Iriarte Gorka, Ferreirós Nerea, Ibarrondo Izaskun, Alonso Rosa Maria, Itxaso Maguregui Miren, Jiménez Rosa Maria
Kimika Analitikoaren Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea/UPV, Bilbo, Basque Country, Spain.
J Sep Sci. 2007 Sep;30(14):2231-40. doi: 10.1002/jssc.200700033.
A simple and fast method for the simultaneous determination of the antihypertensive drug Valsartan and its metabolite in human plasma has been validated. The proposed method deals with SPE, followed by an HPLC separation coupled with fluorimetric and photometric detection. The optimization of the SPE-HPLC method was achieved by an experimental design. The separation was performed on an RP C18 Atlantis 100 mmx3.9 mm column. The mobile phase consisted of a mixture of ACN 0.025% TFA and phosphate buffer (5 mM, pH = 2.5) 0.025% TFA and was delivered in gradient mode at a flow rate of 1.30 mL/min. The eluent was monitored with a fluorescence detector at 234 and 378 nm excitation and emission wavelengths, respectively, and at 254 nm using a photometric detector. The full analytical validation was performed according to the Food and Drug Administration (FDA) 'guidance for industry: bioanalytical method validation' and the recoveries obtained for Valsartan and its metabolite ranged from 94.6 to 108.8%. The validated method was successfully applied to 12 plasma samples obtained from patients under antihypertensive treatment with Valsartan.
已验证一种同时测定人血浆中抗高血压药物缬沙坦及其代谢物的简单快速方法。所提出的方法涉及固相萃取（SPE），随后进行高效液相色谱（HPLC）分离，并结合荧光和光度检测。通过实验设计实现了SPE-HPLC方法的优化。分离在RP C18 Atlantis 100 mm×3.9 mm柱上进行。流动相由乙腈-0.025%三氟乙酸（TFA）和磷酸盐缓冲液（5 mM，pH = 2.5）-0.025% TFA的混合物组成，并以梯度模式在1.30 mL/min的流速下输送。分别使用荧光检测器在激发波长234 nm和发射波长378 nm以及使用光度检测器在254 nm处监测洗脱液。根据美国食品药品监督管理局（FDA）的“工业指南：生物分析方法验证”进行了全面的分析验证，缬沙坦及其代谢物的回收率在94.6%至108.8%之间。该验证方法已成功应用于从接受缬沙坦抗高血压治疗的患者中获得的12份血浆样本。
