Analytical characterization of biomarkers in an optimized novel antidiabetic polyherbal formulation using high-performance thin-layer chromatography and liquid chromatography with tandem mass spectrometry

Background Diabetes mellitus is a chronic health issue that requires novel approaches to treatment and a multimodal approach to prevention. In the treatment of diabetes, a polyherbal formulation is the finest alternative medicine. A polyherbal formulation was developed in-house and evaluated for its antidiabetic potential on streptozotocin-induced diabetes rat. The same extract was now characterized analytically utilizing a variety of methods.Objective The goal of this study was to quantify the biomarkers in a novel antidiabetic polyherbal formulation made in-house with Cinnamonum zeylanicium bark, Eugenia jambolana seeds, Vinca rosea whole plant, and Gymnema sylvestre (GS) leaves, using high-performance thin-layer chromatography (HPTLC) and liquid chromatography with tandem mass spectrometry (LC-MS/MS).Materials and methods Cinnamaldehyde (CIN), gallic acid (GLA), vincristine (VC), vinblastine (VB), and gymnemic acid (GYA) were identified as bioactive components of polyherbal formulation hydroalcoholic extract utilizing HPTLC and LC-MS/MS. Acetonitrile, methanol, and 0.1 percent formic acid was used as mobile phase, chromatographic separation was accomplished in 30 min using a gradient system and a SUNFIRE C18, 250x4.6, 5-mu m analytical column with a flow rate of 1.0 ml/min in LC-MS/MS research. Scanned in a positive mode with a scan speed of 100-2000 AMU/s over a mass range of 20-1974 Da. The electron-spray ionization mode was used, with a source temperature of 150 & DEG;C and a desolvation temperature of 350 & DEG;C. The HPTLC separation was performed using ethyl acetate/acetonitrile/water/formic acid/N-dimethyl formamide 5.5 : 2.5 : 0.5 : 1.0 : 0.5 (v/v) as the mobile phase on precoated silica gel 60 GF254 plates. At room temperature, the plates were developed to a distance of 9.0 cm. CIN, GLA, VC, VB, and GYA plates were scanned and measured at wavelengths of maximum absorption of 259, 287, 342, 355, and 387 nm, respectively. Band size, chamber-saturation duration, migration of the solvent front, slit width, and other experimental parameters were carefully examined, and the optimized chromatographic conditions were chosen.Results LC-MS analysis of the hydroalcoholic extract of the polyherbal formulation revealed the presence of all the five bioactive chemical constituents, CIN, GLA, VC, VB, and GYA. Similarly, the drug samples were satisfactorily resolved with Rf 1.81 & PLUSMN;0.01, 0.05 & PLUSMN;0.01, 0.02 & PLUSMN;0.01, 0.09 & PLUSMN;0.01, and 0.04 & PLUSMN;0.01 for CIN, GLA, VC, VB, and GYA respectively, using HPTLC.Conclusion The importance of combining Ayurvedic formulations with contemporary high-throughput screening techniques will spark new interest in more powerful biocompatible drug leads. The findings of this study lend scientific credence to the therapeutic applications of the polyherbal formulation.