Plasma d-penicillamine redox state evaluation by capillary electrophoresis with laser-induced fluorescence
- 作者：Zinellu ; Angelo ; Carru ; Ciriaco ; Sotgia ; Salvatore ; Deiana ; Luca
- 关键词：CE-LIF ; laser-induced fluorescence capillary electrophoresis ; d-Pen ; d-penicillamine ; 5-IAF ; 5-iodoacetamidofluorescein ; SSA ; 5-sulphosalicylic acid ; TBP ; tri-n-butylphosphine ; d-Penicillamine ; Thiols
- 刊名：Journal of Chromatography B ; Analytical Technologies in the Biomedical and Life Sciences
- 出版年：2004
- 期刊代码：124_15700232
- 类别：ch
- 出版时间：April 25, 2004
- 卷：803
- 期：2
- 页码：299-304
- 文件大小：107 K
摘要
d-Penicillamine (d-Pen) is a thiol drug used in the treatment of Wilson’s disease, rheumatoid arthritis, metal intoxication and cystinuria. We have recently described a new capillary electrophoresis (CE) method to measure physiological thiols, in which separation of total plasma homocysteine, cysteine, cysteinylglycine, glutathione is achieved using the organic base N-methyl-d-glucamine in the run buffer. In this paper, we present an improvement of our method that allows a baseline separation of total plasma d-Pen from the physiological thiols. Moreover, reduced, free and protein-bound forms of drug are measured by varying the order of disulfide reduction with tributylphosphine and proteins precipitation with 5-sulphosalicylic acid (SSA). After derivatization with 5-iodoacetamidofluorescein (5-IAF), samples are separated and measured by capillary electrophoresis with laser-induced fluorescence in an uncoated fused-silica capillary (57cm×75μm i.d.) using a phosphate/borate run buffer pH 11.4. In these conditions, the migration time of d-Pen is about 7min and the time required for each analysis is roughly 10min. The proposed method has been utilized to measure the various forms of the drug in a d-Pen administered Wilson’s disease patient.