Skerjanec A, Tam Y K
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Canada.
J Chromatogr B Biomed Appl. 1995 Jul 21;669(2):377-82. doi: 10.1016/0378-4347(95)00102-o.
The objective of the study was to develop a sensitive and specific assay for studying the pharmacokinetics of a novel calcium antagonist, a benzimidazolyl-substituted tetraline derivative, mibefradil (I) in the dog. The assay involves liquid-liquid extraction of a biological sample, reversed-phase HPLC separation and fluorescence detection (lambda ex = 270 nm and lambda em = 300 nm) of sample components. Each sample was eluted with a mobile phase pumping at a flow-rate of 2 ml/min. The mobile phase composition was a mixture of acetonitrile and aqueous solution (38:62, v/v). The aqueous solution contains 0.0393 M KH2PO4 and 0.0082 M Na-pentanesulphonic acid. The retention times were 10.7 min for I, and 12.2 min for internal standard Ro 40-6792. Calibration curves with concentrations of I ranging from 10 to 500 ng/ml were linear (r2 > 0.99). The detection limit for I was 0.5 ng/ml when 0.5 ml of plasma or urine was used. Intra- and inter-day accuracy and precision were within 10%. The assay was successfully applied to the pharmacokinetic studies of I in dogs.
本研究的目的是开发一种灵敏且特异的分析方法，用于研究新型钙拮抗剂——一种苯并咪唑基取代的四氢萘衍生物米贝拉地尔（I）在犬体内的药代动力学。该分析方法包括对生物样品进行液 - 液萃取、反相高效液相色谱分离以及对样品成分进行荧光检测（激发波长λex = 270 nm，发射波长λem = 300 nm）。每个样品以2 ml/min的流速用流动相洗脱。流动相组成为乙腈和水溶液的混合物（38:62，v/v）。水溶液含有0.0393 M磷酸二氢钾和0.0082 M戊烷磺酸钠。I的保留时间为10.7分钟，内标Ro 40 - 6792的保留时间为12.2分钟。I浓度范围为10至500 ng/ml的校准曲线呈线性（r2 > 0.99）。当使用0.5 ml血浆或尿液时，I的检测限为0.5 ng/ml。日内和日间的准确度和精密度均在10%以内。该分析方法已成功应用于米贝拉地尔在犬体内的药代动力学研究。
