University of Khartoum

Activity- guided Isolation and Structure Determination of Antioxidant and Antidiabetic Compounds from Bauhinia rufescence L.

Activity- guided Isolation and Structure Determination of Antioxidant and Antidiabetic Compounds from Bauhinia rufescence L.

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Title: Activity- guided Isolation and Structure Determination of Antioxidant and Antidiabetic Compounds from Bauhinia rufescence L.
Author: Osman, Wadah Jamal Ahmed
Abstract: Background: Diabetes mellitus is a group of disorders of multiple etiologies resulting from a defect in insulin secretion, insulin action, or both. Diabetes remain one of the world’s major health problems and many repots have been supported the role of oxidative stress in the pathogenesis of both type 1 and type 2 diabetes. This study aimed to evaluate the antidiabetic and antioxidant activity of Bauhinia rufescens a plant used in Sudanese folkloric medicine for treatment of diabetes. It was also aimed to identify isolates and characterize the bioactive antidiabetic and antioxidant compounds using bioactivity guided fractionation followed by High Performance Thin Layer Chromatography (HPTLC) autobiography, Liquid-Mass spectroscopy analysis and nuclear magnetic resonance (NMR). Method: The powdered plant leaves were exclusively extracted using petroleum ether, chloroform and methanol, successively. These extracts were tested in streptozotocin-induced diabetic Wistar rats for antidiabetic as well as DPPH antioxidant potential. Standard Propyl Gallate was used as (+ve) control and the difference in results was considered significant when P < 0.05. The best active extract was fractionated with chloroform, ethyl acetate and butanol along with the remaining aqueous fraction. The fractions are tested for antioxidant and antidiabetic activity using HPTLC autobiography (α amylase and α glucosidase). Further, bioactive compounds were isolated by Preparative Thin Layer Chromatography and identified using LC/MS and NMR spectroscopy.X Results: The methanol extract significantly decreased the blood glucose level in normal fasted rats by 27 %; two hours following administration and decreased the blood glucose level in streptozotocin-treated rats by 48.9%; two hours following oral administration, while chloroform and pet. Ether extracts not significantly decreases the glucose level.Significantly the methanol extract also showed strong antioxidant activity (89%) compared with other extracts; thereforeit was further subjected for fractionation by using chloroform, ethyl acetate, n-butanol and water sequentially, and then tested for their antioxidant activity; where the ethyl acetate fraction was showed good activity compared with other fractions. Bioactivity guided fractionation of Bauhinia rufescensled to isolation and characterization of two compounds with potent antidiabetic and antioxidant activity namely; Quercetin-3-Orutinoside and Kaempferol -3-O-rutinoside. These compounds were isolated by using preparative thin layer c chromatographic techniques (PTLC) and their structures were identified by 1D NMR spectrum (C13and 1HNMR), 2D NMR (COSY, HSQC, HMBC Correlation) and confirmed bycomparing with reported data. LC/MS analysis led to identified more than thirty compounds; which were subjected to in silico assessment against human α-glucosidase and α-amylase enzymes. Insilico assessment showed that more than 14 compounds showed potent activity compared with acarbose (standard anti-diabetic drug). High performance thin layer chromatography (HPTLC) for the first time of the active fractionof the plant revealed the presence of different spots/peaks in different Rf values, corresponding to different compounds.XI Conclusion: This study validated the traditional uses of the evaluated plant in treating diabetes; beside that it was found a good antioxidant. The compounds which were isolated and identified may provide a new lead for more potent analogues for drug discovery. The anti-diabetic potential and antioxidant activity reported in present investigation is reported for the first time on the aerial plant. Further the isolated and characterized compounds from this study are also being reported for the first time. Toxicological studies may further be planned to develop it in the form of new drug for management of diabetes, however its use is available as per traditional claim.
URI: http://khartoumspace.uofk.edu/123456789/27027


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