Rehman Shaheed Ur, Kim In Sook, Kang Ki Sung, Yoo Hye Hyun
Institute of Pharmaceutical Science and Technology and College of Pharmacy, Hanyang University, Ansan, Gyeonggi-do 426-791, Republic of Korea.
College of Korean Medicine, Gachon University, Seongnam, Gyeonggi-do 461-701, Republic of Korea.
J Chromatogr Sci. 2015 Sep;53(8):1322-7. doi: 10.1093/chromsci/bmv014. Epub 2015 Feb 23.
An optimized, sensitive and validated reversed-phase high-performance liquid chromatography (RP-HPLC) method with UV detection is described for simultaneous determination of esculin and its aglycone, esculetin, in rat plasma. After addition of internal standard (chrysin), plasma samples were pretreated by solid-phase extraction and introduced into the HPLC system. Analytes were separated on a RP C18 column with a mobile phase of 0.075% acetic acid in water (solvent A) and 90% acetonitrile in solvent A (solvent B) using gradient elution at a flow rate of 1.0 mL/min. The wavelength for UV detection was set at 338 nm. Calibration curves for esculin and esculetin were constructed over a range of 10-1,000 ng/mL. The developed method was found to be specific, precise and accurate. The method was successfully applied to study the pharmacokinetics of esculin and esculetin in rats. After oral administration of 120 mg/kg, the mean Cmax values were 340.3 and 316.5 ng/mL and the AUClast values were 377.3 and 1276.5 h ng/mL for esculin and esculetin, respectively. The bioavailability of esculin was calculated to be 0.62%.
本文描述了一种经过优化、灵敏且经过验证的反相高效液相色谱（RP-HPLC）法，并采用紫外检测，用于同时测定大鼠血浆中的七叶苷及其苷元七叶亭。加入内标（白杨素）后，血浆样品经固相萃取预处理，然后引入HPLC系统。分析物在RP C18柱上分离，流动相为0.075%乙酸水溶液（溶剂A）和90%乙腈的溶剂A溶液（溶剂B），采用梯度洗脱，流速为1.0 mL/min。紫外检测波长设定为338 nm。七叶苷和七叶亭的校准曲线在10 - 1000 ng/mL范围内构建。所建立的方法具有特异性、精密度和准确性。该方法成功应用于大鼠体内七叶苷和七叶亭的药代动力学研究。口服给药120 mg/kg后，七叶苷和七叶亭的平均Cmax值分别为340.3和316.5 ng/mL，AUC last值分别为377.3和1276.5 h ng/mL。七叶苷的生物利用度经计算为0.62%。
