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

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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

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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.

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 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.
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METHODOLOGY
Sample collection Extraction process
Phytochemical
analysis
Antioxidant activity
Anti haemolytic
activity
Anticariogenic
activity

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Sample collection
Punica granatum Samanea saman
Pisonia alba Psidium guajava

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Extraction process
Preparation of aqueous extract

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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
+ + + +

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Conc.
of
Gallic
acid
in µg/ml
Absorbance
at
760nm
Conc. of
Aqueous
extract
in
µg/ml
Amount of total Phenolic content in terms mg of GAE/g of
extract
25 0.208
50 0.418
75 0.657 100
100 0.896
200 1.114
Punica
granatum
Psidium
guajava
Samanea
saman
Pisonia alba
105 95 80 52.5
Quantitative analysis
Total phenolic content

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Conc.
of
quercetin
in µg/ml
Absorbance
at
517nm
Conc. of
Aqueous
extract
in
µg/ml
Amount of total flavonoid content in terms mg of Quercetin/g
of extract
25 0.12
50 0.31
75 0.45 100
100 0.8
200 2.1
Punica
granatum
Psidium
guajava
Samanea
saman
Pisonia alba
122.5 87.5 72.5 37.5
Total flavonoid content

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Antioxidant activity:
0
10
20
30
40
50
60
70
80
90
100
25 50 100 200
%inhibiton
Concentration µg/ml
DPPH assay
Pisonia alba
Samanea saman
Punica granatum
Psidium guaja
DPPH radical scavenging activity

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Hydrogen peroxide scavenging activity:
0
10
20
30
40
50
60
70
80
90
100
25 50 100 200
Hydrogen peroxide
Pisonia alba
Samanea saman
Punica granatum
Psidium guaja

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0
10
20
30
40
50
60
70
80
90
25 50 100 200
Opticaldensity
concentration µg/ml
Metal chelating
Pisonia alba
Samanea saman
Punica granatum
Psidium guajava
Metal chelating:

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0
10
20
30
40
50
60
70
80
90
100
25 50 100 200
%haemolysis
Concentration µg/ml
Haemolytic acitivity
Pisonia alba
Samanea saman
Punica granatum
Psidium guaja
Haemolytic activity:

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Antimicrobial activity:
Positive
control
Samanea saman Pisonia alba Punica
granatum
Psidium
guajava
Candida
albicans
3mm 2.5mm 1.8mm 0.5mm -
Streptococcus
aureus
5mm 3.6mm 2.7mm 0.2mm -

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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.

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Reference

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