Rapid Screening of Lipase Inhibitors from Ophiopogonis Radix Using High-Performance Thin Layer Chromatography by Two Step Gradient Elution Combined with Bioautographic Method.

Xue Hua,Hui-Jie Hong,Dai-Yan Zhang,Fong Leong,Qiao Liu,Yuan-Jia Hu,Qi Yang,Xiao-Jia Chen

doi:10.3390/molecules27041155

Abstract

In this study, a high-performance thin layer chromatography (HPTLC) method by two step gradient elution with two mobile phases was developed for the simultaneous analysis of seven constituents in Ophiopogonis Radix. The chromatography was performed on silica gel 60 F254 plate with dichloromethane-methanol-ethyl acetate-water (70:25:12:3, v/v/v/v) and dichloromethane-methanol (300:1, v/v) as the mobile phase for two step gradient elution. Then, the HPTLC profiles were observed after derivatization with 10% sulfuric acid in ethanol solution. The obtained HPTLC images were further analyzed by chemometric approaches and the samples could be clustered based on regions and/or growth years, which were two important factors affecting the constituents in Ophiopogonis Radix. Furthermore, five compounds including ophiopogonin D, ophiopojaponin C, ophiopogonin D’, ophiopogonin C’ and methylophiopogonanone B were screened as potential lipase inhibitors from Ophiopogonis Radix by the HPTLC-bioautographic method. The binding modes and interactions between the five compounds and lipase were further explored by molecular docking analysis. The developed HPTLC method could be used for quality control of Ophiopogonis Radix and screening of the potential lipase inhibitors.

Highlights

Obesity, which means abnormal or excessive fat accumulation, is considered as one of the greatest threats to human health
While in Hong Kong Chinese Materia Medica Standards, Thin layer chromatography (TLC) analysis is performed by using dichloromethane-methanol-water (8:2:0.3, v/v/v) as the mobile phase for the identification of ophiopogonin D, a polar saponin [27], but low-polarity components could not be separated under this condition
Our study showed that the growth year was an important factor on the constituents of Ophiopogonis Radix

Summary

Introduction

Obesity, which means abnormal or excessive fat accumulation, is considered as one of the greatest threats to human health. According to the World Health Organization, more than 650 million adults (18 years and older) and over 340 million children and adolescents aged 5–19 were overweight or obese in 2016 [1]. Pancreatic lipase is a key enzyme for the digestion of triacylglycerols, and inhibition of lipase has been the most explored strategy for the treatment of obesity until now. More and more medicinal herbs have been reported to show inhibitory activities against pancreatic lipase, such as Morus alba [5], Atractylodes lancea [6], Forsythia suspensa [7], and Camellia nitidissima [8]. Discovery of lipase inhibitors from herbal medicines may provide a potential alternative for the treatment of obesity

Methods

Results

Conclusion