人参皂苷Rb_3在Caco-2单细胞层模型上的吸收特征研究

目的:研究人参皂苷Rb3在Caco-2单细胞层模型上的吸收特征。方法:人参皂苷Rb3细胞样品经高速离心后取上清液,以乙腈-1mmo·lL-1甲酸铵水溶液(34∶66)为流动相,以人参皂苷Rg2为内标,采用电喷雾离子源(ESI),在负离子条件下以多重离子反应监测模式(MRM)检测。检测离子:m/z1077.7→m/z783.4(人参皂苷Rb3),m/z783.6→m/z475.1(人参皂苷Rg2)。测定透过Caco-2单细胞层的人参皂苷Rb3浓度并计算其表观渗透系数(Papp)。结果:人参皂苷Rb3的线性范围为50～2000ng·mL-1,日间、日内精密度均小于15%。Papp从基顶侧(AP)到基底侧(BL)为3.22×10-6cm·s-1,从BL侧到AP侧为6.0×10-6cm·s-1,其P(BL-AP)/P(AP-BL)比值为1.86。结论:本方法简单、灵敏度高,可用于研究人参皂苷Rb3在Caco-2单细胞层模型上的吸收特征。

Absorptive Characteristics of Ginsenoside Rb_3 in Caco-2 Cell Monolayer Model

OBJECTIVE ： To study the absorptive characteristics of ginsenoside Rb~ in Caco-2 cell monolayer model. METHODS：The ginsenoside Rb3 cell samples underwent high speed centrifugation, then the supernatant was collected and determined by LC-ESI-MS/MS method in which the mobile phase consisted of acetonitrile-1 mmol. Ll ammonium formate water solution （34： 66） with ginsenoside Rg2 as internal standard, and the tandem mass spectrometry was operated in negative electrospray ionization in a multiple reaction monitoring （MRM） mode, with detection ions m/z 1077.7→m/z 783.4 for ginsenoside Rb3 and m/z 783.6→m/z 475.1 for ginsenoside Rg2.The concentration of ginsenoside Rb3 across the Caco-2 cell monolayer model was determined and the apparent permeability coefficient（Papp） of ginsenoside Rb3 was calculated. RESULTS： The calibration curve for ginsenoside Rb3 was linear in the range of 5042 000 ng·mL^-1, with intra-day precision and inter-day precision at less than 15%. P（AP-BL） from apical side （AP） to basolateral side （BL） was 3.22×10^-6cm·s^-1, whereas P（BL-AP） from BL to AP was 6.0×10^-6cm·s^-1,and the ratio of P（BL-AP）/P（AP-BL） was 1.86.CONCLUSION：The LC-ESI/MS/MS method is simple and sensitive, and it is applicable for the study of the absorptive characteristics of ginsenoside Rb3 in Caco-2 cell monolayer model.