DETERMINATION OF PUMPKIN LEAVES EQUIVALENT; AS A MEASURE OF PYHTO TOXICITY ON HUMAN ERYTHROCYTES
Publication Date : 19/05/2020
Pumpkin leaves (Telfairia occidentalis) are extensively used in homes in Nigeria and some West African Countries for soup making and sometimes extracted in water for drinking as medicament for stimulating erythropoiesis. The present study aims at evaluating the in-vitro effect of pumpkin leaves extract on human erythrocytes with an interest in obtaining base data on the phyto-toxicity. Human erythrocytes of the A,B,O. blood groups and Rhesus positive and negative were obtained from blood donors in the University of Benin Teaching Hospital, Benin City. Pumpkin leaves weighing 250g was extract in 250ml cold distilled water to obtain a stock solution of 1gm/ml pumpkin leaves extract. The stock solution was further diluted in sterile test tubes to obtain a concentration range of 10µg/m/, 20µg/m/, 40µg/m/ to 640µg/m/ using cold sterile distilled water. The test proper required the addition of 0.5ml of each pumpkin leaves extracted concentration to a sterile test tube containing 2.5ml of 5% human erythrocyte in sterile physiological saline and allowed to remain at room temperature (27+ 1oc)for 30 minutes. The heamolysis occurring in the tubes was read spectrophometrically (Cornings) at 540nm wavelength along with the neat and saline control. A drop of erythrocyte suspension (0.02ml) from each tube was placed on a clean grease- free slide, covered with a coverslip and examined under the microscope (Olympus) using X10 and X40 objective lenses. Then photomicrographs to show cell structures were obtained using a motic camera with extension lense. The lowest concentration of pumpkin leaves extract to yield heamolysis was 125 µg/m/ irrespective of blood group and Rhesus status. The absorbance reading reflected corresponding increases in the concentration of pumpkin leaves extract using phosphate buffer as negative control and Triton X as positive control (100% heamolysis). The data obtained from this study may have value in determining phytotoxicity with pumpkin leaves.
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