Determination of (3S)-3-hydroxy quinidine for metabolism screening experiments using direct injection capillary electrophoresis and laser-induced fluorescence detection.
Capillary electrophoresis (CE) has been used with collinear laser-induced fluorescence detection (LIF) to determine the amount of (3S)-3-hydroxy quinidine (3OHQ) formed on direct injection of microsomal incubation mixtures. 3OHQ is the CYP 3A4 metabolite of quinidine sulfate (QS) and is therefore useful for metabolism screening studies. The method was validated analytically and tested for its capability of screening for a weak inhibitor of the CYP 3A4 isozyme. A linear calibration was found to provide the best fit for the standard curve with a correlation of 0.9950 and all concentration residuals less than 15%. The percentage relative standard deviations (RSDs) of two controls, 175 and 2250 ng/mL, were 9.29 and 5.68% and the percentage differences from normal (DFN) were 6.87 and -4.37%, respectively. The concentration limit of detection (LOD) for 3OHQ in the incubation matrix was 52.11ng/mL and the mass LOD was approximately 521.1 fg (injection volume 10 nL). The effectiveness of the method to screen for the weak inhibitor erythromycin has been shown by calculating percentage inhibition when incubating with different concentrations of QS. Sensitive detection coupled with the convenience of the direct injection technique makes this an attractive approach for metabolism screening. The small sample size capability of CE will further reduce the quantities of probe drug, microsomes and other reagents required for incubation studies.
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Department of Pharmaceutics, Medical College of Virginia, Virginia Commonwealth University, Richmond, VA 23298-0533, USA.
Karnes
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D000818:Animals; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D019075:Electrophoresis, Capillary / Q000379:methods*; D004791:Enzyme Inhibitors / Q000032:analysis; D007834:Lasers; D008862:Microsomes, Liver / Q000201:enzymology; D006899:Mixed Function Oxygenases / Q000378:metabolism; D011802:Quinidine / Q000031:analogs & derivatives* / Q000378:metabolism*; D051381:Rats; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
D004791:Enzyme Inhibitors; C013569:3-hydroxyquinidine; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C104464:CYP3A protein, human; D051544:Cytochrome P-450 CYP3A; D011802:Quinidine