Free Radical Scavenging Activity of Free Radical Scavenging Activity of Some Malaysian Medicinal PlantsSome Malaysian Medicinal Plants

Muhammad TaherMuhammad Taher

Faculty of Pharmacy, International Islamic University Malaysia

19th CPA-MPS 2007, KUALA LUMPUR

Presentation layoutPresentation layout

IntroductionIntroduction ObjectiveObjective MethodologyMethodology Results and discussionResults and discussion ConclusionConclusion

INTRODUCTIONINTRODUCTION

AntioxidantAntioxidant

““Any substance, when present at low Any substance, when present at low concentrations compared with those concentrations compared with those of an oxidizable substrate, of an oxidizable substrate, significantly delays or prevents significantly delays or prevents oxidation of that substrate “oxidation of that substrate “

Free Radicals in Biological Medicine, 3rd ed. 1999

Antioxidant actionAntioxidant action

The antioxidant can act by the The antioxidant can act by the following mechanisms: reducing following mechanisms: reducing activity, free radical-scavenging, activity, free radical-scavenging, potential complexing of pro-oxidant potential complexing of pro-oxidant metals and quenching of singlet metals and quenching of singlet oxygen. oxygen.

Tissue damage resulting from free Tissue damage resulting from free radical may cause a variety of radical may cause a variety of disorders, including degenerative disorders, including degenerative disorders of the CNS, such as disorders of the CNS, such as Alzheimer’s disease, cancer, Alzheimer’s disease, cancer, atherosclerosis, diabetes mellitus, atherosclerosis, diabetes mellitus, hypertension, AIDS and aginghypertension, AIDS and aging

((Halliwell and Gutteridge, 1998 and and Mantle et al., 2000). ).

Effect of free radicalEffect of free radical

Plant materialsPlant materials

1.1. Andrographis paniculata Andrographis paniculata (leaves),(leaves), 2.2. Averrhoa belimbi Averrhoa belimbi (fruits),(fruits), 3.3. Eurycoma longifolia Eurycoma longifolia (roots),(roots), 4.4. Dipteris lobbiana Dipteris lobbiana (leaves)(leaves), , 5.5. Ficus deltoidea Ficus deltoidea (leaves),(leaves), 6.6. Fagrea obovata Fagrea obovata (roots),(roots), 7.7. Momordica subanguenta Momordica subanguenta (fruits)(fruits),,8.8. Morinda citrifolia Morinda citrifolia (fruits),(fruits), 9.9. Orthosiphon stamineus Orthosiphon stamineus (leaves)(leaves), , 10.10. Smilax myosotiflora Smilax myosotiflora (rhizome) (rhizome)

Traditional UsesTraditional Uses

Aphrodisiac and energy enhancerAphrodisiac and energy enhancer

Eurycoma longifolia, Smilax myosotiflora, Eurycoma longifolia, Smilax myosotiflora, Dipteris lobbiana, Ficus deltoidea and Dipteris lobbiana, Ficus deltoidea and Fagrea obovataFagrea obovata

Antidiabetes mellitusAntidiabetes mellitus

Momordica subanguenta, Orthosiphon Momordica subanguenta, Orthosiphon stamineus, Andrographis paniculata, stamineus, Andrographis paniculata, Averrhoa belimbi and Morinda citrifolia:Averrhoa belimbi and Morinda citrifolia:

Eurycoma longifoliaEurycoma longifolia

Tongkat Ali/Malaysian ginsengTongkat Ali/Malaysian ginseng

Traditional uses: tonic, Traditional uses: tonic, poultice for poultice for headaches, wounds & headaches, wounds & ulcers, febrifuge, ulcers, febrifuge, hepatitis, aphrodisiachepatitis, aphrodisiac

Andrographis paniculataAndrographis paniculataHempedu bumi (‘bile of the earth’), akarHempedu bumi (‘bile of the earth’), akar

cerita cerita

Traditional uses: Roots Traditional uses: Roots and leaves used and leaves used against snake bites, against snake bites, decoction of leaves decoction of leaves against stomach-ache, against stomach-ache, dysentery, typhus and dysentery, typhus and bronchitis, as a bronchitis, as a poultice on swollen poultice on swollen legs, vitiligo, piles and legs, vitiligo, piles and antidiabetesantidiabetes

Orthosiphon aristatusOrthosiphon aristatusMisai kucing, kumis kucing, ruku hutan Misai kucing, kumis kucing, ruku hutan

Traditional usesTraditional uses: : diabetes and kidney diabetes and kidney related diseases, related diseases, diuretic, regulate diuretic, regulate blood pressureblood pressure

Morinda citrifoliaMorinda citrifolianoni, mengkudunoni, mengkudu

Traditional Uses: Traditional Uses: to to treat diabetes, treat diabetes, haemorrhages and haemorrhages and coughs. coughs.

ObjectiveObjective

The research is aimed to study the The research is aimed to study the relationship of traditional usage of relationship of traditional usage of medicinal plants and their free radical medicinal plants and their free radical scavenging activityscavenging activity

METHODOLOGYMETHODOLOGY

1. Extraction1. Extraction

50 g of powdered plants were extracted with 50 g of powdered plants were extracted with EtOH:Water (50:50) (500 mL) and freeze dryedEtOH:Water (50:50) (500 mL) and freeze dryed

2. Phenolic contents was determined by Folin-2. Phenolic contents was determined by Folin-Ciocalteu reagent (McDonald et al., 2001)Ciocalteu reagent (McDonald et al., 2001)

3. Flavonoid contents was determined by aluminum 3. Flavonoid contents was determined by aluminum chloride colorimetric method (Siddhuraju and chloride colorimetric method (Siddhuraju and Becker, 2003)Becker, 2003)

4. Free radical scavenging activity was determined by 2,2-diphenyl-1-4. Free radical scavenging activity was determined by 2,2-diphenyl-1-picrylhydrazyl radical (DPPHpicrylhydrazyl radical (DPPH·· ) scavenging ) scavenging

X= Absorbance of sample at 517 nmX= Absorbance of sample at 517 nm

Y= Absorbance of control at 517 nmY= Absorbance of control at 517 nm

%1001% xY

XScavenging

a. Spectrophotometric assay

b. Thin layer chromatography test (yellow spots against purple background)

RESULTS AND DISCUSSIONRESULTS AND DISCUSSION

Table 1. Percentage of soluble materials Table 1. Percentage of soluble materials extracted from the plants extracted from the plants

Sample (Abbreviation) Local name Yield (% w/w)

Averrhoa belimbi (AB) Belimbing buloh 39.8

Fagrea obovata (FO) Tengkuk biawak 24.3

Dipteris lobbiana(DL) Janggut ali 17.9

Momordica subanguenta (MS) Peria katak 16.6

Andrographis paniculata (AP) Hempedu bumi 14.4

Orthosiphon stamineus (OS) Misai kucing 11.4

Ficus deltoidea(FD) Mas cotek 8.9

Smilax myosotiflora (SM) Ubi jaga 7.9

Morinda citrifolia (MC) Mengkudu 7.7

Eurycoma longifolia (EL) Tongkat ali 2.8

Table 2. Total phenolic and flavonoids contents of Table 2. Total phenolic and flavonoids contents of the samplesthe samples

Sample Total phenolic (mg /g) Total flavonoid (mg/g) % DPPH

Andrographis paniculata

43.58 2.47 19.0

Averrhoa belimbi 40.68 14.88 14.8

Eurycoma longifolia 53.75 3.37 24.0

Dipteris lobbiana 54.92 81.75 92.0

Ficus deltoidea 53.93 42.45 61.1

Fagrea obovata 54.58 74.13 91.6

Momordica subanguenta 48.30 0.15 22.9

Morinda citrifolia 53.75 11.80 46.1

Orthosiphon stamineus 54.54 56.75 87.5

Smilax myosotiflora 54.63 16.19 32.6

Scavenging activityScavenging activity% Scavenging of plant extract

0

20

40

60

80

100

Ap Ab El Dl Fd Fo Ms Mc Os Sm

Sample

Per

cen

t sc

aven

gin

g

% Scavenging

Ap=Andrographis paniculata, Ab=Averrhoa belimbi, El=Eurycoma longifolia, Dl=Dipteris lobbiana, Fd=Ficus deltoidea, Fo=Fagrea obovata, Mo=Momordica subanguenta, Mc=Morinda citrifolia, Os=Orthosiphon stamineus and Sm=Smilax myosotiflora

Flavonoid contents and scavenging activityFlavonoid contents and scavenging activity

Ap=Andrographis paniculata, Ab=Averrhoa belimbi, El=Eurycoma longifolia, Dl=Dipteris lobbiana, Fd=Ficus deltoidea, Fo=Fagrea obovata, Mo=Momordica subanguenta, Mc=Morinda citrifolia, Os=Orthosiphon stamineus and Sm=Smilax myosotiflora, Que= Quercetin

Correlation of flavonoid contents and % scavenging

0

20

40

60

80

100

Ap Ab El Dl Fd Fo Ms Mc Os Sm Que BHA

Sample

Fla

von

oid

co

nte

nt

(mg

/g)

and

sca

ven

gin

g (

%)

Series1

Series2

TLC testTLC test

F. obovata D. lobbianaO. aristatusF. deltoidea

ConclusionConclusion

1.1. There is correlation between There is correlation between flavonoid content and radical flavonoid content and radical scavenging activity. scavenging activity.

2.2. F.obovata, D.lobbianaF.obovata, D.lobbiana and and O.aristatusO.aristatus exhibited highest free exhibited highest free radical scavenging activity.radical scavenging activity.

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