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ACUTE TOXICITY TEST ON RAT USING METHANOL EXTRACT OF THE LEAF OF CARICA PAPAYA: (INTRAVENOUS ROUTE)

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ACUTE TOXICITY TEST ON RAT USING METHANOL EXTRACT OF THE LEAF OF CARICA PAPAYA: (INTRAVENOUS ROUTE)

 

TABLE OF CONTENTS

DEDICATION

ACKNOWLEDGEMENT

ABSTRACT

1          INTRODUCTION AND LITERATURE REVIEW

2          MATERIALS AND METHOD

2.1      PREPARATION OF THE METHANOL EXTRACT

2.2      ANIMALS

2.3      ADMINISTRATION OF THE EXTRACT TO RAT

2.4      DETERMINATION OF LD50

2.5      HISTOPATHOLOGIC EXAMINATION

3          RESULTS

3.1      CLINICAL SIGNS

3.2      LD50

3.3      HISTOPATHOLOGIC FINDINGS

4          DISCUSSION AND CONCLUSION

REFERENCES

ABSTRACT

The acute toxicity of a methanol extract of the leaf of carica papaya (1.2g/ml) was investigated in male rats (150- 227g). In acute toxicity studies carried out in rats, LD 50 of the extract was 3675mg/kg after intravenous administration. Clinical signs of depression (inactivity and anorexia), decrease in motor activity, analgesia and loss of righting reflex were observed.

Histopathologic examination showed inhibition or depletion of the glycogen storage and lipid formation at a dose above 1556, 4 mg/kg.

It was concluded that methanol extract of the leaf of carica papaya when given at a dose of 15556.4 mg (equivalent to 0.24g dried leaf)/kg intravenously does not cause any death or toxic effects in rats.

A further study is required to confirm its efficacy in the treatment of various conditions in folk medicine.

 

 

 

 

 

 

 

 

 

INTRODUCTION AND LITERATURE REVIEW

            The treatment given native doctors and herbalists throughout Nigeria to their patients often includes the administration of exudates of entire plants or extracts of the roots, stems, bark, leaves, fruits, seeds, juice of local plants. Frequently neither the ‘’doctor’’ nor the patient attributes the healing action to the plant itself. The ‘’doctor’’ may say that he has discovered a plant with leaves or roots possessing a spirit is not manifested until he has spoken some magic words or has chanted an incantation over the plant (Oliver 1960).

Apart from the native doctors who treat the more complicated and resistant cases, the more common ailments are treated by ordinary people in the village; in most cases elderly women with some knowledge of the local plats. In a number of cases also, the recovery of the patient may be attributable to his faith or his natural resistance rather than to the ‘’doctors’’ skill. However parts of the plants have been shown to have real therapeutic value (Tang, 1979). Extracts of the roots of Heptacyclum zenkeri have been used as an aphrodisiac and in the treatment of local infections and veneral disease (Duah et al, 1981). Stem bark of Eucalyptus maculate is used generally in the treatment of asthma and chronic bronchitis (Mishra and Misra, 1980).

carica papaya  (pawpaw) is a popular tropical and subtropical fruit. It was originally a native of America but is now extensively grown in all tropical countries including India (Dutta, 1979).

The plants are small, sparsely branched trees with soft wood. The soft wood is formed largely from the phloem that gives the soft large trunck much of its rigidity. The leaves are alternative, digitately lobed or foliated and without stipules. The fruit is a berry and its latex contains papain, an important protease (enzyme) used in tenderizing meat and also for removing biological stains from fibrics (Gledhill, 1972).

The leaf of carica papaya has been used in places like Mexico, Indonesia, India, Africa, Philippines and Ghana under various conditions in folk medicine (Table 1). These conditions include Asthma, Beriberi, Elephantoid growth, Foul wounds, kidney and bladder trouble, Nervours pain and Epithellioma (Tang, 1979)

Several substances have been implicated in the pharmacological effects of carica papaya.  According to

Benzylglucosinolate

Benzyl isothiocyanate

Chloline

Carpaine

Pseudocarpaine

Dehydrocarpaine I

Dehydrocarpaine II

According to Weast et al (1981), the organic constituents of the carica papaya leaf are Ascorbic acid (Vitamin C), B-carotine (Provitamin A) and Carpaine. The presense of the carpaine alkaloid in the leaf of carica papaya is of toxicological interest (Kucera et al, 1974). However two new capaine analogues dehydrocarpaine I and dehydrocarpaine II have been isolated from the alkaloid extract of the dried leaves of carica papaya (Tang, 1979).

Carpaine (C28H50O4N2) is a crystalline alkaloid found in the leaves of carica papaya and a dies built by two identical halves, the carpaic acid molecules (pelletier, 1970). Carpaine was isolated by Greshoff (1890). It has a melting point (M.P) of 1211oc and a boiling point of 215 -2350C (vacuum). It is also soluble in alcohol but only slightly soluble in water (Tang, 1979).

It has been reported that carpaine is essentially a heart poison capable of lowering pulse frequency and depressing contral nervous system (Heary, 1974). Tuffley and williian (1951) had earlier reported that carpaine reduces the contractile power of the heart.

The effect of carpaine on the circulatory function has been recently studied at the medical school University of Hawaii, it was found that carpaine has a dose dependent effect on lowering the blood pressure and heart rate of the rat (Tang 1979).

This study on acute toxicity test will reveal the toxic effects produced by a single dose of a methanol extract of the leaf of carica papaya in rats. The use of rats in this investigation is based on convenience and availability.

Species differences cosntiturte the single largest difficulkty in the interpretation of toxicity testing. In most circumstancves too little is known about the way differing animal species react to toxic chemicals for it to be possible to give any logical reason for choosing one species rather than another (Paget and Barnes, 1964)

All laboratory toxicity studies are directed towards a single objective: the safe use of the candidate substance in man. This objective has never been fully realized because the available laboratory tests are incapable of detecting certain types of toxic manifestations which occur in man.

Since this plant carica papaya is readily available in many localities in Nigeria including Port Harcourt, the possibility of using it as cheap and readily available drug for the above named diseases has to be considered.

As part of the safety evaluation of the leaf of carica papaya this study was undertaken to determine the lethal dose: LD50 (that is the dose that will kill 50% of the rats used) together with doses producing specific toxic effects using the methanol extract of the leaf of carica papaya.

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