Capillary electrophoresis-mass spectrometry coupling versus micro-high-performance liquid chromatography-mass spectrometry coupling: a case study.
Capillary zone electrophoresis (CZE) and micro-high-performance liquid chromatography (mu-HPLC) coupled to electrospray ionisation (ESI) mass spectrometry were compared with respect to their applicability to problems arising in pharmaceutical drug research and development. Both techniques, which are similar with regard to their operational parameters, were coupled to an API III plus triple quadrupole mass spectrometer using laboratory-built interfaces. The results achieved with the two combinations were compared for sensitivity and general applicability to the quantitative analysis of pharmaceuticals in biological fluids. Midazolam, the 8-chloro-6-(2-fluoro-phenyl)-1-methyl-4H-imidazo-[1,5-a][1,4]-benzodiaze pine, and three of its metabolites were used as test compounds, either as standard solution or after sample clean-up from human plasma. Following different sample preparation routes, liquid-liquid extraction or solid-phase extraction, differences in detection limits as well as robustness in CZE or mu-HPLC coupled with ion spray mass spectrometry (IS-MS) were investigated. Detection limits of about 500 pg/ml for the drug and 2 ng/ml for the metabolites were achieved, using 1 ml of human plasma, only when liquid-liquid extraction was used for sample preparation. Sample preparation using the simpler and faster solid-phase extraction route resulted in deterioration of the separation or clogging of the columns. In all cases, when standard solutions or sample extracts were used, CZE-ESI-MS provided both different selectivity and greater sensitivity.
Lausecker
B
B
F. Hoffmann-La Roche, Ltd., Department of Drug Metabolism and Pharmacokinetics, Basel, Switzerland.
Hopfgartner
G
G
Hesse
M
M
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005069:Evaluation Studies as Topic; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008874:Midazolam / Q000032:analysis* / Q000097:blood / Q000378:metabolism; D012016:Reference Values; D012680:Sensitivity and Specificity
D008874:Midazolam