TLC-bioautography-MS-based Identification of Antioxidant, α-Amylase and α-Glucosidase Inhibitory Compounds in a Polyherbal Formulation “Sugreen-120”

Abstract

Background: Sugreen-120 is one of the famous Indian polyherbal formulations used for the treatment of diabetes. Due to a lack of scientific evidence, the present study is aimed at investigating the phytopharmacology of Sugreen-120 concerning its antioxidant and antidiabetic characteristics.Materials and Methods: Total phenols and flavonoid content followed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity was estimated in Sugreen-120. α-Amylase and α-glucosidase inhibitory action of Sugreen-120 was estimated to evaluate its antidiabetic potential. Thin layer chromatography (TLC)-bioautography-MS analysis was performed to determine DPPH free radical, α-amylase and α-glucosidase inhibitory phytoconstituents in Sugreen-120. High performance thin layer chromatography (HPTLC)-based quantitative analysis of Sugreen-120 was performed for the simultaneous separation of caffeic acid and kaempferol. In silico docking analysis was performed to determine the effect of Sugreen-120 metabolites against α-amylase and α-glucosidase enzymes.Results: The results showed that Sugreen-120 is enriched in total phenols and flavonoids and even has good potential to scavenge DPPH free radicals with an inhibitory concentration (IC50) value of 414.59 ± 4.925 µg/mL. In α-amylase and α-glucosidase inhibitory assays, the efficacy of Sugreen-120 was found in a dose-dependent manner and the IC50 values were found as 220.106 ± 1.375 and 441.44 ± 1.992 µg/mL, respectively. TLC-bioautographic analysis showed that 06 constituents were found active against DPPH free radical, 04 constituents active against α-amylase and 03 constituents active against α-glucosidase. A HPTLC quantitative study revealed the content of caffeic acid and kaempferol to be 5.233 ± 0.026 and 16.959 ± 0.036 µg/mg, respectively. In silico docking analysis showed that out of 5 identified metabolites, myricetin, ellagic acid, and kaempferol were found with significant interaction with α-amylase and α-glucosidase proteins. Hence, it can be concluded that Sugreen-120 exhibits not only an antidiabetic effect but also antioxidant potential. It can be a palliative choice and an alternative that can be used for the treatment of diabetes.

Keywords: Sugreen-120; Antidiabetes; Antioxidant; Phytoconstituents; In silico docking analysis