Department of Pharmacy and Biotechnology (FaBiT), University of Bologna, Via Belmeloro 6, I-40126 Bologna, Italy.
Talanta. 2013 Jan 15;103:355-60. doi: 10.1016/j.talanta.2012.10.076. Epub 2012 Oct 29.
D-penicillamine is a thiol drug mainly used for Wilson's disease, rheumatoid arthritis and cystinuria. Adverse effects during normal use of the drug are frequent and may include skin lesions. To evaluate its toxic effects in clinical cases an original method based on high performance liquid chromatography coupled to amperometric detection in a specific biological matrix such as skin has been developed. The chromatographic analysis of D-penicillamine was carried out on a C18 column using a mixture of acid phosphate buffer and methanol as the mobile phase. Satisfactory sensitivity was obtained by oxidizing the molecule at +0.95 V with respect to an Ag/AgCl reference electrode. A chemical reduction of D-penicillamine-protein disulphide bonds using dithioerythritol combined with microwaves was necessary for the determination of the total amount of D-penicillamine in skin specimens. A further solid-phase extraction procedure on C18 cartridges was implemented for the sample clean-up. The whole analytical procedure was validated: high extraction yield (>91.0%) and satisfactory precision (RSD<6.8%) values were obtained. It was successfully applied to skin samples from a patient who was previously under a long-term, high-dose treatment with the drug and presented serious D-penicillamine-related dermatoses. Thus, the method seems to be suitable for the analysis of D-penicillamine in skin tissues.
青霉胺是一种巯基药物，主要用于威尔逊病、类风湿关节炎和胱氨酸尿症。在正常使用药物时，不良反应频繁，可能包括皮肤损伤。为了评估其在临床病例中的毒性作用，开发了一种基于高效液相色谱与特定生物基质（如皮肤）中安培检测相结合的原始方法。青霉胺的色谱分析在 C18 柱上进行，使用酸性磷酸盐缓冲液和甲醇的混合物作为流动相。通过将分子在+0.95 V 下相对于 Ag/AgCl 参比电极氧化，获得了令人满意的灵敏度。为了测定皮肤标本中青霉胺的总量，需要使用二硫苏糖醇对青霉胺-蛋白质二硫键进行化学还原，并结合微波。对 C18 小柱进行进一步的固相萃取程序，对样品进行净化。整个分析程序得到了验证：获得了高提取产率（>91.0%）和令人满意的精密度（RSD<6.8%）值。该方法成功应用于一位长期接受高剂量药物治疗且出现严重青霉胺相关皮肤病的患者的皮肤样本。因此，该方法似乎适用于皮肤组织中青霉胺的分析。
