Simple high-performance liquid chromatography method for the simultaneous determination of ketoconazole and piperine in rat plasma and hepatocyte culture.
Piperine, a major alkaloid of black and long peppers has been reported to act as bioavailability enhancer of several drugs by inhibiting drug metabolising enzymes and/or by increasing oral absorption. Ketoconazole is a well established potent inhibitor of CYP 3A4 and P-glycoprotein. A simple and rapid HPLC method has been developed for the simultaneous analysis of ketoconazole and piperine in rat plasma and hepatocyte culture. Analysis was performed using a Symmetry C18 column (150x4.6 mm, 5 microm) and isocratic elution with 25 mM KH2PO4 (pH 4.5)-acetonitrile (50:50) with a flow-rate of 1 ml/min. Photodiode array detection was used to simultaneously monitor piperine at 340 nm and ketoconazole at 231 nm in a single sample. Calibration plots in spiked plasma, hepatocytes and William's medium E were linear over the range studied (10-2000 ng for both drugs). The detection limits for piperine and ketoconazole are 2 and 4 ng, respectively, and the limits of quantitation are 10 and 12 ng, respectively. Intra- and inter-assay variations were less than 8%.
Bajad
Sunil
S
Pharmacology Division, Regional Research Laboratory, Jammu, India.
Johri
R K
RK
Singh
K
K
Singh
J
J
Bedi
K L
KL
D000470:Alkaloids; D000818:Animals; D052117:Benzodioxoles; D001682:Biological Availability; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D022781:Hepatocytes / Q000737:chemistry*; D007654:Ketoconazole / Q000032:analysis* / Q000097:blood / Q000493:pharmacokinetics; D008297:Male; D010880:Piperidines / Q000032:analysis* / Q000097:blood / Q000493:pharmacokinetics; D053284:Polyunsaturated Alkamides; D051381:Rats; D017208:Rats, Wistar; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D000470:Alkaloids; D052117:Benzodioxoles; D010880:Piperidines; D053284:Polyunsaturated Alkamides; D007654:Ketoconazole; C008922:piperine