Plasma d-penicillamine redox state evaluation by capillary electrophoresis with laser-induced fluorescence
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, cyste...
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Published in | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 803; no. 2; pp. 299 - 304 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
Amsterdam
Elsevier B.V
25.04.2004
Elsevier Science |
Subjects | |
Online Access | Get full text |
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Summary: | 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 (
57
cm×75
μm i.d.) using a phosphate/borate run buffer pH 11.4. In these conditions, the migration time of
d-Pen is about 7
min and the time required for each analysis is roughly 10
min. The proposed method has been utilized to measure the various forms of the drug in a
d-Pen administered Wilson’s disease patient. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 1570-0232 1873-376X |
DOI: | 10.1016/j.jchromb.2004.01.002 |