Simultaneous Determination of 25 Ginsenosides by UPLC-HRMS via Quantitative Analysis of Multicomponents by Single Marker.

Xiujuan Jia,Ye Yuan,Yifa Zhou,Chenxing Hu,Xuepeng Zhu,David M Lubman

doi:10.1155/2021/9986793

Abstract

A method using UPLC-HRMS has been developed for a rapid, simultaneous qualitative and quantitative analysis of twenty-five ginsenosides. Chromatographic separation was achieved on a C18 analytical column with an elution gradient comprising 0.1% aqueous formate/acetonitrile as the mobile phase. HRMS detection acquired full mass data for quantification and fullms-ddms2 (i.e., data-dependent scan mode) yielded product ion spectra for identification. Furthermore, quantitative analysis of multiginsenosides by single marker (QAMS) was developed and validated using a relative correction factor. Under optimal conditions, we could simultaneously separate eight groups of isomers of the 25 ginsenosides. Good linearity was observed over the validated concentration range for each analyte (r2 > 0.9924), showing excellent sensitivity (LODs, 0.003–0.349 ng/mL) and lower limit quantification (LOQs, 0.015–1.163 ng/mL). The LC-MS external standard method (ESM) and QAMS were compared and successfully applied to analyze the ginsenoside content from Panax ginseng roots. Overall, our UPLC-HRMS/QAMS approach provides high precision, stability, and reproducibility and can be used for high-throughput analysis of complex ginsenosides and quantitative analysis of multiple components and quality control of traditional Chinese medicines (TCM).

Highlights

Optimization of ultra-performance LC (UPLC) Conditions. e separation of mixtures of ginsenosides by chromatography is very difficult. is is especially true for the simultaneous separation of eight groups of isomers (Rg1/Rf, Re/Rd/ GXVII, Rh1/F1, Rc/Rb2/Rb3, Notoginsenoside Fe (nFe)/Compound O (CO)/nFd, Rg2/G75/ Rg3/F2, Mc/Compound Y (CY)/Compound Mx (CMx), and Compound K (CK)/Rh2). e use of an appropriate chromatographic column is a key factor for separating ginsenosides. erefore, two chromatographic columns, including HyperSil GOLD C18 (2.1 mm × 100 mm, 1.9 μm) and HyperSil GOLD C18 (2.1 mm × 50 mm, 1.9 μm) columns, were employed and compared as shown in Figure S1 in the Supplementary Materials for comprehensive image analysis
Our results demonstrate that the HyperSil GOLD C18 (2.1 mm × 50 mm, 1.9 μm) column exhibited better resolution and higher peak capacity and could separate ginsenoside isomers Rg3 and G75. erefore, the HyperSil GOLD C18 (2.1 mm × 50 mm, 1.9 μm) column was chosen for this study
Our results demonstrated that relative standard deviation (RSD) measured at different temperatures were less than 1.79%, and at different flow rates were less than 7.60%. is indicates that relative retention time (RTR) is reproducible at different temperatures when the flow rate is constant. us, RTR can be used to accurately locate the chromatographic peak of each analyte

Summary

Introduction

Several methods have been developed to assess ginsenoside content in ginseng. Among these approaches, high-performance liquid chromatography (HPLC) and HPLC-MS are by far the most employed analytical methods [4,5,6,7,8]. Ginsenosides that have been reported were not well separated, leading to inaccurate quantitation. HPLC-MS is a powerful tool for determining ginsenosides in diverse ginseng extracts [9, 10]. Thirteen ginsenosides were simultaneously and quantitatively determined by HPLC-MS [9]. Among the various LC approaches, ultra-performance LC (UPLC) has provided better separations over short periods of time and increased sample throughput and sensitivity [11]. Among different MS analyzers, Orbitrap has provided much better accuracy, precision, and greater mass resolution [12]

Methods

Results

Conclusion