Anti oxidants for Cosmetics Dr. Panvipa Krisdaphong Mae Fah Luang University by Dean School of Cosmetic Science.

Anti oxidants for CosmeticsAnti oxidants for Cosmetics

Dr. Panvipa KrisdaphongDr. Panvipa Krisdaphong

Mae Fah Luang UniversityMae Fah Luang University

by

DeanDean

School of Cosmetic Science School of Cosmetic Science

* Outer beauty:Outer beauty:

Anti oxidantsAnti oxidants

Shining healthy hair and a clear radiant complexion.

The beauty that shines from inside.

Younger than your chronological age.

* Inner beauty:Inner beauty:

* Lasting beauty:* Lasting beauty:

Oral CosmeticsOral Cosmetics

Topical CosmeticsTopical Cosmetics

Topical Cosmetics &Topical Cosmetics & Oral CosmeticsOral Cosmetics

Aging

1. Genetic level – DNA

2. Cellular level - Free radical

RadicalRadicalToxic-chemicalToxic-chemical

SunSun

X-rayX-ray

PollutantPollutant

SmokeSmoke

DietDietDrugsDrugs

External SourceExternal Source

Anti-oxidationAnti-oxidation

Radical TheoryRadical Theory

1) Free radical source1) Free radical source

Immune destroy Immune destroy virus/bacterialvirus/bacterial

Activities enzymeActivities enzyme EnergyEnergy

Anti-oxidationAnti-oxidation

2) Dangerous of Free radical2) Dangerous of Free radical

Damage to cellsDamage to cells Damage to tissueDamage to tissue

Change protein structureChange protein structure Protein synthesis errorProtein synthesis errorAnti-oxidationAnti-oxidation

Free radical

Atom & molecule with one or more unpaired electron

- Reactive Oxygen Species (ROS)

1. Superoxide radical (O•2)

2. Hydrogen peroxide (H2O2)

3. Hydroxy radical (•OH) **

1. Superoxide radical (O•2)

Attack in body : - Enzyme

- Cell membrane : “lipid peroxidation”

O•2 + Unsaturated fatty acid lipid free radical

brack down to cell2. Hydrogen peroxide (H2O2)

Attack in body : - Nucleus (attack DNA : protein cross link)

3. Hydroxy radical (•OH) **

** Most potent oxidant

** Major causes of the change we see in aging

Attack in body : - Enzyme

- Proteins (attack DNA : most injurious to body overall)

- Carbohydrates

- Lipid

Hydroxy radicalPeroxide

all Reactive Oxygen Species

Hydroxy radicalPeroxideSuperoxide

Lipid peroxidation

Peroxide

Nucleus DNA attack

Cell membraneMitochondria**

Peroxisome

Cellular proteins

All reactive oxygen species (ROS)

Cell                                                                                      

Hydroxy radicalPeroxide

all Reactive Oxygen Species

Hydroxy radicalPeroxideSuperoxide

Lipid peroxidation

Peroxide

Nucleus DNA attack

Cell membraneMitochondria**

Peroxisome

Cellular proteins

All reactive oxygen species (ROS)

Cell                                                                                      

MitochondriaFunction : - Produce energy by oxidation.

- Produce free radicals - 1º sites for free radical damage

Ref : The free radical theory of aging , Nathan C. Nelson, Department of Physics, Ohio State University, Columbus, OH 43201

Source of oxidation

• Topical Cosmetic

• Oral Cosmetic

Anti oxidants for CosmeticAnti oxidants for CosmeticAnti oxidants for CosmeticAnti oxidants for Cosmetic

Topical CosmeticTopical CosmeticTopical CosmeticTopical Cosmetic

• PomegranatePomegranate

• Giant curcumaGiant curcuma

• RhubarbRhubarb

Botanical nameBotanical name : : Punica granatum Punica granatum L.L. Family name Family name : : PUNICACEAE PUNICACEAE Thai name Thai name : : Tab-tim (Tab-tim (ทั�บทั�มทั�บทั�ม) ) Chinese nameChinese name : : Shi Liu Pi Shi Liu Pi Part use Part use : : FruitsFruitsOrigin Origin : : Southwest Asia.Southwest Asia.Compound Compound :: Polyphenolic compoundPolyphenolic compound

PomegranatePomegranate

Traditional used :

1. Diarrheal remedy 1. Diarrheal remedy 2. Anthelmintic remedy2. Anthelmintic remedy

1. Anti-oxidant 1. Anti-oxidant 2. Anti-bacterial2. Anti-bacterial

Action :Action :

Antioxidant activity of pomegranate fruit extract and its Antioxidant activity of pomegranate fruit extract and its anthocyanidins: delphinidin, cyanidin, and pelargonidin anthocyanidins: delphinidin, cyanidin, and pelargonidin

MethodMethod : In vitro (rat brain) : In vitro (rat brain)

Examined Examined : ESR technique* with spin trapping : ESR technique* with spin trapping (Free radical scavenging activities)(Free radical scavenging activities)

ResultResult : - Pomegranate extract can : - Pomegranate extract can exhibited scavenging activity exhibited scavenging activity against against ..OH and OOH and O2 2

.. --..

- Anthocyanidins scavenged O- Anthocyanidins scavenged O22

·-·- in a in a dose-dependent dose-dependent

- The - The IDID5050 values** values** of delphinidin, cyanidin, and of delphinidin, cyanidin, and

pelargonidin were 2.4, 22, and 456 ų M pelargonidin were 2.4, 22, and 456 ų M

*ESR = Electron Spin Resonance (found that superoxide anion free radicals in mitochondria)** ID50 values = dose of inhibition

Ref : Noda Y, Kaneyuki T, Morl A and Packer L., Antioxidant activity of pomegranate fruit extract and its anthocyanidins: delphinidin, cyanidin, and pelargonidin. J Agric Food Chem. 2002 Jan 2 ; 50(1) : 166-71.

Antioxidant activityAntioxidant activity

Pomegranate (Punica granatum L.) fruits are widely consumed Pomegranate (Punica granatum L.) fruits are widely consumed as juice (PJ). The potent antioxidant and anti-atherosclerotic activities of as juice (PJ). The potent antioxidant and anti-atherosclerotic activities of PJ are attributed to its polyphenols including punicalagin, the major PJ are attributed to its polyphenols including punicalagin, the major fruit ellagitannin, and ellagic acid (EA)fruit ellagitannin, and ellagic acid (EA)

Sample Sample Antioxidant activityAntioxidant activity

Pomegranate Juice Pomegranate Juice ++++++++Total Pomegranate Tannin Total Pomegranate Tannin +++ +++Punicalagin Punicalagin ++ ++Ellagic acid Ellagic acid + +

Antioxidant activityAntioxidant activity

In vitro antiproliferative, apoptotic and antioxidant activities of punicalagin, ellagic acid and a total pomegranate tannin extract are enhanced in combination with other polyphenols as found in pomegranate juice. Seeram NP, Adams LS, Henning SM, Niu Y, Zhang Y, Nair MG, Heber D. Center for Human Nutrition, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA. [email protected]

Successive petroleum ether, chloroform, methanol and Successive petroleum ether, chloroform, methanol and waterextracts of waterextracts of Punica granatum Punica granatum Ž . were tested in vitro for their Ž . were tested in vitro for their antibacterial activity. antibacterial activity.

MethodMethod : In vitro (Agar dilution): In vitro (Agar dilution) Test onTest on : : Staphylococcus aureus MTCC 737

Escherichia coli MTCC 723 Klebsiella pneumoniae MTCC 109 Proteus ulgaris MTCC 1771 Bacillus subtilis MTCC 441 Salmonella typhi MTCC 537

Ref : Antibacterial activity of Punica granatum D. Prashanth, M.K. Asha, A. Amit Microbiology Laboratory, Research & Deelopment Centre, Natural Remedies Pt. Ltd., Plot No. 5B, Veerasandra Indl. Area, Hosur Road, Bangalore 561 229, India

Antibacterial activityAntibacterial activity

ResultResult : : Antibacterial activity of extractives from Antibacterial activity of extractives from Punica granatum Punica granatum fruit *fruit *

Tested material Tested material Minimum inhibitory concentration (mg/ml)Minimum inhibitory concentration (mg/ml)

Conclusion Conclusion : : All the tested extracts showed some antibacterial All the tested extracts showed some antibacterial activities which were significant for the methanolic extract, especially activities which were significant for the methanolic extract, especially against against P. ulgaris** P. ulgaris** and and B.subtilis***B.subtilis***..

** P. ulgaris : (urinary tract infections, infant diarrhea, respiratory infections)

*** B.subtilis : A common soil bacterium (illness causing diarrhea, nausea, vomiting)

S. aureus S. aureus E. coli E. coli K. pneumoniae P. ulgaris B. subtilis S. typhiK. pneumoniae P. ulgaris B. subtilis S. typhi

Petroleum ether extract Petroleum ether extract 12 12 12 12 12 12 12 12 12 12 1212Chloroform extract Chloroform extract 12 12 12 12 25 25 12 12 6 6 1212Methanol extract Methanol extract 6 6 12 12 12 12 1.5 1.5 6 6 1212Water extract Water extract 25 25 50 Not active 12 50 Not active 12 25 2525 25Chloramphenicol (Standard) 0.0025 0.005 0.005 0.005 0.005 0.0025Chloramphenicol (Standard) 0.0025 0.005 0.005 0.005 0.005 0.0025

*All determinations were done in triplicate.*All determinations were done in triplicate.

Ref: Pomegranate juice flavonoids inhibit low-density lipoprotein oxidation and cardiovascular diseases: studies in atherosclerotic mice and in humans. Aviram M, Dornfeld L, Kaplan M, Coleman R, Gaitini D, Nitecki S, Hofman A, Rosenblat M, Volkova N, Presser D, Attias J, Hayek T, Fuhrman B.Lipid Research Laboratory, Technion Faculty ofMedicine, Rappaport Family Institute for Research in the Medical Sciences, Rambam Medical Center, Haifa, Israel, 31096. [email protected]

Cardiovascular

MethodMethod : : in vitro in vitro - Effect of - Effect of Pomegranate on macrophage oxidative statusPomegranate on macrophage oxidative status. .

- Macrophages were incubated without (control) or with - Macrophages were incubated without (control) or with increasing conc. of pomegranate for 90 min at 37 C. increasing conc. of pomegranate for 90 min at 37 C.

Measurement Measurement :: Cellular oxidative stress following cell incubationCellular oxidative stress following cell incubation.. - Cellular fluorescence was determined (cytometry analysis)- Cellular fluorescence was determined (cytometry analysis)

As cellular oxidative stress influences both scavenger As cellular oxidative stress influences both scavenger receptor-mediated uptake of Ox-LDL and cellular cholesterol biosynthesis.receptor-mediated uptake of Ox-LDL and cellular cholesterol biosynthesis.

Result Result :: Pomegranate shown to Pomegranate shown to reduces cholesterol reduces cholesterol accumulationaccumulation..

- inhibit macrophage foam cell formation - inhibit macrophage foam cell formation - inhibit development of atherosclerotic lesions- inhibit development of atherosclerotic lesions

0 12 25 50 75

Pomegranate Juice Concentration (mmol of total polyphenols/L)Pomegranate Juice Concentration (mmol of total polyphenols/L)

300

250

200

150

100

50

0

C

ellu

lar

Oxi

dat

ive

Str

ess

Cel

lula

r O

xid

ativ

e S

tres

s(M

ean

Flu

ores

cen

ce I

nte

nsi

ty)

(Mea

n F

luor

esce

nce

In

ten

sity

)

(Control)(Control)

ConclusionConclusion : Demonstrates that upon in vitro incubation of : Demonstrates that upon in vitro incubation of macrophages with Ppomegranate, cellular uptake of Ox-LDL and cellular macrophages with Ppomegranate, cellular uptake of Ox-LDL and cellular cholesterol biosynthesis were cholesterol biosynthesis were significantly reduced parallel to a significant significantly reduced parallel to a significant reduction in cellular oxidative stress.reduction in cellular oxidative stress.

These effects were possibly responsible for the observed These effects were possibly responsible for the observed reduction in foam cell formation and atherosclerotic lesionreduction in foam cell formation and atherosclerotic lesion attenuation by attenuation by Pomegranate consumption .Pomegranate consumption .

Pomegranate reduces oxidative stress in macrophagesPomegranate reduces oxidative stress in macrophages

Application Application

Skin care product :Skin care product :

- Anti aging products - Anti aging products - Anti bacterial products- Anti bacterial products

Rhubarb

Botanical Name : Rheum Officinale Baill..

Common Name : Rhubarb

Part Used : Root, Rhizomes

Rhubarb Originated

This plant originated from the Asian such as

* Thailand

* China

ACTIVE INGREDIENTS

Anthraquinones

1,8-dihydroxyanthraquinone; R1=H, R2=H

Aaloe-emodin; R1=H, R2=CH2OH

Chrysophanol; R1=H, R2=CH3

Rhein; R1=H, R2=COOH

Emodin; R1=H, R2=CH3

Physicion; R1=OCH3, R2=CH3

The active ingredients in rhubarb are anthraquinnone such as rhein

Analysis of HPLC

In Cosmetics

The Rhubarb extract is use as

whitening products

Antioxidant Phenolic Constituents in Root of Antioxidant Phenolic Constituents in Root of Rheum officinaleRheum officinale

Method Method : : in vitroin vitro Radical scarvenging activities (RSA)Radical scarvenging activities (RSA)Determine byDetermine by : : Sample + ABTSSample + ABTS++ (radical compound)* (radical compound)*Detection by : Detection by : AbsorbanceAbsorbance

* 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid)* 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid)

AntioxidantAntioxidant

Ref:Cai Y, Sun M, Xing J, Corke H. Antioxidant phenolic constituents in roots of Rheum officinale and Rubia cordifolia: structure-radical scavenging activity relationships.J Agric Food Chem 2004 Dec 29;52(26):7884-90.

Hydroxyanthraquinone from roots of R. officinaleHydroxyanthraquinone from roots of R. officinale

NameName Content (%) Content (%) Radical scarvenging activities Radical scarvenging activities (mM)(mM)

Anthraquinone glycoside 6.8Anthraquinone glycoside 6.8 0.172 ± 0.0030.172 ± 0.003Aloe emodinAloe emodin 1.5 1.5 0.173 ± 0.0010.173 ± 0.001RheinRhein 1.9 1.9 0.174 ± 0.0010.174 ± 0.001EmodinEmodin 2.6 2.6 0.172 ± 0.0020.172 ± 0.002R. officinale Crude extract Alcoholic extractR. officinale Crude extract Alcoholic extract 88.6 ± 6.488.6 ± 6.4

AntioxidantAntioxidant

Ref:Cai Y, Sun M, Xing J, Corke H. Antioxidant phenolic constituents in roots of Rheum officinale and Rubia cordifolia: structure-radical scavenging activity relationships.J Agric Food Chem 2004 Dec 29;52(26):7884-90.

Result

MethodMethod :: in vitro in vitro MeasurementMeasurement : Determination of tyrosinase inhibitory activity : Determination of tyrosinase inhibitory activity

was performed by the dopachrome method* was performed by the dopachrome method* using L-DOPA as the substrate.using L-DOPA as the substrate.

ResultResult : : Rhubarb extract had 28.03 % tyrosinase inhibition**Rhubarb extract had 28.03 % tyrosinase inhibition** at 0.5 mg/ml.at 0.5 mg/ml.

*Dopachrome = intermidiate substances in the melanin biosynthesis**The potent tial tyrosinase inhibitor would show decrease dopachrome absorbtion

Tyrosinase inhibitory activityTyrosinase inhibitory activity

Evaluation of Rheum extract cream Evaluation of Rheum extract cream for whitening productsfor whitening products

1. 1. Ethical committee to ask for Ethical ClaranceEthical committee to ask for Ethical Clarance

2. 2. Seletion subjectsSeletion subjects

- Male or FemaleMale or Female- Age 20-35 yearsAge 20-35 years- No history of skin disease such as eczema or psoriasisNo history of skin disease such as eczema or psoriasis- No use the whitening products belong 6 monthNo use the whitening products belong 6 month- No use the whitening products between the testNo use the whitening products between the test

3. 3. Study designStudy design

DoseDose :: 3 % rhubarb extract cream3 % rhubarb extract creamOftenOften :: Twice dailyTwice dailyDurationDuration :: 8 weeks8 weeks * Compare with cream base* Compare with cream base

4. 4. Determination of skin conditionDetermination of skin condition

Before applicationBefore application :: Melanin valueMelanin valueAfter applicationAfter application :: Melanin value (Every 2 week Melanin value (Every 2 week

during the period of 8 weeks)during the period of 8 weeks)

5. 5. The The in vivoin vivo results results

ResultResult :Increase in parameter L*after application:Increase in parameter L*after application Positive depigmenting effectPositive depigmenting effect ConclusionConclusion :Rhubarb creams (3 % ww) significantly :Rhubarb creams (3 % ww) significantly

decreased skin color intensity compare decreased skin color intensity compare with controlwith control

L*parameter L*parameter - For clearity;from dark to light - For clearity;from dark to light - If the L*parameter is high, the skin color is light.- If the L*parameter is high, the skin color is light.

““Evaluation of the color of the treated area”Evaluation of the color of the treated area”

Chromameter® measurementChromameter® measurement

L :L : Clarity (dark to light) Clarity (dark to light)

a :a : Green to Red spectrum Green to Red spectrum

b :b : Blue to Yellow spectrum Blue to Yellow spectrum

These parameters are These parameters are exploited through the exploited through the calculation of the Individual. calculation of the Individual.

Typological Angle (ITA), Typological Angle (ITA), which defines the degree of which defines the degree of skin pigmentationskin pigmentation

The higher L parameter and the ITA are, the lighter the skin The higher L parameter and the ITA are, the lighter the skin

50

52

54

56

58

60

62

64

66

70

68

72

74

76

78

0

80

2 4 6 8 10 14 16 18 20 22 24 26 28 12

b* parameter

L* parameter

very light

light

medium

dark (dark)

very dark

5 5 °

41°

28°

10°

ITA

Principle Principle

““The chromameter converts colors into a digital code”The chromameter converts colors into a digital code”

Calculation formulasCalculation formulas

Variations (Variations () of the colorimetric parameters L*, b* and ) of the colorimetric parameters L*, b* and the ITA° were calculated in arbitrary units according to the the ITA° were calculated in arbitrary units according to the following formula:following formula:

Variations (Variations () = Ti - T0 ) = Ti - T0

Ti : value of the colorimetric parameters on the treated zone Ti : value of the colorimetric parameters on the treated zone

on D28 and D56on D28 and D56

T0: value of the colorimetric parameters on the treated zone T0: value of the colorimetric parameters on the treated zone

on D0on D0

Oral CosmeticOral CosmeticOral CosmeticOral Cosmetic

• ResveratrolResveratrol• Black paperBlack paper• EmblicEmblic• beta glucanbeta glucan

ResveratrolResveratrol

Source :Source : Bread fruitsBread fruits

Function:Function:

Dose:Dose:

• Anti-oxidantAnti-oxidant• Anti-cancerAnti-cancer• CardioprotectiveCardioprotective• Potential cholesterol lowering effectPotential cholesterol lowering effect

30 - 50 mg day30 - 50 mg day

Compound: Compound: Polyphenolic compoundPolyphenolic compound

Cardioprotective effectCardioprotective effect

• Inhibition of platelet aggregation

• Promotion of vasodilation by enhancing the production of nitric oxide

• Inhibition of inflammatory enzymes

Ref 1: Ray PS, Maulik G, Cordis GA, et al. The red wine antioxidant resveratrol protects isolated rat hearts from ischemia reperfusion injury. Free Rad Biol Med. 1999; 27:160-169Ref2:Pace-Asciak CR, Hahn S, Diamandis EP, et al. The red wine phenolics trans-resveratrol and quercetin block human platelet aggregation and eicosanoid synthesis: implications for protection against coronary heart disease. Clin Chim Acta. 1995; 235:207-219.

Anti oxidationAnti oxidationAntioxidant (on blood platelet function)Antioxidant (on blood platelet function)

It is a potent antioxidant, reactive oxygen species scavenger It is a potent antioxidant, reactive oxygen species scavenger and metal chelators. and metal chelators.

Resveratrol reduces lipid peroxidation, oxidation and Resveratrol reduces lipid peroxidation, oxidation and nitration of platelet and plasma proteins.nitration of platelet and plasma proteins.

Lipid Lipid peroxidationperoxidation

Nitration of Nitration of platelet platelet

Plasma Plasma proteinsproteins

Platelet recruitmentPlatelet recruitmentThrumbus formationThrumbus formation

Cardiovascular Cardiovascular diseasedisease

Oxidation of Resveratrol Oxidation of Resveratrol

(-)(-)inhibitinhibit

Inhibition of platelet aggregation

Method : in vitro induce platelet aggregation by collagen, thrombin, and ADP*

Dose : 10-1000 µmol/l resveratrolResult

InducerInducer % Platelete aggregation% Platelete aggregation

Control Resveratrol Resveratrol Resveratrol AspirinControl Resveratrol Resveratrol Resveratrol Aspirin (10 µmol/l) (100 µmol/l) (1000 µmol/l) (10 µmol/l) (100 µmol/l) (1000 µmol/l) (10 µmol/l) (10 µmol/l)

Thrombin Thrombin 82.982.912.5 64.0 12.5 64.0 13.913.9a a 43.6 43.6 6.36.3bb 30.6 30.6 5.25.2bb 52.5 52.5 4.64.6bb

ADP ADP 64.8 64.8 5.1 5.1 55.2 55.2 6.26.2aa 46.2 46.2 9.39.3bb 15.5 15.5 2.32.3b b 48.8 48.8 6.36.3bb

Collagen Collagen 74.0 74.0 4.0 4.0 60.6 60.6 4.84.8aa 47.2 47.2 7.17.1bb 26.9 26.9 4.34.3bb 43.9 43.9 4.74.7bb

aap<0.05, p<0.05, bbp<0.01, compared with control group.p<0.01, compared with control group.*ADP : Adenosin diphosphate*ADP : Adenosin diphosphate

Ref: Effects of red wine and wine polyphenol resveratrol onplatelet aggregationin vivo and in vitro ZHIRONG WANG1, YUANZHU HUANG1, JIANGANG ZOU1, KEJIANG CAO1, YINAN XU1 and JOSEPH M. WU21Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University, Nanjiang 210029, China;2Department of Biochemistry and Molecular Biology and Brander Cancer Research Institute,New York Medical College, Valhalla, NY 10595, USAReceived October 22, 2001; Accepted November 12, 2001

Vascular ProtectionVascular Protection

Resveratrol can protecting the vascular walls Resveratrol can protecting the vascular walls from from oxidation, inflammation, platelet aggregation, oxidation, inflammation, platelet aggregation, and thrombus formation.and thrombus formation.

In this review, we focus on the mechanism of In this review, we focus on the mechanism of resveratrol cardiovascular benefic effects. resveratrol cardiovascular benefic effects.

Ref : Resveratrol: preventing properties against vascular alterations and ageing.Delmas D, Jannin B, Latruffe N.University of Burgundy, Laboratory of Molecular and Cell Biology, Dijon, France.

Anti-cancer and Immune stimulatingAnti-cancer and Immune stimulating

Preliminary test are findings of some resveratrol-related Preliminary test are findings of some resveratrol-related anti-cancer and immune-stimulating effects. anti-cancer and immune-stimulating effects.

MethodMethod :: in vitroin vitro studies studies

ConclusionConclusion :: 1. Resveratrol can inhibit tumor initiation, 1. Resveratrol can inhibit tumor initiation, promotion and progression. promotion and progression.

2. inhibit DNA synthesis and ribonucleotide 2. inhibit DNA synthesis and ribonucleotide reductase in mammalian cells. reductase in mammalian cells.

Botanical nameBotanical name : : Piper nigrumPiper nigrum Linn. Linn.

Family nameFamily name : : PIPERACEAEPIPERACEAE

Part usedPart used : : SeedSeed

OriginOrigin : : Jungles of southwestern AsiaJungles of southwestern Asia

Black paperBlack paper

Antioxidant efficacyAntioxidant efficacy

Ref : Vijayakumar RS, Surya D, Nalini N.Department of Biotechnology, Annamalai Nagar, Tamilnadu, India.

MethodMethod :: In vivoIn vivo SubjectSubject :: 30 male rats.30 male rats.DurationDuration :: 10 weeks.10 weeks.ResultResult :: Supplementation with black pepperSupplementation with black pepper or piperine lowered TBARSor piperine lowered TBARS And CD levels and maintained SOD, CAT,And CD levels and maintained SOD, CAT, GPx, and GSH levels to near those GPx, and GSH levels to near those of control rats.of control rats.ConclusionConclusion :: Supplementation with black pepper orSupplementation with black pepper or the active principle of the active principle of black pepper, piperine,black pepper, piperine, can reduce high-fat diet induced oxidativecan reduce high-fat diet induced oxidative stress to the cells.stress to the cells.

Botanical Name Botanical Name :: Emblica officinalisEmblica officinalis Family name Family name :: EUPHORBIACEAE EUPHORBIACEAECommon name Common name :: Emblic EmblicThai nameThai name :: Ma-kham-pom Ma-kham-pom Part use Part use :: Fruits Fruits

Emblica officinalisEmblica officinalis

It grows in tropical and subtropical parts of China, It grows in tropical and subtropical parts of China, India, Indonesia and India, Indonesia and ThailandThailand

DescriptionDescription

Traditional medicine :Traditional medicine : Treat broad spectrum of disorders Treat broad spectrum of disorders

•TonicTonic

•RejuvenativeRejuvenative

•AstringentAstringent

•AntipyreticAntipyretic

•DiureticDiuretic

•Anti-tussive Anti-tussive

•ExpectorantExpectorant

•Anti-diarrhealAnti-diarrheal

• Ascorbic acidAscorbic acid

• Flavonoids Flavonoids

• TanninsTannins

• Alkaloids Alkaloids

• DiterpeneDiterpene

• TriterpeneTriterpene

• Benzenoid (Gallic acid)Benzenoid (Gallic acid)

CompositionComposition

In vitroIn vitro

- Anti oxidant - Anti oxidant

- Anti inflammatory- Anti inflammatory

- Anti bacterial - Anti bacterial

- Anti fungal - Anti fungal

- Anti viral - Anti viral

- Anti mutagenic- Anti mutagenic

In vivoIn vivo

- Ulcer protective activityUlcer protective activity

- Anti diabetic activityAnti diabetic activity

- Reduce lipid levelReduce lipid level

- Protective liver injuryProtective liver injury

- Immuno-modulatoryImmuno-modulatory

- Anti-oxidantAnti-oxidant

Scientifically published dataScientifically published data

Active CompoundActive Compound

HO

HO

HO

OH

O

O

HO OHOH

HOOO

OO

OH

HO

HO

O

R1R2

O

C

OH

HO

HO

Gallic acidGallic acid Hydrolyzable tanninHydrolyzable tannin

Anti-oxidation of EmblicAnti-oxidation of Emblic

EnvironmentEnvironment

Reactive Oxidation Species Reactive Oxidation Species (ROS) (ROS) OO••

22, H, H22OO22 ,, ••OHOH

Damage of the cellDamage of the cell

Free radicalsFree radicalsOxidative StressOxidative Stress

Emblic extractEmblic extract

Anti-oxidationAnti-oxidation

(-) SOD, Catalase etc.(-) SOD, Catalase etc.

Ref : Sabu MC, Kunttan R. Antidiabetic activity of medical plant and its relation ship with their antioxidant activity. J Ethanopharmacol 2002;81:155-60

ActivityActivity : : Anti-oxidantAnti-oxidant

MethodMethod :: In vitroIn vitro

MaterialMaterial :: Methanolic extractMethanolic extract

RResesultult :: The concentration needed for the The concentration needed for the inhibition of hydroxyl radical inhibition of hydroxyl radical scavenging were 155.5 mg/ml, scavenging were 155.5 mg/ml, and that for superoxide scavenging and that for superoxide scavenging activity were found to be 6.5 mg/mlactivity were found to be 6.5 mg/ml

Scientifically published dataScientifically published data

Ref : Characterizing the antioxidant activity of amla(Phyllanthus emblica) extractS. M. Khopde†, K. Indira Priyadarsini†,H. Mohan†, V. B. Gawandi†,J. G. Satav‡, J. V. Yakhmi†, M. M. Banavaliker**, M. K. Biyani** and J. P. Mittal†,*,#Chemistry Group, and ‡Radiation Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India**Ajanta Pharma Ltd, Kandivali, Mumbai 400 067, India#Also with the Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore 560 064, India

ActivityActivity : : Anti-oxidantAnti-oxidantMethodMethod :: In vivoIn vivoTest on Test on :: rat liverrat liverDoseDose :: 5-500 mg/ml 5-500 mg/ml ResultResult :: EmblicEmblic extracts are able to fully extracts are able to fully

suppress tumer cell after suppress tumer cell after 4 days.4 days.

The antioxidant activity of the extract was found The antioxidant activity of the extract was found to be concentration-dependent.to be concentration-dependent.

Scientifically published dataScientifically published data

Antioxidant ActivityAntioxidant Activity

Method Method :: In vitro In vitro (DPPH method)(DPPH method) Determined :Determined : free radical scavenging by measuring antioxidant free radical scavenging by measuring antioxidant

activity (DPPH)activity (DPPH)

Detection Detection :: UV Spectrophotometer and ESR Spectrometer UV Spectrophotometer and ESR Spectrometer

Sample Sample :: 3 fraction3 of3 fraction3 of E. offcinalis E. offcinalis fruit extract, gallic acid fruit extract, gallic acid and ascorbic acidand ascorbic acid

The ability of three fractions ofThe ability of three fractions of E. offcinalis E. offcinalis fruit fruit

extract, gallic acid and ascorbic acid for antioxidant activityextract, gallic acid and ascorbic acid for antioxidant activity

In vitroIn vitro Antioxidant Activity in Antioxidant Activity in Emblica offcinalis Emblica offcinalis Extract against DPPH and measured by Extract against DPPH and measured by UV SpectrophotometerUV Spectrophotometer

E. officinalisE. officinalis hexane extract hexane extract617.84 617.84 g/mlg/mlE. officinalisE. officinalis ethyl acetate ethyl acetate extractextract 2.072.07 g/mlg/mlE.officinalisE.officinalis methanol extract methanol extract

3.82 3.82 g/mlg/mlAscorbic acidAscorbic acid 4.34 4.34

g/mg/mGallic acidGallic acid 2.48 2.48 g/mlg/ml

CompoundCompound IC 50 IC 50

Result Result (antioxidation)(antioxidation)

In vitroIn vitro Antioxidant Activity in Antioxidant Activity in Emblica offcinalis Emblica offcinalis Extract against DPPH and measured by Extract against DPPH and measured by ESR SpectrometerESR Spectrometer

E. officinalisE. officinalis hexane extract hexane extract 750.58 750.58 g/mlg/mlE. officinalisE. officinalis ethyl acetate ethyl acetate extractextract 2.662.66 g/mlg/mlE.officinalisE.officinalis methanol extract methanol extract

6.02 6.02 g/mlg/mlAscorbic acidAscorbic acid 8.028.02

g/mg/mGallic acidGallic acid 3.673.67 g/mlg/ml

CompoundCompound IC 50 IC 50

Result Result (antioxidation)(antioxidation)

Conclusion Conclusion (antioxidation)(antioxidation)

Three fractions of Three fractions of E. officinalisE. officinalis extract, gallic acid and extract, gallic acid and ascorbic acid showed ascorbic acid showed antioxidant activity against free radical antioxidant activity against free radical (DPPH)(DPPH) both in measure by UV Spectrophotometer and ESR both in measure by UV Spectrophotometer and ESR

spectrometer.spectrometer.

The The antioxidant activity. antioxidant activity.

Ethyl acetate ext. Ethyl acetate ext. gallic acid gallic acid methanol ext. methanol ext. ascorbic acid ascorbic acid hexane ext. hexane ext.

Skin lightening Skin lightening

50

52

54

56

58

60

62

64

66

70

68

72

74

76

78

0

80

2 4 6 8 10 14 16 18 20 22 24 26 28 12

b* parameter

L* parameter

very light

light

medium

dark (dark)

very dark

5 5 °

41°

28°

10°

ITA

Evaluation of Evaluation of Emblica officinalisEmblica officinalis extract cream extract cream

Method Method :: In vivo In vivo

Subjects Subjects :: 20 subjects (20-60 yr.) 20 subjects (20-60 yr.)

Instrument Instrument :: Chromameter CR 300 Chromameter CR 300

Sample Sample :: 3 % 3 % Emblica officinalis Emblica officinalis extract cream extract cream

The clinical efficacy of The clinical efficacy of E. officinalisE. officinalis formulations were formulations were evaluated in a placebo-controlled trial using cream base as control. evaluated in a placebo-controlled trial using cream base as control.

ResultResult

The facial skin conditions were evaluated pre and post The facial skin conditions were evaluated pre and post application, determined L- value.application, determined L- value.

E. officinalisE. officinalis extract at extract at 3 % w/w showed lightening 3 % w/w showed lightening effect start at 4 th week.effect start at 4 th week.

L-value

L*parameter - For clearity;from dark to light (If the L*parameter is high, the skin color is light.)

- Beta glucan is polysaccharide derived from the cell wall of bacterial, fungal, yeasts and cereals.

- Beta glucan has many structure depend on source.

Beta-glucan

“Beta glucan of yeast (Saccharomyces serevisae) is mainly of 1,3/1,6-β-D-glucose”

Structure of Beta-glucan

Application• Immunostimulation

• Cholesterol reduction

• Vaccine adjuvants

• Cosmetics for skin repair

• Improving food products

• Feed supplements

Application• Anti oxidantion

• Anti inflammation

Application of beta glucan for Immunostimulation

Application of beta glucan for Immunostimulation

Effect of oral administration b-glucan on cytokines

JANA (2002)

Application of beta glucan for Immunostimulation

0

200

400

600

800

1000

1200

IL-2 IFN-g TNF-a

Cytokine

Cyto

kin

e level (

pg

/ml)

Control

beta-glucan - 6-wk-old BALA/c mice.

- Tumor cells were injected into abdominal wall.

- Adding beta-glucan (28.4 mg/kg) for 21 days.

IL-2

(Interleukin 2 )

IFN-

(Interferon-γ)

IFN-

(Interferon –

00.5

11.5

22.5

33.5

44.5

55.5

66.5

77.5

88.5

99.510

1 2 3 4

To

tal c

ho

leste

rol:

HD

L c

ho

leste

rol

Mean changes in ratio of total to HDL-cholesterol concentrations during supplementation with beta-glucan

Am J Cli Nutr (1999)

128Baseline 7 Time (wk)

Application of beta glucan for cholesterol reduction

15 obese men ages 20 to 60 with high cholesterol

Supplement with 7.5 g of beta-glucan in orange juice twin daily for eight weeks

Application of beta glucan for Cosmetics for skin repair

Davis (1992 )and Donzis (1993) were used beta glucan that purified from yeast cell to cosmetic composition. The products were reduced skin irritation, lesions, cracks and dryness.

Greene (2001) was applied beta-glucan to cosmetic to for the protection of skin from damage caused by ultraviolet radiation.

Application of beta glucan for improving food products

New value products to improve health (Wheatcroft et al., 2002)- Yoghurt - Fruit juice- Salad dressing - Many food products

Improving propertied food products (Thammakiti et al., 2004)

- Thickening

- Water-holding agent

- Emulsifying stabilizer

- Fat replacer