Jangid Arvind G, Tale Rajesh H, Vaidya Vikas V
School of Chemical Sciences, Swami Ramanand Teerth Marathwada University, Nanded, India.
Biomed Chromatogr. 2012 Jan;26(1):95-100. doi: 10.1002/bmc.1632. Epub 2011 Apr 13.
A single, simple and selective method for simultaneous estimation of amiloride and hydrochlorothiazide in human plasma was validated using triamterine and hydrochlorothiazide (13)C,d2 as internal standard. The compounds were separated on a reverse-phase column with an isocratic mobile phase consisting of 2 mm ammonium acetate pH 3.0 and acetonitrile (30:70, v/v) and detected by tandem mass spectrometry with positive/negative ion mode. The analytes and internal standards were extracted from plasma using simple solid phase extraction. The ion transitions recorded in multiple reaction monitoring mode were m/z 230.1 → 116.0 for amiloride, m/z 254.1 → 237.1 for internal standard, triamterine in positive mode and m/z 296.1 → 204.9 for hydrochlorothiazide, m/z 299.2 → 205.8 for internal standard, hydrochlorothiazide (13)C,d2 in negative ion mode. Linearity in plasma was observed over the concentration range 0.1-10 ng/mL for amiloride and 5.0-500.0 ng/mL for hydrochlorothiazide. The mean recovery was 41.1 and 81.5% for amiloride and hydrochlorothiazide respectively. The coefficient of variation of the assay was less than 11.2 and 5.2% for amiloride and hydrochlorothiazide, respectively, and the accuracy was 89.0-98.1 and 96.6-102.9% for amiloride and hydrochlorothiazide, respectively. The validated method can be applied to the pharmacokinetic study of amiloride and hydrochlorothiazide.
采用氨苯蝶啶和氢氯噻嗪（13）C，d2作为内标，验证了一种用于同时测定人血浆中阿米洛利和氢氯噻嗪的单一、简单且选择性的方法。化合物在反相柱上分离，等度流动相由2 mM pH 3.0的醋酸铵和乙腈（30:70，v/v）组成，并通过串联质谱在正/负离子模式下检测。使用简单的固相萃取从血浆中提取分析物和内标。在多反应监测模式下记录的离子跃迁为：正模式下，阿米洛利的m/z 230.1 → 116.0，内标氨苯蝶啶的m/z 254.1 → 237.1；负离子模式下，氢氯噻嗪的m/z 296.1 → 204.9，内标氢氯噻嗪（13）C，d2的m/z 299.2 → 205.8。血浆中阿米洛利的线性范围为0.1 - 10 ng/mL，氢氯噻嗪的线性范围为5.0 - 500.0 ng/mL。阿米洛利和氢氯噻嗪的平均回收率分别为41.1%和81.5%。该测定法的变异系数对于阿米洛利和氢氯噻嗪分别小于11.2%和5.2%，准确度对于阿米洛利和氢氯噻嗪分别为89.0 - 98.1%和96.6 - 102.9%。经验证的方法可应用于阿米洛利和氢氯噻嗪的药代动力学研究。
