Quality Control Method (UPLC-PDA) of Ajuga parviflora Benth. and Its Antiadipogenic Effect on Differentiated Preadipocytes

Abstract

Introduction

Ajuga parviflora is traditionally used for fever, diabetes, and digestive problems. Currently, Ultraperformance liquid chromatography-Photodiode array (UPLC-PDA) method to determine ajugasterone C, cyasterone, and vanillic acid in A. parviflora was focused to develop and validate a method for quality control perspective. Further, the evaluation of antiadipogenic and antidiabetic potential of A. parviflora was also targeted.

Methods

UPLC-PDA method was developed and validated for targeted compounds as per International Council on Harmonisation guidelines. The validated method was used to determine marker compounds in A. parviflora extracts (Ethanol: EtOH, 50% EtOH, and water; leaves and roots). Further, UPLC-ELSD was used to determine free sugars in samples. Moreover, the antiadipogenic effect of A. parviflora extracts was examined on 3T3-L1 murine preadipocytes cell line. Cells were subjected to various dosages of leaves and root extracts, and the extent of lipid accumulation was evaluated. Furthermore, cells were treated with different extracts prior to hydrogen peroxide exposure, and then the effects of A. parviflora treatments on oxidative stress, cell survival, and insulin sensitisation were assessed.

Results

The validated UPLC-PDA method was found reproducible to determine ajugasterone C, cyasterone, and vanillic acid. These compounds were found in all the samples. Biologically, leaves and roots extract of A. parviflora drastically suppressed adipogenesis by lowering intracellular lipid accumulation in dose-dependent manner. They improved insulin sensitivity by promoting glucose uptake and protected cellular health from oxidative damage by reducing reactive oxygen species generation and reversing apoptosis.

Conclusions

Findings suggested that ethanol extract of leaves exhibited potent antiadipogenic properties and UPLC-PDA will be a reproducible method to assess quality of A. parviflora and its derived products.