Metabolism of Ginsengside Rb1 in Rat Liver Microsomal in vitro by HPLC-MS and Cocktail Probe Substrats Method†

doi:10.7503/cjcu20150307

Abstract

Abstract:

The effect and metabolisms of ginsenoside Rb1 were studied on cytochrome P450s(CYP450). The induction or inhibition effects of CYP450 enzymes were evaluated by investigating the changes of probe drug in concentration. The metabolites of ginsenoside Rb1 including ginsenoside Rd, ginsenoside F2 and the oxidation of ginsenoside Rb, were found and identified in CYP450 enzyme with RRLC-MS/MS. The difference between the experimental group and control group was not significant except CYP1A2 group, indicating that ginsenoside Rb1 has more induced effect to the CYP1A2 group than the others. Ginsenoside Rb1 promoted reaction and speed up the metabolism of probe drug. The method can effectively and easily analyze the impact on the enzyme and possible metabolites.

Key words: Ginsenoside Rb1, Cytochrome P450 enzyme, Liquid chromatogram-mass spectrum(LC-MS), Metabolism in vitro(Ed.: N, K)

CLC Number:

O657.6

TrendMD:

WANG Yibo, XIAO Dan, LI Xiaoyu, DAI Yulin, YUE Hao, LIU Shuying. Metabolism of Ginsengside Rb1 in Rat Liver Microsomal in vitro by HPLC-MS and Cocktail Probe Substrats Method^†[J]. Chem. J. Chinese Universities, 2015, 36(10): 1894.

Figures/Tables 4

Table 1 Regression equations and correlation coefficients of four probe drugs(Mean±SD, n=3)*

Probe drug	Liner range/(ng·mL^-1)	Regression equation	R²
Phenacetin	8—9000	y=(0.003351±0.000342)c+(0.027752±0.0164271)	0.9968±0.0028
Omeprazole	8—9000	y=(0.003957±0.000569)c+(0.064631±0.0284354)	0.9987±0.0117
Carbamazepine	8—9000	y=(0.003527±0.004322)c+(0.104952±0.1797514)	0.9942±0.0056
Phenytoin	8—9000	y=(0.013345±0.002689)c+(0.088254±0.0290461)	0.9904±0.0134

Fig.1 Mass spectra of positive ion mode about the four probe drugs(A) Blank liver microsomal incubation system; (B) blank liver microsomal incubation system+phenytoin+omeprazole+carbamazepine+phenacetin; (C) liver microsomal incubation system with ginsengside Rb1 added to.

Fig.2 Effects on the cytochrome P450s activities of ginsengside Rb1*P<0.05 vs. blank group; **P<0.05 vs. control group.

Fig.3 Chromatogram(A) and mass spectra(B—E) of ginsengside Rb1 and its metabolites in liver microsome(A) Chromatogram of ginsengside Rb1 and its metabolites in liver microsome. Peak Ⅰ: ginsengside Rb1; peak Ⅱ: m/z=1123.5900; peak Ⅲ: ginsengside Rd; peak Ⅳ: ginsegnside F2; (B) mass spectra of Ⅰ; (C) mass spectra of Ⅱ; (D) mass spectra of Ⅲ; (E) mass spectra of Ⅳ.

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