1

In vitro Antioxidant, anticariogenic and haemolytic activity of leaf extract of medicinal plants against two oral pathogens

Sreejith P S (15MSB0045), Anju G (15MSB0062), Majesh Mathew (15MSM0070).

Research Guide: Dr. Devi Rajeswari V

Assistant Professor (Sr),School of Biosciences and Technology,

VIT University, Vellore-14.

12SETMSB0106

2

INTRODUCTION

Free radicals play a vital role in most major health problems like cancer, rheumatoid arthritis, cardiovascular diseases,

alzeihmer’s disease and other neurodegenerative disorders. Antioxidant that scavenges these free radicals proves to be

beneficial for these disorders as they prevent damage against cell proteins, lipids and carbohydrates.

Antioxidant activity includes free radical scavenging capacity, inhibition of lipid peroxidation, metal ion chelating ability

and reducing capacity.

Antioxidants are molecules that inhibit the initiation of oxidation chain reactions thereby preventing damage to human

body cells. At present, synthetic antioxidants are available such as butylated hydroxyanisole (BHA) and butylated

hydroxytoluene (BHT) but they were proven to be toxic for human beings.

Medicinal plants containing polyphenols have been reported for antioxidant and other pharmacological activities.

Plant based natural antioxidants are at most interest worldwide because of non toxic nature.

3

Haemolysis has long been used to measure free radical damage and counteraction by antioxidants. It is useful for screening

for oxidising or antioxidising agents . Several herbal secondary metabolites such as flavonoids have been found to protect

cells from oxidative damage. These compounds have been evidenced to stabilise RBC membrane by scavenging free

radicals and reducing lipid peroxidation.

Medicinal plants are the rich source of medicinally important compounds and since ancient time, plants and plant derived

products are used as medicine in traditional and folk medicinal system.

Plant-derived natural products such as flavonoids, terpenes, alkaloids, anthraquinones, saponins, tannins, steroids, lactones

and volatile oils received considerable attention in recent years due to their diverse pharmacological properties, including

cytotoxic and chemo-preventive effects.

Using plant for medicinal purposes dated back to prehistory and people of all continents have this old tradition. Until today,

plant-based medicine continues to play a key role in healthcare systems in many regions worldwide.

.

4

METHODOLOGY

Sample collection Extraction process

Phytochemical analysis

Antioxidant activity Anti haemolytic activity

5

Sample collection

Punica granatum Samanea saman

Pisonia alba Psidium guajava

6

Extraction process

Preparation of aqueous extract

7

Phytochemical analysis

Qualitative analysis

Phytochemical Qualitative Analysis

Samanea saman Pisonia alba Punica granatum Psidium guajava

Alkaloids Dragon drop test + - - +Carbohydrates Benedict’s test + + + +

Proteins Biuret test - - + +Glycoside Borntrager’s test + + + +Saponins Foam test + + - -Steroids Salkowski’ s test + + + +Phenolic Lead acetate test + + + +Tannins Ferric chloride

test+ + + +

Flavonoids Sodium hydroxide test

+ + + +

8

Conc. of Gallic acid in µg/ml

Absorbanceat760nm

Conc. of Aqueousextract in µg/ml

Amount of total Phenolic content in terms mg of GAE/g of extract

25 0.20850

0.418

75 0.657 100100 0.896200 1.114

Punica granatum

Psidium guajava

Samanea saman

Pisonia alba

105 95 80 52.5

Quantitative analysis

Total phenolic content

9

Conc. of quercetinin µg/ml

Absorbanceat517nm

Conc. of Aqueousextract in µg/ml

Amount of total flavonoid content in terms mg of Quercetin/g of extract

25 0.1250

0.31

75 0.45 100100 0.8200 2.1

Punica granatum

Psidium guajava

Samanea saman

Pisonia alba

122.5 87.5 72.5 37.5

Total flavonoid content

10

Antioxidant activity:

25 50 100 2000

102030405060708090

100DPPH assay

Pisonia albaSamanea samanPunica granatumPsidium guaja

Concentration µg/ml

%in

hibi

ton

DPPH radical scavenging activity

11

Hydrogen peroxide scavenging activity:

25 50 100 2000

10

20

30

40

50

60

70

80

90

100

Hydrogen peroxide

Pisonia albaSamanea samanPunica granatumPsidium guaja

12

25 50 100 2000

102030405060708090

Metal chelating

Pisonia albaSamanea samanPunica granatumPsidium guajava

concentration µg/ml

Opti

cal d

ensit

y

Metal chelating:

13

25 50 100 2000

102030405060708090

100 Haemolytic acitivity

Pisonia albaSamanea samanPunica granatumPsidium guaja

Concentration µg/ml

%ha

emol

ysis

Haemolytic activity:

14

Antimicrobial activity:

Positive control

Samanea saman Pisonia alba Punica granatum

Psidium guajava

Candida albicans

3mm 2.5mm 1.8mm 0.5mm -

Streptococcusaureus

5mm 3.6mm 2.7mm 0.2mm -

15

Conclusion:

•In this study total phenolic and flavanoid content of extracts were determined. The radical scavenging activity was

evaluated by DPPH assay, Hydrogen peroxide and metal chelating assays were performed. The result of extracts against the

tooth decaying organisms Candida albicans, and Streptococcus pyogenes will be assessed by visualizing the presence or

absence of inhibition zone and measuring diameter of zone of inhibition.

•Hemolytic activity of the four different extracts was screened against normal human erythrocytes. The four diferent

extracts that showed high antihaemolytic activity in our study, Pisonia alba, Samanea saman, Punica granatum and

Psidium guajava. Hence, it may be considered as safe to human erythrocytes and it serve as easily accessible sources of

natural antioxidants for the pharmaceutical industry.

16

Reference

THANK YOU

17