Validation Of Rp-Hplc Uv Method for Determination Ketoconazole in Rabbit Plasma: an Application to The Pharmacokinetic Study
Penulis/Author
VIVIANE ANNISA (1); Prof. Dr. Teuku Nanda Saifullah Sulaiman, S.Si., M.Si., Apt. (2); Prof. Dr. apt. Akhmad Kharis Nugroho, S.Si., M.Si. (3); Prof. Dr. apt. Agung Endro Nugroho, S.Si., M.Si. (4)
Tanggal/Date
20 2023
Kata Kunci/Keyword
Abstrak/Abstract
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 reversed-
phase 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.
