目的 建立同时测定丹参红花提取液中3种有效成分丹参素、羟基红花黄色素A(HSYA)、迷迭香酸含量的高效液相色谱-二极管阵列检测器(HPLC-DAD)方法;以丹参红花为模型药物,对微透析技术应用于中药复方药动学研究进行探索。方法 利用微透析法获得透析液,高效液相色谱-二极管阵列检测器(HPLC-DAD)梯度洗脱测定透析液中待测成分含量,计算丹参红花体外回收率,考察丹参素、羟基红花黄色素A、迷迭香酸在丹参提取液、红花提取液和丹参红花提取液透析媒介中回收率变化情况,并考察浓度、流速对回收率的影响。结果 3种待测成分在30 min内达到良好分离,线性关系良好(r>0.999 0,n=6)。当灌流速度为1.0 μL·min^-1时,混合标准液中丹参素钠、羟基红花黄色素A、迷迭香酸的回收率依次为 11.05%,5.81%,6.73%,随着透析媒介中物质数量的增多,特定成分的回收率有下降趋势,在相同浓度下回收率随流速增大而减少,但与透析媒介中该成分的浓度无关。结论 微透析技术可以用于丹参红花复方药动学研究。本实验建立的HPLC-DAD可作为微透析研究中丹参红花回收率的测定方法。微透析实验应考虑影响回收率因素,选择合适的实验条件,以方便微透析检测。

Objective To develop an HPLC-DAD method for the simultaneous quantitation of three effective constituents(salvianic acid A, hydroxysafflor yellow A and rosmarinic acid) in Danhong extract; traditional Chinese medicine and taking Danhong as a model drug to explore the feasibility of carrying out pharmacokinetic studies of traditional Chinese medicines by using microdialysis technique. Methods The samples were obtained by microdialysis method and determined by HPLC-DAD gradient elution to calculate the in vitro recovery rate(RR) of specific components. The difference of RRs of salvianic acid A, hydroxysafflor yellow A and rosmarinic acid in the dialysis media of Salvia extract, Safflower extract and Danhong extract, and the effects of flow rate and concentration on RR were studied. Results Good separation of salvianic acid A, hydroxysafflor yellow A and rosmarinic acid was achieved within 30 min, and the three constituents showed good linear relationship(r＞0.999 0, n=6). When the flow rate was 1.0 μL·min^-1, the RRS of salvianic acid A sodium, hydroxysafflor yellow A and rosmarinic acid in the mixed standard solution were 11.05%, 5.81%, and 6.73%, respectively. Along with the increase of the number of substances in the dialysis medium, the RRs of the specific components showed a downward trend. The RR reduced with increasing flow rate in the same dialysis medium, but was independent of the concentration of the component. Conclusion The microdialysis technique can be used in pharmacokinetic studies of traditional Chinese medicines. The developed HPLC-DAD method in this study can be used for determining the RR of Danhong in the microdialysis study. The factors influencing RR should be considered when choosing appropriate conditions for the microdialysis experiment, thus to make a convenient determination.