Pharmacological Evaluation and Phytochemical characterization

of Aegiceras corniculatum (Linn.) Blanco (Family-Myrsinaceae) Bark

Presented byVaskor BalaRoll: 021113

Pharmacy Discipline

Phytochemical analysis of the extract of this plant indicated the presence of alkaloids, steroid, flavonoids, tannin, saponins and absence of gums and reducing sugar type of compounds. The general toxicity was investigated on brine shrimp (Artemia salina) & showed the most prominent cytotoxic activity (LC50 = 5.5 µg/ml and LC90 =9.3 µg/ml). At a dose of 250 & 500 mg/kg exhibited significant (p<0.001) inhibition of writhing reflex (59.8% and 74.52% respectively) with a dose dependant manner which were compared to the standard drug Diclofenac sodium at the dose of 25 mg/kg of body weight. When tested for its antidiarrhoeal effects on castor oil induced diarrhoea in mice, it increased mean latent period and decreased the frequency of defecation significantly at the oral dose of 250 and 500 mg/kg body weight (P<0.01) comparable to the standard drug Loperamide at the dose of 50 mg/kg of body weight.

Abstract

Plant Preview

Botanical features

Bengali KholshiBengali Kholisa English Kajla

Regional name

Small tree or shrub, up to 4 m high, spreading duffused branched, glabrous, bark almost smooth, reddish brown or dark grey Tap roots are well developed, much branched, horizontally spreading.Fruit capsule, 2.0 cm - 9.0 cm long, completely curved with pointed apex. 

Botanical Name: Aegiceras corniculatum (Linn.) Blanco

Family: Myrsinaceae

Traditional Use: Its fruits and seeds are used as fish-poison; barks and leaves are used in the treatment of asthma, diabetes, rheumatism, etc. (Balasooriya et al; 1982).

DACB acc. no. 31381 Fig: Stem of A. corniculatum

Literature Review

Pharmacological Literature Review:

A. corniculatum afforded 5-O-methylembelin, which was toxic to fish (Tilapia nilotica) and retarded the growth of the fungus Pythium ultimum (Venkateswara, K. et al: 1961).

Phytochemical Literature Review:

The following chemicals are isolated form A. corniulatum 2-methoxy-3-nonylresorcinol, 3,7-dihydroxy-2,5-diundecylnaphthoquinone, 2,7-dihydroxy-8-methoxy-3,6-diundecyldibenzofuran-1,4-dione, 2,8-dihydroxy-7-methoxy-3,9-diundecyldibenzofuran-1,4-dione, and 10-hydroxy-4-O-methyl-2,11-diundecylgomphilactone (Xu et al; 2004).

Wahidullah et al; 2002, reveals oleanane triterpenoid oligoglycoside (corniculatonin) and Gomez et al; 1989, 5-O-methylembelin form A. corniculatum

A. Corniculatum contains a saponin in 1.5% in the fruits and 0.5% in the leaves and barks. Hydrolysis yielded sapogenin named kanjalagin (C30H52O3) (Kincl et al;1956).

Phytochemical Study

Method:Trease et al; (1983) and Myers (1982), Method was adopted for phytochemical anlalysis.Preparation of bark extract: The extract of the bark of the plant was prepared by using 80% ethanol as solvent . The yield of the extract was 15.66% (47gm form 300 gm).

Result

Extract Steroids Alkaloids Tannins Gums Flavonoids SaponinsReducing

suger

Ethannol + + + - + + -

+ Presence - No reaction

Test sample

Conc. (g/ml

)

Test1

Test 2

Average Alive

shrimp

% mortality

LC50

(g/ml)

LC90

(g/ml)

Ethanolic

extract

1 10 9 9.5 5

5.5 9.3

2 8 8 8 20

4 7 6 6.5 35

6 4 5 4.5 55

8 2 3 2.5 75

10 0 0 0 100

Brine shrimp lethality assay

Method:The Meyer et al; (1982) method was adopted for the study of the toxicity of A. corniculatum bark.

Table: Result of Brine Shrimp lethality bioassay

Materials:100mg of dried ethanolic extract of A. corniculatum was taken in 10 ml

volumetric flask and volume was adjusted by DMSO and water (6:4). The concentration of this solution was 10 μg/μl.

0

10

20

30

40

50

60

70

80

90

100

0 1 2 3 4 5 6 7 8 9 10

Conctration

% M

ota

lity

(g/ml)

Fig: Effect of A. corniculatum extract on brine shrimp

Brine shrimp lethality assay

Analgesic Activity Study Method:Whittle, (1982) method that is acetic acid induced writhing method was followed for the analgesic study of A. corniculatum bark.

Study Design:Experimental animals were randomly selected and divided into four groups

denoted as group-I, group-II, group-III, group- IV consisting of 5 mice in each group. Each group received a particular treatment i.e. control, positive control

and the two doses of the extract.

Animal group Mean writhing ± SE

%Protection

Control 34±1.48 -

Diclofenac(25mg/kg)

13.6±1.37* 60

Extract(250 mg/kg)

21.8±1.12* 35.88

Extract(500 mg/kg)

18.8±1.36* 44.71

Table: Statistical evaluation of the Analgesic Activity

SD- Standard deviation, SE- Standard error ,* = P Value<0.001

Analgesic Activity Study

Fig: Effect of crude extract of bark of A. corniculatum on acetic acid induced writhing effect of mice.

Method: The castor oil induced antidiarrhoeal method was adopted form Galvez et al; (1993). The animals were all screened initially by giving 0.3 ml of castor oil and only those showing diarrhoea were selected for the final experiment. The animals were divided into control, positive control and test groups containing five mice in each group.

Antidiarrhoeal Activity Study

Group Dose (oral) Mean Latent Period ±SE t-test(P-value)

IControl (1% tween 80)

10 ml/kg 0.684 ±0.84 -

IIPositive Control (Loperamide) 50 mg/kg 1.52 ±0.257 3.09

(p<0.01)

IIIEthanol extract 250 mg/k) 0.758 ±0.075 1.21

(p<0.3)

IVEthanol extract 500 mg/kg 0.944 ±0.093 1.42

(p<0.2)

Table: Effect of A. corniculatum on the latent period of castor oil induced Diarrhoeal episode in mice

GroupsPeriod

of study (hr)Mean no. of

stoolsS.E.

t-test(p value)

IControl

1 2.6 0.51 --

2 4.8 0.67 --

3 3.6 0.68 --

4 3.4 0.87 --

II(PositiveControl)

1 0.2 0.20 4.38b

2 0.8 0.37 5.23a

3 1.8 0.37 2.32c

4 1.2 0.20 2.47c

IIIExtract of Aegiceras

corniculatum (250mg/kg)

1 2.0 0.41 0.91f

2 3.7 0.52 1.3f

3 2.3 0.61 1.43d

4 2.5 0.32 1.00f

IVExtract of Aegiceras

corniculatum (500mg/kg)

1 1.6 0.51 1.39d

2 3.0 0.32 2.42c

3 1.8 0.45 2.21e

4 2.0 0.63 1.30f

Values of t-test. (n=5); fp<0.3, ep<0.1, dp<0.2, cp< 0.05, bp< 0.001, ap< 0.01 vs. control

Table: Effect of Aegiceras corniculatum on castor oil induced diarrhoea in mice at the doses of 250 and 500 mg/kg-body weight

Fig: Effect of crude extract of bark of A. corniculatum on castor oil induced diarrhea of mice. Each bar represents mean latent period (4hr).

Fig: Effect of Loperamide and A. corniculatum on castor oil induced diarrhea in mice through the observation period (4hr)