A rapid, simultaneous and quantitative analysis of 26 ginsenosides in white and red Panax ginseng using LC–MS/MS

Abstract A quantitative analysis of ginsenoside is very important for ginseng studies because each ginsenoside shows different medical activity and metabolic pathway. In this study, a rapid, simultaneous, and quantitative analysis of 26 ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, Rg2(R), Rg2(S), Rg3(S), Rg3(R), Rg5, Rg6, Rh1(R), Rh1(S), Rh2(R), Rh2(S), F1, F2, F3, F4, K, Mc, PPT(S), XVII, and Y) in white, and red Panax ginseng was established using multiple reaction monitoring (MRM) mode on ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC–MS/MS). The mobile phase of water and methanol containing 0.1% formic acid and HSS T3 C18 analytical column was used for the chromatographic separation. The four sets of stereoisomers were successfully separated within a 26-min run time, eluting the S-isomer faster than the R-isomer with higher concentration. The ginseng extract was diluted by 100, 400 and 8000 times to fit in the calibration range and quantitated by the standard addition method. Matrix matched calibration by mixing 64 µL of the ginseng extract with 16 µL of the standard solution was used for compensating the matrix effect. Such quantitation methodology using dilution, standard addition and matrix matching resulted in precise and unambiguous quantitation of 26 ginsenosides in ginseng products. Major ginsenosides were observed at relatively higher concentrations in red Panax ginseng and the Mc was detected and quantitated for the first time in this study. The comprehensive quantitation system established in this study will contribute to quality evaluation, breeding and culturing, and quantitative metabolomics study of ginseng.