Validation Of Rp-Hplc Uv Method for Determination Ketoconazole in Rabbit Plasma: an Application to The Pharmacokinetic Study

The validated method for determining ketoconazole in plasma rabbit is not yet reported. The HPLC-UV method is simple, rapid, cost-effective, sensitive, and only requires a small blood sampling. The chromatographic system used a mobile phase consist of NaH2PO4:Acetonitrile (30:70) and stationary phase as a reversedphase C18 column (250 x 4.6 mm, 5 μm) at a flow rate of 1 ml/ min and detection wavelength of 240 nm, and the retention time of about 5 minutes for ketoconazole and 11 min for itraconazole as internal standard. The peak of ketoconazole can separate from other peaks and has no interference from the diluent, indicating this method was selective to detect ketoconazole. The calibration curve presented linearity in the 0.05-8 μg/ml with R2=0.9969, which showed good linearity. Precision and accuracy of the method were obtained. The result is 9.47 diff and 10.13-12.08 RSD for LLOQ and 0.59-3.94 diff and 1.82-13.56 RSD <20 for low, medium, and high levels. The LLOQ in this method is 0.05 μg/mL. Plasma stability under storage in a freezer (-200C) for three days was studied. The validated analytical method was successfully applied to determine the pharmacokinetics parameter of KTZ after a single oral administration. © 2023 Society of Pharmaceutical Sciences of Ankara (FABAD). All rights reserved.