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Development and validation of a reversed-phase liquid chromatographic method for analysis of D4T (Stavudine) in rat
J Contreras1, H Ma González, R Menéndez
1Department of Analytical Chemistry, HPLC and Pharmacokinetics Laboratory, National Center for Scientific Research, PO Box 6412/6414, Havana, Cuba. contreras@diramic.cneuro.edu.cu
Abstract:
An improved and simplified high-performance liquid chromatographic (HPLC) method at UV detection 265 nm is presented for the determination of d4T in rat plasma. The mobile phase consists of methanol-distilled water-acetic acid in the 23:77:0.2 (v/v) ratio. Quantification is achieved by the peak-area ratio method with reference to the internal standard. This paper presents linearity, accuracy, precision, limit of quantification and limit of detection, specificity-selectivity and sample stability data. Based on the intra and inter-day validation, all coefficients of variation (CV) were found less than 15%. The assay is sufficiently rapid and sensitive and was applied in a pharmacokinetic study in rats.
