak forest

(1800–2000 m), Oak-mixed forest (1500–

2500 m) and Oak-Abies mixed forest (2700–

3114 m) however, some patches are occupied

by pine and scrub forest along with grassy

slopes.

Fig. 1 Map showing the study area.

Fig. 2 Plant parts being preferred for medicine by local inhabitants.



Methodology

Extensive field surveys were made in the

study area during the years 2009–2011 for the

survey of the vegetation and ethno-medicinal

uses. A structured questionnaire was used to

collect data on local name of plants, uses, parts

used and mode of application. Ethno-medicinal

information on plants was collected through

interviewing local communities. The

informants were medicine-men (Vaidhyas),

peasants, shepherds, priests and village

headmen. To determine the authenticity of

information collected during field work,

repeated verification of data from different

informants was done. Thus, only the specific

and reliable information, cross-checked with

informants has been incorporated in the present

study. Recorded plant species were identified

with the help of Garhwal University Herbarium

(GUH), Herbarium of the Botanical Survey of

India, Northern Circle, Dehradun (BSD) and

regional Floras (Duthie, 1906; Osmaston, 1927;

Rau, 1961; Naithani, 1984-85; Gaur, 1999).

The availability status of plants such as

abundant, common and uncommon was given

based on their occurrence in the study area.

RESULTS AND DISCUSSION

The study revealed 140 medicinal plant

species belonging to 64 families in the Lohba

range of the Kedarnath Forest Division,

Garhwal Himalaya. The availability status and

ethno-medicinal uses of the plant species are

presented in Table 1. The recorded species

diversity represents trees (23 species), shrubs

(37), herbs (69) and climbers (11).

The families, Asteraceae (15 species),

Lamiaceae (10), Rosaceae (10), Solanaceae (5),

Liliaceae (4), Orchidaceae (4), Ranunculaceae

(4), Rutaceae (4), Acanthaceae (3), Apiaceae

(3), Caesalpiniaceae (3), Cucurbitaceae (3),

Euphorbiaceae (3), Rubiaceae (3) and

Zingiberaceae (3) were major representations,

whereas Artemisia (3 species), Cassia (3),

Anaphalis (3), Asparagus (2), Berberis (2),

Ficus (2), Geranium (2), Mentha (2),

Polygonatum (2), Rumex (2), Salvia (2) and

Swertia (2) were the genera with more than one

species being used (Table 1). As per the plant

parts, root/rhizome of 54 species was used to

cure different ailments (Fig. 2), followed by

leaves (53 species) and whole plant (30

species). Plant parts are being used to cure

cold, cough, fever, stomach disorders, joints

pain, eye diseases, healing of cuts and wounds,

toothache etc.

Table 1. Diversity, availability status and ethno-medicinal uses of plant species in the Lohba

range of Kedarnath Forest Division (KFD), Garhwal Himalaya.

S.

No.

Botanical

Name

Local

Name

Family

Elevation (m

asl)

Life

Form1 Availability

Status2 Ethnomedicinal uses

1 Abies pindrow

Royle

Raisul Pinaceae 2500–3000 T + Bark extract is given in

cough and bronchitis.

2 Achyranthes

aspera L.

Lich

kura

Amaranthaceae 1300–2300 S ++ Root infusion is given in

fever. Leaf extract is given to

women to facilitate delivery.

Plant decoction is given in

dropsy and bronchitis.

3 Aconitum

balfourii Staf

Meetha

jari

Ranunculaceae 2900–3000 H + Root paste is used in snake-

bite and also applied over

chest in chest pain.

4 Acorus

calamus L.

Bauj Araceae 1600–2200 H ++ Extract of rhizome is given in

gastric troubles.



5 Adhatoda

zeylanica

Medikus

Basinga Acanthaceae 1200–1400 S + Root bark is given in fever.

Young twigs used for cough

and cold. Leaf juice with

honey is given in cough and

fever.

6 Aesculus

indica (Colebr.

ex Cambess)

Hook.

Pangar Hippocastanaceae 2000–2500 T + Root juice is given in bowl

complaints.

7 Ageratum

conyzoides L.

Gunrya Asteraceae 1300–2000 H ++ Root paste is applied on

sores, cuts and various skin

diseases.

8 Agrimonia

pilosa

Ledebour

Kuriya Rosaceae 1200–2500 H ++ Plant decoction is given in

cough and diarrhea.

9 Ajuga

bracteosa

Wallich ex

Benth.

Neelkanthi Lamiaceae 1200–1500 H ++ Leaf extract is given in fever.

Plant extract used as a tonic.

10 Anagallis

arvensis L.

Jonkmari Primulaceae 1200–2500 H ++ Externally applied in dropsy.

11 Anaphalis

adnata

Wallich ex

DC.

Kabash,

Bugla

Asteraceae 1300–2500 H ++ Leaf paste is applied on cuts

and wounds.

12 Anaphalis

busua (Buch.-

Ham. ex

D.Don) DC.

Bugla,

Buglya

Asteraceae 1600–2200 H ++ Leaf juice is applied on

bruises, wounds and cuts.

13 Angelica

glauca Edgev.

Choru Apiaceae 2900–3000 H + Root is given with tea in

cough and cold.

14 Arisaema

tortuosum

(Wallich)

Schott.

Bag-

mungari

Araceae 1600–2200 H ++ Paste of tuber is applied on

the burns and cuts.

15 Artemisia

capillaris

Thunb.

Jhirun Asteraceae 1300–2300 H ++ Stem and leaf juice is given

in fever and constipation.

Root power used in

stomachache

16 Artemisia

nilagrica

(Clarke) Pamp.

Kurnja Asteraceae 1600–2200 S ++ Plant juice is given in

intestinal worms and

externally applied on cuts

and wounds.

17 Artemisia

roxburghiana

Wallich ex

Besser

Kurnja Asteraceae 1600–2200 S + Plant extract is given in fever

and also used in skin

diseases.

18 Asclepias

curassavica L.

---- Asclepiadaceae 1300–1500 S ++ Latex applied on cuts and

wounds.

19 Asparagus

adscendens

Buch.-Ham. ex

Roxb.

Jhirni Liliaceae 1500–2200 S ++ Tuberous roots with honey

are used in dysentery.

20 Asparagus

racemosus

Willd

Jhirni Liliaceae 1300–1500 S ++ Roots extract is given in

fever.



21 Barleria

cristata L.

Kala-

bansa

Acanthaceae 1200–2300 S ++ Leaves are crushed, mixed

with seeds of black pepper

and given orally in dyspepsia.

22 Berberis

aristata DC.

Kirmor Berberidaceae 1700–3000 S +++ Stem and root juice is applies

in ophthalmic infections.

23 Berberis

asiatica Roxb.

ex DC.

Kirmor Berberidaceae 1200–2500 S ++ Stem bark and root juice used

in eye afflictions. Infusion of

root given in fever.

24 Bergenia

ciliata

(Haworth)

Sternberg

Silpari Saxifragaceae 2200–2400 H +++ Root paste is given in fever,

urinary and renal troubles.

25 Bidens pilosa

L.

Kumra Asteraceae 1300–2700 H ++ Vegetable of the plant useful

in skin ailments.

26 Boehmeria

regulosa

Wedd.

Genthi Urticaceae 1200–1400 T + Paste of bark is applied on

the bone fracture.

27 Boennighause

nia albiflora

(Hook.)

Reichb ex

Meisn.

Upaniy

a-Ghas

Rutaceae 1200–1700 H ++ Leaf paste is applied on cuts

and wounds.

28 Boerhavia

diffusa L.

Pundera Nyctaginaceae 1200–1700 H + Roots chewed as tonic. Leaf

extract used in eye diseases.

Plant infusion is given in

asthma and bronchitis.

29 Brugmansia

suaveolens

(Humb. &

Bonpl. ex

Willd.)

Berchtold & J.

S. Presl

Dhatura Solanaceae 1200–1400 S + Leaf, flower and seed paste is

applied on joints pain.

30 Buddleja

asiatica Lour.

---- Buddlejaceae 1300–1600 S + Leaf juice is applied on skin

eruption.

31 Callicarpa

macrophylla

Vahl.

Daiya Verbenaceae 1200–1500 S ++ Stem bark is used in skin

ailments.

32 Cannabis

sativa L.

Bhang Cannabaceae 1300–2100 S +++ Leaves boiled with butter are

taken in vomiting. Leaf juice

poured in ear in ear pain.

Leaves are used as an

intoxicating agent.

33 Cassia

occidentalis L.

Taror Caesalpiniaceae 1200–1400 S + Leaves used in skin disease.

Decoction of roots is given in

dropsy. Leaf and root paste

useful in piles and

ringworms.

34 Cassia tora L. Chakuda Caesalpiniaceae 1200–1400 S + Leaves and seeds are used in

skin diseases, cuts, wounds

and bone fracture.



35 Cassia fistula

L.

Amaltas Caesalpiniaceae 1200–1400 T + Fruits are given to women to

expel the placenta. Fruit pulp

is given in asthma, bronchitis

and skin diseases.

36 Cedrus

deodara

(Roxb. ex

D.Don) G.Don

Deodar Pinaceae 1400–2200 T ++ Paste of bark is externally

applied on piles and arthritis.

37 Centella

asiatica (L.)

Urban

Brahmi Apiaceae 1400–2400 H + Leaves paste is applied

externally in skin diseases.

38 Cinnamomum

tamala (Buch.-

Ham.) Nees &

Ebermaeir

Dalchini Lauraceae 1300–1700 T ++ Leaves paste is used in throat

irritation.

39 Cissampelos

pareira L.

Parha Menispermaceae 1200–3000 Cl ++ Root juice is given to

children in dyspepsia,

diarrhea, stomach ache and

colic.

40 Citrus

aurantifolia

(Christmann)

Swingle

Kagzi-

nimbu

Rutaceae 1300–2200 S ++ Leaf decoction inhaled in

headache, cold and fever.

41 Clematis

buchananiana

DC.

Laguliya Ranunculaceae 1200–3000 Cl ++ Decoction of leaves and roots

is applied in scabies.

42 Coccinia

grandis (L.)

Voigt

Kandaroi Cucurbitaceae 1200–2000 Cl ++ Root paste is applied on the

pelvic region in suppressed

urination.

43 Colebrookia

oppositifolia

J.E. Smith

Binda Lamiaceae 1300–2000 S ++ Leaf paste applied on cuts

and wounds.

44 Cotoneaster

microphyllus

Wallich ex

Lindley

Bani Rosaceae 1300–2800 S ++ Root paste is applied on cuts

and wounds.

45 Curcuma

longa L.

Haldi Zingiberaceae 1400–2100 H ++ Rhizome paste is applied on

cuts and wounds. It is also

used as antiseptics.

46 Cuscuta

europaea L.

Akas-

laguli

Cuscutaceae 1600–2300 Cl +++ Plant extract used in skin

diseases.

47 Cynodon

dactylon (L.)

Persoon

Doob Poaceae 1300–3000 H ++ Roots are taken in fever and

internal injury.

48 Cynoglossum

glochidiatom

Wallich ex

Benth.

Likh-

kura

Boraginaceae 1500–2000 H +++ Root paste is applied on

sores.

49 Datura metel

L.

Dhatura Solanaceae 1200–1400 H ++ Leaf and Seed paste is

applied on sores and used as

a massage in arthritis.

50 Desmodiun

elegans DC.

Chamliya Fabaceae 1500–2700 S ++ Root decoction is given in

renal disorder.



51 Dicliptera

roxburghiana

Nees

Kuthi Acanthaceae 1300–2600 S +++ Leaf paste applied on

wounds, check bleeding.

52 Dioscorea

bulbifera L.

Geithi Dioscoreaceae 1400–1900 Cl ++ Root and fruit paste is

applied on burns and

wounds.

53 Duchesnea

indica

(Andrews)

Focke

Kaphliya Rosaceae 1400–2200 H ++ Leaf juice is given in

diarrhea.

54 Echinops

cornigerus

DC.

Kandara Asteraceae 1200–2000 H ++ Root juice is taken in urinary

trouble and fever.

55 Eupatorium

adenophorum

Sprengel

Guyajhar Asteraceae 1300–3000 H ++ Crushed leaves applied on

cuts and wounds.

56 Euphorbia

royleana

Boissier

Sulla Euphorbiaceae 1800–2000 S ++ Latex is used as an antiseptic

on cuts and wounds.

57 Ficus palmata

Forsk.

Bedu Moraceae 1300–2000 T ++ Fruits are used in digestive

disorders. Latex is applied on

the pimples.

58 Ficus religiosa

L.

Peepal Moraceae 1300–1500 T ++ Root bark, young shoots and

fruit decoction is given in

sexual weakness in men.

Infusion of bark is given in

constipation.

59 Foeniculum

vulgare (L.)

Miller

Saup Apiaceae 1700–2000 H + Seed paste is applied on

mouth during the teething of

child.

60 Fumaria

indica

(Haussknecht)

Pugsley

Kherua Fumariaceae 1300–2200 H ++ Plant decoction is given in

fever and suppressed

urination.

61 Geranium

nepalense

Sweet

---- Geraniaceae 1600–2200 H + Roots are used as an

antiseptic. Root extract is

given in lever troubles and

fever.

62 Geranium

wallichianum

D. Don ex

Sweet

Ratanjot Geraniaceae 2200–3000 H + Root paste is applied on

wounds and cuts.

63 Girardinia

diversifolia

(Link.) Friis.

Bhaiska

nali

Urticaceae 1500–3000 H ++ Leaf juice is given in

gonorrhea.

64 Gnaphalium

hypoleucum

DC.

Bukhil Asteraceae 1300–2500 H ++ Juice of stems and leaves is

given in bruises and cuts.

65 Hedychium

spicatum

Buch.-Ham. ex

J. E. Smith

Ban-

haldi

Zingiberaceae 2000–2300 H ++ Rhizome powder is used in

asthma.

66 Holoptelea

integrifolia

(Roxb.)

Planchon

Papri Ulmaceae 1700–1900 T ++ Bark decoction is used in

rheumatic pain.



67 Hypericum

oblongifolium

Choisy

Phioli Hypericaceae 1500–2000 S ++ Plant juice is given in

diarrhea and intestinal

worms.

68 Inula

cuspidata

(DC.) C.B.

Clarke

---- Asteraceae 1300–2500 S ++ Root juice is given in urinary

trouble.

69 Ipomoea nil

(L.) Roth

Sulkairi Convolvulaceae 1300–2300 Cl ++ Decoction of seed is given in

fever and constipation. Seed

paste is applied in urticaria.

70 Jasminum

humile L.

---- Oleaceae 1200–2500 S ++ Root juice is given to

children in stomach troubles.

71 Juglans regia

L.

Akhor Juglandaceae 1300–2300 T ++ Leaves used as fungicide and

insecticide and bark used in

bone fractures.

72 Leucas lanata

Benth.

Niras Lamiaceae 1300–2600 H ++ Plant extract is given in

cough.

73 Lyonia

ovalifolia

(Wallich)

Drude

Anyar Ericaceae 1400–3000 T +++ Leaves juice is applied in

eczema.

74 Mallotus

philippensis

(Lam.) Muell.-

Arg.

Ruina Euphorbiaceae 1200–1300 T ++ Bark juice is given to

children in diarrhea and

dysentery. Paste of fruit

powder is applied externally

on cuts, wounds. Root and

seed paste is applied on sores

and skin eruptions.

75 Malva

verticillata L.

---- Malvaceae 1400–2000 H + Root paste is given in

whooping cough.

76 Martynia

annua L.

Bichu Martyniaceae 1200–1800 S + Fruit powder is given in cold

and cough.

77 Melia

azedarach L.

---- Meliaceae 1200–1400 T + Decoction of bark used in

gonorrhea, bark paste is

applied on skin eruptions.

Infusion of heart wood is

given in asthma. Leaves,

fruits and seeds are useful in

skin diseases

78 Mentha

arvensis L.

Paudina Lamiaceae 1200–2300 H ++ Plant extract used in

vomiting and indigestion.

79 Mentha

piperita L.

Pepermint Lamiaceae 1200–2300 H ++ Plant extract used in

indigestion.

80 Micromeria

biflora Buch.-

Ham. ex

D.Don

Ban

ajwain

Lamiaceae 1300–3200 H +++ Crushed leaves inhaled in

cold and sinusitis. Leaf

extract with milk is given in

gastroenteritis.

81 Mukia

maderaspatan

a (L.) M.

Roemer

Guliya-

Kakri

Cucurbitaceae 1300–2100 Cl ++ Fruit paste is used in urinary

disorder and vomiting.

82 Murraya

koenigii (L.)

Sprengel

Karipatta Rutaceae 1200–1400 S + Bark, leaves and roots are

used as insecticide.



83 Nasturtium

officinale

R.Br.

---- Brassicaceae 1200–2500 H ++ Plant juice is given in

diarrhea.

84 Nicotiana

plumbaginifoli

a Viviani

Bantamb

aku

Solanaceae 1200–1400 H + Leaf juice is applied on sores.

85 Nicotiana

rustica L.

Tambaku Solanaceae 2000–3000 H ++ Leaves juice is applied on

cuts and wounds.

86 Oberonia

falconeri

Hook.f.

---- Orchidaceae 2000–2300 H + Plant extract is given in

diarrhea and bronchitis.

87 Ocimum

tenuiflorum L.

Tulsi Lamiaceae 1200–1600 H + Plant used in fever, cold,

cough, colitis, urinary

troubles and vomiting.

88 Papaver

somniferum L.

Post Papaveraceae 2900–3000 H + Milky juice obtained from

immature capsules is given to

children in fever, dysentery

and cholera.

89 Parnassia

nubicola

Wallich ex

Royle

Phutkya Saxifragaceae 2200–2400 H ++ Plant extract is used to

stimulate vomiting.

90 Peperomia

tetraphylla

(Forster f.)

Hook. & Arn.

Tirpirya Piperaceae 1500–3000 H +++ Leaf paste is applied on

wounds and burns.

91 Perilla

frutescens (L.)

Britton

Bhangje

era

Lamiaceae 1500–2100 H + Plant extract or power of

dried plant parts used for

cold, cough, bronchitis and

uterine ailments; leaf paste

applied on arthritis.

92 Phoebe

lanceolata

(Nees) Nees

Kaula Lauraceae 1400–1700 T ++ Fruits paste is used against

wounds and sores.

93 Pholidota

articulata

Lindley

Jewanti Orchidaceae 2000–2300 H + Plant extract is used as tonic.

94 Phyallanthus

emblica L.

Aola Euphorbiaceae 1200–1400 T + Fruits are used in digestive

disorders and fruit juice

useful in leucorrhoea. Fruit

powder is given in fever

95 Polygonatum

multiflorum

(L.) Allioni

---- Liliaceae 1200–1400 H + Decoction of rhizome is

given in urinal disorders

96 Polygonatum

verticillatum

(L.) Allioni

---- Liliaceae 1400–2300 H + Root paste is applied on

wounds.

97 Populus ciliata

Wallich ex

Royle

Pupular Salicaceae 1300–2600 T ++ Leaf juice is used as blood

purifier and stimulant.

98 Potentilla

fulgens

Wallich ex

Hook.

Bajrdanti Rosaceae 2300–3000 H ++ Plant juice is applied on

mouth in tooth ache.

99 Prinsepia

utilis Royle

Bhainkal Rosaceae 1300–3000 S +++ Seed oil is applied in joint

pain and arthritis.



100 Prunus

cornuta

(Wallich ex

Royle) Steudel

Jamnoi Rosaceae 2400–3000 T ++ Bark decoction is given in

diarrhea.

101 Punica

granatum L.

Darim Punicaceae 1300–1700 T + Fruit juice is used in cough

and diarrhea.

102 Pyrus pashia

Buch.-Ham. ex

D.Don

Melu Rosaceae 1200–2900 T +++ Ripened fruits are used in

digestive disorders. The fruit

is crushed with teeth and

juice is forced into the eyes

of cattle in cataract and

injuries.

103 Ranunculus

arvensis L.

Chambul Ranunculaceae 1200–3000 H +++ Leaves paste is applied in

skin diseases.

104 Raphanus

sativus L.

Muli Brassicaceae 1200–2500 H ++ Cooked leave and leave juice

is given in jaundice.

105 Reinwardtia

indica

Dumortier

Phuenli Linaceae 1200–2200 S ++ Leaf juice is applied on cuts

and wounds.

106 Rhododendron

arboreum

Smith

Burans Ericaceae 1200–3000 T +++ Flower juice is given in hart

troubles and diseases. Young

shoot poisonous to cattle.

107 Rosa brunonii

Lindley

Kunja Rosaceae 1400–2000 S ++ Leaf juice used in cuts,

wounds. Dried flower

powder used in diarrhea

108 Rosa sericea

Lindley

Dhurku

nja

Rosaceae 2700–3000 S + Flower juice is applied

externally in eye diseases.

109 Rubia manjith

Roxb. ex

Fleming

Majeithi Rubiaceae 1300–3000 Cl +++ Stem used as an antidote to

snake bite; roots are used as a

tonic; flower extract used in

bacillary dysentery.

110 Rubus niveus

Thunb.

Kali

Hinsar

Rosaceae 1200–2200 S ++ Root decoction is given in

diarrhea. Juice of young

shoots and roots is given in

stomachache.

111 Rumex

hastatus

D.Don

Almor Polygonaceae 1300–2300 H +++ Leaf extract applied on cuts

and wounds to check

bleeding.

112 Rumex

nepalensis

Sprengel

Khoya Polygonaceae 1300–3000 H ++ Leaf juice is given in

digestive disorders.

113 Salvia lanata

Roxb.

Pakhuliya Lamiaceae 1300–3000 H ++ Flower paste is given in cold

and cough.

114 Salvia

nubicola

Wallich ex

Sweet

Ganiya Lamiaceae 2500–2700 H ++ Root paste is given in cold

and cough.

115 Sarcococca

saligna

(D.Don)

Muell.-Arg.

Peruli Buxaceae 1300–3000 S ++ Leaf paste is applied on joint

pain.

116 Satyrium

nepalens

D.Don

---- Orchidaceae 1600–2100 H + Plant extract is used in

diarrhea.



117 Sida

rhombifolia L.

Bhiunli Malvaceae 1200–2100 S + Leaves and root bark used in

gonorrhea.

118 Smilax aspera

L.

Kukrdar Smilacaceae 1400–2400 Cl +++ Root paste mixed with

mustard oil is applied in

arthritis and joint pain.

119 Solanum

nigrum L.

Makoi Solanaceae 1200–2300 H ++ Plant extract is given in liver

trouble, piles and dysentery.

Fruits are useful in diarrhea.

120 Solidago

virgaurea L.

---- Asteraceae 1800–3000 H ++ Plant juice is applied on cuts

and in scabies.

121 Sonchus asper

(L.) Hill

Dudhi Asteraceae 1300–2200 H ++ Plant juice is used as blood

purifier.

122 Spermadictyon

sauveolens

Roxb.

Padera Rubiaceae 1400–2600 S +++ Leaf juice is applied on cuts,

wounds and sores.

123 Stellaria

media (L.)

Villars

Badyalu Caryophyllaceae 1200–1700 H ++ Plant paste is externally

applied on burns, wounds

and boils.

124 Stephania

glabra (Roxb.)

Miers

Gindaru Menispermaceae 2000–2500 Cl + Root juice is given in fever

and dysentery.

125 Swertia

angustifolia

Buch.-Ham. ex

D.Don

Chirota Gentianaceae 2100–3000 H ++ Seed powder is given in

cough and asthma.

126 Swertia

chirayita

(Roxb. ex

Fleming)

Karsten

Chirota Gentianaceae 2100–3000 H + Seed powder is given in

cough and asthma.

127 Symplocos

paniculata

(Thunb.) Miq.

Lodh Symplocaceae 1500–2500 T +++ Decoction of bark is given in

diarrhea.

128 Tanacetum

dolichophyllu

m (Kitamura)

Kitamura

Dhoop Asteraceae 2800–3000 H ++ Plant juice is given to

children to expel intestinal

worms.

129 Taraxacum

officinale

Weber

Kanphu

liya

Asteraceae 1300–2000 H ++ Latex is applied over corns

and warts.

130 Taxus baccata

L.

Thuner Taxaceae 2500–3000 T + Bark paste is applied in

headache and bone fracture.

131 Thalictrum

foliolosum DC.

Mamira Ranunculaceae 1200–2200 H ++ Root powder mixed with

honey is given in fever,

dyspepsia and piles.

132 Trichosanthus

tricuspidata

Lour.

Indraiyan Cucurbitaceae 1400–2300 Cl ++ Root juice is given as emetic.

133 Valeriana

wallichii DC.

Sumaya Valerianaceae 1400–2600 H ++ Root paste is applied in

muscular pain.

134 Vanda cristata

Lindley

---- Orchidaceae 1600–2100 H + Plant paste is applied in bone

fractures.



135 Verbascum

thapsus L.

---- Scrophulariaceae 1200–2500 H ++ Plant extract used in

bronchitis and asthma. Seeds

are used as sedative.

136 Viburnum

grandiflorum

Wallich ex

DC.

Ghenu Caprifoliaceae 2800–3000 T ++ Bark decoction is used in

hepatic trouble.

137 Viola pilosa

Blume.

Kaura Violaceae 1200–2100 H ++ Decoction of plant useful in

fever and bronchitis. Root is

used as an emetic. Leaf juice

is applied on cuts and

wounds.

138 Vitex negundo

L.

Siwain Verbenaceae 1300–1800 S + Leaves, roots and fruits

pastes are applied in arthritis.

139 Zanthoxylum

armatum DC.

Timru Rutaceae 1400–1800 S ++ Leaves and fruits chewed for

mouth wash and tooth care.

Seed paste is applied on teeth

in toothache.

140 Zingiber

officinale

Roscoe

Adu Zingiberaceae 1300–1900 H ++ Used in cold, cough and

stomach troubles.

Abbreviations used: 1. H = herb, S = shrub, T = tree, Cl = climber, 2. +++ = Abundant, ++ =

Common, + = Uncommon

The present study indicates that the area

harbors a high diversity of medicinal plants.

Out of 140 plant species, 17 were abundant, 83

common and 40 uncommon to this area.

Species like Abies pindrow, Aconitum balfourii,

Adhatoda zeylanica, Aesculus indica, Angelica

glauca, Artemisia roxburghiana, Boehmeria

regulosa, Boerhavia diffusa, Brugmansia

suaveolens, Buddleja asiatica, Centella

asiatica, Foeniculum vulgare, Geranium

nepalense, Geranium wallichiana, Malva

verticillata, Martynia annua, Melia azedarach,

Murraya koenigii, Nicotiana plumbaginifolia,

Oberonia falconeri, Ocimum tenuiflorum,

Perilla frutescens, Pholidota articulata,

Polygonatum multiflorum, Polygonatum

verticillatum, Punica granatum, Rosa sericea,

Satyrium nepalens, Sida rhombifolia,

Stephania glabra, Swertia chirayita, Taxus

baccata, Vanda cristata, Vitex negundo etc. are

uncommon to this area and being threatened

due to unplanned exploitation. The inhabitants

revealed abundance of many of these species in

the past, which has got restricted now to certain

patches. If immediate steps for their sustainable

utilization and conservation are not taken, these

species may reach to the status of threatened in

the area.

CONCLUSION

It is evident from the investigation that the

local people have great familiarity with the

plants of their ambient environment which has

immense importance in advancement of

modern sustainable technology. The occurrence

of a number of economically important

medicinal plant species demands the

conservation of these species as the day-to-day

need of forest resources particularly for

medicine has increased the pressure in the area

and may lead to reduction of these species.

Therefore, there is a need to develop adequate

strategy and action plan for the conservation

and management of habitats and species.

ACKNOWLEDGEMENTS

The authors are grateful to the inhabitants

of the Lohba range of the Kedarnath Forest

Division (KFD) for providing the information

about the ethno-medicinal uses of the plant

resources.



REFERENCES

Badoni AK, Badoni K (2001). Ethnobotanical

heritage. In: Kandari OP, Gusain OP

(eds) Garhwal Himalaya: Nature,

Culture and Society Transmedia, Media

House, Srinagar, Garhwal, pp 127–147.

Duthie JF (1906). Catalogue of plants of

Kumaon and of the adjacent portions of

Garhwal and Tibet based on the

collections made by Strachey and

Winterbottom during the years 1846–

1849. London. Reprint 1994, Bishan

Singh Mahendra Pal Singh, Dehradun.

Gaur RD (1999). Flora of the District Garhwal,

North West Himalaya (With

Ethnobotanical Notes). Trasmedia:

Srinagar (Garhwal), Uttarakhand, India.

Gaur RD, Semwal JK, Tiwari JK (1984). A

survey of high altitude medicinal plants

of Garhwal Himalaya. Bull. Medic.

Ethnobot. Res., 3: 102–116.

Gaur RD, Tiwari JK (1987). Some little known

medicinal plants of Garhwal Himalaya:

An ethnobotanical study. In:

Leeuwenberg AGM (ed). Medicinal and

Poisonous Plants of the Tropics Pudoc,

Wageningen, Netherlands, pp 139–142.

Gaur RD, Tiwari JK, Negi KS (1985). Plants

used for magico-religious practices by

tribals of Garhwal Himalayas.

Proceedings of 2nd Annual Workshop

on MAB Projects. Dept. of

Environment, New Delhi, pp 88–95.

Hodges S, Bennett BC (2006). The

ethnobotany of Pluchea carolinesis

(Jacq.) G. Don (Asteraceae) in the

Botanicas of Miami, Florida. Econ.

Bot., 60(1): 5–84.

Maikhuri RK, Nauityal S, Rao KS, Saxena KG

(1998). Role of medicinal plants in the

traditional health care system: a case

study from Nanda Devi Biosphere

Reserve. Current Science, 75(2): 152–

157.

Naithani BD (1984-85). Flora of Chamoli. 2

Vols. Botanical Survey of India,

Howrah.

Negi CS, Nautiyal S, Dasila L, Rao KS,

Maihkuri RK (2002). Ethnomedicinal

Plant Uses in a Small Tribal

Community in a Part of Central

Himalaya, India. J. Hum. Ecol., 14(1):

23–31.

Negi KS, Tiwari JK, Gaur RD, Pant KC

(1993). Notes on ethnobotany of five

districts of Garhwal Himalaya, U.P.,

India. Ethnobot., 5: 73–81.

Osmaston AE (1927). A Forest Flora for

Kumaun. Government Press, Allahabad.

Reprint 1990, Bishan Singh Mahendra

Pal Singh, Dehradun.

Rau M A (1961). Flowering plants and ferns of

north Garhwal, Uttar Pradesh, India.

Bull. Bot. Surv. India, 3: 215–251.

Semwal DP, Kala CP, Bhatt AB (2010).

Medicinal Plants and Traditional Health

Care Knowledge of Vaidyas, Palsi and

Others: a Case Study from Kedarnath

Valley of Uttarakhand, India. Medicinal

Plants, 2(1): 51–57.

Tiwari JK (1986). Medicinal Plants of Garhwal

Himalaya: An Ethnobotanical Survey.

Unpublished D. Phil. Thesis University

of Garhwal, Srinagar Garhwal (U.P.),

India.



Tiwari JK, Radha Ballabha, Tiwari P (2010a).

Ethnopaediatrics in Garhwal Himalaya,

Uttarakhand, India (Psychomedicine

and Medicine). New York Science

Journal, 3(4): 123–126.

Tiwari JK, Radha Ballabha, Tiwari P (2010b).

Diversity and Present Status of

Medicinal Plants in and around Srinagar

Hydroelectric Power Project in Garhwal

Himalaya, India: Needs for

Conservation. Researcher, 2(2): 50–60.

Tripathi G (2002). Indigenous Knowledge and

Traditional Practices of Some

Himalayan Medicinal Plants. In:

Samant SS, Dhar U, Palni LMS (eds)

Himalayan Medicinal Plants Potential

and Prospects, Gyanodaya Prakashan,

Nainital, pp 151–156.

Tripathi S, Varma S, Goldey P (2000). Using

plants for health: indigenous knowledge

in health care in a tribal region of Bihar,

India. Int. J. Sust. Develop. & World

Ecol., 7: 321–332.

WHO (2002). WHO Traditional Medicine

Strategy 2002–2005. World Health

Organization, Geneva.

Source of Support: Nil Conflict of Interest: None Declared




ANTICATARACT ACTIVITY OF ERVATAMIA CORONARIA LEAF

EXTRACT ON CHEMICALLY INDUCED CATARACTOGENESIS IN RATS

Rathnakumar K

1*, Jaikumar S

2, Duraisami R

3, Sengottuvelu S

4

1, 2

Department of Ophthalmology, Sri Lakshminarayana Institute of Medical Sciences, Pondicherry, India 3, 4

Department of Pharmacology, Nandha College of Pharmacy and Research Institute, Erode- 638052

*Corresponding Author: Email: [email protected]


ABSTRACT

The anticataract activity of the Ervatamia coronaria leaf extract was evaluated against

naphthalene induced cataract in Wistar albino rats. Thirty rats in five groups were used for the study.

Ervatamia coronaria leaf extract at dose levels (200 and 400 mg/kg) respectively and Vitamin E

(50 mg/kg) were used as standard drugs while liquid paraffin was used for control. The test drugs

were administered simultaneously with naphthalenefor 25 days. Naphthalene (0.5 g/kg for first 3

days and 1 g/kg thereafter for a period of 25 days) was used to induce cataract. The percentage of

cataract incidence and opacity index were examined using ophthalmoscope. Naphthalene produced a

marked mature cataract and an increase in the opacity index at various stages. The extract treated

animals showed decrease in the onset and maturation of cataract against naphthalene challenge. From

the results it was concluded that Ervatamia coronaria leaf extract protected the cataract maturation

induced by naphthalene and it exhibited anticataract activity.

KEYWORDS: Ervatamia coronaria, Cataract, Naphthalene

Research article

Cite this article: Rathnakumar K, Jaikumar S, Duraisami R and Sengottuvelu S (2013), ANTICATARACT ACTIVITY

OF ERVATAMIA CORONARIA LEAF EXTRACT ON CHEMICALLY INDUCED

CATARACTOGENESIS IN RATS, Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 213–218




INTRODUCTION

Cataract is opacity of the lens that

interferes with vision, and is the most frequent

cause of visual impairment worldwide,

especially for the elderly, because the incidence

of cataracts increases with increasing age. It is

the leading cause of blindness and contributes

to 50% of blindness worldwide (Haque and

Gilani, 2005). The only present remedy for

cataract is surgery. The opacity of lens occurs

as a result of oxidation which is augmented by

the free radical generation. The intensity of

opacification of lens can be reduced by the

antioxidants which scavenge the generation of

free radicals. Previous studies confirmed that

diet rich in vitamins, carotenoids and

flavonoids may reduce the cataract intensity

(Bunce et al., 1990). Normal levels of the anti-

oxidant’s defense mechanism are not sufficient

for the eradication of free radical induced

injury. Therefore, the administration of

antioxidants from a natural origin has a

promising role to play. Several antioxidants of

plant materials have been experimentally

proven and widely used as more effective

agents against oxidative stress.

Ervatamia coronaria Stapf Local name :

Adukkunandiyavattai (In Thamizh Language)

(Synonym: Tabernaemontana divaricata)

belongs to the family Apocynaceae, is a

glabrous, evergreen tree indigenous to India

and is cultivated in gardens for its ornamental

and fragrant flowers.

This species has been extensively

investigated and a number of chemical

constituents such as alkaloids (Pawelka and

Stoeckigt., 1983), triterpenoids, steroids

(Sharma and Cordell., 1988), flavonoids

(Daniel and Sabnis., 1978) and phenolic acids

(Henriques et al., 1996) were isolated from

leaves, roots and stems of the plant.

In Indian traditional system of medicine the

plant material is widely used as a purgative,

tonic to the brain, spleen and the liver (Kirtikar

and Basu., 1975). Also used in the treatment of

cancer, wounds and inflammations (Kirtikar

and Basu., 1975). The plant extract was also

found to possess analgesic, antipyretic,

vasodilator and CNS depressant effects

(Taesotikul., 1989), antispasmodic, hypotensive

activity (Dhar et al., 1968), anti-inflammatory

(Henriques et al., 1996), uterine stimulant

effect (Da Sil Va et al., 1984), cytotoxic

(Yamamoto et al., 1997) and anti oxidant

activity (Malaya Gupta et al., 2004).

Traditionally the plant is also used in

ophthalmic disorders. Hence the present study

was aimed to evaluate the anti-cataract activity

of the leaf extract of E. coronaria.


Drugs and chemicals

Naphthalene and vitamin E were obtained

from SD fine chemicals, Mumbai, India. All

other drugs and chemicals used in the study

were of analytical grade.

Plant material

Leaves of Ervatamia coronaria were

collected from outskirts of of Erode,

Tamilnadu. Authentication has been done by

Prof. V. S. Kumar, Scientists (F) and Head of

the Office, Tamilnadu Agriculture University,

Coimbatore (Tamilnadu).The voucher

specimen (No.: BSI/ SRC/ 5/ 23/ 12-13/ Tech.

816) has been deposited in the herbarium for

future references.

Preparation of extract

The leaves were washed with fresh water to

remove adhering dirt and foreign particles. The

leaves were shade dried, crushed and grinded to

get coarse powder. The coarse powder was then

placed with 90% ethanolic solution in a round

bottomed flask. 500 g of the coarse powder of

the leaves of Ervatamia coronaria in 1.0 liter

of 90% ethanolic solution were macerated for 7

days. The mensturm was collected,

concentrated by vacuum distillation and then

air dried in an evaporating dish till constant

weight was obtained.



Animals

Wistar albino rats of either sex weighing

150–200 g were used for this study. The

animals were placed randomly and allocated to

treatment groups in polypropylene cages with

paddy husk as bedding. Animals were housed

at a temperature of 24 ± 2oC and relative

humidity of 30–70%. A 12:12 light: day cycle

was followed. All the animals were allowed to

free access to water and fed with standard

commercial pelleted chaw (M/s.Hindustan

Lever Ltd., Mumbai). All the experimental

procedures and protocols used in this study

were reviewed by (IAEC) Institutional Animal

Ethics Committee (932/a/06/CPCSEA) of Sri

Lakshminarayana Institute of Medical

Sciences, Pondicherry and were in accordance

with the guidelines of the IAEC.

Experimental protocol

Experimental model of cataractogenesis

was induced in rats by feeding naphthalene

(Umamaheswari et al., 2011) at a dose of

0.5 g/kg orally for the first 3 days followed by

1 g/kg thereafter for a period of 25 days.

Animals were divided into 5 groups consisting

of six animals each. Group I received liquid

paraffin (5 ml/kg b.w. orally) and served as the

solvent control. Group II received naphthalene

(0.5 g/kg b.w., orally for first 3 days and 1 g/kg

thereafter for 25 days) and served as the

cataract control. Group III received the

standard drug vitamin E at a dose of 50 mg/kg

b.w., orally along with naphthalene and served

as the positive control. Groups IV and V

received the E. coronaria leaf extract orally at a

dose of 200 and 400 mg/kg b.w. respectively

simultaneously with naphthalene. All the test

drugs were administered for a period of 28

days.

Examination of the eyes

The eyes of the rats were examined using

an ophthalmoscope for morphological changes

in the lens. Examination was performed after

dilatation of the pupil with 1% tropicamide

solution. Cataract formation was scored

according to different stages. Stage 1: Clear

normal lens, Stage 2: Peripheral vesicles, Stage

3: Peripheral vesicles with cortical opacities,

Stage 4: Diffuse central opacities, Stage 5:

Opacity involving the entire lens (Mature

cataract). Cataract formation was considered

complete (stage 5) when the red fundus reflex

was no longer visible through any part of the

lens and the lens appeared dull white to the

naked eye. Percentage incidence of cataract

was calculated using the following formula

(Vats et al., 2004).

% Incidence = No of animals in each stage × 100

Total no. of animals

Opacity index was calculated using the following formula (Fukushi et al., 1980),

Opacity Index = No. of eyes in each stage × Stage of the eye

Total no. of eyes

RESULTS

Anticataract activity of Ervatamia

coronaria leaf extract on naphthalene induced

cataractogenesis in rats was studied and the

percentage incidences of cataract and opacity

index were observed. Effect of E. coronaria

leaf extract on % incidence of cataract on 28th

day in naphthalene induced cataract rats are

shown in table. 1.

Ophthalmic examinations of the normal

control lenses in the eyes l were normal

throughout the duration of experimental period.

The animals treated with naphthalene showed

varying degree of cataractogenic changes as

indicated by about 72.54% in stage 4 and

45.66% in stage 5 on the 28th

day of drug

treatment. At the end of drug treatment, the

animals treated with the extract at a dose of 200

mg/kg b.w. showed 50% of animals in stage 2,



18.22% in stage 3 and 45.16% in stage 4

cataract, whereas the group treated with the

extract at a dose of 400 mg/kg b.w. showed

50% of animals in stage 2, 25.11% in stage 3

and 18.14% in stage 4 cataract. Vitamin E the

standard drug showed 18.19% of animals in

stage 1, 60.23% in stage 2 and 35.32% in stage

3. None of the animals treated with the E.

coronaria leaf extract and the standard drug

vitamin E showed stage 5 mature cataract at the

end of the experiment.

Table 1. Effect of Ervatamia coronaria leaf extract on % incidence of cataract on 28th

day in

naphthalene induced cataract rats

Drug Treatment

% Incidence of Cataract

Stage 1 Stage 2 Stage 3 Stage 4 Stage 5

Normal Control

Liquid Paraffin

(10 ml/kg)

100

0

0

0

0

Cataract Control

Naphthalene

(1 g/kg)

0

0

0

72.54

45.66

Vitamin E

(50 mg/kg)

18.19

60.23

35.32

0

0

Ervatamia

Coronaria Extract

(200 mg/kg)

0

50.00

18.12

45.16

0

Ervatamia

Coronaria Extract

(400 mg/kg)

0

50.00

25.11

18.14

0

Table 2. Effect of Ervatamia coronaria leaf extract on opacity index in naphthalene induced

cataract rats

Drug Treatment

Opacity Index

4th

day 7th

day 14th

day 21st day 28

th day

Normal Control

Liquid Paraffin

(10 ml/kg)

0

0

0

0

0

Cataract Control

Naphthalene

(1 g/kg)

0.45

0.62

2.47

3.55

4.98

Vitamin E

(50 mg/kg)

0

0

0.18

0.72

1.55

Ervatamia

Coronaria Extract

(200 mg/kg)

0

0

2.24

2.55

1.72

Ervatamia

Coronaria Extract

(400 mg/kg)

0

0

1.54

1.22

1.56



Effect of E. coronaria leaf extract on

opacity index in naphthalene induced cataract

rats are shown in table 2. Treatment with

naphthalene showed an increase in the opacity

index from 0.45 on the 4th

day, 0.62 on the 7th

day, 2.47 on the 14th

day, and 3.55 on the 21st

day followed by complete opacification

(opacity index 4.98) on the 28th

day. The

groups treated with the E. coronaria leaf

extracts at a dose of 200 and 400 mg/kg b.w.

showed a decrease in opacity index (1.72 and

1.56 respectively) when compared to

naphthalene control. There was a marked

reduction in opacity index (1.55) of the vitamin

E treated group when compared to the

naphthalene control.

DISCUSSION

Cataract is a visual impairment that occurs

due to the opacification of crystalline lens. It

affects around 17 billion people worldwide,

although incidence of cataracts is increasing

day by day among the elderly persons. Still

today except surgery no other effective

treatments have been successfully developed

for cataract (Piyush Patel et al., 2012). From a

public health perspective, it is important to

identify the risk factors that affect the

development and progression of cataract.

Ervatamia coronaria possess significant amount

of flavonoids and a potent antioxidant has been

studied against naphthalene induced cataract in

rats. The cataract induced by naphthalene

treatment in rats were confirmed by varying

degree of cataractogenic changes and an

increase in opacity index with complete

opacification at the end of the 4th

week. The

free radical involvements in the generation of

cataract have been evidenced (Kothadia et al.,

2011). The in vitro antioxidant and free radical

scavenging activities of E. coronaria has

already been reported (Malaya Gupta et al.,

2004). The anti cataract activity possessed by

E. coronaria leaf extract may be due to the

presence of flavonoids the well known free

radical scavengers.

CONCLUSION

It can be concluded that the Ervatamia

coronaria leaf extract possess

anticatractogenesis activity against naphthalene

induced cataract in rats. According to the

results obtained from the study it may be

inferred that E. coronaria reversed the cataract

induced by naphthalene in rats. To the best of

our knowledge this is the first report about in

vivo activity of E. coronaria in a specific eye

disorder and seems to raise some concern about

the traditional indication of this species. A

certainly further study needs to be carried out

in order to prove its actual mechanism of

action.

REFERENCES

Bunce GE, Kinoshita J, Horwitz J (1990).

Nutritional factors in cataract. Annu

Rev Nutr. 10: 233–254.

Da Sil Va NH, De Melo AA, Casado MMCV,

Hendriques AT, Wandscheer DC, Da

Sil Va OE (1984). Indole alkaloids with

potential endocrine activity. Rev. Inst.

Antibiot. Univ. Fed. Pernambuco.

Recife. 22: 27–32.

Daniel M, Sabnis SD (1978).

Chemotaxonomical studies on

Apocynaceae. Indian J Exp Biol. 16:

512–513.

Dhar ML, Dhar MM, Dhawan BN, Mehrotra

BN, Ray C (1968). Screening of Indian

plants for biological activity: Part - I.

Ind J Exp Biol. 6: 232–247.



Fukushi S, Merola LO, Kinoshita JH (1980).

Altering the course of cataracts in

diabetic rats. Invest Ophthamol Vis Sci.

19: 313–315.

Haque SE, Gilani KM (2005). Effect of

Ambroxol, Spirulina and Vitamin E in

naphthalene induced cataract in female

rats. Ind J Physiol Pharmacol. 49: 57–

64.

Henriques AT, Melo AA, Moreno PRH, Ene

LL, Henriques JAP, Schapoval EES

(1996). Ervatamia coronaria: Chemical

constituents and some pharmacological

activities. J Ethnopharmacol. 50: 19–

25.

Hsu YT (1967). Study on the Chinese drugs

used as cancer remedy. J Southeast

Asian Res. 3: 63–66.

Kirtikar KR, Basu BD (1975). Indian

Medicinal Plants, V 2, Bishen

Mahendra Pal Singh Dehradun, India.

842–844.

Kothadia AD, shenoy AM, Shabaraya AR,

Rajan MS, Viradia UM, Patel NH,

(2011). Evaluation of cataract

prevention action of phycocyanin. Int J

Pharm Sci and Drug Res. 3: 42–44.

Malaya Gupta, Mazumdar UK, Gomathi P,

Sambath Kumar R (2004). Antioxidant

and Free Radical Scavenging Activities

of Ervatamia coronaria Stapf. Leaves.

Iran J Pharm Res. 2: 119–126.

Pawelka KH, Stoeckigt J (1983). Indole

alkaloids from cell suspension cultures

of Tabernaemontana divaricata and

Tabernaemontana iboga. Plant Cell

Rep. 22: 105–107.

Piyush Patel, Nurudin Jivani, Shailesh

Malaviya, Tushar Gohil, Yagnik

Bhalodia (2012). Cataract: A major

secondary diabetic complication. Int

Cur Pharm J. 1: 180–185.

Sharma P, Cordell GA (1988). Heyneanine

hydroxyindolenine, A new indole

alkaloid from Ervatamia coronaria var.

plena. J Nat Prod. 51: 528–531.

Taesotikul T, Panthong A, Kanjanapothi D,

Verpoorte R, Scheffer JJC (1989).

Hippocratic screening of ethanolic

extracts from two Tabernaemontana

species. J Ethnopharmacol. 27: 99–106.

Umamaheswari M, Asokkumar K, Lalitha V,

Sivashanmugam AT, Subhadradevi V

(2011). Anticataract and antioxidant

activities of Citrus aurantium L. peel

extract against naphthalene induced

cataractogenesis in rats. J Phar Res. 4:

680–682.

Vats V, Yadav SP, Biswas NR, Grover JK

(2004). Anticataract activity of

Pterocarpus marsupium bark and

Trigonella foenum-graecum seeds

extract in alloxan diabetic rats. J

Ethnopharmacol. 93: 289–294.

Yamamoto T, Takahashi H, Sakai K,

Kowithayakorn T, Koyano T(1997).

Screening of Thai plants for anti-HIV-1

activity. Natural Med. 51: 541–546.





STUDIES ON TRADITIONAL HERBAL PEDIATRICS PRACTICES IN

JAISINGHPUR, DISTRICT KANGRA (HIMACHAL PRADESH, INDIA)

Rawat Dhiraj S1, Kharwal Anjna D

2

1Department of Botany, R.K.M.V. – Shimla, Himachal Pradesh, India

2 Department of Botany, Govt. Degree College- Dharamshala. Himachal Pradesh, India



ABSTRACT

Plants are the basis of life on earth and are central to people‟s livelihood. Glimpses of our

knowledge in ethnomedicine are available in vedic texts and there is an inextricable link between

indigenous culture and biodiversity as areas of high biodiversity are often found on indigenous

community‟s lands. The local communities and rural populace of Jaisinghpur is highly dependent on

nature for meeting their healthcare needs and has a repository of accumulated experience and

knowledge of prevailing vegetation of the region. Medical ethno–botany forms a major part of

medicinal aspects of aboriginal child care. 70 % of world population uses herbal traditional remedies

in treatment of sick and injured children. Indigenous herbal practices related to child–care provide

invaluable knowledge and aid in making best use of natural resources as it is dynamic in

dissemination and scientific in indigenous experimentation. Present study includes 21 plants (15

dicots and 6 monocots) belonging to 16 families used as herbal remedies for child–care, while 2

plant spp. are used along with other plant resources in herbal preparations. Among various plants,

fruits of 7 species (32%), seeds of 6 species (29%), leaves of 5 species (24%), peduncle, bark and

rhizome of one plant (5%) each is used predominantly for child–care by the rural populace of the

study area. Mostly, the people of age groups 41–60 years (AG–3) and >60 (AG–4) years mostly

ladies, are aware of these herbal practices. Local communities not only use these plants but also care

for their conservation and protection; thus contributing towards sustainable development.

KEY–WORDS: Ethnobotany, Biodiversity, Child–care, Herbal remedies, ODA (Observed density

& availability), Phenological pattern, TIV (Total importance value), Pediatrics.

ABBREVIATIONS: (AG– Age group/s)

Research article

Cite this article:

Rawat Dhiraj S, Kharwal Anjna D (2013), STUDIES ON TRADITIONAL HERBAL PEDIATRICS

PRACTICES IN JAISINGHPUR, DISTRICT KANGRA (HIMACHAL PRADESH, INDIA), Global J

Res. Med. Plants & Indigen. Med., Volume 2(4): 219–230

mailto:*[email protected]



INTRODUCTION

The term “Ethno–botany” was coined by

J.W. Harshberger in 1895, meaning “the study

of plants used by primitive and aboriginal

people” (Anonymous, 1895). Since then, the

subject has been variously defined and

interpreted by different workers as its discipline

began to follow multidisciplinary approach

combining a diversity of knowledge bases and

methods through the use of anthropological

methods (Robbins et al., 1916; Schultes &

Reis, 1995). Plants are the basis of life on earth

and are central to people‟s livelihood. Glimpses

of our knowledge in ethno–medicine are

available in vedic texts (Jain, 1987).

Undeniably, there is an inextricable link

between indigenous culture and biodiversity as

areas of high biodiversity are often found on

indigenous community‟s lands and in their

water bodies (Alcorn, 1996). The 15

th session

of the General Assembly of IUCN held in

Christchurch, New Zealand, in October 1991,

recognized the importance of the cultural

heritage of mankind and the role of traditional

cultures in conservation of nature (McNeely &

Pit, 1985). Agenda 21 of the Rio Earth Summit

(1992) stated that indigenous people have a

vital role in environmental management and

development because of their knowledge and

traditional practices. Ethno–botanical

information in the form of folklore is passed

through generations in certain restricted and

remote habitations (Chauhan, 1999; Choudhary

et al., 2008; Ganesan, 2008; Saini, 1996).

“Jaisinghpur” (592m), one of the tehsils of

district Kangra in Himachal Pradesh has

common boundaries with districts Mandi and

Hamirpur (Fig. 1). The word “Jaisinghpur” is

derived from the name of a famous king Raja

Jai Singh who is believed to be a great warrior

of “Rajgir” dynasty. Still the name of the

legislative constituency is “Rajgir” (reserved

for S.C.) which comprises most of the areas of

tehsil “Jaisinghpur”. The town “Jaisighpur” is

located on the bank of river “beas” with a

population of 1,273 while the population of

tehsil is 58,623. Tehsil “Jaisinghpur” is full of

natural water resources and is a combination of

greenery and water, thus, has given the tehsil a

distinctive look, located at an altitude between

500–1800 m above MSL between

31°53′55′′N/76°35′58′′E latitudes. The area is a

combination of the plains and the hills and

blessed with remarkable natural beauty and

high ranges of Dhauladhar mountains at the

backdrop with tops remain snow covered for

most part of the year. The natives are the

Kangri people and the local language is

„Kangri‟. The majority of the people are

„Hindu‟. Traditional dresses of men are „kurta‟,

„pyjamas‟ with a woolen jacket in winter.

Women generally wear „salwaar‟, „kameez‟

along with „chunni‟ („chaddru‟ in local

language). Maize, wheat and paddy are the

main staple foods of the rural populace and the

villagers are very fond of butter, milk, curry

preparations and pickles.

The place unfolds four broad seasons with

winters spreading generally from December–

February, summers from March–June, rainy

season extending from July–September with

landscape becoming lush green, and autumns

from October–November. Agriculture is the

main stay of the inhabitants of the area. Soil

varies from sandy loams to clay. The agro–

climatic conditions favour the growth of food

crops such as wheat, paddy, maize, potatoes,

etc. Agricultural operations are carried out in

two spells. Spring crops popularly called as

Rabi („Harri‟) comprise of wheat, barley, gram

and oil seeds (linseed) whereas autumn crops

(Kharif / „Savani‟) are maize, paddy, pulses,

spices and potatoes (Balokhra, 2002). The area

on the bank of river “beas” is highly fertile and

is famous for vegetable cultivation. This area is

known as “shukdi ka bag”. The region is also

famous for its “holi” and “dussehra” festivals.

“Chaugan” of “Jaisinghpur”, “Harsi”, “Laddi”,

“Lambagaon”, “Ashapuri” temple, “Nagban”

and “Sai” are the places of interest.

The local communities and rural populace

of Jaisinghpur is highly dependent on nature for

meeting their healthcare needs. The rural

populace of the region has a repository of

accumulated experience and knowledge of

prevailing vegetation of the region. They have



a deep belief on their nature folklore medicines

for remedies and they rely exclusively on their

herbal cure. Recently, considerable attention

has been paid to utilize eco–friendly plant

based products for the prevention and care of

different human diseases. In India over 6,000

plants are in use in traditional folk and herbal

medicine, which constitutes about 75% of the

medical needs of third world countries

(Rajshekharan, 2002). Similarly, herbal

medicines for infants and child–care are not

exceptions in the study area. The women folks

of the region play a vital role in use and

mobilization of biodiversity based knowledge

system. Medical ethnobotany forms a major

part of medicinal aspects of aboriginal child

care. 70 % of world population uses herbal

traditional remedies in treatment of sick and

injured children (Pearn, 2005).

Fig 1. Map of tehsil Jaisinghpur (district Kangra, H.P.) showing main locations visited for

survey.

There is an inextricable nexus between

aboriginal men, women and land in which they

live. Aboriginal women in traditional

communities use a sophisticated botanical

material media in the treatments of sick and

injured children. Drugs and medicaments used

in treatments are obtained from various plant

parts usually as fresh preparations in the form

of infusions, macerations, decoctions etc. and

are rarely stored (Ganesan, 2008). The

multipurpose and broad–spectrum use of plants

to treat symptoms and symptom complexes in

context of preventive medicine for child care is

the heart of discussion. Internationally, one of

the best works is that of Pearn, 2005, which

throws a light on traditional pediatric practices

in Australia and the work of Allen & Hatfield,

2004 which emphasized on ethnic studies of

Britain and Ireland and that of Salah &

Nyunda, 2012 which emphasized on pre-natal

care. The information on floristic and ethno–

botanical studies related to child–care in India

is scattered meager (Babu, 1998; Borthakur,

1993; Choudhary et al., 2008; Ganesan, 2008;

Goyal et al., 2011; Joshi, 1989; Pal et al., 2000;

Qureshi, 2007; Rajshekharan, 2002; Robbins et

al., 1916; Saini, 1996; Sen et al., 2008).

Ethnobotanical information on child care in

Himachal Pradesh (H.P.) and district Kangra is

meagre in literature (Chauhan, 1999; Dhiman,

1976). Inspite of rich floristic diversity and

cultural values, nothing is available in literature

on the floristic and ethnobotanical information

of the region (study area). Keeping this in

SIGNS

1- Lambagaon block

-Locations

Source

www.google.co.in

http://www.google.co.in/



mind, the present study had been undertaken

with the following objectives – (a) to collect

and identify the plant spp. used for child–care

along with their flowering and fruiting seasons

(b) to study the phonological pattern of

collected plants (c) to calculate total

importance value (TIV) and observed density

availability (ODA) of plant spp. (d) to know

about the effective age groups involved in

herbal practices.

Since time immemorial, the Himalaya has

influenced the life and culture of the diverse

ethnic communities living all along the length

of its mountainous chain. Keeping this in mind

it is pertinent to document this knowledge for

future generations.

METHODOLOGY

Intensive ethnobotanical exploration were

undertaken in the rural pockets of tehsil

Jaisinghpur, district Kangra (H.P.). The villages

selected for study are: Draman (900 m),

Dhupkyara (720 m), Laddi (1290 m), Nee

(910 m), Langa (845 m), Arth (835 m), Bhedu

(790 m), Bhaati (750 m), Jalag (840 m),

Nahlana (785 m), Saped (810 m), Nakki

(930 m), Tamru (920 m), Bhodi (890 m), Hadot

(915 m), Kosri (760 m), Ropari (750 m), Suan

(750 m), Tarapad (925 m), Tikri (620 m),

Kamanda (630 m), Dwata (980 m), Sai

(800 m), Harsi (550 m), Kathla (590 m) and

Bardama (690 m). The field tours were planned

in such a way so as to collect the

ethnobotanically interesting species used for

infants and child–care either in flowering or

fruiting stage. Herbarium of collected plants

was prepared following Jain & Rao, 1978. For

a better understanding of local beliefs, habits

and uses of plants, different categories of

people like family heads, healers, old

experienced and knowledgeable informants,

especially old ladies were repeatedly

interviewed. Specific questions based upon

Proforma designed by Jain & Rao, 1978 were

asked and the resultant information was

recorded in the ethno-botanical field notebook

along with the name of locality and local name.

Botanical identification of the selected species

was first done with the help of regional floras

(Chauhan, 1999; Chowdhery & Wadhwa,

1984; Dhiman, 1976; Hooker, 1897).

For more information three basic

approaches were adopted following Phondani

et al., 2010:

An interview based approach– Questions

from informants on infant and child–care plants

mainly from old experienced people, especially

old ladies as they were more aware of the

child–care plants.

An inventory based approach– An

inventory based approach is followed on

following questions:

Whether whole plant or plant parts are

used?

Which age group is more aware to these

herbal remedies?

The season of flowering and fruiting

TIV (Total importance value) of these

plants

The density of plants in the region

Whether the plant is used for one

disease or for more than one disease?

An interactive discussions approach with

communities–

How to use plants?

Are they better than market products or

not?

Are they used singly or in combination?

Are all the plants or plant parts

available in nature or some of them are

taken from market?

While collecting the plant specimens, their

uses related to child–care and their local names

were also ascertained and recorded carefully in

the field notebook with the assistance of local

informants. The data were verified in different

regions among the interviewers and showing

the same plant sample, and even with the same

informants on different occasions. Ethno–

botanical lore was considered valid if at least

three informants made similar comments. Four

age groups (AG) were investigated i.e. AG–1

(0–20 years), AG–2 (21–40 years), AG–3 (41–



60years) and AG–4 (>60 years) to find out the

impact of ethno–botanical lore. Men and

women of local communities are interviewed

separately to find out the gender based herbal

knowledge.

Phenological pattern of the plants were

observed to find out the seasonal variation

while ODA (Observed density availability) was

observed according to Sood et al., 2012 in

which plants were classified into abundant,

considerable and rare extent. Nomenclature of

these taxa were confirmed from Bennet, 1986

and Wielgorskaya,1995.

Economic valuation of all the presently

recorded ethno–botanical species was also

carried out to calculate the total importance

values (TIV) on the sum basis of parameters

like life cycle strategy, periodicity of use, habit,

availability throughout year and uses as per

detailed methodology outlined by Belal &

Springuel, 1996.

OBSERVATIONS

The local communities of tehsil Jaisinghpur

of district Kangra (H.P.) use 21 plant spp. in 18

different types of herbal practices related to

child–care. These local communities are a rich

repository of traditional knowledge, so a

sincere effort has been made to get the

information on these herbal practices which

are:

Bark of Ficus religiosa is cut into pieces

and to this seeds of “puthkanda” Achyranthes

bidentata are added along with peduncles of

“challi” maize (Zea mays). The mixture is

burned to ash. Ash is mixed with borax

(Suhaga) and honey. Half teaspoon is

recommended thrice a day (one week) for

bronchitis, cough and congestion. (AG–3, 4).

1–2 small holes are created in unripe fruits

of “rada” (Randia dumetorum) with the help of

thorns of the same and these holes are filled

gently with the milk (latex) of Ficus palmata.

These fruits are sealed with kneaded “kanak”

wheat flour and finally the sealed fruits are

roasted in a fire–place “chullah”. Powder of

roasted fruits along with honey (½ teaspoon

twice a day for a week) is highly recommended

against bronchitis, cough and congestion. (AG–

3, 4).

Leaves of “Kouru” Roylea cinerea are

crushed with the help of a clean pastel and

mortar. Juice is filtered with a fine clean cloth

piece and 1–2 drops of it are used twice as

nasal drops for three consecutive days. It is

considered good for cough. (AG–3, 4).

A longitudinal fine cut is made into

“chhuara” (Fruit of Phoenix dactylifera) and

the seed is taken out. The left out cavity so

created, is filled up with powdered seeds of

“chhoti ellaichi” (Elettaria cardamomum) and

the fruit is sealed with thread. It is roasted in a

fire–place “chullah”. ½ of the fruit is taken

with milk at bed time for 1–2 months for

checking frequent urination in infants and

children. (AG–3, 4).

Rhizomes of “barain” Acorus calamus are

washed with water and its paste along with

honey is taken thrice a day for a week against

cold, cough and bronchitis. (AG–2, 3).

4–5 small stones having smooth surface are

thoroughly washed and are dumped at fire–

place “Chullah” for 20–30 min. About 100 ml

of drinking water is taken in a clean bowl and 1

tablespoon full of “ajwain” seeds

(Trachyspermum ammi) is added to it. Hot

stones are taken out from fireplace with the

help of “chimtah”– a tong like household

article and ash is removed with the help of

cotton cloth. The stones are dipped in water

taken in the bowl. The fluid in the bowl is

filtered and the filtrate so obtained is locally

known as “gitt– juanhe” as the word “gitt” is

used for stones and “juanhe” for ajwain

(Trachyspermum ammi) in local „Kangri‟

language. 2–3 teaspoons of “gitt–juanhe” is

prescribed thrice a day for 3–4 days to check

stomach ailments in infants. (AG–4).

Fruits of “jaiphal” (Myristica fragrans) are

rubbed on a clean stone with water and the

paste so obtained is prescribed for children to

check cough and congestion thrice a day for 4



days and the same is also applied over nose for

the same. (AG–2, 3).

Decoction of “ajwain” seeds

(Trachyspermum ammi) is highly prescribed for

infants and children for stomachache. Usually

5–10 tablespoons are given to infants and

children after every 2–3 hours until relief.

(AG–1, 2, 3, 4).

Decoction of aerial parts (leaves, stem,

inflorescence and even seeds) of “tulsi”

(Ocimum sanctum) is highly prescribed for

couch and fever. (AG–2, 3, 4).

The fruits of “harad” (Terminalia chebula)

are rubbed over a clean stone or in “kundi"– a

mortar type household article and a pinch of its

paste along with lukewarm water is prescribed

thrice a day against constipation until relief.

(AG–2, 3, 4).

Decoction of “kadwi–saunf ” seeds

(Foeniculum vulgare) is prescribed for infants

and children for stomachache. Usually 1–2

tablespoons are given to infants. (AG–1, 2, 3,

4).

Decoction of “chhoti ellaichi” (Elettaria

cardamomum) along with sugar is considered

good for lung ailments. (AG–3, 4).

Fruit poultice of “dodey” (Sapindus

mukorosii) is made and a thick layer of it is

applied over a clean cotton cloth and this cloth

is tied around mumps and considered one of the

best traditional remedy against it. (AG–3, 4).

Seeds of “til” (Sesamum orientale) along

with jaggery and “soya” (Anethum graveolens

L.) are used to make traditional “laddu” which

are given to infants to check bed wetting.

Similarly ripe dried fruits of “Chhuara”

(Phoenix dactylifera) are prescribed for the

same. (AG–3, 4).

Onion “pyaz” (Allium cepa) juice is mixed

with mustard “sarson” (Brassica campestris L.)

oil. 1–2 drops thrice a day (for a week) is

recommended for ear itching. (AG–2, 3, 4).

Patients exposed to the smoke on burning

of “naule ra lingna” (aerial parts of Verbascum

thapsus) are considered to have relief from

measles. (AG–4).

Pieces of leaves of “kwarya” (Aloe vera)

are heated gently. Each piece is cut gently into

two pieces to expose the gel of leaves. Each

piece singly or in combination with “haldi” i.e.

turmeric powder (Curcuma angustifolia Roxb.)

is used against muscle pull. (AG– 1, 2, 3, 4).

“Jaiphal” Myristica fragrans” is rubbed

over a soft clean stone and the paste of it is

applied over cotton plugs. These cotton plugs

are placed in mustard (“sarson”– Brassica

campestris) oil taken in an earthen pot “diya”

and enlightened with fire. The pot is placed in

the corner of a dark room and a brass plate is

placed on it for 3–4 hours. The deposited

carbon on brass plate is collected. It is mixed

with cow‟s ghee and small quantity of

“jaiphal” paste. The obtained product is locally

called as “kajjal” which is applied in eyes to

check any infection. It is also used for healthy

and beautiful eyes. (AG– 3, 4).

RESULTS & DISCUSSION:

The study of ethno–medical systems and

herbal medicines as therapeutic agents of a

paramount importance in addressing health

problems of traditional communities and third

world countries as well as industrialized

societies (Rajshekharan, 2002; Saini, 1996).

The present study yielded interesting data

which provides information of the 21 plants

used for child–care in tehsil Jaisinghpur of

district Kangra (Himachal Pradesh). The plants

are used in 18 different herbal practices of the

region in which these plants are used either

singly or in combination. These plants are

arranged in alphabetical order; with their local

name, flowering and fruiting seasons, ODA

(Observed density & availability) and part /

parts used as in table 1.

Present study includes 21 plants (16 dicots

and 5 monocots) (Table 1), used as herbal

remedies for child–care belonging to 16

families. The predominant families are



Liliaceae with 3 spp, Moraceae, Apiaceae and

Poaceae with 2 spp each. Ficus with 2 spp is

the dominant genus. Among various plants,

fruits of 7spp (32%), seeds of 6 spp (29%),

leaves of 5 spp (24%), peduncle, bark and

rhizome of one plant (5%) each is used

predominantly for child–care by the rural

populace of the study area (Figure 2). 12 plant

spp are used singly while 5 plant spp. are used

in combination. One plant i.e. Ellettaria

cardamomum is used along with sugar for lung

ailments.

ODA (Observed Density Availability)

reveals that 8 plant spp are in abundant extent

while 9 spp. are in considerable extent. One

cultivated plant (Gossypium arboreum) is in

rare extent and the cotton (seed surface hairs)

of this can be purchased from the market so

that the plant resource can be used in a

sustainable manner in the study area.

Phenological pattern of plants suggest that most

of the plants are in flowering and fruiting stage

during rainy, summer and spring seasons (Fig.

3).

Fig.2. Various plant parts used in herbal remedies in tehsil Jaisinghpur of district Kangra

(H.P.).

Fig. 3. Phenological pattern of ethnobotanical plants in tehsil Jaisinghpur of district Kangra

(H.P.).

Table 1

Plant parts in herbal remedies

Fruits

Seeds

Leaves

Bark

Rhizome

Peduncle

29%

24%

5% 5%5%

32%

0

2

4

6

8

10

12

14

Winter Spring Summer Rainy

No

. o

f p

lan

ts

Seasons

Phenological pattern



Table -1 List Of Plants Used as Herbal Remedies for Child–Care in Tehsil Jaisinghpur of District Kangra

(Himachal Pradesh)

ODA – Observed Density Availability

+ + + – Abundant

+ + – Considerable extent

+ – Rare extent

S.N. Botanical Names and Family Local Names Flowering & Fruiting

Seasons

ODA Part/s

Used

1. Achyranthes bidentata Blume (Amaranthaceae) Puthkanda Mar.–Oct. + + + Seeds

2. Acorus calamus L. (Liliaceae) Barain June–July + + Rhizome

3. Allium cepa L. (Liliaceae) Pyaz Dec.–Mar. + + Scales

4. Aloe vera (L.) Webb. & Berthel. (Liliaceae) Kwarya Throughout year + + Leaves

5. Brassica campestris L. (Brassicaceae) Sarson Oct.–Mar. + + + Seeds

6. Foeniculum vulgare Mill. (Apiaceae) Kadwi Saunf Oct.–Mar. ++ Seeds

7. Ficus palmata Forsk. (Moraceae) Dhuda, Phegda April–Sept. + + + Latex

8. Ficus religiosa L. (Moraceae) Peepal Mar.–Oct. + + + Bark

9. Gossypium arboreum L. (Malvaceae) Kapaa Jul.–Sept. + Seed–hairs

10. Ocimum sanctum L. (Lamiaceae) Tulsi Aug.–Sept. + + + Aerial Parts

11. Randia dumetorum (Retz.) Lam. (Rubiaceae) Rada April–Sept. + + Fruits

12.

Roylea cinerea (D. Don) Baill. (Verbenaceae) Kouru Mar.–Nov. + + Leaves

13.

Sapindus mukorosii Gaertn. (Sapindaceae) Dodan, Reetha April–Sept. + + + Fruits

14. Sesamum orientale L. (Pedaliaceae) Til July–Sept. + + Seeds

15. Terminalia chebula Retz. (Combretaceae) Harad April–June + + + Fruits

16. Trachyspermum ammi Sprague (Apiaceae) Ajwain Oct.–Mar. + + Seeds

17. Verbascum thapsus L. (Scrophularaceae) Naule ra

leengna

Jan.–April + + Aerial parts

18. Zea mays L. (Poaceae) Challi June–Sept. + + + Peduncle

19. Phoenix dactyllifera L. (Arecaceae) Khajur From market Fruits

20. Myristica fragrans Houtt. (Myristicaceae) Jaiphal From market Fruits

21. Elettaria cardamomum (L) Maton

(Zingiberaceae)

Chhoti–ellaichi From market Fruits



Table 2 - Total importance value (TIV) of ethnobotanical plants in tehsil Jaisinghpur of District Kangra

(Himachal Pradesh)

S.N. Botanical Names and Family Life

cycle

strategy

Periodicity

of use

Habit Availability

throughout

year

Uses TIV

(%)

1. Achyranthes bidentata Blume

(Amaranthaceae)

2 2 2 3 2 55

2. Acorus calamus L. (Liliaceae) 2 3 2 2 4 65

3. Allium cepa L. (Liliaceae) 2 2 2 2 3 55

4. Aloe vera (L.) Webb. & Berthel.

(Liliaceae)

2 2 2 3 3 60

5. Brassica campestris L. (Brassicaceae) 2 2 2 2 2 50

6. Foeniculum vulgare Mill. (Apiaceae) 2 3 2 3 3 70

7. Ficus palmata Forsk. (Moraceae) 4 2 4 4 2 80

8. Ficus religiosa L. (Moraceae) 4 2 4 4 2 80

9. Gossypium arboreum L. (Malvaceae) 3 1 3 1 1 40

10. Ocimum sanctum L. (Lamiaceae) 3 3 2 2 3 65

11. Randia dumetorum (Retz.) Lam.

(Rubiaceae)

4 2 3 3 2 70

12. Roylea cinerea (D. Don) Baill.

(Verbenaceae)

4 2 3 3 3 75

13. Sapindus mukorosii Gaertn.

(Sapindaceae)

4 2 4 2 3 75

14. Sesamum orientale L. (Pedaliaceae) 2 2 2 2 2 50

15. Terminalia chebula Retz.

(Combretaceae)

4 3 4 2 4 85

16. Trachyspermum ammi Sprague

(Apiaceae)

2 3 2 3 3 70

17. Verbascum thapsus L.

(Scrophularaceae)

2 2 2 2 2 50

18. Zea mays L. (Poaceae) 1 2 2 1 2 40



Fig. 4. Herbal practices in different age groups in Tehsil Jaisinghpur.

Out of discussed 18 herbal practices, 3

herbal practices were known to the population

of 0–20 years i.e. age group1 (AG–1); while 9

to age group 2 of 21–40 years (AG–2). Mostly,

the people of age groups 41–60 years (AG–3)

and >60 years were aware to 17 practices while

2 practices are restricted to age group 4 (AG–4)

i.e. > 60 years (Fig. 4). From this it is crystal

clear that these practices are mostly restricted

to AG–3 and AG–4 while few of them are

restricted to old ladies i.e. this knowledge is

fast depleting in younger generations so it is

pertinent to document this invaluable eco–

friendly herbal remedies. Gender wise analysis

reveals that 14 practices (5–18) are known to

both men and women of the study area while

first 4 practices (1–4) are restricted only to

women of different communities of the region

(Fig.4).

Two plant spp. i.e. Curcuma angustifolia

roxb. (powdered rhizome) and Anethum

graveolens L. (seeds) are used with other plant

resources in some of the herbal practices. These

plants are also cultivated in the study area and

can be purchased from the market. Due to the

utmost importance of these plants in the region,

local communities not only use these plants but

also care for their conservation and protection;

thus contributing towards sustainable

development.

Statistically, the total importance value

(TIV) reveals that Terminalia chebula tops the

list with TIV of 85%. Ficus reliogosa, Ficus

palmata have 80% TIV while Roylea cinearea

and Sapindus mukorosii have TIV of 75%.

Gossypium arboretum and Zea mays have

lowest TIV of 40% with respect to medicinal

values for child–care. (Table II)

CONCLUSION

Indigenous herbal practices related to

child–care provide invaluable knowledge and

aid in making best use of natural resources as it

is dynamic in dissemination and scientific in

indigenous experimentation. In the modern

days of technological advancement, this

knowledge is falling prey to the lure of

modernization and urbanization. Negligible

efforts have been undertaken to understand the

scientific basis of the knowledge. It is

recommended that the documentation of

indigenous herbal practices should be included

in the curricula of environment and sustainable

development as a cross cutting issue.

0

2

4

6

8

10

12

14

16

18

No. of herbal

practices

AG-1 AG-2 AG-3 AG-4 >60 only

Age groups

Herbal practices in different age groups



REFERENCES

Alcorn, JB (1996). Is biodiversity conserved by

indigenous people? In: Ethnobiology in

HumanWelfare. SK Jain (Ed.), New

Delhi: Deep Publ., pp 233–238.

Allen, DE & Hatfield, G (2004). Medicinal

plants in folk tradition : An ethnobotany

of Britain & Ireland. UK: Timber Press.

Anonymous (1895). Some new ideas, The

plants cultivated by aboriginal people

and how used in primitive commerce.

Philadelphia: The Daily Evening

Telegraph, vol. 64, no 134.

Babu R (1998). Mother & Child Care. J.

Amruth 2(6) : 98–102.

Balokhra, JM (2002). The Wonderland

Himachal Pradesh. New Dehli: H.G.

Publication.

Belal, Ahmed E & Springuel I (1996).

Economic value of plant diversity in

arid environments. Nature & Resources

32 (1): 33–39.

Bennet, SSR (1986). Name Changes in

Flowering Plants of India and Adjacent

Regions. Dehradun, India: Triseas Publ.

Borthakur, SK (1993). Native phytotherapy for

child and women diseases from Assam.

Ethnobotany 5 (87): 87–91.

Chauhan, NS(1999). Medicinal and Aromatic

Plants of Himachal Pradesh. New

Delhi: Indus Publ. Co.

Choudhary, K, Singh, M & Pillai, U (2008).

Ethnobotanical survey of Rajasthan –

An uptade. Am Eurasian J. Bot. 1(2):

38–45.

Chowdhery, H J & Wadhwa , BM (1984).

Flora of Himachal Pradesh. Calcutta:

Vol. 1–3. Bot. Surv. India.

Dhiman, DR (1976). Himachal Pradesh Ki

Vanoshdhiya Sampada. Dharamsala,

H.P.: Imperial Printing Press.

Ganesan, S (2008). Traditional oral care

medicinal plants survey of Tamil Nadu,

Natural Product Radiance 7: 166–172.

Goyal, M, Sasmal, D &Nagori, BP (2011).

Review on medicinal plants used by

local community of Jodhpur district of

Thar desert. Int. J. Pharmacology 7:

333–339.

Hooker, JD (1872–1897). The Flora of British

India. Allahabad: Vol. I–VIII, Lalit

Mohan Basu.

Jain, SK & Rao, RR (1977). A Handbook of

Field and Harbarium Methods. New

Delhi: Today‟s & Tomorrow‟s Printers

& Publ.

Jain, SK (1987). A Manual of Ethnobotany.

Jodhpur : Sci. Publ.

Joshi, P (1989). Herbal drugs in Rajasthan–

from childbirth to child–care, Ethnobot.

1:77–87.

McNeely, JA & Pit, D (Eds.) (1985). Culture

and Conservation, The Human

Dimension in Environmental Planning.

London, New York, Sydney: Croom

Helm.

Pal, DC, Guha, A & Sen, R (2000). Medicinal

Plants and Plant Products Used in

Children Diseases among Aboriginal in

India. In: Ethnobotany and Medicinal

Plants of Indian Subcontinent. JK

Maheshwari (Ed.), Jodhpur: Scientific

Publishers.

Pearn, J (2005). The world‟s largest surviving

pediatric practices: some themes of

aboriginal medical ethnobotany in

Australia. J. Paediatr. Child Health

41(5–6): 284–290.



Phondani, PC, Maiknuri, RK, Rawat, LS,

Farooquee, NA, Kala, CP,

Vishwakarma, SCR, Rao, KS &

Saxena, KG (2010). Ethnobotanical

uses of plants among the Bhotiyal tribal

communities of Niti valley in Central

Himalaya, India. Ethnobot. Res. &

Application 18 : 233–244.

Qureshi, RA, Ghufran, MA, Gilani, SY,

Sultane, K & Ashraf. M (2007).

Ethnobotanical survey of selected

medicinal plants of Sudhan gali and

Ganga chotti hills, district Bagh,

Kashmir. Pak. J Bot. 39 (1) : 2275–

2283.

Rajshekharan, PE (2002). Herbal medicine, in

world of science. Employment News

(21–27), 3.

Robbins, WW, Herrin, JP & Freire –Marreco,

B (1916). Ethnobotany of the Tewa

Indians, Bureau of American

Ethnology. Washington, D.C. : Bulletin,

no 55. Smithsonian Institutions.

Saini VK (1996). Plants in the Welfare of

Tribal Women and Children in Certain

Areas of Central India. In:

Ethnobiology in Human Welfare. SK

Jain (Ed), New Delhi: Deep. Publ., pp

140–144.

Salah, AM & Nyunda, LA (2012). Effect of

Ruellia praetermissa extracts on

erythropoiesis in pregnant women.

GJMRI. 1(8): 309–314.

Schultes, RE & Reis, SV (1995). Ethnobotany

– Evolution of a Discipline.

London:Chapman.

Sen, S, Chakraborty, R, Biplab D & Devanna ,

N (2008). An ethnobotanical survey of

medicinal plants used by ethnic people

in west and south district of Tripura,

India, J. For. Res. 22 (3) : 417–426.

Sood, S K, Rawat, D, Rawat, S & Kumar, S

(2012). A Handbook of Wild Edible

Plants. Jaipur: Pointer publishers.

Wielgorskaya, T (1995). Dictionary of Generic

Names of Seed Plants. Dehradun:

B.S.M.P.S.





LIVER ENZYMES AND ITS ASSOCIATION WITH AGE AND SEX IN

SICKLE CELL ANAEMIA PATIENTS AND HAEMOGLOBIN

S TRAIT CARRIERS.

Chuku L C1, Chinaka N C

2

1, 2 Department of Biochemistry, University of Port Harcourt, P.M.B. 5323, Choba, Port Harcourt, Rivers

State, Nigeria

*Corresponding Author: [email protected]


ABSTRACT

The activities of the liver enzymes, alanine transaminase (ALT), aspartate transaminase (AST)

and alkaline phosphatase were compared between males and females of various age groups (0–25+)

during sickle cell crisis and in steady state. Results show that enzyme activities increased during

painful crisis. The difference in activities of the enzymes in normal homozygous HbAA and

heterozygous (HbAS) blood was not statistically significant (p ≥ 0.05). The activities of the enzyme

increased with age in all the genotypes studied. There was no significant difference (p ≥ 0.05) in the

activities of ALT, AST and ALP between males and females for all the age groups studied.

KEY WORDS: Alanine transaminase, alkaline phospatase, anaemia, aspartate transaminase,

genotype, heterozygous, homozygous, sickle cell.

Research article

Cite this article:

Chuku L C, Chinaka N C (2013), LIVER ENZYMES AND ITS ASSOCIATION WITH AGE AND SEX

IN SICKLE CELL ANAEMIA PATIENTS AND HAEMOGLOBIN S TRAIT CARRIERS., Global J

Res. Med. Plants & Indigen. Med., Volume 2(4): 231–237



INTRODUCTION

Sickle cell anaemia is a form of sickle cell

disease in which both abnormal genes are for

the formation of HbS (Konotey-Ahulu, 1974).

Sickle cell haemoglobin (HbS) is the major

abnormal haemoglobin. Its solubility is

sufficiently altered to produce a serious disease

when present in the homozygotes (White, et al.,

1978). Sickle cell anaemia (SCA) has been a

great clinical problem in the history of West

Africans since the 7th

century. The disease was

first described by a Chicago physician, J.B.

Herrick (Herrick, 1910). Herrick observed

peculiar elongated and sickled-shaped red cells

in a case of severe anaemia of an ailing West

Indian student.

The term “sickle cell trait” is used to

describe a person who has inherited one normal

haemoglobin gene (A) from one parent and one

abnormal gene (S) from the other parent. Sickle

cell disease arose as a result of mutation which

caused a single amino acid substitution (valine

for glutamic acid) at the sixth position of the β-

globin chain of the haemoglobin molecule

(Acquaye, et al., 1981). This change has an

adverse effect on the haemoglobin molecule.

Hence, the research was undertaken to study

any variation that might occur in the enzymes

in normal and sickle cell subjects over a wide

age range and sexes (male and female).


Materials, test kits and equipments used

were of laboratory standards. Blood from

normal and heterozygous individuals was

obtained at the University of Port Harcourt

Teaching Hospital and LABMEDICA

laboratory, Port Harcourt. Samples were then

grouped according to their ages and sex.

The normal subjects (HbAA and HbAS)

were aged between 0–70 years and of both

sexes. It was not possible to obtain HbSS blood

from patients beyond 20 years of age. Blood

was also collected from HbSS patients who

were in haemolytic crisis.

Electrophoresis

In all cases, blood samples from different

individuals were genotyped. Different

molecular species of Hb were separated from

each other by electrophoresis at pH 8.4 on

cellulose acetate. The separated haemoglobin

bands were then stained by Ponceau S dye and

identified by comparison with known

haemoglobin standards separated stained in the

same manner.

Different types of Hb were separated from

one another by electrophoresis on cellulose

acetate paper. Separation was clear at the buffer

pH of 8.4. Sixty four (64) blood samples were

separated into 24 AA, 16 AS and 24 SS.

Assay of enzyme activity

Alanine transaminase (ALT) activity: By

Ratman and Frankel, (1957).

Alanine transaminase activity was

estimated by the procedure described in the

RANDOX kit for the determination of alanine

transaminase activity in serum or plasma at

546 nm. The optical density (O.D) of the

reaction mixture was taken colorimetrically at

546 nm as a measure of the enzyme activity.

Aspartate transaminase (AST) activity: By


Aspartate transaminase catalyses the

reaction (transamination) between α-

ketoglutarate and L-aspartate. The oxaloacetate

formed in the reaction reacts with 2, 4-

dinitrophenylhydrazine (DNPH), which in

alkaline medium gives a red-brown colour.

This is measured in a colorimeter at 546 nm.

This measures the activity of aspartate

transaminase. The enzyme activity was

measured by a procedure described in the

RANDOX GOT kit.

Alkaline phosphatase (ALP) activity: By


Alkaline phosphatase (ALP) hydrolyses p-

nitrophenyl phosphate in alkaline conditions to



yield phosphate and p-nitrophenol. The

nitrophenol is yellow in colour and absorbs

maximally at 405 nm. The intensity of the

colour due to p-nitrophenol during a fixed time

is measured at 405 nm colometrically and is

proportional to the ALP activity in the sample.

This was estimated by the procedure described

in the “BESSEY LOWRY” colour method kit

for the determination of ALP activity in plasma

or serum at 420 nm.

RESULTS

Enzyme Activity

The effect of age on ALT, AST and ALP

activities of HbAA, HbAS and HbSS subjects

are shown in tables 3.1 as well as their steady

and crisis state as shown in table 3.2.1, 3.2.2

and 3.2.3 respectively.

The effect of sex on ALT, AST and ALP

activities in HbAA and HbAS subjects are is

illustrated in table 3.3.1 respectively. The effect

of sex on AST and ALP activities of HbSS and

its crisis state is shown on table 3.3.2 and 3.3.3

respectively.

DISCUSSION

Alanine transaminase (ALT):

The activity of ALT of sickle cell subjects

was found to be higher than normal. There was

however no significant difference (p ≥ 0.05) in

the specific activity of ALT for normal (AA

and AS) subjects. It was also observed that the

activity of ALT did not increase steadily with

age and did not also depend on sex. Serum

ALT is highly variable in sickle cell disease.

Values obtained for subjects between 11–25

years were far lower than those obtained for

subjects between 6–10 years although higher

than the values obtained for subjects between

0–5 years. This was observed in all the

genotypes.

In crisis and steady state, ALT activity in

HbSS increased with age (11–20 years) in

steady state.

However, with the age range studied there

was no significant difference (p ≥ 0.05) in

mean enzyme activity values when compared

with normals. The pattern could change in

older subjects, but for age 0–25 years at least

hepatic dysfunction based on this enzyme alone

could not be definitely deduced (see tables 3.1,

3.2.1).

Table 3.1: Effects of age on ALT, AST, and ALP activities in HbAA, HbAS and HbSS subjects.

ALT

Activty

(µL)

AST

Activty

(µL)

ALP

Activty

(µL)

Age

(yrs)

AA AS SS AA AS SS AA AS SS

0-5

n = 5

5.3 ± 1.0 5.5 ± 1.2 30.1 ± 2.0 7.5 ± 0.2 15.75 ± 0.4 19.75 ± 0.6 137.66 ± 23.1 102.8 ± 18.2 148.75 ± 14.9

6-10

n = 5

11.7 ± 0.8 13.2 ± 0.9 36.9 ±1.8 13.33 ± 0.4 14.70 ± 0.3 10.5 ± 0.5 100.33 ± 15.2 102.8 ± 12.6 126.0 ± 8.49

11-15

n = 5

7.8 v 1.2 7.5 ± 1.0 26.8 ± 1.4 5.5 ± 0.2 11.0 ± 0.1 31.0 ± 1.9 66.75 ± 6.7 70.0 ± 5.6 88.7 ± 26.6

16-20

n = 5

7.2 ± 0.6 10.5 ± 0.8 43.5 ± 1.2 9.67 ± 0.7 14.5 ± 0.2 39.1 ± 2.0 40.8 ± 13.3 49.5 ± 0.71 72.4 ± 13.6

21-25

n = 5

8.0 ± 1.0 7.5 ± 0.9 12.0 ± 0.2 13.5 ± 0.4 31.5 ± 0.7 28.0 ± 2.8

>25

n = 5

8.0 ± 1.0 8.8 ± 1.0 10.25 ± 0.7 10.67 ± 0.5 29.8 ± 2.4 31.0 ± 16.2

n = number of samples analysed.

Values are Mean ± SD.



Table 3.2.1: Effect of age on ALT activity in HbSS subjects (Crisis and steady state).

ALT Activity (µL)

Age (yrs) Steady state Age (yrs) Crisis

0–5

n = 5

15.3 ± 0.5 0–5

n = 3

30.0 ± 0.7

6–10

n = 4

15.3 ± 0.6 6–10

n = 3

36.8 ± 1.4

11–15

n = 4

23.7 ± 0.4 11–15

n = 3

26.8 ± 0.4

16–20

n = 5

28.4 ± 0.2 16–20

n = 3

43.5 ± 0.8



Table 3.2.2: Effect of age on AST activity in HbSS subjects (Crisis and steady state).

AST Activity (µL)

Age (yrs) Steady state Crisis

0–5

n = 5

19.75 ± 0.6 30.0 ± 0.7

6–10

n = 5

10.5 ± 0.5 36.8 ± 1.4

11–15

n = 5

31.0 ± 0.4 26.8 ± 0.4

16–20

n = 5

33.2 ± 0.6 43.5 ± 0.8



Table 3.2.3: Effect of age on ALP activity in HbSS subjects (Crisis and steady state).

ALP Activity (µL)

Age (yrs) Steady state Crisis

0–5

n = 5

148.75 ± 14.9 164.4 ± 4.3

6–10

n = 5

126.0 ± 8.49 155.6 ± 7.6

11–15

n = 5

88.7 ± 26.6 150.0 ± 9.9

16–20

n = 5

72.4 ± 13.6 119.0 ± 4.4





Table 3.3.1: Effect of sex on ALT, AST and ALP activities in HbAA and HbAS subjects

ALT Activity AST Activity ALP Activity

Sex AA AS Sex AA AS Sex AA AS SS

Males

n = 15

6.8 ±

0.4

7.6 ±

0.4

Males

n = 5

9.0 ±

4.7

13.0 ±

4.0

Males

n = 5

57.5 ±

3.9

68.0 ±

5.9

125.6 ±

3.8

Females

n = 9

9.0 ±

0.52

8.0 ±

0.4

Females

n = 9

10.0 ±

6.85

12.0 ±

5.6

Females

n = 9

56.8 ±

2.9

62.0 ±

4.6

121.3 ±

2.9



Table 3.3.2: Effect of sex on AST and ALP activity in HbSS subjects (Crisis and steady state).

AST Activity ALP Activity

Sex Steady State Sex Crisis Sex Steady state Crisis

Males

n = 5

23.8 ±

13.54

Males

n = 8

43.88 ± 11.56 Males

n = 5

125.6 ± 3.8 144.6 ± 2.7

Females

n = 5

23.5 ±

12.18

Females

n = 7

35.14 ± 6.36 Females

n = 5

10.0 ± 6.85 157.3 ± 7.8



Aspartate tarnsaminase (AST):

The level of AST activity in sickle cell

anaemia (HbSS) subjects was higher than

normal. In HbAS there was a slight decrease in

activity as age increased. For HbSS subjects,

AST activity was high at age 15. Subjects in

the age bracket 6–10 years exhibited a decrease

in AST activity when compared to those

between 0–5 years.

In steady and crisis situation, AST activity

was highest in HbSS subjects when compared

to HbAA and HbAS subjects. However, during

crisis a higher activity was observed for all age

groups studied which appears to be no age

dependent trend in AST activity for HbSS

subjects under crisis situation with mean values

for each age group increase progressively (see

table 3.2.2). AST activity in steady state

increased with age up to age 20. Subjects in

steady state between 6–10 years exhibited a

low AST activity when compared to subjects

between 0–5 years. However, there was no

significant difference (p ≥ 0.05) in AST activity

in HbSS subjects between 11–15 years and 16–

20 years (see table 3.1).

On the effect of sex, statistical analysis

showed no significant difference (p≥ 0.05)

between both sexes in AST activity for HbAA,

HbAS and HbSS (crisis and steady state)

subjects.

Alkaline phosphatase (ALP):

From table 3.1 above, ALP activity of

heterozygous (AS) is not significantly different

(p ≥ 0.05) from that of homozygous (AA)

subjects. As shown in table 3.2.3, ALP activity

increased during crisis compared to steady state

condition.

The effect of ALP activity in normal (AA)

and heterozygous (AS) subjects decreased with

age significantly (p ≤ 0.05), while a steady

decrease in ALP activity was observed with

increase in age for HbSS subjects. However,

the difference in activity between the various

age groups is statistically significant (p ≤ 0.05).



Table 3.3.1 shows the effect of sex on ALP

activity of normal and sickle cell subjects. The

mean value for each group did not depend on

sex. There was no significant difference (p ≥

0.05) in ALP activity between both sexes.

Table 3.3.2 shows the effect of sex on ALP

activity in HbSS subjects in steady state and

painful crisis. Sex did not seem to affect the

enzyme activity whether in steady state or

crisis.

In this work, ALP had the highest levels of

activity, especially in growing children – in

both normal and sickle cell subjects. This may

be as a result of its involvement in the

calcification of bone and teeth. Also, raised

serum alkaline phosphatase accompanies

rickets of various etiologies.

CONCLUSION

In conclusion, the ALT and AST activities

in sickle cell disease as regards age are raised

from childhood to adolescence and that in the

steady state of sickle cell, there were elevations

which were further increased during painful

crisis. The sexes may not lead to any major

hepatic dysfunction, but should be considered

as a diagnostic parameter in sickle cell

management.

REFERENCES

Acquaye, C., Wilchek, M. and Gorecki, M.

(1981). Strategies for tackling sickle

cell disease. Trends in Biochemical

Sciences, 146–148.

Attah, E.B. (1975). Pathology of sickle cell

anaemia. Dokita. 7:19–21.

Dacie, J.V. (1960). The haemolytic anaemia,

congenital and acquired anaemia. (2nd

ed.). J.A. Churchill Ltd. Part 1. London:

200–230.

Dean, J. and Schechter, A.N. (1978). Sickle cell

anaemia. Molecular and cellular basis

of therapeutic approaches. The New

England Journal of Medicine. 229:753–

755.

Ekeke, G.I. and Ibeh, G.O. (1990). Liver

function enzymes and serum inorganic

phosphate levels in sickle cell disease.

2: 6–9.

Emmel, V.E. (1917). A study of the

erythrocyte in the case of severe

anaemia with elongated and sickle

shape red blood corpuscles. Arch.

Intern. Med. 20: 586.

Hahn, E.V. and Gillepspie, G.E.G. (1927).

Sickle cell anaemia. Arch. Intern. Med.

39: 233–236.

Herrick, J.B. (1910). Peculiar elongated and

sickle shaped red blood corpuscle in a

state of severe anaemia. Arch. Intern.

Med. 6: 517–521.

Konotey-Ahulu, F.I.D. (1974). The sickle cell

diseases, clinical manifestations

including the “sickle crisis”. Arch.

Intern. Med. 133: 611–619.

Lehmann, H. and Raper, A.B. (1949).

Distribution of sickle cell trait in

Uganda, and its ethnological

significance. Nature, 164: 494–495.



Nygren, A. (1967). SGOT in chronic

alcoholism. Act. Med. Scand. 182–383.

Smith, C.H. (1972). Blood disease of infancy

and childhood (3rd

ed.). C.V. mosby and

Co., St. Louis, 353–449.

Sofowora, A.O. (1993). Medicinal plants and

traditional medicine in Africa. 2nd

ed.

Ibadan: Spectrum books limited; pp

150.

White, A., Handler, P., Smith, E.L., Hill, R.L.

and Lehmann, I.R. (1978). Principles of

Biochem., 6th

ed. 947–1003.





CHELIDONIUM MAJUS L. - A REVIEW ON PHARMACOLOGICAL

ACTIVITIES AND CLINICAL EFFECTS

Biswas Surjyo Jyoti1*

1Department of Zoology, Midnapore College, Midnapore, West Bengal, India-721101



ABSTRACT

Chelidonium majus L. (Papaveraceae) is a plant that has been used for centuries in treating many

diseases in European and Asian countries. Crude extracts from various parts of the plant contain

isoquinoline alkaloids. The alkaloids derived from C. majus have not yet much studied; however,

some reports are available on toxicity studies of alkaloids of this plant. In such a scenario there is

need for understanding its therapeutic potential and its toxic actions. This review summarizes

scientific findings and suggests areas where further research is needed.

KEY WORDS: Chelidonium majus, alkaloids, pharmacology, antioxidant

Review article

Cite this article:

Biswas S J (2013), CHELIDONIUM MAJUS: A REVIEW ON PHARMACOLOGICAL ACTIVITIES

AND CLINICAL EFFECTS., Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 238–245




INTRODUCTION:

Chelidonium majus commonly known as

swallow-wort, rock poppy or greater celandine

belongs to Family-Papaveraceae. This plant is

distributed across the globe viz. Europe, Asia,

North America and in northwest Africa,

particularly in soils rich in nitrogen. The name

‘Chelidonium’ came from Chelidon-a greek

word which means swallow bird, as the plant

begins to flower when the swallows return. The

plant is widely regarded for its therapeutic

potential in Western and Asian countries

particularly in Chinese traditional medicine and

homeopathy. Crude extracts of C. majus and

isolated compounds exhibit numerous biological

activities (Colombo and Bosisio, 1996; Gilca et

al., 2010). Though many diseased conditions

even today are being treated with C. majus both

in traditional and homeopathic medical systems

but it has some self limitations therefore, its

therapeutic efficacy needs critical evaluation.

The current review summarizes scientific

findings of other investigators on C. majus and

suggests areas where further

investigations/research is needed.

Uses in traditional medicine systems

In many European, Asian and African

countries C. majus latex was used for bile and

liver disorders, for treatment of warts, corns,

eczema and solid tumors. It has traditionally

being used to treat liver diseases, gastric ulcer,

tuberculosis, skin eruptions and oral infections.

In Chinese traditional medicine and in

homeopathy C. majus is used to treat blockage of

blood circulation, to relieve pain edema and

jaundice.

Phyto-constituents (Figure. 1)

Extracts of Chelidonium has been found to

contain three types of benzyl isoquinoline

alkaloids viz. protoberberine, protopine,

benzophenanthredine. Sanuinarine and

chelerythrine are the prominent compounds

obtained from roots while coptisine, chelidonine

and berberine are obtained from the aerial parts

(Colombo and Bosisio, 1996). Other constituents

include malic, citric, gentisic, and hydrobenzoic

acids. It also contains hydroxycinnamic acid

derivatives, sparteine, saponin, carotenoids,

chelidocystatin and flavonoids.

Figure. 1 Chemical structure of Phyto-constituents

PHARMACOLOGICAL ACTIVITIES

Hepatoprotective effects

It has been demonstrated that Chelidonium

majus favourably modulates carbon tetrachloride

induced toxicity in rats. The treatment with C.

majus considerably reduced the number of

necrotic cells and decreased the activities of

transaminases and bilirubin (Mitra et al., 1992;

Mitra et al., 1996). Biswas et al. (2008) have

reported that ethanolic whole plant extract of

Chelidonium majus, has been tested for its

possible anti-tumor, hepato-protective and anti-

genotoxic effects in p-dimethylaminoazobenzene

(p-DAB) induced hepatocarcinogenesis in mice

through multiple assays: cytogenetical,

biochemical, histological and electron

microscopical. Data of several cytogenetical

endpoints and biochemical assay of some

toxicity marker enzymes at all fixation intervals

and histology of liver sections through ordinary,



scanning and transmission electron microscopy

at certain fixation intervals were critically

analyzed. The results suggest anti-tumor, anti-

genotoxic and hepato-protective effects of the

plant extract, showing potentials for use in

cancer therapy. Chung et al., (2004)

demonstrated that C. majus enhances nitric oxide

and TNF-α production via NF kappa B activation

in mouse.

Effects on enzymes

Mazzanti et al., (2009) reported that there

was a significant reduction in glutathione level

and SOD activity in liver after high oral dose of

C. majus. It was reported by others that C. majus

has a strong antioxidant activity as revealed from

FRAP assay (Then et al., 2003). Biswas et al.,

(2008) also reported that LPO and transaminases

activity reduced significantly after treatment with

C. majus extract against p-DAB induced

hepatocarcinogenesis.

Antimicrobial, antiviral and antiparasitic

effects

The modulatory effect of C. majus extract

against virus was evaluated in various in vitro

and in vivo studies C. majus showed

antimicrobial effect on gram positive bacteria

and on Candida albicans (Lendfeld et al., 1981).

Crude extracts of several alkaloids extracted

from C. majus exhibited antimicrobial, antiviral

and antifungal properties (Lozyuk, 1977;

Gerencer, et al., 2006; Parvu et al., 2008; Meng

et al., 2009; Monavari et al., 2012). Growth of

Alternaria, Aspergillus flavus, Candida albicans,

Rhizopus orizae and Scopulariopsis was

inhibited by berberine at 10–25 μg/ml

concentration (Mahajan et al., 1982). Ma et al.,

2000 demonstrated that chelidonine,

dihydrochelerythrine and dihydrosanguinarine

isolated from C. majus roots have activity

against Cladosporium herbarum at 4-10 μg/ml

concentration. It has been experimentally proved

that compounds (8-hydroxydihydro-

sanguinarine, dihydro-sanguinarine, dihydro-

chelerythrine, 8-hydroxydihydro-chelerythrine)

isolated from aerial parts of the plant showed

anti-bacterial effect against methicillin resistant

Staphylococcus aureus (Zuo et al., 2008).

Alkaloids extract showed antiviral efficacy

against human adenoviruses type 5 and 12,

herpes simplex virus, and RNA polio virus (Zuo

et al., 2008; Horvath et al., 1983; Kery et al.,

1987). Zhu and Ahrens (1982) investigated that

berberine successfully controlled the intestinal

secretion enhanced by E. coli enterotoxin, the

effect of which was dose dependent and it may

be due to quaternary ammonium group which is

responsible for anti-bactericidal property of

berberine and protoberberine was found active

against reverse transcriptase enzyme of RNA

tumor viruses. Chelidocystatin decreases the

activity of cysteine proteinases but further in

depth research are necessary especially in vivo

conditions.

Cardiovascular effects

Sanguinarine has been involved in

suppression of angiogenesis by inhibition of

VEGF signaling, this has been experimentally

proved in pig granulosa cells and in porcine

endothelial cells (Basini et al., 2007).

Immuno-modulatory activity

Immuno-modulatory properties of C. majus

have been investigated by Song et al., 2002,

where he obtained a protein bound to

polysaccharide from water extracts of the plant,

(CM-Ala) which showed mitogenic activity on

spleen, bone marrow cells, it also increased the

number of granulocyte macrophage colony

forming cells, further it suppressed immune

response locally by decreasing epidermal

Langerhans cells (Song et al., 2002). It has been

demonstrated that C. majus extract improved

overall humoral and cellular immunity response

and decreased incidence of recurrences of

tonsillitis in children (Khmel’nitskaia et al.,

1998).

Anti-inflammatory and Analgesic activity

Stylopine is a major component of leaf of C.

majus and it suppresses NO and PGE2 production

in macrophages by inhibiting iNOS and COX 2

expressions. It has been demonstrated that 5 and

12 lipoxygenase were inhibited by sanguinarine

and chelerythrine because these enzymes are



involved in leukotriene B4 and 12

hydroxyeicosatetranoic acid syntheses. As

compared to chelerythrine, sanguinarine showed

higher anti-inflammatory activity due to different

oxygen electron donating constituents (Lendfeld

et al., 1981). It has been reported that C. majus

extract increases TNF α production due to NF κB

production. It has also been reported that Ukrain

induces depolarization of mitochondrial

membrane potential and activates caspase in

Jurkat T lymphoma cell model (Habermehl et al.,

2006).

Choleretic effects

Vahlensieck et al., (1995) used phenolic and

alkaloid fractions of C. majus for their choleretic

activity using perfused rat livers. He

demonstrated that total extract induced choleresis

i.e. the bile flow was significantly elevated and

the amount of the bile was more than double the

quantity. Though it was not ascertained which

fraction of the extract was responsible for the

increased bile flow.

Effects on reproductive systems

The feeding of ethanolic extract of C. majus

showed that it could combat the spermatotoxic

effects to some extent in induced p-DAB induced

carcinogenesis. As benzophenanthridine

alkaloids have marked nucleophilic properties,

they might intercept the reactive metabolites;

thereby preventing their attack on nucleophilic

sites on DNA, and hence blocking adduct

formation (Vavreckova et al., 1996 a, b). Further

it has been suggested that many enzymatic

functions are essential for the normal integrity

and function of testis i.e. synthesis, development

and maintenance of normal sperm. Therefore, the

protective role of C. majus on sperm head could

also be attributed to its regulatory effect on

protein metabolism and repair activities in the

germinal cells (Biswas and Khuda-Bukhsh,

2002).

Antihyperglycemic and Hypoglycemic activity

Berberine an isoquinoline alkaloid obtained

from C. majus is used widely in China to reduce

blood glucose, in type II diabetes. Xuan et al.,

(2011) reported that berberine inhibits

mitochondria function and decreases intracellular

ATP in streptozotocin induced diabetes in rats.

This leads to a reduction in transcription factors

such as FoxO1, SREBP1, and ChREBP. As a

result, expression of gluconeogenic genes

(PEPCK and G6Pase) and lipogenic gene (FAS)

decreases. These molecular changes represent a

signaling pathway for improvement of fasting

glucose in the berberine treated diabetic rats (Xia

et al., 2011).

Anti-cancer efficacy

The anti-leukaemic activity of protoberberine

alkaloids has been reported and Smekal et al.

(1984) demonstrated that sanguinarine

intercalates partially as well as totally into the

DNA double helix. It has been demonstrated by

circular dichroism that the spectrum of DNA is

similar to ethidium binding to DNA. C. majus

had antiproliferative effect on human

keratinocyte cell lines (Vavreckova et al., 1996a,

b). Berberine intercalation to DNA might be due

to the planes of intercalated molecules which lie

parallel to those of purine-pyrimidine pairs. An

important constituent berberine has been shown

to interact with nucleic acids by various optical

methods. It was tested that administration of

350 μg/kg of protopine intra-peritoneally

inhibited very less regression of Erlich

carcinoma and application of 50 μg/kg b.w. of

chelidonine regressed sarcoma 180 (Sokoloff,

1968). UkrainTM

an anticancer drug whose

components include most of C. majus

compounds exerts multiple effects on cancer cell

lines (Cordes et al., 2003). Several reports have

been obtained in both animal and human models

regarding anticancer efficacy of Ukrain, against

various types of induced stomach carcinogenesis,

induced hepatocarcinogenesis, in patients

suffering from pancreatic cancer, Kaposis

sarcoma (Kim et al., 1997; Biswas and Khuda-

Bukhsh, 2002; Lohninger et al., 1996; Gansauge

et al., 2002; Ernst and Schmidt, 2005).

Chelidonine inhibits telomerase in tumor cells

strongly and may provide a basis for probable

anticancer agent also this alkaloid arrest mitosis

as a result of inhibition of tubulin polymerization

and activation of SAPK/JNK pathway.



Central Nervous system

An alkaloid obtained from C. majus,

thiophosphoric acid has been tested on rodents

regarding it action on CNS, it was found that it

depresses spontaneous motor activity; it seems to

stimulate dopaminergic system and depresses the

serotoninergic system (Kleinrok et al., 1992).

CLINICAL STUDIES:

Dysentry or gastroenteritis

Ardjah (1991) studied action of celandine on

upper abdominal symptoms in human subjects

such as cholinergic and spasmolytic effects using

panchelidon®. In case of patients with

postcholecystectomy 29 patients out of 35

showed clear improvement. A similar study

using 21 patients with dyspeptic complaints with

alcohol toxic liver parenchyma damage, 20

reported improvement after two weeks of

treatment. Limited numbers of clinical studies

have been carried out with total extracts in

patients with epigastric complications and the

sample size was small and definite conclusion

could not be ascertained from the study.

Periodontal effects

Benzophenanthridine alkaloids are routinely

used for the treatment of periodontal diseases,

Boulware et al., (1985) investigated that

sanguinaria extract was helpful in lowering of

volatile sulphur present in the oral cavity.

Southard et al., (1987) reported that

benzophenanthridine alkaloids act as an anti-

caries thereby preventing tooth decay.

Radioprotective effects

Song et al., 2003 demonstrated that extracts

of C majus have certain radioprotective effects.

Cytotoxic effects

There were spontaneous reports of adverse

drug reactions associated with C. majus

preparation. Incidences of hepatotoxicity have

been reported by several authors (Moro et al.,

2002, Kaminsky et al., 2006). It has been

reported that C. majus showed cytotoxicity

towards lymphoma cells and murine cell lines. In

some countries Complementary Evaluation

Committee recommended that products

containing alkaloids obtained from C. majus

must have a warning label and it should be

administered under medical supervision only.

CONCLUSION

We have reported hepatoprotective ability

of C. majus crude extract and various potencies

of it in induced hepatocarcinogenesis. It would

be prudent to investigate its constituents singly

and in combination, how they modulate

pathological changes and which form is more

potent or effective. Time of collection of plant

materials, place of collection, extraction

procedures, and its storage might affect its

active compounds both quantitatively and

qualitatively. The information summarizes

here concerning C. majus is intended to serve

as a reference to researchers involved in ethno-

pharmacological research.

ACKNOWLEDGEMENT

Grateful acknowledgements are made to

Professor A. R Khuda-Bukhsh, Department of

Zoology, University of Kalyani and Dr. Prabir

De, Scientist, CCMB, Hyderabad for

encouragements.

REFERENCES

Amoros M, Fauconnier B, Girre L (1977).

Propriétés antivirales de quelques

extraits de plantes indigenes. Annals

Pharm Françaises. 35:371–376.

Ardah H (1991). Therapeutic aspects of

functional symptoms of the upper

abdomen in biliary tract diseases

Fortschr Med.Med. 109 (S 115): 2–8.



Basini G, Santini S, Bussolati S, Grasselli F

(2007). The plant alkaloid sanguinarine

is a potential inhibitor of follicular

angiogenesis. J. Reproduc. Dev.

53(3):573–79.

Biswas SJ, Bhattacharjee N, Khuda Bukhsh AR

(2008). Efficacy of a plant extract

(Chelidonium majus L) in combating

induced hepatocarcinogenesis in mice.

Food Chem. Toxicol 46(5):1474–87.

Biswas SJ, Khuda-Bukhsh AR (2002). Effect

of a homeopathic drug, Chelidonium, in

amelioration of p-DAB induced

hepatocarcinogenesis in mice. BMC

Complement. Altern. Med. Vol. 2/4: 1–

16.

Boulware RT, Southard GL, Yankell SL

(1985). Sanguinaria extract, a new

agent for the control of volatile sulphur

compounds in the oral cavity. J Soc.

Cosmetic Chem. 36(4):297–302

Chung HS, An HJ, Jeong HJ, Won JH, Hong

SH, Kim HM (2004). Water extract

isolated from Chelidonium majus

enhances nitric oxide and tumor

necrosis factor alpha production via

nuclear factor kappa B activation in

mouse peritoneal macrophages. J.

Pham. Pharmacol. 56(1): 129–134.

Colombo M, Bosisio E (1996).

Pharmacological activities of

Chelidonium majus L. (Papaveraceae).

Pharmacol Res. 33(2): 127–134

Cordes N, Blaese MA, Plasswilm L, Rodemann

HP, Van Beuningen D (2003).

Fibronectin and laminin increase

resistance to ionizing radiation and the

cytotoxic drug Ukrain in human tumour

and normal cells in vitro. Int. J. Radiat.

Biol. 79:709–20.

Ernst E, Schmidt K (2005). Ukrain – a new

cancer cure? A systematic review of

randomized clinical trials. BMC Cancer

5: 69.

Gansauge F, Ramadani M, Pressmar J,

Gansauge S, Muehling B, Stecker K,

Cammerer G, Leder G, Beger HG

(2002). NSC-631570 (Ukrain) in the

palliative treatment of pancreatic

cancer. Results of a phase II trial.

Langenbecks Arch. Surg. 386:570–4.

Gerencer M, Turecek PL, Kistner O, Mitterer

A, Savidis-Dacho H, Barrett NP (2006).

In vitro and in vivo anti-retroviral

activity of the substance purified from

the aqueous extract of Chelidonium

majus L. Antiviral Res. 72 (2): 153–6.

Gilca ML, Gaman E, Panait I, Stoian, Atanasiu

V (2010). Chelidonium majus- an

intergrative review: Traditional

knowledge versus modern findings.

Forsch Komplementmed 17(5): 241–8.

Habermehl D, Kammerer B, Handrick R, Eldh

T, Gruber C, Cordes N, Daniel PT,

Plasswilm L, Bamberg M, Belka C,

Jendrossek V (2006). Proapoptotic

activity of Ukrain is based on

Chelidonium majus L alkaloids and

mediated via a mitochondrial death

pathway. BMC Cancer. 6:14.

Horvath J (1983). Antiviral effect of

Chelidonium extracts. Proc. Int. Congr.

Chemother. 9:124/106–124/110.

Kaminsky V, Lootsik M, Stoika R (2006).

Correlation of the cytotoxic activity of

four different alkaloids, from

Chelidonium majus (greater celandine),

with their DNA intercalating properties

and ability to induce breaks in the DNA

of NK/Ly murine lymphoma cells.

Cent. Eur. J. Biol. 191: 2–15.

Kery RY, Horvath J, Nasz I, Verzar-Petri G,

Kulcsar G, Dan P (1987). Antiviral

alkaloid in Chelidonium majus L. Acta

Pharmaceu. Hungerica. 57: 19–25.



Khmel’nitskaia NM, Vorob’ev KV, Kliachko

LL, Ankhimova ES, Kosenko VA,

Tymova EV, Mal’seva GS, Medvedev

EA (1998). A comparative study of

conservative treatment schemes in

chronic tonsillitis in children. Vest.

Otorinolaringol. 4: 39–42.

Kim DJ, Ahn. B, Han BS, Tsuda H. (1997).

Potential preventive effects of

Chelidonium majus L. (Papaveraceae)

herb extract on glandular stomach

tumor development in rats treated with

N-methyl- N0-nitro-N-nitrosoguanidine

(MNNG) and hypertonic sodium

chloride. Cancer Lett. 112: 203–8.

Kleinrok Z, Jagiello-Wojtowicz E, Matuszek B,

Chodkowska A. (1992). A basic central

pharmacological properties of

thiophosphoric acid alkaloid derivatives

from Chelidonium majus L. Pol. J.

Pharmacol. Pharm. 44: 227–239.

Lendfeld J, Kroutil M, Marsalek E, Slavik J,

Preininger V, Simanek V (1981).

Isolation, chemistry and biology of

alkaloids from plants of the

papaveraceae: anti-inflammatory

activity of quaternary

benzophenanthridine alkaloids from

Chelidonium majus. Planta Medica

43(2): 161–5.

Lohninger A, Korsh OB, Melnyk A (1996).

Combined therapy with Ukrain and

chemotherapy in ovarian cancer (case

report). Drugs Exp. Clin. Res. 22: 259–

62.

Lozyuk LV (1977). Antiviral properties of

some compounds of plant origin.

Mikrobiol. Zh (Kiev). 39: 343–8.

Ma WG, Fukushi Y, Tahara S, Osawa T

(2000). Fungitoxic alkaloids from

Hokkaido Papaveraceae. Fitoterapia.

71(5): 527–34.

Mahajan VM. (1982). Antimycotic activity of

berberine sulphate: an alkaloid from an

Indian Medicinal herb. Sabouraudia 20:

79–81.

Mazzanti G, Sotto A Di, Franchitto A,

Mammola CL, Mariani P, Mastrangelo

S, Menniti-Ippolito F, Vitalone A.

(2009). Chelidonium majus is not

hepatotoxic in Wistar rats, in a 4 weeks

feeding experiment. J. Ethnopharmacol.

126 (3): 518–24.

Meng F, Zuo G, Hao X, Wang G, Xiao H,

Zhang J, Xu G (2009). Antifungal

activity of the benzo[c]phenanthridine

alkaloids from Chelidonium majus Linn

against resistant clinical yeast isolates.

J. Ethnopharmacol. 125(3): 494–6.

Mitra S, Gole M, Samajdar K, Sur RK,

Chakraborty BN (1992)

Antihepatotoxic activity of

Chelidonium majus. Int J

Pharmacognosy 30:125–8.

Mitra S, Sur RK, Roy A, Mukherjee AS

(1996). Effect of Chelidonium majus L.

on experimental hepatic tissue injury.

Phytother Res. 10: 354–356.

Monavari SH, Shahrabadi MS, Keyvani H,

Bokharaei-Salim F (2012). Evaluation

of in vitro antiviral activity of

Chelidonium majus L. against herpes

simplex virus type-1. African. J.

Microbiol. Res. 6(20): 4360–64.

Moro PA, Cassetti F, Giugliano G, Falce MT,

Mazzanti G, Menniti-Ippolito

F,Raschetti R, Santuccio C

(2009).Hepatitis from Greater celandine

(Chelidonium majusL.): review of

literature and report of a new case. J

Ethnopharmacol.124(2):328–32.



Parvu M, Parvu AE, Cranium C, Barbu-

Tudoran L, Tamas M (2008).

Antifungal activities of Chelidonium

majus extract on Botrytis cinerea in

vitro and ultrastructural changes in its

conidia. J. Phytopath. 156(9): 550–2.

Smekal E, Kubova N, Kleinwachter V (1984).

Interaction of benzophenanthridiene

alkaloid sanguinarine with DNA. Stud.

Biophys. 101:125–32.

Sokoloff B (1968). Oncostatic and oncolytic

factors present in certain plants.

Oncology 22(1): 49–60.

Song JY, Yang HO, Pyo SN, Jung IS, Yi SY,

Yun YS. (2002). Immunomodulatory

activity of protein-bound

polysaccharide extracted from

Chelidonium majus. Archives.

Pharmacol. Res. 25(2):158–164.

Song JY, Yang HO, Shim JY, Ji-Yeon-Ahn,

Han YS, Jung IS, Yun YS (2003).

Radiation protective effect of an extract

from Chelidonium majus. Int. J.

Hematol. 78(3):226–32.

Southard GL, Parsons LG, Thomas LG,

Woodall IR, Jones BJ. (1987) Effect of

sanguinaria extract on development of

plaque and gingivitis when

supragingivally delivered as a manual

rinse or under pressure in an oral

irrigator. J. Clin. Periodontol. 14(7):

377–80.

Then M, Szentmihalyi K, Sarkozi A, Varga IS

(2003). Examination of antioxidant

activity in greater celandine

(Chelidonium majus L.) extracts by

FRAP method. Acta Biol. Szeged

4791– (4):115–7.

Vahlensieck U, Hahn R, Winterhoff H,

Gumbinger HG, Nahrstedt A, Kemper

FH (1995). The effect of Chelidonium

majus herb extract on choleresis in the

isolated perfused rat liver. Planta Med.

61:267–271.

Vavreckova C, Gawlik I, Muller K.1996b.

Benzophenanthridine alkaloids of

Chelidonium majus: 2. Potent inhibitory

action against the growth of human

keratinocytes. Planta Med. 62 (6):491–

4.

Vavreckova C, Gawlik I, Muller K. 1996a.

Benzophenanthridine alkaloids of

Chelidonium majus:1 Inhibition of 5-

and 12-lipoxygenase by a non redox

mehanism. Planta Med. 62 (5):397–401.

Xia X, Yan J, Shen Y, Tang K, Yin J, Zhang Y,

Yang D, Liang H, Ye J, Weng J (2011).

Berberine improves glucose metabolism

in diabetic rats by inhibition of hepatic

gluconeogenesis. PLoS One. 3;

6(2):e16556.

doi:10.1371/journal.pone.0016556.

Zhu B, Ahrens FA (1982). Effect of berberine

on intestinal secretion mediated by

Escherichia-coli heat stable enterotoxin

in jejunum of pigs. American J. Vet.

Res. 43(9):1594–8.

Zuo GY, Meng FY,. Hao XY, Zhang YL,

Wang GCH,. Xu. GL (2008).

Antibacterial alkaloids from

Chelidonium majus Linn

(Papaveraceae) against clinical isolates

of methicillin-resistant Staphylococcus

aureaus. J. Pharm. Pharmaceut. Sci.

11(4): 90–4.





A DETAILED PHARMACOGNOSTICAL EVALUATION ON LEAF OF

OLAX SCANDENS ROXB.

Naik Raghavendra1*, Borkar Sneha D

2, Harisha C R

3, Acharya R N

4

1,2P G Scholar, Department of Dravyaguna, IPGT & RA, Gujarat Ayurved University, Jamnagar, Gujarat,

India 3Head, Pharmacognosy Laboratory, IPGT & RA, Gujarat Ayurved University, Jamnagar, Gujarat, India

4Associate professor, Department of Dravyaguna, IPGT & RA, Gujarat Ayurved University, Jamnagar,

Gujarat, India



ABSTRACT

Leaves of Olax scandens Roxb. (Olacaceae) are edible and used for cure of headache. A detailed

pharmacognostical character of its leaf is lacking. In the present study, its leaves evaluated for their

morphological, microscopical and quantitative microscopic characters following standard

procedures. The overall study showed the leaves are 3.5–9 × 2.5–3.2 cm in size, reticulate venation,

petiole 0.2–0.5 cm. Leaves showed the presence of trichomes, collenchyma, and vascular bundles.

Powder microscopy of the dried leaves shows paracytic stomata of lower epidermis, epidermal cells

of upper epidermis, and unicellular trichomes of epidermis, rosette and prismatic crystals of calcium

oxalate.

KEY WORDS: Olax scandens, microscopy, quantitative methods, stomata.

Research article

Cite this article:

Naik R, Borkar S D, Harisha C R, Acharya R N (2013), A DETAILED PHARMACOGNOSTICAL

EVALUATION ON LEAF OF OLAX SCANDENS ROXB., Global J Res. Med. Plants & Indigen. Med.,

Volume 2(4): 246–253




INTRODUCTION

Plant and various plant products are being

used by human and animals either directly or

indirectly, since the existence of life, for food,

medicine, clothing, shelter etc. Olax scandens

Roxb. (Olacaceae), known as Badru, is a shrub

or small tree, distributed throughout tropical

India Flowers white, 6–7.5 mm long, in short

racemes; Fruit yellow, or orange fleshy,

subglobose, 0.8–1.5 cm in diameter, more than

half enclosed in accresent calyx. (Saxena H.O,

1995). Leaves of Olax scandens Roxb.

(Olacaceae) is being used by tribal people of

Odisha for medicinal and food purposes

(Tribhubana Panda et al., 2007). Decoction of

stem bark is taken internally to cure fever and

cough, (Veeramuthu et al., 2006) (Kirtikar &

Basu, 2003), boiled leaves fomentation is

applied to cure headache (Anonymous, 1990).

Review of literature reveals that its leaves have

not been studied in detail for

pharmacognostical characters, which is an

essential parameter for identification of a crude

drug (Anonymous, 1999). Hence, the present

study was undertaken to establish certain

botanical standards for identification and

standardization of O. scandens leaf.

MATERIALS AND METHODS:

Collection and preservation of the sample

Leaves of Olax scandens Roxb. were

collected from its natural habitat, Balangir,

Odisha, during September 2012 and identified

with the help of botanical texts and flora

(Saxena H.O, 1995). A sample specimen was

deposited to Pharmacognosy lab (SPECIMEN

NO- PHM 6062/21/09/2012) for future

references. The leaves were washed, shade

dried, powdered, sieved through 80 no. mesh

and preserved in an air-tight glass vessel. For

microscopical evaluation, fresh sample was

preserved in a solution prepared from 70%

ethyl alcohol : glacial acetic acid : formalin

(AAF) in the ratio of 90:5:5 (Johnson

Alexander Donald, 1940).

Morphological study.

The morphological study includes size,

shape, apex, margin, venation, base, petiole,

surface, color of leaves of O. scandens.

Microscopical study:

Microscopical examinations were carried

out by taking transverse section of petiole, leaf

through midrib (Khandelwal K.R et al., 1996),

type and distribution of stomata, epidermal cell

and trichomes (Anonymous, 1999) following

standard guidelines.

Quantitative microscopy:

Quantitative microscopy was carried out to

determine epidermal cell number, stomatal

number, stomatal index and size of the stomata

(Wallis, 1985).

Powder microscopy:

Dried leaf powder was studied following

standard procedures (Trease GE et al., 2002).

The micro photographs were taken by using

Carl zeiss trinocular microscope.

RESULT AND DISCUSSION:

Morphological study:

Macroscopic investigation showed that the

leaves are alternate, exstipulate, petiolate,

petiole 0.2–0.5 cm, somewhat twisted, base

equal, leaf measures about 3.5–9 × 2.5–3.2 cm,

dark green above, light green below, ovate to

lanceolate, margin simple, apex obtuse, strong

midrib, 6–7 pairs of nerves with reticulate

venation. (Plate A1–2).

Microscopical study:

Petiole:

The T.S of petiole is half-moon shaped in

outline. It showed, outer single layer of

epidermis with numerous simple, horn shaped

trichomes, followed by cortex, endodermis,

pericyclic fibers, phloem and centrally located

xylem. (Plate A 3–8).



PLATE A

3. T.S through petiole

4. Rosette crystal & tannin content

5. Epidermal layer with trichomes and

rosette crystal

6. Prismatic crystals & tannin

1. Morphology of leaf

2. Measurement of leaf

7. T.S showing cortex and vascular bundle

8. Pericyclic fiber, phloem and xylem.



Epidermis is single layered, barrel shaped,

compactly arranged cells, filled with yellow

colored material. Some of the epidermal cells

showed simple, unicellular, horn shaped

trichomes. Epidermis is covered with thick

cuticle.

Cortex is made up of 8–10 layers of loosely

arranged parenchyma cells. Lower side of the

parenchyma cells filled with rosette crystals,

prismatic crystals of calcium oxalate and some

tannin contents as compared to the upper side.

Some of the cells contained oil globules.

Endodermis is Inner to the cortex, single

layered, somewhat elongated thin walled cells

forming endodermis. Around the endodermis

4–6 layers of pericyclic fibers forming a ring

like structure covering the vascular bundle.

Vascular bundles are open and bi-collateral

type. Phloem present around the xylem with

some phloem fibers and sieve elements forming

a ring like structure. The metaxylem facing

towards lower epidermis and protoxylem facing

towards upper epidermis. Xylem bundles were

separated by uniserrate medullary rays along

with xylem parenchyma and fibers.

T.S Through mid rib

The T. S of leaf showed upper and lower

epidermis with mesophyll tissue having upper

pallisade and lower spongy parenchyma cells.

Section through midrib showed centrally

located vascular bundle covered with ground

tissue. On the lower side of the transverse

section 1–3 layers of collenchymatous cells

were present. (Plate B1–4)

Epidermis was Single layered, barrel

shaped epidermal cells both on upper and lower

epidermis with unicellular trichomes.

Epidermis was covered with cuticle. Trichomes

were more in lower epidermis than upper

epidermis. Stomata found only at the lower

epidermis.

Mesophyll tissue was differentiated into

two layers. Upper 1–2 layers of compactly

arranged pallisade parenchyma with oil

globules, and rich in chloroplast pigments.

Rarely with some rosette crystals of calcium

oxalate. Lower 3–5 layers of spongy

parenchyma cells, loosely arranged with air

spaces and loaded with prismatic, rosette

crystals of calcium oxalate.

Section through mid-rib showed a large

vascular bundle located at the centre, 1–3

layers of collenchyma tissue present at the

lower epidermis surrounding the ground tissue.

Ground tissue was made up of thin walled

compactly arranged parenchyma cells heavily

loaded by rosette crystals, prismatic crystal of

calcium oxalate and some oil globules.

Inner to the ground tissue, single layered,

somewhat elongated thin walled cells forming

endodermis. Inner to the endodermis 4–5 layers

of pericyclic fibers forming a ring like structure

covering the vascular bundle.

Vascular bundle was open and bi-collateral

in type. Phloem’s present around the xylem

with some phloem fibers and sieve elements

formed a ring like structure. The metaxylem

facing towards lower epidermis and protoxylem

facing towards upper epidermis. The xylem

bundles were separated by uniserrate medullary

rays along with xylem parenchyma and fibers.

Surface study of epidermis: (Plate B5–8)

Surface study of epidermis was carried out

to determine type and distribution of stomata,

epidermal cell and trichomes.

Stomata were absent in the upper epidermis

and consists only wavy epidermal cells. Some

of the trichomes and cicatrix were also

observed.

The lower epidermis consists of numerous

stomata of paracytic type. Some of the

trichomes and cicatrix were also observed.



PLATE-B

1. T.S through mid rib

2. Prismatic crystals

3. Vascular bundles with ground tissue

4. Xylem and phloem

5. Lower epidermis with 6. Stomata & cicatrix with

paracytic stomata epidermal cells

7. Upper epidermis with 8. Upper epidermis with trichome

cicatrix



PLATE- C

1. Lower epidermis with stomata 2. Measurements of stomata

3. Simple trichome 4. Crystal fibers

5. Prismatic crystal 6. Epidermal cells

7. Paracytic stomata 8. Annular & spiral vessels



TABLE 1: Quantitative microscopy of O. scandens leaf

Sr. No Parameter Result

1 Type of the stomata Paracytic

2 Length of the stomata 29.99 μm

3 Width of the stomata 22.43 μm

4 Outline of the stomata 673.19 μm2

5 Number of the stomata 12

6 Number of epidermal cells 24

7 Stomatal index 33

Quantitative microscopy: (Plate C1–2)

Quantitative microscopy of leaves was

carried out to determine epidermal cell number,

stomatal number, stomatal index and size of the

stomata.

The stomatal number, stomatal index,

stomatal size, epidermal cell size were

calculated by trial and error method (by taking

3–5 successive readings. Mean value was taken

into consideration.) Results are tabulated in

table – 1

Powder microscopy (Plate C3–8)

Powder microscopy of the dried leaf

powder was carried out following standard

guidelines. Organoleptic characters showed the

presence of greenish color with leafy odour and

bitter taste.

Microscopic characters

Diagnostic characters of powder

microscopy showed the paracytic stomata from

lower epidermis, epidermal cells of upper

epidermis, unicellular, simple horn shaped

trichomes from epidermis, rosette and prismatic

crystals of calcium oxalate from mesophyll and

ground tissue. Annular and spiral vessels from

vascular bundles, some of the brown content

(tannin) from ground tissue, crystal fibers and

lignified fibers.

CONCLUSION

Leaf of Olax scandens Roxb. (Olacaceae)

can be identified on the basis of key

microscopical characters like paracytic stomata,

unicellular, simple horn shaped trichomes,

rosette and prismatic crystals of calcium

oxalate, bi collateral vascular bundles, annular

and spiral vessels, tannin, crystal fibers and

lignified fibers. The quantitative surface

microscopy study showed 24 numbers of

epidermal cells, and stomatal index 33. These

observed parameters could be useful to

establish certain botanical standards for

identification and standardization of O.

scandens leaf.

ACKNOWLEDGEMENT

The authors are thankful to Director,

IPGT&RA, Gujarat Ayurved University,

Jamnagar and Department of AYUSH, for

providing financial support and other facilities

to carry out the research work. We express our

thankfulness to Mr. B. N. Hota, Rtd. DFO,

Govt. of Odisha; Mr. Pareswar Sahoo

Pharmacognosy expert; Mr. Malaya Das, Forest

Range Officer, Govt. of Odisha and other

traditional healer who helped us during drug

collection at Gandhamardan Hills, Balangir and

Bargarh, Odisha.



REFERENCES

Anonymous, (1990), Glimpses of medico-

botany of Bastar district, Madhya

Pradesh, CCRAS publication, pp. 114.

Anonymous, (1999), The Ayurvdic

Pharmacopoeia of India, Govt. of India

publication New Delhi, 1st edition, Vol.

I, Appendix 2.

Asia, Pvt. Ltd. and W.B. Saunders Company

Ltd., Edition 15, pp. 32, 33, 95–Johnson

Alexander Donald, (1940), Plant Micro

technique. New York, London,

Maccgrow Hill Book Company, pp.

105.

Khandelwal K.R., Kokate C.K., Gokhale S.B.

(1996). Practical pharmacognosy

techniques and experiments. Nirali

Prakashan, Pune, pp. 10–39.

Kirtikar & Basu, (2003), Indian Medicinal

Plants, Bishen Singh Mahendra Pal

Singh, Dehra Dun Vol. I, pp 568.

Saxena H.O, (1995), The Flora of Orissa,

Regional Research Laboratory,

Bhubaneshwar, 1st edition, pp. 288.

Trease GE, Evans WC (2002), Trease and

Evans Pharmacognosy, Harcourt brace

& Co. 99, 512, 547.

Tribhubana Panda, Rabindra N Pandhy, (2007),

Sustainable food habits of the hill

dwelling kandha tribe in kalanandi

district of Orissa, Indian journal of

traditional maedicine, vol. 6(1), pp.

103–105.

Veeramuthu Duraipandiyan, Muniappan

Ayyanar, Savarimuthu Ignacimuthu,

(2006), Antimicrobial activity of some

ethnomedicinal plants used by Paliyar

tribe from Tamil Nadu, India, BMC

Complementary and Alternative

Medicine, 6:35 doi:10.1186/1472-6882-

6-35

Wallis T E. (1985), Textbook of Pharmacognosy,

London Churchill Publication, pp.572–82

Source of Support: Department of AYUSH,

Govt. of India

Conflict of Interest: None Declared

Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 3 | March 2013 | 254–258



A CLINICAL EVALUATION ON RUJAKARA MARMA WITH SPECIAL

REFERENCE TO PAIN THRESHOLD

Benjwal Shobha1*

1Asst. Prof. Rachana Sharir Dept., M.S.M. Institute of Ayurveda, Khanpur Kalan, Sonipat Haryana



ABSTRACT

Marmas are vulnerable spots, constituting the essential aspect of surgico-anatomical knowledge.

While defining the type of marmas based on the effect or prognosis of the trauma, Acharya Sushruta

has classified five types of marmas. Among them he has appreciated pain as a residual effect of

trauma in Rujakara marma. This study was planned to analyze the tissues which were responsible for

pain in Rujakara marma & to evaluate pain-threshold relation with this marma. The study involved

60 healthy individuals who volunteered. According to anatomical sites, in Group I was taken to

evaluate pain threshold at 8 (Eight) Rujakara marmas (vital spots which have pain due to trauma)

sites of body and In Group II the same 60 individuals were taken to measure pain threshold at sites

other than Rujakara marmas. The results have shown that eight sites of Rujakara marma has

different types of fibrous Scleratogenic tissues having high pain threshold noceceptive impulses,

which might be the probable reason for Rujakara marmas to have a high pain-threshold in

comparison to other sites of the body.

KEYWORDS: Ruja, Painthreshold, noceceptive impulse

Research article

Cite this article:

Benjwal S (2013), A CLINICAL EVALUATION ON RUJAKARA MARMA WITH SPECIAL

REFERENCE TO PAIN THRESHOLD, Global J Res. Med. Plants & Indigen. Med.,

Volume 2(4): 254–258



INTRODUCTION

The Ayurvedic Science of Marma is itself a

treatise on Surgico-anatomical learning. The

concept of marma is a great contribution of

Sushruta, who mentioned 107 vital points in

various parts of the body, which should be

carefully dealt with during surgery & should

always be protected from injury, as the essence

of life (prana) rest in them (Sushruta 200 B.C).

Though general definition of Marma signifies

that every marma is the confluence of five

types of tissue, namely mamsa (muscle) Sira

(vessels), Snayu (ligaments) Asthi (bone) &

Sandhi (joints) (Charaka 200 B.C); but it is

evident from the description of injuries that the

traumatic effect or prognosis entirely depends

on the predominance of the tissue type at the

marma. Sushruta has classified Parinam

prakar marmas (residual effect of trauma) into

sadhyo pranhara (death on the spot after

trauma) Kalantara pranhara,(death occuring

after a short period post-trauma) Vishalyghana

(death occuring after removal of foreign body

from a traumatic wound) Vaikalyakara (there

will be a stable deformity in the body structure

post-trauma) & Rujakara. The vitiation of

Rujakara marma creates only the feeling of

pain and there is no condition of death or

morbidity (Sushruta 200 B.C). There are eight

points of Rujakara marma in the body out of

which Gulfa (Ankle joint) and Manibandh

(wrist joint) are sandhi & Kurchshira (brush

like structure) are of Snayu predominance. The

Rujakara marmas possess properties of Agni

(Fire) & Vayu (Air) Mahabhutas both of which

causes pain as a residual effect of trauma

(Sushruta 200 B.C). The perception of pain due

to trauma depends upon many factors like pain

receptors in the skin (mechanical, Chemical &

Thermal receptors) & rate of tissue damage

(Snell S.Richard, 1992). The differentiation

without discrimination was assessed on the

basis of gradation of pain-threshold & in the

present work this criterion was adopted for

differentiating tissues at different anatomical

sites to understand ‘Rujakara marma’ in a

better way. Hence a study was planned to

differentiate out the tissues responsible for

Rujakara marma and to study the pain

threshold at Rujakara marma sites & compare

it with pain threshold in other body parts.

MATERIALS

The research work was conducted on 60

healthy individuals selected from Indira Gandhi

Girls Hostel, P.G.Hostel of State Ayurvedic

College, Lucknow. Some of the observations

have been made by the researcher itself at the

neighbourhood. All the 60 healthy individuals

were taken in two groups:

Group I-Measurement of Pain threshold at site

of Rujakara marma sthana

Group II- Measurement of Pain threashold 5

cms proximal to the site of Rujakara marma

sthana

S.No Inclusion Criteria Exclusion Criteria

1 Individual between age of 18 – 40 years Individual below 18 or above 40 years.

2 Either Sex Pregnant women

3 Having no disease Any medical surgical co-morbidity

4 If taking any medicine

For this study a Proforma was prepared.

The information regarding name, age, sex and

occupation were noted down. The blood

pressure, temperature, respiratory rate & pulse

rate were also recorded. For the purpose of

measuring pain threshold following instruments



were used: Sphygmomanometer, Specially

designed blunt conical wooden object, Scale,

Stethoscope, Watch. (River.J, 1999).

METHOD

Measuring of Pain Threshold:

The subjects were asked to sit on a chair in

erect posture placing forearm in supinated

position on a table. A specially designed blunt

conical wooden object with cuff of

sphygmomanometer was kept on the site of

Rujakara marma i.e. at the wrist joint (for

manibandh marma) & just medial to the

tubercle of scaphoid at the palmar surface (for

kurchashira marma) (Solanki J.C.1982).

Thereafter, cuff was wrapped & air was

gradually pumped to produce, pressure pain. As

soon as the subject complained the pain, the

pressure necessary to produce that degree of

pain was recorded in terms of mm Hg. These

readings were utilized as the parameter of pain-

threshold. For the measurement of pain

threshold in lower extremities the wooden

conical object was placed on Ankle joint (For

Gulfa marma) & at the cross of line drawn

horizontally on the planter surface by joining

the medial conversity of medial cuniform bone

and base of the 5th

metatarsal bone and

vertically following the junction of 1st and 2

nd

toe (For Kurcha Shira marma) (Solanki

J.C.1982). For accuracy three readings were

taken at the same place at a 10 min interval. For

comparative studies, measurement of pain

threshold was done on other body parts i.e. 5

cm proximal to Rujakara marma site. Average

of all these readings of pain-threshold and

observations were statistically calculated.

Table-1 Statistical analysis of Pain Threshold in Group I and Group II

S.n

o Group I Group II ‘t’

value

‘p’

value

Name of

Rujakara

marma

Mean value

of pain

threshold

S.D 5cm proximal

to Rujakara

marma

Mean

value of

pain

threshold

S.D

1. Manibandh

(Wrist joint)

132.02 ± 46.23 Manibandh

(Wrist joint)

128.99 ± 48.58 18.52 <0.001

2. Gulfa Marma

(Ankle Joint)

151.71 ± 52.07 Gulfa Marma

(Ankle Joint)

118.25 ± 49.60 13.30 <0.001

3. Kurcha shira

(Upper

extremity)

130.27 ± 48.64 Kurcha shira

(Upper

extremity)

91.78 ± 45.68 7.14 <0.001

4. Kurcha shira

(lower

extremity)

170.65 ± 68.88 Kurcha shira

(lower

extremity)

120.95 ± 60.18 9.68 <0.001



Table-2 Comparative Assessment of Pain threshold in Group I and Group II

S.No. Name of the Site of Pain Threshold Mean value of Pain Threshold S.D

1. Rujakara marma 136 ± 6.23

2. Control site of Rujakara marma 128.99 ± 48.85

t value = 4.237

p value = <0.001

RESULTS

Table-1 indicates that the average pain

threshold at Manibandha (wrist joint) was

190.36 mm Hg with a SD of ± 56.40 and

124.22 mm Hg with a SD of ± 46.60 was found

at a site, 5 cm proximal from Manibandha

Marma (wrist joint). Comparative assessment

was statistically significant with ‘t’ value of

18.52 and p value < 0.001. The average of pain

threshold at Gulfa (ankle joint) was 151.71 mm

Hg with a S.D. of ± 52.07 and for sites other

than Gulfa (ankle joint) i.e., 5 cm proximal to

the Gulfa Marma (ankle joint) was lower with a

mean value 118.25 mm Hg with a S.D of

± 49.60. The difference observed was highly

significant, with ‘t’ value being 13.30 and p

value <0.001. The mean value of pain threshold

was 130.27 mm Hg with a S.D. of ± 48.48 and

for site other than Kurchshira Marma of Upper

extremity while it was 91.78 mm Hg with a

S.D. of ± 45.68 for the other sites on the palm

i.e. 5 cm distal from the Kurchashira Marma of

Upper extremity. The comparative assessment

shows the ‘t’ value being – 7.149 and p value <

0.001, which is highly significant. The mean

value of pain threshold at Kurchshira Marma

of lower extremity was 170.65 mm Hg with a

S.D. of ± 68.88 and other site on planter

surface 5 cm distol to Kurchshira Marma of

lower extremity while it was 120.95 mm Hg

with a S.D. of ± 60.18. The comparative

assessment shows the value were statistical

significant with a ‘t’ value of 9.685 and p

value < 0.001. In Table-2 the mean value of

pain threshold at Rujakara marma site was

136.02 mm Hg with S.D. of ± 46.23. The mean

value of pain threshold of control group of

Rujakara marma was 128.88 mm Hg with S.D

of ± 48.85. So it was observed that the average

pain – threshold of control site of Rujakara

marma was lower than the average pain

threshold at Rujakara marma sites. The

comparison between the two groups is highly

significant with a ‘t’ value of 4.237 and p value

< 0.001. The mean value of pain threshold at

Rujakara marma site was 136.02 mm Hg with

S.D. of ± 46.23. The mean value of pain

threshold of control group of Rujakara marma

was 128.88 mm Hg with S.D ± 48.85. So it was

observed that the average pain – threshold of

control site of Rujakara marma was lower than

the average pain threshold at Rujakara marma

sites. The comparison between the two groups

is highly significant with a ‘t’ value of 4.237

and p value <0.001.

DISCUSSION

Based on the result of vitiation of marma

sites, the marmas are divided into 5 types in

which the Rujakara marma belongs to least

morbidity (Sushruta, 200 B.C). Ruja (pain) is a

expression of body related to damage of tissue.

This is a psychosomatic phenomenon which is

different in every individual (www.iasp-

pain.org), which has been made the criteria in

this study to differentiate one site of tissue to

another site of tissue. This phenomenon has

been used as a basic tool by school of Sushruta

to classify Marmas (vital spot) on the basis of

result of the traumatic effect. All the tissues



such as Mansa (Muscle) Sira (vessels) Snayu

(ligament) Asthi (bone) Sandhi (joints) carry

noceceptor a biological sensor which is related

to noxious stimuli which is cast by mechanical

(direct trauma) biological irritation & thermal

The nonceceptive receptors are specialized

nerve endings in skin and deep tissues and

unlike other sensory receptors they are

activated at high threshold by a range of

potentially damaging stimuli (Grey’s, 1980).

This reveals that school of sushruta has

concerned nonceceptive pain as a criterion for

Rajukar marma. There are eight Rajukar

marmas being comparatively high

nonceceptive pain-threshold which also

indicates that it belongs to sclerotogenous pain

rather dermatogenic pain. The sclerotogenic

pain pattern is selective of ligament, tendon,

disc, periosteum & apophysial joint (Grey’s

1980) the anatomical places of Rujakara

marma carry one or more than one type of

sclerotogenous tissue like manibandh (wrist

joint) and gulfa (ankle joint) carry abundant

ligaments and kurch shira consists largely

tendons and all these have sclerogenatic pain

receptors having high pain threshold. (Snell S

Richard 1992). This discussion lead to

Sushruta’s observation in the form of Rujakara

marma that the vital parts are precisely of

sclerotogenous type of nonceceptive pain

receptor, bearing high threshold as compared to

other places where tendon, ligament & other

fibrous structures are lesser in quantum. The

Group II are largely consisting of muscle &

vascular tissue which are definitely having

lesser pain-threshold as compared to Group I.

CONCLUSION

The classified Rujakara marmas according

to school of Sushruta anatomically placed at

eight sites carrying different fibrous

sclerotogenic tissues, have comparatively high

threshold of noceceptive impulse. This

observation further draws the attention to the

subject of inquiry that whyManibandha (wrist)

has the highest pain threshold than Kurchashira

of upper extremity. This study opens the gate of

further research to prove these specific

observations made in present study.

REFERENCES:

Charak (200B.C) Chakrapani Tika

commentary by Tripathy Brahmanand

(2002) on Charak Samhita 6th

edition,

published by Chaukhambha Subharti

Prakashan 1999, Varanasi, Vol. II

Chikitsasthan Chapter-26th

page 720–

734.

Grey Henery (1980) Grey’s Anatomy by Peter

L Williams and Roger Warwick 36th

edition, Churchill Livingstone,

Published by Jarrold and Sons Ltd.

Snell.S.Richard (1992) Clinical Anatomy for

Medical student 4th

edition Chapter-9,

page- 487–495.

Solanki J.C, A Study on Rujakara marma with

Special Reference ot their anatomical &

surgical significance. Thesis M.D

Lucknow University, 1982.

Source: //www..iasp-pain.org/. Basic concept

of Pain physiology.

Sushruta (200 B.C.) Sushruta Samhita hindi

commentary by Ghanekar B.G.(1999),

14th

edition, published by Mehar Chand

Luxmi Chand Publication, Chapter-6th

page-184–189.

River J (1999) Adolescence pain measurement,

pain threshold. Journal of paediatrics

75(4):244–248





PHARMACOGNOSTICAL AND PRELIMINARY PHYTOCHEMICAL

INVESTIGATIONS ON DIFFERENT PARTS OF BULBOPHYLLUM

NEILGHERRENSE WIGHT. -AN ORCHID USED IN FOLK MEDICINE.

Kumari Harshitha1*

, Nishteswar K2, Harisha C R

3

1Ph D Scholar, Department of Dravyaguna, IPGT&RA, GAU, Jamnagar.Gujarat, India

2Professor and HOD, Department of Dravyaguna, IPGT&RA, GAU, Jamnagar. Gujarat, India

3Head, Pharmacognosy, IPGT&RA, GAU, Jamnagar. Gujarat, India

*Corresponding author: [email protected], Ph: 8460832302.


ABSTRACT

Bulbophyllum neilgherrense Wight. is an epiphyte belonging to the family Orchidaceae found

growing on medium to large sized host trees endemic to the forests of Western ghats which is yet to

be scientifically explored. A few published reports are available with regards to its medicinal uses

and scientific studies. In the present study, systematic pharmacognostic evaluation of leaf, stem and

root of the plant were carried out with respect to macroscopy, microscopy and preliminary

phytochemical screening. Macroscopic study detailed the structure of leaf, stem and root.

Microscopic study demonstrated the presence of mucilage cells and calcium oxalate crystals in leaf;

oil globules, tannin and mucilage cells in stem; velamen tissue, passage cells and lignified

parenchyma cells in root. Qualitative phytochemical investigations showed the presence of alkaloids,

tannin and phenol in methanol extract of stem and root whereas in leaf, constituents like alkaloids,

saponin glycosides, tannin, phenols and reducing sugar were observed. The morphological,

histological and phytochemical investigations reported in the paper may become supportive to

establish the authenticity of the plant simultaneously giving a wide scope for further pharmacological

and clinical researches.

KEY WORDS: Bulbophyllum neilgherrense, leaf, stem, root, pharmacognosy, phytochemical

studies.

Research article

Cite this article:

Kumari H, Nishteswar K, Harisha C R (2013), PHARMACOGNOSTICAL AND PRELIMINARY

PHYTOCHEMICAL INVESTIGATIONS ON DIFFERENT PARTS OF BULBOPHYLLUM

NEILGHERRENSE WIGHT. - AN ORCHID USED IN FOLK MEDICINE.,

Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 259–269




INTRODUCTION

Bulbophyllum neilgherrens Wight.

belonging to Orchidaceae family, commonly

known as Pottlekai in Kannada (Bhat

Gopalkrishna, 2003), Kalmel pullurvi in

Malayalam is abundantly available in Western

ghats (Rajendran A et al., 1997) and sparsely

distributed in Eastern ghats (Jadhav S N, 2003).

Forests of Karnataka, mainly Udupi (Bhat

Gopalkrishna, 2003), Belgaum (Gamble J.S,

2011), Malabar (Hooker J D, 1885) are the

places from where the species can be traced.

The plant is endemic to South India, which

occurs in plains and in higher elevations up to

900 m (Abraham and Vatsala, 1981).

The special characteristic of the plant is the

presence of pseudobulb for the preservation of

water and nutrients. Pseudobulbs are 3–3.5 cm

long and 2 cm across, smooth, green, four

angled. Progressing yellowing of pseudobulbs

is observed on ageing. Leaves 10–15cm long,

2–3 cm broad, coriaceous, elliptic to broadly

oblong, obtuse at apex, base narrowed tapering

into short petiole attached to the pseudobulb

(Bhat Gopalkrishna, 2003, Abraham and

Vatsala, 1981). Scape stout, from the base of

the pseudobulb, sheathed at the base, jointed

and with bract-like sheaths at the joints.

Flowers in racemes, petals small, pale yellow,

lip purple (Theodore cooke, 2006).

Karyomorphological study of this orchid

species has shown the chromosomal number to

be 19 (Abraham and Vatsala, 1981). The plant

is used by the folk people for restoration of

adolescence and as tonic in the form of juice

extracted from pseudobulb (Hossain MM,

2011). The paste prepared from pseudobulb and

leaf is consumed along with cow’s milk to treat

leucoderma (Rajendran A et al., 1997). In

certain regions of Karnataka, various parts of

this orchid are used by the villagers in the

management of heart diseases (Kumari

Harshitha, 2011).

Scarce information is available regarding

the phytochemical and pharmacological

profiles of this drug. The methanolic extract of

leaf showed the presence of flavonoids and

cyanogenic glycosides (Maridass M et al.,

2008). In vitro study of ethanol extract of leaf

and pseudobulb showed pronounced

antibacterial effect as compared to the effect

produced by chloroform and aqueous extracts

(Priya K et al., 2005).

No information is available regarding the

microscopical characters of the plant. In this

regard, pharmacognosy including transverse

section, powder microscopy, histochemical

tests of leaf, stem and roots with their

preliminary phytochemical investigations were

carried out with a view to establish purity and

standard of the sample.

MATERIAL AND METHODS

Plant material

Whole plant was collected from its natural

habitat in Puttur TQ, Karnataka, India during

the month of April 2012. The botanical identity

was confirmed by the botanist Dr. K.

Gopalakrishna Bhat, Professor of Botany

(Rtd.), Poorna prajna college, Udupi, India. A

herbarium specimen was preserved in the

Pharmacognosy lab, IPGT&RA, Gujarat

Ayurved University, Jamnagar with voucher

specimen number 6025\2012.

Macroscopical evaluation (Kokate et al.

2005):

The sample was cleaned and macroscopic

evaluation of whole plant was carried out. The

leaf, stem and root were then separated and

individual macroscopic characters like size,

shape, texture were noted in detail.

Microscopical evaluation (Trease and Evans

2009, Wallis 1985):

Free hand sections of leaf, stem and root

were taken and washed with chloral hydrate

solution. Sections were first observed in

distilled water then stained with phloroglucinol

and conc. HCl. Powder microscopy of shade-

dried powder was also carried out.

Photomicrographs were taken by Carl zeiss

trinocular microscope.



Histochemical tests (Krishnamurty 1988):

Thick sections were treated with various

reagents to locate chemical constituents i.e.

Tannin, mucilage, lignin and calcium.

Pharmaceutical evaluation (Anonymous,

2000):

Physicochemical parameters and

preliminary phytochemical investigations were

conducted on shade dried powders of leaf, stem

and root. The presence or absence of different

phyto-constituents in aqueous & methanol

extracts was detected.

RESULTS

Macroscopical characters- [Fig 1]

Bulbophyllum neilgherrens Wight. is an

epiphytic rhizomatous orchid with greenish

angled pseudobulbs bearing a single leaf at its

apex. Roots arise from the base of pseudobulb.

Scape is longer than the leaf which emerges

from the base of the pseudobulb. It is sheathed

at the base and sheath is also seen at the joints.

Inflorescence is raceme, drooping with many

flowers. Fruit is a capsule, green, globular-

elongate, 2 cm long, 5 angled with minute

seeds. [Fig 1A & 1B]

Description of leaf- Leaves are 8 cm–15 cm

long, 2–3 cm broad, coriaceous, elliptic to

broadly oblong, flattened, succulent, obtuse at

apex with narrow base. Midrib is prominent in

the ventral surface where as grooved on the

dorsal surface. Leaf margins are simple. Leaf

blade has parallel venation, tapers into short

petiole attached to the pseudobulb. A single

leaf emerges from the top of each pseudobulb.

[Fig 1C]

Description of stem- Horizontally creeping

stout somewhat rhizomatic, measuring about 6–

8 × 0.2–0.3 cm, rounded with slight ridges and

grooves, hard, light brown in colour with dark

brown scaly rings formed at successive

intervals. Nodal region is bulged, flattened

gives cone shaped structure inside the bulb.

Tuft of roots arise from the lower side of the

node. [Fig 1D]

Description of root- Tuft of thin roots arise

from the nodal region of the stem (adventitious)

measuring about 6–10 cm long, greenish to

light brown in colour, covered with delicate

fibrous type of absorptive tissue called

velamen, which is dead and perforated. When

the root becomes dry velamen part detaches

from the thin wiry central part which is

strongly attached to the stem. [Fig 1E]



Microscopical characters-

Leaf [Fig 2]

Leaf was isobilateral in nature. The upper

and lower epidermis was undifferentiated,

mesophyll tissue is filled with chloroplast

pigments consisting number of secretory cells.

Vascular bundles were centrally located. Upper

epidermis was single layered with compactly

arranged barrel shaped cells. At the midrib

portion the barrel shaped cells were

morphologically modified into tangentially

elongated compactly arranged cells resulting in

the formation of motor cells or hinge cells.

These epidermal cells were covered with thick,

ridged cuticle. Lower epidermis has only the

barrel shaped cells without motor cells, with

thick cuticle [Fig 2A]. Both the epidermis

possesses sunken stomata. Mesophyll was

spirally thickened near the epidermis with

banded parenchyma cells. Rest of the

mesophyll tissue was filled with

undifferentiated isodiametric parenchyma cells

containing numerous choloroplast, and was

compactly arranged. The mesophyll also

consists of some mucilage cavities. The

mesophyll also consists of raphide idioblast. In

midrib portion there was a centrally located

large vascular bundle and smaller vascular

bundles are passing through the main nerves

[Fig 2B]. Vascular bundle consists of phloem

towards lower epidermis, xylem towards upper

epidermis with few xylem elements with xylem

parenchyma and its fibres, where as phloem

with few sieve elements and fibres [Fig 2C].

Some of the mesophyll parenchyma consists of

reddish brown colour contents. The thick

cuticle and sunken stomata shows the

xerophytic nature of the plant, where as the

parenchyma cells, large mucilage cells show

the hydrophytic nature of the plant.

Powder microscopy: Diagnostic characters of

powder showed anisocytic stomata in epidermis

[Fig 2D], fibers from vascular bundles [Fig

2E], acicular crystals of calcium oxalate [Fig

2F], raphides, mucilage cells [Fig 2G], annular

thickened parenchyma cells, tannin from

mesophyll tissue and epidermal cells in surface

view.

Stem [Fig 3]

TS of stem showed the outer epidermis is

followed by hypodermis and the ground tissue

[Fig 3A]. Epidermis is made of single layered

compactly arranged barrel shaped cells without

intercellular spaces and is covered with thick

cuticle. Hypodermis made of 6–7 layers of

compactly arranged lignified sclerenchyma

cells seen below epidermis [Fig 3B].

Endodermis is made of abruptly placed barrel

to uneven shaped cells with thick lignified cells

followed by ground tissue. Ground tissue is

consisting of outer thin walled parenchymatous

zone, then pericyclic fibre zone and the central

region with scattered vascular bundles. Ground

tissue occupied 2/3rd

portion of the section

lying beneath the endodermis. 5–7 layers of

loosely arranged parenchyma cells without

intercellular spaces are seen consisting oil

globules, tannin [Fig 3C]. Some of the inner

parenchyma cells were lignified and pitted. 2–3

layers of lignified pericyclic fibres forming a

ring like structure were noticed. Number of

vascular bundles are scattered all over the zone.

Vascular bundles in peripheral zone are larger

in size than those in the central zone. Vascular

bundles are collateral and closed, each bundle

surrounded by a sheath which is more

conspicuous towards upper and lower side of

the bundle. The Vascular bundle consists of

xylem and phloem. Xylem consisting 2–3 large

metaxylem and small protoxylem with

tracheids and lysogenous cavity is present inner

to the protoxylem. Phloem consists of

conspicuous sieve tubes and companion cells.

Phloem parenchyma is not found. Mostly the

ground tissue is parenchymatous; some of the

lignified pitted parenchyma surrounds the

vascular bundles. Rest of the parenchyma cells

were also lignified and are thick walled [Fig

3D].

Powder microscopy: Powder showed the

presence of acicular crystals, tannin, fragments

of annular vessel, tracheids with oil globules

[Fig 3E], fibers [Fig 3F], fragments of

parenchyma cells, annular pitted vessel[Fig 3G]

and lignified parenchyma cells [Fig 3H].







Root [Fig 4]

TS of root showed outer velamen followed

by single layer of exodermis, leading into a

wide zone of cortex. Inner to it there was

circularly arranged single layer of endodermis

followed by pericycle and the vascular bundles

with alternatively arranged xylem and phloem,

forming central large pith [Fig 4A]. The outer

layer of the root showed velamen tissue. These

are dead cells, variously elongated, thick

walled and compactly arranged. Exodermis is

the outermost layer of the cortex, tangentially



arranged, thick walled and suberized. Few cells

which are unthickened are passage cells.

Beneath the exodermis many layers of (5–6)

loosely arranged parenchyma cells, consisting

chloroplast pigments, along with some air

chambers are present. Some of the parenchyma

cells have raphides, oil globules, starch grains

or yellowish brown tannin material [Fig 4B,

Fig 4C]. Endodermis is single layered with

compactly and circularly arranged barrel

shaped cells forming a ring, followed by single

layer of thin walled pericycle. Some of the

endodermal cells opposite to phloem are

thickened. Vascular bundles form polyarch,

radially arranged and exarch. Xylem vessels

with metaxylem towards pith and protoxylem

towards the endodermis. Xylem alternate with

the phloem, xylem consists of xylem

parenchyma and tracheids. The xylem

parenchyma is angular. Phloem consists of

sieve tube and companion cells. Pith is the

central most part of the root being angular

parenchymatous, thick walled and lignified

[Fig 4D].

Powder microscopy: Diagnostic characters of

powder showed the presence of lignified fiber

[Fig 4E], oil globules [Fig 4F], tannin and

starch grains [Fig 4G], acicular crystals,

tracheids [Fig 4H], mucilage containing cell,

fragments of pitted vessel [Fig 4I].

Histochemical tests-

The results of various histochemical tests

conducted on the leaf, stem and root powder

are depicted in Table I.

Pharmaceutical evaluation

The physicochemical parameters and

qualitative analysis results are enumerated in

Table II and Table III respectively.

Table: I showing Histochemical test results

Table: II showing results of physicochemical parameters

Sl. no Parameters Leaf Stem Root

1 Foreign Matter Nil Nil Nil

2 Loss on Drying % w/w 5.844 10.009 5.638

3 Total Ash Content% w/w 6.05 0.499 2.497

4 Acid Insoluble Ash % w/w 0.1 0.0998 0.4995

5 Water Soluble Extractive Value % w/w 33 3.7 4.7

6 Alcohol Soluble Extractive Value % w/w 5.9 6.8 3.7

7 PH 5 5 5

Sl.

no

Reagent Observation Characteristics Result

Leaf Stem Root

1. Phloroglucinol+Conc.

HCl

Red Lignified cells ++ ++ ++

2. Iodine Blue Starch grains − − ++

3. Phloroglucinol+Conc.

HCl

Dissolved Calcium oxalate

crystals

++ ++ ++

4. Fecl3 solution Dark blue to

black

Tannin cells ++ ++ ++

5. Ruthenium red Red Mucilage ++ ++ ++



Table : III showing results of phytochemical evaluation

Sl. no Parameters Leaf Stem Root

1 Alkaloids (M.E.)* + + +

2 Alkaloids (W.E.)* − − −

3 Saponin Glycosides (W.E.) + − −

4 Tannins &Phenols (M.E.) + + +

5 Tannins &Phenols (W.E.) + − −

6 Flavanoid (M.E.) − − −

7 Steroids (M.E.) − − −

8 Reducing sugar(W.E.) + − −

* M.E- Methanol extract; W.E- Water extract

DISCUSSION

B. neilgherrens Wight. is an epiphytic

orchid with stout rhizome modified into

pseudobulb to store moisture in excess so that it

can survive in unfavorable seasons. The plant

usually found growing in humid conditions of

moist deciduous and evergreen forests on tree

trunks of Anacardium occidentale (Cashew),

Syzigium cumini (Jamun) or other medium

sized to large trees. A very few indigenous

medicinal claims have been recorded on leaves

and pseudobulb of the plant. Limited number of

scientific studies has been reported on this

species with regard to structural, chemical,

pharmacological or clinical evaluation.

Pharmacognostical study reveals that the

leaf is isobilateral. Epidermis has some motor

cells and the mesophyll is undifferentiated with

spirally thickened parenchyma cells, which

shows that the cells are able to retain water for

long duration. The motor cells help the leaf to

roll due to the changes in their turgidity there

by reducing the stomatal transpiration under

xeric conditions. Presence of mucilage cells,

calcium oxalate crystals, sunken stomata and

the absence of trichomes are the important

characters of leaf. Oil globules and tannin

containing cells, calcium oxalate crystals,

mucilage containing cells are the important

characters found in rhizomatic stem. Velamen

tissues in roots during dry weather remains

filled with air and during rain quickly absorb

water. Passage cells serve as channels for flow

of water absorbed by the velamen. Lignified

parenchyma cells in the pith region of the root

are the other special characters. These may help

the species to adapt itself in the stress of

climatic variations.

Microscopic study of the plant powder

shows the presence of large quantities of fibers,

oil globules, tannin, and mucilage containing

cells. Histochemical tests of leaf, stem and root

shows the presence of lignin, tannin, calcium

and mucilage. Starch was detected only in root.

The physicochemical parameters shows that

loss on drying is more in stem as compared to

leaf and root which shows that higher moisture

content is in stem followed by leaf. Total ash is

more in leaf followed by root and less in stem.

Water soluble extractive value is more than five

times higher than alcohol soluble extractive

value in leaf showing the presence of more

water soluble constituents. Root also shows

higher extractive value in water where as stem

has higher extractive value in alcohol. PH

value

of leaf, stem and root did not show any

difference.

Among the qualitative assessment,

alkaloids were detected in methanolic extracts

of all three parts of the plant taken for

investigation, where as it could not be detected

in water extract. Stem and root shows the

presence of tannin and phenols only in

methanol extract while leaf contains tannin and

phenols in both water and methanol extract.

Water soluble extractives being highest in leaf,

also shows the presence of saponin glycosides

and reducing sugar in water extract.



Natural products with reservoirs of

structural and chemical entities will have

definite therapeutic relevance. Tannins are

reported as potential antiviral, antibacterial,

antiparasitic and hypolipidemic (Tannin.2013),

Pengelly Andrew 2004). Tannin and saponin

containing drugs demonstrated anti diabetic

activity

(Akhlaghi Farideh et al., 2012,

Pengelly Andrew 2004). Saponins, tannin and

phenols exhibit antibacterial activity (Doughari

JH et al., 2007). Research studies also

supported the view that phyto-constituents like

phenols, saponins, tannins, alkaloids exhibit

antioxidant, adaptogenic and antimicrobial

activities (Sukh Dev, 2006).

CONCLUSION

The studies carried out on the sample not

only established the appropriate data that may

be utilized for identification, but also

established the purity and standard of the plant

sample. Based on the reported phyto-

constituents some more pharmacological as

well as clinical studies may be carried out for

producing a proper scientific validation of the

folk orchid Bulbophyllum neilgherrense wight.

ACKNOWLEDGEMENT

The authors acknowledge Dr. K.

Gopalakrishna Bhat, Professor of Botany (Rtd.)

for his help in identification of this orchid

species and its authentication, Villagers of

Sullia TQ for sharing their knowledge and

experience on the plant.

REFERENCES

Abraham and Vatsala. (1981), Introduction to

Orchids. Tropical Botanic Garden and

Research Institute, Pp 533.

Akhlaghi Farideh, Rajaei Ziba, Mousa-Al-Reza

Hadjzadeh, Mehrdad Iranshahi and

Mahadi Elizabeth. (2012),

Antihyperglycemic effect of Asafoetida

(Ferula assafoetida Oleo gum resin) in

streptozotocin- induced diabetic rats,

World Applied Sciences Journal, 17 (2):

157–162.

Anonymous. (2000), Protocol for testing of

Ayurveda, Siddha and Unani

medicines.Ghaziabad: pharmacopoeial

laboratory for Indian medicines,

Department of Ayush, ministry of

health and family welfare, Government

of India.

Bhat Gopalakrishna K. (2003), Flora of Udupi.

Indian naturalist, Pp 913.

Doughari JH, Pukuma MS, De N. (2007),

Antibacterial effects of Balanites

aegyptica L. Drel. and Moringa

oleifera Lam. on salmonella typhi, Afr J

Biotechnol;6:2212–5.

Gamble J.S. (2011), Flora of the presidency of

Madras, Shiva offset press, Dehradun.

Pp 2017.

Hooker J D (1885), The Flora of British India,

L. Reeve & Co., Ltd.D-2, vol5, Pp 910.

Hossain M M. (2011), Therapeutic orchids:

traditional uses and recent advances —

An overview, Fitoterapia (82) pp 104.

Jadhav S N. (2003), Medicinal plants

conservation and sustainable utilization

project, EPTRI-ENVIS news letter,

vol.9 No.2.

Kokate C K, Purohit AP, Gokhale SB (2008)

Pharmacognosy (42nd

Edn), Nirali

Prakashan, Pune, pp 99.



Krishnamurty K.V. (1988), Methods in the

plant histochemistry. Vishwanadhan Pvt

Limited, Madras, Pp 1–77.

Kumari Harshitha. (2011), personal

communication with villagers of Sullia

TQ, Karnataka, India.

Maridass M, Zahir Hussain M I, Raju G.

(2008), Phytochemical survey of

orchids in the Thirunelveli hills of

South India, Ethnobotanical leaflets 12:

705–12.

Pengelly Andrew. (2004), Constituents of

Medicinal Plants. CABI publishing, Pp

31.

Priya K and Krishnaveni C. (2005),

Antibacterial effect of Bulbophyllum

neilgherrense Wt. (Orchidaceae). An

invitro study. Ancient science of life,

Vol XXV (2).

Rajendran A, Rama Rao N, Ravikumar K and

Henry A N. (1997), Some medicinal

orchids of Southern India. Ancient

Science of Life, Vol. No. 17 (1).

Sukh Dev. (2006), A selection of prime

Ayurvedic plant drugs Ancient-modern

concordance. Anamaya Publishers, Pp

501.

Tannin. (2013). Wikipedia, the free

encyclopedia.Retrieved February, 2013,

from

http://en.wikipedia.org/wiki/Tannin.

Theodore cooke. (2006), Flora of the

presidency of Bombay, Bishen singh

Mahendrapal Singh, Dehradun, Vol 2,

Pp 1083.

Trease G. E and Evans W.C. (2009),

Pharmacognosy, 16th

Ed. Saunders,

Elsevier. Pp 537–562.

Wallis T.E. (1985), Text book of

Pharmacognosy, 5th

Ed, CBS

Publishers, New Delhi, 1985. Pp.571–

578.


http://en.wikipedia.org/wiki/Tannin




A COMPARATIVE ACUTE TOXICITY EVALUATION OF ASHOKA

KSHEERAPAKA PREPARED FROM TWO DIFFERENT SPECIES OF

SARACA (S. ASOCA & S. THAIPINGENSIS)

Chavan Sulakshan S1*, Gamit R V

2, Ashok B K

3, Shukla V J

4, Das P

5, Ravishankar B

6

1Ph D Scholar in Ayurvedic Pharmacology, IPGT&RA, Gujarat Ayurveda University, Jamnagar, Gujarat,

India 2Laboratory Assistant, IPGT&RA, Gujarat Ayurveda University, Jamnagar.Gujarat, India

3Drug discovery lab, R & D, Himalaya drug company, Bangalore, Karnataka, India

4Head, Pharmaceutical Chemistry Lab, IPGT & RA, Gujarat Ayurveda University, Jamnagar, Gujarat, India

5Chairman, The Science Foundation For Tribal & Rural Resource Development, Bhubaneswar, Odisha, India

6Director, SDM Research Centre for Ayurveda and Allied Sciences, Kuthpady, Udupi, Karnataka, India



ABSTRACT

Ashoka (Saraca asoca) is an important Ayurvedic drug for treating gynecological disorders.

Hence it is economically important. There were reports that it has become quite scarce in several

localities and reported to be threatened in North Eastern Region of India. Ashoka bark widely

adulterated with other barks & or from same genus of different species. Ashoka Ksheerapaka is one

among clinical formulations of the plant Ashoka. Literature review revealed that no toxicity studies

have been undertaken on this formulation especially on Ashoka Ksheerapaka made from S. asoca &

S. thaipingensis. Because of this the present study was designed to evaluate Ashoka Ksheerapaka

made from two species for acute toxicity in Wistar strain albino rats as per OECD (Organization for

Economic Co-operation and Development) guideline 425 with 2000 mg/kg as limit test. On 1st day

test formulations were administered & observed for any toxicity changes for next 14 days. On 15th

day serum biochemical and hematological parameters were estimated. In all the three groups normal

weight gain was observed. Both the formulations did not produce any mortality up to the dose of

2000 mg/kg on oral administration. S. asoca increased the Monocyte percentage & Blood Sugar

Level (BSL) significantly, while in S. thaipingensis significant increase in Monocyte percentage &

significant decrease in the Platelet count in comparison to Normal Control group.The significant

increase in Monocyte percentage is in accordance with the property of estrogens to mediate its effect

through estrogen receptor in monocytes.

KEY WORDS: Ashoka, Ashoka Ksheerapaka, Acute toxicity, Saraca asoca, Saraca thaipingensis

Research article

Cite this article:

Chavan Sulakshan S, Gamit R V, Ashok B K, Shukla V J, Das P, Ravishankar B (2013), A

COMPARATIVE ACUTE TOXICITY EVALUATION OF ASHOKA KSHEERAPAKA PREPARED

FROM TWO DIFFERENT SPECIES OF SARACA (S. ASOCA & S. THAIPINGENSIS),

Global J Res. Med. Plants & Indigen. Med., Volume 2(4): 270–277



INTRODUCTION

Ashoka [Saraca asoca (Roxb.) Willd.] is

one of the most important Ayurvedic drug for

the treatment of various feminine disorders

especially in menorrhagia (Kirtikar & Basu,

2001). The word Ashoka means “without

sorrow”, a reference to reputation of it‟s bark

for keeping a woman healthy and youthful

(Shashikant Patwardhan, 2013). The natives

and traditional healers of Chhattisgarh use Sita-

Ashoka (the name given to Saraca asoca)

mainly in treatment of gynecological disorders

(Nalin, 2005). Its bark is bitter, astringent and

sweet in taste. It has stimulating effect on

endometrial and the ovarian tissue. It is useful

in internal bleeding, hemorrhoids, ulcers,

uterine affections, menorrhagia especially due

to uterine fibroids, meno-metrorrhagia,

leucorrhoea and pimples (Govind Das, 1970).

Dried stem bark of Saraca asoca (Roxb.)

Willd. is a genuine drug collected from wild or

cultivated trees, found in Central and Eastern

Himalayas, Western Ghats and Deccan

(Anonymous, 1986).

Number of studies has shown the

adulteration of Ashoka bark with barks of other

trees, but less on the trees of the same genus.

Ashoka bark is widely adulterated with barks of

Polyalthia longifolia, ocassionally bark of

Ashoka is mixed with Rohitaka bark

(Aphanamixis polystachya (Wall.) R.Parker)

and Caesalpinia pulcherrima (L.) Sw (Pradhan

P et al., 2009). Raw material from wild is

mostly collected by local people. There are

high and unintentional chances for a mistaken

identity of various other species of the same

Genus in the name of Ashoka. The drug from

the same genus but of different species is

difficult to identify. Such close resemblances of

Saraca asoca plant is observed with Saraca

thaipingensis. So it is imperative to carry out

the toxicological study of these drugs (spp.)

before being used in therapeutics. Since ancient

times, Ashoka is being used in Ayurvedic

preparations but comprehensive data on

majority of them is not available. Till date no

reports on the toxicological study on Ashoka

Ksheerapaka (a medicament prepared with

milk, water & plant drug) made from S. asoca

& S. thaipingensis Prain. are available. Hence,

this study was designed to evaluate Ashoka

Ksheerapaka made from two species of Saraca

i.e. S. asoca & S. thaipingensis for acute

toxicity in Wistar strain albino rats.

MATERIAL AND METHODS

Plant material

The dried bark of S. asoca and S.

thaipengensis were procured from Orissa from

authentic source and also correct identification

was made in Pharmacognosy laboratory

attached to the Institute. The herbarium

samples of these two species were deposited in

the laboratory (S. asoca- voucher specimen

no.6024 & S. thaipingensis- voucher specimen

no.6023). Dried barks were coarsely powdered

and stored in dry air tight container.

Preparation of Ksheerpaka

The Ksheerapaka (a medicament prepared

with milk, water & plant drug) was prepared

according to Acharya Sharangdhara by taking

one part drug material, adding cow‟s milk 8

times of bark then adding water 32 times of

bark, (i.e. 1:8:32). Ashoka bark powder was

weighed. Then, in stainless steel pot weighed

coarsely powdered bark of Ashoka was taken as

one part, then eight times of the bark cow milk

was added & 32 times of bark, water was added

in it. Then it was boiled till only milk remained

& water was evaporated (Tripathi Brahmanand,

2004).

Animals

Twenty Wistar strain female albino rats,

weighing 160 ± 20 g were taken from the

animal house attached to the institute (Institute

of PG Teaching and Research in Ayurveda,

Jamnagar). They were housed in polypropylene

cages with stainless steel cover meshes, at 22 ±

3°C with relative humidity of 50–60 %, on a 12

h natural day and night cycle. They were fed

with Amrut brand rat pellet feed supplied by

Pranav Agro Industries and with tap water ad

libitum. The experiments were carried out in



accordance with the norms of the Institutional

Animal Ethics Committee (IAEC), after

obtaining its permission (IAEC -04/09-10/PhD-

1).

Study protocol

Acute oral toxicity study for both the

samples was carried out as per OECD

(Organization for Economic Co-operation and

Development) guideline 425 with 2000 mg/kg

as limit test. Out of twenty animals 6 animals

were allotted to normal control (NC) group. In

both the test drug groups (i.e. S. asoca & S.

thaipingensis), single animals were dosed in

sequence usually at 48 h intervals. Using the

default progression factor, doses were selected

from the sequence 175, 550, and 2000 mg/kg

(because no estimate of the substance‟s

lethality was available, dosing was initiated at

175 mg/kg) as recommended in OECD

Guidelines 425. Food, but not water was

withheld for overnight before the experiment

and further 2 h after administration of test drug.

As there was no mortality observed even at

2000 mg/kg, additional 4 more animals were

dosed with 2000 mg/kg and observed for 14

days with different parameters. The animals

were observed continuously for 6 hours after

the dosing. The careful cage side observation

was done without disturbing the animal

attention and at the end of every hour the

animals were individually exposed to open

arena for recording the behavioural changes.

On 14th

day evening the rats were kept in

metabolic cages for fasting. On 15th

day body

weight of each animal was recorded. Blood was

collected by supra-orbital puncture with the

help of micro capillary tubes under mild ether

anesthesia for estimation of serum biochemical

and hematological parameters.

To estimate haematological parameters

0.08 ml blood was mixed with 0.02 ml of

EDTA (33.33 mg/ml) and fed to the auto

analyzer (Sismes KX-21, Trans Asia). The

parameters measured were; Total WBC count,

differential leucocyte count, Total RBC count,

haemoglobin content, PCV, MCV, MCH,

MCHC and platelet count.

For estimation of biochemical parameters,

serum was separated from collected blood and

requisite quantity of serum was fed to the auto

analyzer (Fully automated Biochemical

Random Access Analyzer, BS-200; Lilac

Medicare Pvt. Ltd., Mumbai) which was

automatically drawn in to the instrument for

estimating different parameters. Biochemical

parameters like blood sugar (BSL) (Pennock

CA et al., 1973), serum cholesterol (Roeschlau

P et al., 1974), serum triglyceride (Fossati P &

Prencipe L, 1982), HDL cholesterol, blood

urea, serum creatinine (Slot C, 1965), serum

glutamic pyruvic transaminase (SGPT) (Burtis

CA & Ashwood ER, 1999), serum glutamic

oxaloacetic transaminase (SGOT) (Tietz NW,

1995), serum total protein (Tietz NW, 1986),

serum albumin and serum globulin (Doumas

BT, 1972), serum alkaline phosphatase

(Wilkinson JH, 1969), total billirubin

(Pearlman PC & Lee RT, 1974), uric acid

(Kabasakalian P, 1973), were estimated.

Statistical analysis

The results are presented as Mean ± SEM.

The generated data were analyzed by

employing student‟s t test for unpaired data.

One way ANOVA was also employed with

Dunnets‟ multiple t test (DMTT) as post-hoc

test. For this purpose Sigma-stat software

(version 3.1) was employed.

RESULTS

In all the three groups normal weight gain

was observed (Table no. 1). Data pertaining to

the effect of test drug on WBC related

parameters are given in Table no.2. Both the

test drugs increased the Monocyte percentage

significantly in comparison to Normal Control

group. While a non-significant increase in Total

WBC count, Neutrophils percentage &

decrease in Lymphocyte percentage,

Eosinophils percentage was observed.



Table - 1 : Effect of test drug on Body weight (BW)

Group Body weight at different time duration % change in (BW)

Initial (g) On 7th

day (g) Final (g)

NC 161.67 ± 5.85 171.00 ± 6.98 173.00 ± 4.31 07.01 ↑

S. asoca 166.40 ± 8.45 174.40 ± 3.49 188.00 ± 4.20 12.98 ↑

S. thaipingensis 162.80 ± 7.79 172.40 ± 4.45 180.80 ± 7.97 11.06 ↑

(Data: Mean ± SEM, ↑ :- Increase, ↓ :- Decrease)

Table – 2 : Effect of test drug on WBC related parameters

WBC related

Parameter

Group

NC S. asoca S. thaipingensis

Total

WBC(/cumm)

5660.00 ± 647.77 6880.00 ± 561.61 (21.55↑) 7060.00 ± 553.72 (24.73↑)

Neutrophils(%) 22.40 ± 3.61 24.60 ± 3.83 (9.82↑) 26.00 ± 3.85 (16.07↑)

Lymphocytes(%) 71.40 ± 3.25 68.40 ± 3.96 (4.20↓) 67.20 ± 3.74 (5.88↓)

Eosinophils(%) 03.80 ± 0.58 03.60 ± 0.25 (5.26↓) 03.60 ± 0.25 (5.26↓)

Monocytes(%) 02.40 ± 0.25 03.40 ± 0.25* (41.67↑)

03.20 ± 0.20

* (33.33↑)

(Data: Mean ± SEM, The values in parenthesis are the percentage change in comparison to Normal Control

group. ↑ :- Increase, ↓ :- Decrease, * :- P< 0.05 by student‟s „t‟ test for unpaired data)

Table – 3 : Effect of test drug on RBC and Platelet related parameters

RBC and Platelet

related Parameter

Group


Total RBC

count(10e6/µl)

7.58 ± 0.19 7.57 ± 0.18 (0.05↓) 7.66 ± 0.26 (1.11↑)

Haemoglobin (g %) 14.24 ± 0.32 14.80 ± 0.57 (3.93↑) 13.90 ± 0.33 (2.39↓)

P.C.V. (%) 44.34 ± 1.14 44.22 ± 0.77 (0.27↓) 44.18 ± 1.13 (0.36↓)

MCV (fl) 58.50 ± 0.53 58.44 ± 0.91 (0.10↓) 57.76 ± 0.75 (1.26↓)

MCH (pg) 18.82 ± 0.20 18.78 ± 0.25 (0.21↓) 18.18 ± 0.28 (3.40↓)

MCHC (g/dl) 32.14 ± 0.45 32.14 ± 0.22 (0.00) 31.48 ± 0.24 (2.05↓)

Platelet count (10e3/µl) 1049.60 ± 67.01 1056.20 ± 62.54

(0.63↑)

663.00 ± 102.64*†

(36.83↓)


group. ↑ :- Increase, ↓ :- Decrease, * :- P< 0.05 by student‟s „t‟ test for unpaired data, † :- P< 0.05 by

ONE WAY ANOVA & Dunnet‟s Multiple „t‟ test as post-hoc test.)

Table – 4 : Effect of test drug on BSL and lipid profile

Parameter Group


BSL (mg/dl) 80.20 ± 4.76 100.00 ± 5.02*†

(24.69↑)

94.80 ± 5.72 (18.20↑)

S.Cholesterol (mg/dl) 65.20 ± 5.32 63.40 ± 2.16 (2.76↓) 67.40 ± 7.33 (3.37↑)

S. Triglyceride (mg/dl) 89.25 ± 6.47 92.20 ± 3.73 (3.31↑) 106.25 ± 7.16 (19.05↑)

HDL Cholesterol (mg/dl) 38.80 ± 2.31 36.80 ± 2.75 (5.15↓) 33.00 ± 4.01 (14.95↓)


group. ↑ :- Increase, ↓ :- Decrease, * :- P< 0.05 by student‟s „t‟ test for unpaired data, † :- P< 0.05 by ONE WAY ANOVA & Dunnet‟s Multiple „t‟ test as post-hoc test.)



Table – 5 : Effect of test drug on serum biochemical parameters

serum biochemical

Parameter

Group


Blood Urea(mg/dl) 54.60 ± 2.93 50.60 ± 1.50 (7.33↓) 65.60 ± 5.19 (20.15↑)

S.Creatinine(mg/dl) 0.62 ± 0.05 0.56 ± 0.07 (9.68↓) 0.58 ± 0.06 (6.45↓)

S.G.P.T.(IU/L) 42.60 ± 3.86 47.60 ± 4.62 (11.74↑) 36.40 ± 3.96 (14.55↓)

S.G.O.T.(IU/L) 126.20 ± 8.00 147.80 ± 11.16 (17.12↑) 121.60 ± 10.52 (3.65↓)

Total Protein(g/dl) 7.90 ± 0.19 7.80 ± 0.15 (1.27↓) 7.66 ± 0.39 (3.04↓)

Albumin(g/dl) 4.38 ± 0.07 4.38 ± 0.16 (0.00) 4.12 ± 0.26 (5.94↓)

Globulin(g/dl) 3.52 ± 0.12 3.42 ± 0.10 (2.84↓) 3.54 ± 0.31 (0.57↑)

A/G ratio 1.26 ± 0.03 1.26 ± 0.07 (0.00) 1.20 ± 0.13 (4.76↓)

Alkaline Phosphatase(IU/L) 134.60 ± 32.19 192.40 ± 19.51 (42.94↑) 169.60 ± 38.65 (26.00↑)

Bilirubin(T)(mg/dl) 0.38 ± 0.05 0.34 ± 0.04 (10.53↓) 0.34 ± 0.12 (10.53↓)

Uric Acid(mg/dl) 0.88 ± 0.09 1.26 ± 0.17 (43.18↑) 1.08 ± 0.20 (22.73↑)

(Data: Mean ± SEM, The values in parenthesis are the percentage change in comparison to Normal Control group. ↑ :- Increase, ↓ :- Decrease)

The effect of test drugs on RBC related

parameters is shown in Table no. 3. Both the

test drug treated groups did not show any

significant changes in RBC related parameters.

Test drug B decreases the Platelet count

significantly in comparison to NC group, while

the test drug S. asoca did not show any

significant changes.

Effect of test drug on BSL, lipid profile is

presented in Table no. 4. Animals from S.

asoca group exhibit significant elevation in the

BSL while non-significant up & downs were

observed in lipid profile in comparison to NC

group. Test drug S. thaipingensis did not

produce any significant changes in these

parameters.

Both the test drugs did not produce any

significant change in Blood Urea, S. Creatinine,

S.G.P.T., S.G.O.T. activity, Total Protein,

Albumin, Globulin, A/G ratio, Alkaline

Phosphatase activity & Bilirubin. (Table no. –

5)

DISCUSSION

Ashoka, which is an economically

important plant, has become quite scarce in

several localities and is reported to be

threatened in North Eastern Region of India

(Sharma PC et al., 2005; Alok Sharma, 2008).

Hence, it has become the target for adulteration

of its bark. A drug is taken for the beneficial

effect so it must not produce any toxicological

changes in the recipient‟s body. But till date no

study has been reported on the safety aspect of

Ashoka Ksheerapaka of S. asoca and S.

thaipingensis. 15 days after drug

administration, no mortality was observed

hence both the drugs are not lethal even at the

limited test dose. Normal weight gain shows

there are no serious effects on the body weight

which was common side effect in synthetic

estrogenic compound which were mostly used

in menstrual disorders (Olinda rola, 2012).

Both the drugs elevate the WBC count non-

significantly and no significant changes were

observed in Neutrophils percentage in

comparison to NC group values i.e. both the

drugs restricted the increase in WBC count and

Lymphocyte percentage which was observed

significantly in earlier studies on oral

contraceptive pills. Further, Monocyte

percentage in S. asoca group (41.64%) & S.

thaipingensis group (33.33%) was significantly

increased which is also in accordance with

property of estrogens, this finding suggests that

test drugs modulate the monocyte numbers and

its effect may be mediated through estrogen

receptor in monocytes (Sajida SH et al., 2006).



Both the test groups did not produce any

significant change in RBC related parameters

i.e. RBC count, Hb, MCV, MCH, MCHC.

Platelet count in S. thaipingensis group was

decreased significantly (36.83%), while in S.

asoca group no significant changes were

observed in comparison to NC group values. It

can be suggested that observed no significant

effect on Hb in both the test drugs was in

accordance with the earlier studies on oral

contraceptive pills (Sajida SH et al., 2006).

In S. asoca group significant increase in

BSL (24.69%) was observed in comparison to

NC group value while on lipid profile no

significant changes were observed. S.

thaipingensis did not produce any significant

changes on glycemic control (BSL) value and

on lipid profile. S. asoca lowers S. cholesterol

(2.76%) and HDL cholesterol (5.15%) non-

significantly in comparison to NC group

values. Both the test formulations did not

produce any significant changes in Blood urea,

S. creatinine, SGPT, SGOT, Total protein,

Albumin, Globulin, A/G ratio, Alkaline

phosphatase, Bilirubin levels. This indicates

that they do not have any toxicological

implication for acute administration even at

very high dose levels.

CONCLUSION

Both the species of Saraca i.e. S. asoca &

S. thaipingensis are safe at limited test dose

when administered orally in the form of

Ksheerapaka. However further toxicological

evaluation like chronic toxicity studies etc. are

required to provide complete safety profile.

Both the drugs modulate the Monocytes

percentage which may produce its estrogenic

effect by affecting the estrogenic receptor on

Monocytes which may be evaluated only after

detailed efficacy related study on these plants.

ACKNOWLEDGEMENTS

The authors are thankful to the authorities

of IPGT and RA, Gujarat Ayurved University

for providing facilities to carry out the research

work. One of the authors extends his deep

gratitude to Director General, CCRAS, New

Delhi & Dr. Sudesh Gaidhani, Dy. Director

(Pharmacology) CCRAS, New Delhi, for

providing fellowship.

REFERENCES

Alok Sharma, Shanker C, Lalit Kumar Tyagi,

Mahendra Singh, Ch.V.Rao (2008).

Herbal Medicine for Market Potential in

India: An Overview. Academic J. Plant

Sci; 1 (2): 26–36.

Anonymous (1986). The Ayurvedic

Pharmacopoeia of India, part I, Vol. I.

New Delhi: Ministry of Health and

Family Welfare, Department of

AYUSH, Govt. of India; pp. 14

Burtis CA, Ashwood ER, (eds) (1999). Tietz

Textbook of Clinical Chemisry, 3rd

edition, Philadelphia PA: Moss DW,

Henderson AR; pp. 652.

Doumas BT, Arends RL, Pinto PC (1972). In:

Standard methods of clinical chemistry.

Chicago: Academic Press; Vol 7. pp.

175–89.

Fossati P, Prencipe L (1982). Serum

triglycerides determined

colorimetrically with an enzyme that

produces hydrogen peroxide. Clin

Chem; 28:2077–80.

Govind Das (1970). Bhaishajya Ratnavali,

Streerogadhikara, 5/17–23, 114–116.

In: Pandit Shree Lalchandraji Vaidya

(ed.). Delhi: D.P.B. publications; pp.

698, 699, 704.



Kabasakalian P, Kalliney S, Wescott A (1973).

Determination of uric acid in serum,

with use of uricase and tribromophenol-

aminoantipyrine chromogen. Clin

Chem; 19:522.

Kirtikar KR, Basu BD (2001). Indian

Medicinal Plants. In: Blatter E, Caius J,

Mhaskar KS (eds), 2nd

ed. Dehra Dun:

Bishen Singh Mahindra Pal Singh

Publishers; vol 9. pp. 3068–3069.

Nalin (2005). Coment on PlantFiles: Red

Saraca, Ashoka Tree, SorrowlessTree

Saraca declinata cited, retrieved on

23.01.2013 from:

http://davesgarden.com/guides/pf/go/67

517/

Olinda rola (2012). Estrogen Side Effects – Do

You Know Them? retrieved on

17.08.2012 from:

http://www.healthguidance.org/entry/28

52/1/Estrogen-Side-Effects---Do-You-

Know-Them.html

PC Sharma, MB Yelne, TJ Dennis (eds)

(2005). Database on medicinal plants

used in Ayurveda, Vol. 3, New Delhi:

Documentation and Publication

Division, Central Council for Research

in Ayurveda & Siddha; p76–87.

Pearlman PC, Lee RT (1974). Detection and

measurement of total bilirubin in serum

with use of surfactants as solubilising

agents. Clin Chemi; 20:447.

Pennock CA, Murphy D, Sellers J, Longdon KJ

(1973). A comparison auto analyzer

method for the estimation of glucose in

blood. Clin Chim Acta; 48:193–201.

Pradhan P, Joseph L, Gupta V, Chulet R, Arya

H, Verma R, Bajpai A. (2009). Saraca

asoca (Ashoka): A Review. J. Chem.

Pharm. Res.; 1(1):62–71.

Roeschlau P, Bernt E, Gruber WA (1974).

Enzymatic determination of total

cholesterol in serum. Jour clinical Chem

clinical Biochem; 12:226.

Sajida SH, Sanaa MT, Amjad FA (2006). The

effect of oral contraceptive pills on

haematological indices, Tikrit Medical

Journal; 12(1): 65–69.

Tripathi Brahmanand (2004). Sharangdhar

samhita,Sharangadhara, Madhyama

khanda, Kwathadikalpana, 2/161. In:

Dr. Brahmanand Tripathi (ed.).

Varanasi: Chaukhamba Surabharati

Prakashan; pp. 159.

Shashikant Patwardhan (2013). Medicinal

Herb: Ashoka [Saraca Indica], retrived

on 23.01.2013 from: http://ayurveda-

foryou.com/ayurveda_herb/ashok.html

Slot C (1965). Plasma creatinine determination.

A new and specific Jaffe reaction

method. Scand J Clin Lab Invest;

17:381–87.

Tietz NW (1986). Text book of Clinical

Chemistry. W.B. Saunders; 579.

Tietz NW (1995). Clinical guide to laboratory

tests, 3rd

ed. Philadelphia, PA: WB

Saunders; pp. 76.

Wilkinson JH, Boutwell JH, Winsten S (1969).

Evaluation of a new system for kinetic

measurement of serum alkaline

phosphatase. Clin Chem; 15:487–95.


http://davesgarden.com/guides/pf/go/67517/

http://davesgarden.com/guides/pf/go/67517/

http://www.healthguidance.org/entry/2852/1/Estrogen-Side-Effects---Do-You-Know-Them.html



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