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Quantitative analysis of bucillamine in blood using high‐performance liquid chromatography–mass spectrometry technique
Author(s) -
Beaudry Francis,
Proulx Dave,
Furtado Milton
Publication year - 2004
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.393
Subject(s) - chromatography , chemistry , tris , hydroxymethyl , mass spectrometry , protein precipitation , derivatization , quantitative analysis (chemistry) , analytical chemistry (journal) , organic chemistry , biochemistry
A fast and sensitive method has been developed and validated for the determination of bucillamine in human blood by derivatizing the free sulfhydryl groups with isobutyl acrylate (IA), by APCI‐LC/MS/MS. The collected blood sample was immediately mixed with a mixture of IA and 0.05 m Tris(hydroxymethyl)aminomethane hydrochloride (Tris–HCl) buffer, pH 9.2, to stabilize the sulfyhydryl moieties. The derivatized samples were then extracted by protein precipitation, evaporated, reconstituted and injected using an LC‐APCI/MS/MS instrument. Separation was achieved using a C 18 analytical column and a gradient mobile phase within a chromatographic run time of 5 min. A quadratic (weighted 1/concentration 2 ) relationship was observed during validation over a concentration range of 0.4–40 µg/mL with a correlation value of r ≥ 0.9966. The inter‐batch precision and accuracy at low, medium and high concentrations were 8.1, 8.4 and 7.3%; 113.3, 104.9 and 103.9%, respectively, and the intra‐batch precision and accuracy at low, medium and high concentrations were 7.7, 5.4 and 2.7%; 105.1, 111.9 and 113.2%, respectively. Copyright © 2004 John Wiley & Sons, Ltd.