Quantitative analysis of bucillamine in blood using high-performance liquid chromatography-mass spectrometry technique.
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 C18 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 microg/mL with a correlation value of r > or = 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.
Beaudry
Francis
F
University of Montréal, Faculty of Veterinary Medicine, Department of Veterinary Biomedecine, St-Hyacinthe, Quebec, Canada J2S 7C6. francis.beaudry@umontreal.ca
Proulx
Dave
D
Furtado
Milton
M
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003545:Cysteine / Q000031:analogs & derivatives* / Q000097:blood* / Q000737:chemistry; D004355:Drug Stability; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D012680:Sensitivity and Specificity
D000894:Anti-Inflammatory Agents, Non-Steroidal; D003545:Cysteine; C026535:bucillamine