Hsieh J Y, Lin C, Matuszewski B K, Dobrinska M R
Department of Drug Metabolism, Merck Research Laboratories, West Point, PA 19486, USA.
J Pharm Biomed Anal. 1994 Dec;12(12):1555-62. doi: 10.1016/0731-7085(94)00096-4.
LC assays utilizing fully automated sample preparation procedures on Zymark PyTechnology Robot and BenchMate Workstation for the quantification of hydrochlorothiazide (HCTZ) in human plasma and urine have been developed. After aliquoting plasma and urine samples, and adding internal standard (IS) manually, the robot executed buffer and organic solvent addition, liquid-liquid extraction, solvent evaporation and on-line LC injection steps for plasma samples, whereas, BenchMate performed buffer and organic solvent addition, liquid-liquid and solid-phase extractions, and on-line LC injection steps for urine samples. Chromatographic separations were carried out on Beckman Octyl Ultrasphere column using the mobile phase composed of 12% (v/v) acetonitrile and 88% of either an ion-pairing reagent (plasma) or 0.1% trifluoroacetic acid (urine). The eluent from the column was monitored with UV detector (271 nm). Peak heights for HCTZ and IS were automatically processed using a PE-Nelson ACCESS*CHROM laboratory automation system. The assays have been validated in the concentration range of 2-100 ng ml-1 in plasma and 0.1-20 micrograms ml-1 in urine. Both plasma and urine assays have the sensitivity and specificity necessary to determine plasma and urine concentrations of HCTZ from low dose (6.25/12.5 mg) administration of HCTZ to human subjects in the presence or absence of losartan.
已经开发出利用Zymark PyTechnology机器人和BenchMate工作站上的全自动样品制备程序对人血浆和尿液中的氢氯噻嗪（HCTZ）进行定量的液相色谱（LC）分析方法。在等分血浆和尿液样品并手动添加内标（IS）后，机器人对血浆样品执行缓冲液和有机溶剂添加、液-液萃取、溶剂蒸发和在线LC进样步骤，而BenchMate对尿液样品执行缓冲液和有机溶剂添加、液-液和固相萃取以及在线LC进样步骤。色谱分离在Beckman辛基超球柱上进行，流动相由12%（v/v）乙腈和88%的离子对试剂（血浆）或0.1%三氟乙酸（尿液）组成。柱流出物用紫外检测器（271 nm）监测。使用PE-Nelson ACCESS*CHROM实验室自动化系统自动处理HCTZ和IS的峰高。该分析方法在血浆中2-100 ng/ml和尿液中0.1-20 μg/ml的浓度范围内得到了验证。血浆和尿液分析方法均具有在有或无氯沙坦存在的情况下，测定低剂量（6.25/12.5 mg）HCTZ给予人体受试者后血浆和尿液中HCTZ浓度所需的灵敏度和特异性。
