Kumar Arvind, Dwivedi Surya Prakash, Prasad Tulika
School of Biotechnology, IFTM University, Moradabad, India.
Advanced Instrumentation Research & Facility (AIRF) and Special Centre for Nano Sciences (SCNS), Jawaharlal Nehru University, New Delhi, India.
Front Pharmacol. 2019 Jul 23;10:810. doi: 10.3389/fphar.2019.00810. eCollection 2019.
A new, simple, sensitive, selective, rapid, and high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for simultaneous quantification of Olmesartan and hydrochlorothiazide in human plasma. Simple liquid-liquid extraction procedure was applied for plasma sample pretreatment using a mixture of diethyl ether and dichloromethane, as an extraction solution. Analytes were separated on UNISOL C18 150*4.6 mm, 5 µm column using methanol, and 2 mM ammonium acetate pH 5.5 (80:20, v/v) as a mobile phase and detected by electrospray ionization in the multiple reaction monitoring (MRM) mode. The mass transition ion pairs were followed in negative ion mode as m/z 445.20 → 148.90 for Olmesartan; m/z 451.40 → 154.30 for Olmesartan D and m/z 295.80 → 205.10 for hydrochlorothiazide; m/z 298.90 → 206.30 for hydrochlorothiazide C D. The method showed excellent linearity ( > 0.99) over the concentration range of 5.002-2,599.934 ng/ml for Olmesartan and from 3.005 to 499.994 ng/ml for hydrochlorothiazide. Precision (% CV) and accuracy (% bias) for Olmesartan were found in the range of 3.07-9.02% and -5.00-0.00%, respectively. Precision (% CV) and accuracy (% bias) for hydrochlorothiazide were found in the range of 3.32-8.21% and 1.99-3.80%, respectively. This as developed novel and high-throughput liquid-liquid extraction bioanalytical method has substantial innovative value with the benefits of cost effectiveness, good extraction efficiency, shorter analysis run time, low organic solvent consumption, and simpler procedure over the previously reported solid-phase extraction method. The application of this method in pharmacokinetic studies was further demonstrated successfully through a bioequivalence study conducted on healthy human subjects, following oral administration of combined formulation of Olmesartan medoxomil and hydrochlorothiazide in fixed-dose tablet.
已开发并验证了一种新的、简单、灵敏、选择性好、快速且高通量的液相色谱 - 串联质谱（LC-MS/MS）方法，用于同时定量测定人血浆中的奥美沙坦和氢氯噻嗪。采用简单的液 - 液萃取程序，以乙醚和二氯甲烷的混合物作为萃取溶液对血浆样品进行预处理。分析物在UNISOL C18 150*4.6 mm、5 µm柱上进行分离，以甲醇和2 mM pH 5.5的醋酸铵（80:20，v/v）作为流动相，并通过电喷雾电离在多反应监测（MRM）模式下进行检测。在负离子模式下，奥美沙坦的质量转移离子对为m/z 445.20 → 148.90；奥美沙坦D的为m/z 451.40 → 154.30；氢氯噻嗪的为m/z 295.80 → 205.10；氢氯噻嗪C D的为m/z 298.90 → 206.30。该方法在奥美沙坦浓度范围为5.002 - 2,599.934 ng/ml以及氢氯噻嗪浓度范围为3.005 - 499.994 ng/ml内显示出良好的线性（>0.99）。奥美沙坦的精密度（%CV）和准确度（%偏差）分别在3.07 - 9.02%和 -5.00 - 0.00%范围内。氢氯噻嗪的精密度（%CV）和准确度（%偏差）分别在3.32 - 8.21%和1.99 - 3.80%范围内。这种新开发的高通量液 - 液萃取生物分析方法具有显著的创新价值，与先前报道的固相萃取方法相比，具有成本效益高、萃取效率好、分析运行时间短、有机溶剂消耗低以及程序更简单等优点。通过对健康人体受试者口服奥美沙坦酯和氢氯噻嗪固定剂量片剂联合制剂后进行的生物等效性研究，进一步成功证明了该方法在药代动力学研究中的应用。
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