Validated HPTLC Method for Detection of Kaempferol in <i>Moringa oleifera</i> Leaf Extract and Investigation of its Antidiabetic Potential by <i>In Vitro</i> and <i>In Silico</i> Studies

Background: Exploring traditional treatments offers valuable foundations for the innovation of new pharmaceuticals for the management of diabetes mellitus and its complications. Validation of plant extract by suitable chromatographic techniques is a part of standardization procedures. Moringa oleifera is one such traditionally used plant to be explored. Aim: The study explores the antidiabetic potential of M. oleifera through both in vitro and in silico explorations along with HPTLC method development and validation for estimation of kaempferol in M. oleifera extract. Method: Plant extract was prepared by maceration method using ethanol: water (70:25) and followed by Soxhlet(95% ethanol). Pharmacognostical analysis was conducted. In silico and in vitro antidiabetic and antioxidant studies were conducted, as well as quality control analysis was done using HPTLC method the mobile phase used was toluene: ethyl acetate: formic acid: methanol: (6:3:0.3:1v/v/v/v). Results: HPTLC analysis showed the presence of kaempferol, and the method was validated as per ICH guidelines. According to reports on molecular docking studies, several phytocompounds inhibited when porcine pancreatic α-amylase (PPA) complexed with a carbohydrate inhibitor (PDB ID: 4W93).The hydroalcoholic extract was discovered to have the ability to inhibit α-amylase, with IC50 values of 57.38588±1.92 μg/mL and 32.51564±1.59 μg/mL for standard acarbose, respectively. Conclusion: Overall, this research provides comprehensive data for the estimation of kaempferol in Moringa oleifera by HPTLC and presents valuable evidence on its antidiabetic potential both in vitro and in silico.