INHIBITORY EFFECT OF ACETONE FRACTION OF Lonchocarpus cyanescens ON ALPHA AMYLASE AND ALPHA GLUCOSIDASE

Introduction: Diabetes mellitus is a metabolic disorder characterized by high blood sugar levels. Inhibiting enzymes like alpha-amylase and alpha-glucosidase is a key strategy to control hyperglycemia. Lonchocarpus cyanescens is a medicinal plant with potential antidiabetic properties that warrants scientific evaluation. The major aim of this research is to ascertain and provide scientific information on the antidiabetic properties of Lonchocarpus cyanescens utilizing alpha amylase and alpha glucosidase inhibitory assay All materials used were of high quality which includes alpha amylase, alpha glucosidase, distilled water, ethanol, acetone, hexane. Methodology involves detrmining the antidiabetic properties of the plant extract utilizing alpha amylase and alpha glucosidase inhibitory assay. The potential for Lonchocarpus cyanescens extract to reduce hyperglycemia and perform antidiabetic functions was determined. Alpha-amylase inhibition activity of each fraction was determined by the method of Worthington 1993. An aliquot of 500 microliter of the extract (0.1–0.4 mg/ml) and 500 microliters (0.02M) of sodium phosphate buffer (pH 6.9 with 0.006M NaCl) containing 0.5 mg/ml of alpha-amylase will be mixed together and incubated for 10 min at room temperature.Afterwards, 500 microliters of 1% starch solution prepared with 0.02M sodium phosphate buffer (pH 6.9 with 0.006M NaCl) will be added and incubated in a water bath at 25°C for 10 minutes.The reaction mixture will be stopped by adding 1.0 ml (96 mM) of Dinitro salicylic acid.The mixtures in the test tubes will be incubated in boiling water in a water bath for 5 minutes and then cooled for alpha amylase. Then for alpha glucosidase Alpha-glucosidase activity of each fraction will be determined by the method of Apostolidis et al., 2007.The substrate solution, p-nitrophenyl- glucopyranoside (pNPG), was prepared in 0.02M phosphate buffer, pH 6.9. 1000 microliter of alpha-glucosidase was incubated with 500 microliters of different concentrations of the extract for 10 minutes at 25°C. An aliquot (500 microliter) of freshly prepared phosphate-buffered p- nitrophenyl-glucopyranoside (5 mM) solution will be added. The reaction mixture will be incubated at 25°C for 5 minutes and stopped by adding 2 ml of 0.1 Na₂CO₃. The alpha- glucosidase activity was determined by measuring the absorbance at 405 nm using a spectrophotometer. Absorbance reading assay was carried out and statistical analysis was also carried out to checkn for satistical significance. A p-value for alph amylase was found out to be [ p=0.0321] and for alpha glucosidase the p –value was recorded to be [p=0.0002] in this assay. In conclusion, Lonchocarpus cyanscens possess antidiabetic properties and can serve for several medical purposes.