三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1的HPLC定量分析及其肠黏膜渗透性研究

Quantitative analysis of notoginsenoside R1,ginsenoside Rg1 and ginsenoside Rb1 by HPLC and their intestinal mucosal permeability

目的通过前期建立的3种体外肠黏液渗透模型[纯化黏蛋白渗透模型(PIM)、人工肠黏液渗透模型(AIM)及大鼠肠黏液渗透模型(RIM)],研究三七中主要单体成分三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1在肠黏液层的渗透作用,以期为三七皂苷类成分的口服吸收及应用提供实验依据。方法采用高效液相色谱(HPLC)法建立三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1的定量分析方法,并对其线性范围、精密度、稳定性、重复性、加样回收率进行考察;分别测定不同浓度三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1溶液在3种肠黏液渗透模型上的表观渗透系数(Papp),分析比较其肠黏液渗透作用。结果建立了三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1的HPLC定量分析方法,精密度、稳定性、重复性、加样回收率均符合要求。3种皂苷成分在PIM、AIM、RIM模型的渗透性结果显示:三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1在3种模型上的渗透作用随着药物浓度的升高均有不同程度的增加;在PIM中,与人参皂苷Rb1和人参皂苷Rg1相比,三七皂苷R1在不同浓度下的Papp均显著增加(P<0.05);在AIM中,20g·L-1浓度下的Papp为三七皂苷R1>人参皂苷Rb1>人参皂苷Rg1(P<0.05);在RIM中,20 g·L-1浓度下的Papp为人参皂苷Rg1>三七皂苷R1>人参皂苷Rb1(P<0.05)。结论不同浓度的三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1在3种肠黏液渗透模型中的稳定性、重现性较好,且随着药物浓度的上升渗透性也随之增加;水溶性较强且分子量相对较小的三七皂苷R1在脂类成分较少的PIM和AIM中渗透作用较强,而脂溶性较强的人参皂苷Rg1在RIM中渗透作用较强。

Objective To investigate the effects of notoginsenoside R1,ginsenoside Rg1 and Rb1 on intestinal mucus permeation by using three in vitro intestinal mucus permeation models[purified mucin infiltration model(PIM),artificial intestinal mucus infiltration model(AIM)and rat intestinal mucus infiltration model(RIM)]in order to provide experimental basis for the oral absorption and application of notoginseng Radix Et Rhizoma.Methods Quantitative analysis methods of notoginsenoside R1,ginsenoside Rg1 and Rb1 were established by high performance liquid chromatography(HPLC),and their linear range,precision,stability,reproducibility and sample recovery rate were investigated.The apparent permeability coefficients(Papp)of different concentrations of notoginsenoside R1,ginsenoside Rg1 and ginsenoside Rb1 solutions on three intestinal mucus permeation models were determined and analyzed to compare their intestinal mucus permeation effects.Results The HPLC methods for the quantitative analysis of notoginsenoside R1,ginsenoside Rg1 and ginsenoside Rb1 were successfully established.The precision,stability,reproducibility and recovery all met the requirements.The permeability results of three kinds of saponins in PIM,AIM and RIM models showed that the permeability of notoginsenoside R1 and ginsenoside Rg1 and ginsenoside Rb1 in the three models increased in different degrees with the increase of drug concentration.In PIM,notoginsenoside R1 showed a significant increase in the permeation rate compared with ginsenoside Rb1 and ginsenoside Rg1 at different concentrations(P<0.05).In AIM,the permeation rate at a concentration of 20 g·L-1 was PappR1>PappRb1>PappRg1(P<0.05).In RIM,the permeation rate at a concentration of 20 g·L-1 was PappRg1>PappR1>PappRb1(P<0.05).Conclusions Notoginsenoside R1,ginsenoside Rg1 and ginsenoside Rb1 with different concentrations have good stability and reproducibility in three intestinal mucus permeation models,and the permeability increases with the increase of drug concentration.Notoginsenoside R1 with strong water solubility and relatively low molecular weight is more permeable in PIM and AIM with less lipids,while ginsenoside Rg1 with strong fat solubility is stronger in RIM.