presentation on my project work

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My presentation

Assalamu alaikum

Presented By: Matric No:P-121004

Session: Spring ‘12-Spring ’16

DEPARTMENT OF PHARMACYInternational Islamic University Chittagong

Evaluation of in-vitro antioxidant and in-vitro antimicrobial activity of methanolic extract of

Mimosa invisa (leaves)

Project title :

Description :Mimosa invisa is a fast-growing a scrambling climber,

which can form dense thickets in a short span of time. It is an annual, although behaves as a perennial. Leaves are bright green, alternate, each leaf with about 20 pairs of small leaflets, opposite, 6-12 mm long and 1.5 mm wide, sensitive to disturbance.

Distribution : Native to South and Central America. Its found in Indian

sub continent, Malaysia, Hong kong, Indonesia, Sri lanka , Philippines. It is also found in forest of Chittagong , Coxsbazar , Rajshahi and Bandorban .

Plant literature

Plant taxonomy :Kingdom PlantaePhylum Spermatophyta

Subphylum AngiospermaeClass DicotyledonesOrder FablesFamily FabaceaeGenus Mimosa

Species Mimosa invisa

Figure: Mimosa invisa

Local name :Sada sorminda patha , boro lozzaboti , sada lozzaboti

.Traditional uses :The plant’s root paste with goats milk is used in treatment of insanity

. It is also used to treat impotency , skin disease , inflammation ,dysentery and burning sensation .

Literature review :1. Faruque (2007) reported that the plant of Mimosa invisa is used

to treat insanity.2. Faruque (2007) also reported that it is used to treat impotency.3. Rahman (2014) reported that skin disease is treated by this plant.4. Meenatchisundaram S(2009) examined that Mimosa pudica

which is another member of this family has anti venom activity.

Literature review revealed that the plants of, Mimosa invisa are used in different tribes as well as some areas of Rajshahi and Bandorban without scientific basis or safety concerns. Determination of its antioxidant and antimicrobial activities will provide supportive evidence in favor of continuous usage.

The objectives of my project work is In-vitro antioxidant activity assay In-vitro antimicrobial activity assay

Aim of study :

Collection and proper identification of the plant Preparation of plant materialsExtraction In-vitro antioxidant assay In-vitro antimicrobial assay

Outline of the study :

Plant materials were collected from the forest of Bandarban district, Bangladesh and authenticated by Dr. Shaikh Bokhtear Uddin, Professor, Department of Botany, University of Chittagong.

Plant selection and proper identification :

The plant materials were collected in fresh condition. Then these are cut into small pieces to make it suitable for grinding purpose and dried. The materials are grinded into coarse powder with the help of a grinder and stored in an air tight container for further use

Preparation of plant materials :

The coarse powder was extracted with Methanol. The powder

was submerged in a clean flat bottomed glass container, for few days with occasional shaking and stirring. The content was then filtered through several means, e.g. cotton, clothe, filter paper etc. to get the pure extract

Extraction process :

DPPH scavenging assay Iron reducing assay Determination of total phenol content Determination of total flavonoid content

In-vitro antioxidant assay :

Concentration (µgm/ml)

% of scavenging of ascorbic acid

% of scavenging of methanolic

extract of Mimosa invisa

20 91.517 42.09110 88.560 33.1925 60.544 26.046

2.5 37.198 20.3891.25 29.494 14.863

Data for DPPH scavenging assay :

0 5 10 15 20 250

10

20

30

40

50

60

70

80

90

100

f(x) = 1.35491827956989 x + 16.8155833333334R² = 0.932946408151683

f(x) = 3.29826559139784 x + 35.9010416666668R² = 0.778440644938047

DPPH scavenging

% of scavenging of AA Linear (% of scavenging of AA )% of scavenging of MMI Linear (% of scavenging of MMI)

Calibration & scattering chart for DPPH scavenging assay :

IC 50 of ascorbic acid IC 50 of extract 4.275 24.513

Chart for IC50 value’s comparison in DPPH scavenging assay :

0

5

10

15

20

25

30

IC 50 of AA

IC 50 of MMI

Concentration Absorbance of extract

Absorbance of ascorbic acid

1000 0.808 1.878

500 0.510 1.295

250 0.331 0.905

125 0.194 0.625

62.5 0.081 0.398

Data for iron reducing assay :

Scattering chart of absorbance of Ascorbic acid & extract :

0 200 400 600 800 1000 12000

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

absorbance of MMIabsorbance of AA

Concentration(µgm/ml)

Absorbance of galic acid

1000 1.98

500 0.520

250 0.266

125 0.152

62.5 0.084

Determination of total phenol assay :

0 100 200 300 400 500 6000

0.51

1.52

2.5

f(x) = 0.00393934328358215 x + 0.0330348258706422R² = 0.998669748560243

Calibration curve of Gallic acid

Absorbance of Gallic Acid Linear (Absorbance of Gallic Acid)

Abso

rban

ce

Sample solution(µg/ml)

Weight of dry extract per mlm(gm)

Absorbance GAE conc.C(µg/ml)

GAE conc.C(mg/ml)

TPC as GAE,A=(Cv)/m

(µg/ml)

Mean± SEM

1000 0.001 0.262 76.33 0.08 38.17 36.56

1000 0.001 0.241 69.33 0.07 34.67 ±

1000 0.001 0.254 73.67 0.07 36.83 1.02

Concentration (µgm/ml) Absorbance of quercetin 100 0.99550 0.34425 0.196

12.5 0.116

Determination of total flavonoid content :

0 20 40 60 80 100 1200

0.2

0.4

0.6

0.8

1

1.20.995

0.344

0.1960.116

0

f(x) = 0.009782 x − 0.036625R² = 0.972319089707755

Calibration curve of Quercetin

Absorbance of Quercetin Linear (Absorbance of Quercetin)

Abso

rban

ce

Sample solution(µg/ml)

Weight of dry extract per mlm(gm)

Absorbance QE conc.C(µg/ml)

QE conc.C(mg/ml)

TFC as QE,A=(C*V)/m(µg/ml)

Mean± SEM

1000 0.001 0.513 57.6 0.0576 28.8 28.10

1000 0.001 0.488 55.1 0.0551 27.55 ±

1000 0.001 0.496 55.9 0.0559 27.95 0.37

Chart for the determination of total flavonoid content :

The plant extract shows a significant IC50 values in compare to standard ascorbic acid were 24.513µgm/ml and 4.275µgm/ml in DPPH scavenging assay.The plant extract also showed a significant iron reducing activity .Total phenol content of the plant extract was 36.56±1.02 µgm/ml and the total flavonoid content was 28.10±0.37 µgm/ml. From above observation it appears that that the plant extract has a moderate antiradical activity.

Discussion :

The antimicrobial activity was assayed by disc diffusion method by using about 11 microorganism and kenamycin(30µgm/disc) as standard .

In-vitro antimicrobial assay :

The zone of inhibition at different concentration against several microorganism in compare with standard kenamycin (30 µgm/disc) was shown in given column chart :

Staph

ylococ

cus au

reus

Shige

lla dy

sentry

bacillu

s azat

fumass

K. pn

eumon

ia

Psedum

onas a

erugin

osa

Vibro

choler

a

Escher

chia c

oli

Salmon

ella th

ypi

Bacill

us cer

eus

Lactob

acillus

cocci

Lactob

acillus

coryi

formis

0

5

10

15

20

25

30

35

40

45

1000 800 500 Kenamycin (30µgm/disc)

In this antimicrobial screening highest inhibitory activity were noticed against the growth of Bacillus cereus with the zones of inhibition 30mm at 1000µg/disc. In conclusion, the extract showed significant zone of inhibition against Bacillus cereus,Vibro cholera,Shigella dysentery, Eschericia coli and Salmonella typhi at higher concentration.

Discussion :

Based on the present investigation , it can be assumed that the methanolic extract of Mimosa invisa (leaves) have antiradical and antimicrobial activity .Further investigation are required to isolate the active constituents responsible for the observed effects and to elucidate the possible mechanisms of action responsible for those activities of this plant extract.

Conclusion:

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