Ivanova Violeta, Zendelovska Dragica, Stefova Marina, Stafilov Trajce
Institute of Chemistry, Faculty of Science, Sts. Cyril and Methodius University, POB 162, 1000 Skopje, Republic of Macedonia.
J Biochem Biophys Methods. 2008 Apr 24;70(6):1297-303. doi: 10.1016/j.jbbm.2007.09.009. Epub 2007 Oct 6.
A simple, rapid and precise HPLC method has been developed for the assay of verapamil in human plasma. The clean up of the plasma samples was tested using several adsorbents for solid-phase extraction and best recovery was obtained using mixed-mode cartridges (HLB - hydrophilic-lipophilic balance) ranging between 94.70 and 103.71%. HPLC separation was performed with isocratic elution on Lichrospher 60 RP-select B column (250 mm x 4 mm I.D., 5 microm particle size). The mobile phase was 40% acetonitrile and 0.025 mol/L KH2PO4 with pH 2.5 at flow rate of 1 mL/min. Diltiazem was used as internal standard and the detection wavelength was 200 nm. The calibration curves were linear in the range of 10-500 ng/mL. The developed method is convenient for routine analysis of verapamil in human plasma.
已开发出一种简单、快速且精确的高效液相色谱(HPLC)方法用于测定人血浆中的维拉帕米。使用多种吸附剂进行固相萃取以测试血浆样品的净化效果,使用混合模式柱(HLB - 亲水亲脂平衡)获得了最佳回收率,回收率在94.70%至103.71%之间。在Lichrospher 60 RP-select B柱(250 mm×4 mm内径,5微米粒径)上采用等度洗脱进行HPLC分离。流动相为40%乙腈和0.025 mol/L磷酸二氢钾,pH 2.5,流速为1 mL/min。地尔硫卓用作内标,检测波长为200 nm。校准曲线在10 - 500 ng/mL范围内呈线性。所开发的方法便于对人血浆中的维拉帕米进行常规分析。