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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1.0μL·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 establish a high performance liquid chromatography-diode array detector (HPLC-DAD) method for the simultaneous determination of three active ingredients, danshensu, hydroxysafflor yellow A (HSYA) and rosmarinic acid, Salvia safflower as a model drug, the application of microdialysis technology in traditional Chinese medicine pharmacokinetic study to explore. Methods The dialysate was obtained by microdialysis and the content of analyte in the dialysate was determined by HPLC-DAD gradient elution. The in vitro recovery of Danshen Honghua was calculated. The effects of Danshensu, Yellow pigment A, rosmarinic acid in Salvia extract, safflower extract and Danshen Safflower extract dialysis medium changes in the recovery rate, and study the concentration and flow rate on the recovery rate. Results The three analytes were well separated within 30 min with a good linear relationship (r> 0.999 0, n = 6). When the perfusion rate was 1.0 μL · min-1, the recoveries of sodium danshensu, hydroxysafflor yellow A and rosmarinic acid in mixed standard solutions were 11.05%, 5.81% and 6.73%, respectively. With the dialysis medium The increase of the number of substances, the recovery rate of specific components have a downward trend, the same concentration of the recovery rate decreases with the increase of flow rate, but with the dialysis medium concentration of the ingredient has nothing to do. Conclusion Microdialysis technology can be used for the pharmacokinetics of Compound Danshen Honghua. The HPLC-DAD established in this experiment can be used as a method for the determination of the recovery rate of safflor yellow in microdialysis. Microdialysis experiments should consider the impact of recovery factors, select the appropriate experimental conditions to facilitate the detection of microdialysis.