Kuśmierek Krzysztof, Bald Edward
Department of Environmental Chemistry, University of Lodz, 163 Pomorska Str., 90-236 Lodz, Poland.
Anal Chim Acta. 2007 May 2;590(1):132-7. doi: 10.1016/j.aca.2007.03.025. Epub 2007 Mar 18.
d-Penicillamine and tiopronin are drugs widely used for the treatment of many diseases. Because of the relatively high frequency of side effects to these compounds, some of which are dose-related, drug monitoring in urine samples during treatment is advisable. In this paper, we describe a simple method for the determination of tiopronin and d-penicillamine in human urine. The method was based on derivatization with 2-chloro-1-methylquinolinium tetrafluoroborate followed by ion-pairing reversed-phase liquid chromatography separation and ultraviolet-absorbance detection. 2-S-quinolinium derivatives of thiols were detected at 355 nm. The derivatization was optimized in terms of pH and time of the reaction. Baseline separation was achieved on an analytical Zorbax SB C-18 (5 microm, 150 mm x 4.6 mm) column with a mobile phase consisting of pH 2.0 0.09 mol L(-1) trichloroacetic acid buffer (component A) and acetonitrile (component B) pumped at 1.0 mL min(-1). Gradient elution was used: 0-4 min, 12% B; 4-8 min, 12-40% B; 8-12 min, 40-12% B. The d-penicillamine and tiopronin standards added to the urine show that the response of the detector is linear within the range studied, from 1 to 200 micromol L(-1) urine. The imprecision ranges for tiopronin and d-penicillamine were within 1.61-8.24% and 2.92-10.60%, respectively. The analytical accuracy for determined compounds was from 97.24 to 109.39%. The lower limits of detection and quantitation were 0.5 micromol L(-1) and 1.0 micromol L(-1) urine, respectively. This method can be used for routine clinical monitoring of the title thiol-drugs. Cysteine can be measured concurrently, if needed.
青霉胺和硫普罗宁是广泛用于治疗多种疾病的药物。由于这些化合物的副作用发生率相对较高，其中一些与剂量相关，因此在治疗期间对尿液样本进行药物监测是可取的。在本文中，我们描述了一种测定人尿中硫普罗宁和青霉胺的简单方法。该方法基于用四氟硼酸2-氯-1-甲基喹啉鎓进行衍生化，然后进行离子对反相液相色谱分离和紫外吸收检测。硫醇的2-S-喹啉鎓衍生物在355nm处检测。对衍生化反应的pH和时间进行了优化。在分析型Zorbax SB C-18（5μm，150mm×4.6mm）柱上实现了基线分离，流动相由pH 2.0的0.09mol L(-1)三氯乙酸缓冲液（A组分）和乙腈（B组分）组成，流速为1.0mL min(-1)。采用梯度洗脱：0-4分钟，12% B；4-8分钟，12-40% B；8-12分钟，40-12% B。添加到尿液中的青霉胺和硫普罗宁标准品表明，在所研究的1至200μmol L(-1)尿液范围内，检测器的响应呈线性。硫普罗宁和青霉胺的不精密度范围分别在1.61-8.24%和2.92-10.60%之间。所测化合物的分析准确度为97.24%至109.39%。检测限和定量下限分别为0.5μmol L(-1)尿液和1.0μmol L(-1)尿液。该方法可用于对上述硫醇类药物进行常规临床监测。如有需要，可同时测定半胱氨酸。
