PHYTOCHEMICAL SCREENING AND BIOACTIVITY OF PETROLEUM

ETHER AND ETHANOL EXTRACT FROM THE STEM BARK OF

Dichapetalum filicaule.

INTRODUCTION

The use of Plants extracts and phytochemicals

Isolated from them provide the foundation to modern

pharmaceutical compounds.

FAMILY: DICHAPETALACEAE

PLANT : Dichapetalum filicaule

• In the world distribution it is found in Cote d’Ivoire and Ghana

• Ghana: Yamoransa and Boi-Tano forest reserve.

SOME USES OF THE FAMILY DICHAPETALACEAE

Medicinal use; for the treatment of jaundice, diarrhoea ,viral hepatitis and rheumatism

General purpose

• Used as rodenticides.

• Arrow poison.

• Carving utensils eg. bowls ,plates.

• Chewing sticks for cleaning teeth.

AIM

TO DETERMINE THE CLASSES OF COMPOUNDS PRESENT IN THE PETROLEUM ETHER AND ETHANOL EXTRACTS FROM THE STEM BARK Dichapetalum filicaule AND TO CARRY OUT BIOACTIVITY TEST ON THE TWO CRUDE EXTRACT USING BRINE SHRIMP LETHALITY TEST.

Statements of the problem

• Does the stem bark of Dichapetalum contain Phytochemicals such as Alkaloids, Anthraquinones and Anthracene Derivatives, Flavonoids and Leuco-anthocyanins, Cardiac glycosides, Tannins ,Terpenoids, Saponins?

• Do the crude extracts show activity toward brine shrimp? If they do ,to what extent?

Objectives of the study

• To carry out petroleum ether and ethanol Extraction of the stem bark of D. fillicaule

• To determine the Phytochemicals present in each extract.

• To carry out Brine Shrimp Lethality test to ascertain the extent of toxicity of the extracts.

LITERATURE REVIEW

Scientific Investigation into the Dichapetalaceae started way back in 1940’s on the toxic species, eg. D. cymosum and D. toxicarium. These toxic constituents have been proved to be mainly fluorinated organic compounds ,particularly fluorocarboxylic acids. To be specific Fluoroacetate

• In this Department much work has been done on the species: D.bateri and D. madagascariense. Work on D. madagascariense has led to the isolation and characterization of a novel and unique class of triterpenoids, the Dichapetalins.so far 13 Dichapetalins namely Dichapetalin A-M have been isolated.

• Bioactivity test revealed that among the Dichapetalins,Dichapetalin A and M showed high activity towards Brine Shrimp with Dichapetalin M being more potent.

• Further tests revealed that Dichapetalin A has Anti-cancer activity which increases in vitro and decreases in vivo. Both Dichapetalins A and M did not exhibit appreciable selectivity in anti-HIV activity. Generally the Dichapetalins are not known for anti-microbial activities.

THE DICHAPETALIN SKELETON

COLLECTION AND IDENTIFICATION OF PLANT MATERIAL

EXTRACTION OF PLANT MATERIALS

PHYTOCHEMICAL SCREENING

BRINE SHRIMP LETHALITY TEST

METHODOLOGY

Results and Discussion

EXTRACTS ALKALOIDS A AND A

F AND L-A

C. GLYCOSIDES TANNINS SPN TT

Ethanol - + - + + + +

Petroleum ether - - - + - + +

TABLE 1 : RESULTS OF PHYTOCHEMICAL SCREENING OF CRUDE EXTRACTS

KEY

+ =Present ; - = Absent ;A and A = Anthraquinones and Anthracene derivatives ; F and L-A = Flavonoids and Leuco-Anthocyanins ; C.= Cardiac ; SPN = saponins ;TT = Terpenoids

CONCENTRATION(µg/ml) NUMBER OF BRINE SHRIMP KILLED LARVAE KILLED

1 2 3 TOTAL

10 7 8 0 25

1 7 7 6 20

0.1 4 5 4 13

0.01 3 3 2 8

0.001 1 2 0 3

Control 0 0 0 0

TABLE 3 : PETROLEUM ETHER EXTRACT

CONCENTRATION(µg/ml)

NUMBER OF BRINE SHRIMP LARVAE KILLED

1 2 3 TOTAL

10 0 0 0 30

1 9 9 9 27

0.1 6 5 7 18

0.01 4 3 4 11

0.001 2 3 2 7

Control 0 0 0 0

TABLE 3 : ETHANOL CRUDE EXTRACT

A GRAPH OF PROBIT OF MORTALITY AGAINST LOG10 OF CONCENTRATION OF PETROLEUM ETHER CRUDE EXTRACT

A GRAPH OF PROBIT OF MORTALITY AGAINST LOG10 OF CONCENTRATION OF ETHANOL CRUDE EXTRACT

Extracts LC50(µg/ml)

Ethanol 0.02

Petroleum ether 0.32

TABLE 4: BIOASSAY RESULTS OF CRUDE ECTRACTS FROM THE STEM BARK OF D. filicaule

EXTRACTS LC50(µg/ml)

THIS WORK D. barteri (stem) D. Madagascariense (roots)

Petroleum ether 0.32 >0<1 0.041

TABLE 5 : COMPARISON OF LC50 VALUES

• Phytochemical screening was carried according to standard procedures, from the

results above ; saponins ,anthraquinones/ anthracene derivatives, cardiac glycosides,

terpenoids and tannins were present in the ethanol crude extract ,same was recorded

for the petroleum ether with the exception of tannins and anthraquinones

/anthracene derivatives which were absent.

• Bioactivity test carried out indicated that both petroleum ether and ethanol

extracts showed activity towards Brine shrimp recording LD50 0.32µg/ml and

0.02µg/ml respectively.’

CONCLUSION

Phytochemicals are present in the petroleum ether and ethanol crude extracts and the crude extracts showed activity towards Brine shrimp with the ethanol extract being more potent.

RECOMMENDATION • Further work should be done to isolate the

compounds in the two extracts and characterised

• Bioactivity test should be carried out on each isolated compound.

• Anti-HIV activity test should be carried out on each isolated compound

• Anti-cancer activity test should be carried out on each isolated compound.

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Assesssment of Bioactivity of India Medicinal Plants Using Brine Shrimp( Artemia salina): International Journal of Applied Science and Engineering (3,2 ).125-134.

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ACKNOWLEDGEMENT

• ALMIGHTY GOD

• DR. MARY ANTI CHAMA

• REV. DR GBEWONYO

• DR. DORCAS OSEI-SARFO

• MR. BOB ESSIEN

• MR.GODWIN DZIWORNU AKPEKO

• MR. GYASI HAYFORD

• MR.DANIEL KONADU

THANK YOU