FPSE-HPLC-PDA Method for the Determination of Twelve Azole Antimicrobial Drug Residues in Human Plasma and Urine

A novel fabric phase sorptive extraction-high performance liquid chromatography-photodiode array detection (FPSE-HPLC-PDA) method has been developed for the simultaneous extraction and analysis of twelve azole antimicrobial drug residues possessing a wide range of logKow values (from 0.4 to 6.70) that include ketoconazole, terconazole, voriconazole, bifonazole, clotrimazole, tioconazole, econazole, butoconazole, miconazole, posaconazole, ravuconazole, and itraconazole in human plasma and urine samples [1]. The selected azoles were resolved using a Luna C18 column in gradient elution mode within 36 minutes. The analytical method was calibrated and validated in the range from 0.1 to 8 g/mL for all the drug compounds. Blank human plasma and urine were used as the sample matrix for the analysis; while benzyl-4-hydroxybenzoate was used as the internal standard (IS). The limit of quantification of the FPSE-HPLC-PDA method was found as 0.1 g/mL and the weighted-matrix matched standard calibration curves of the drugs showed a good linearity up to a concentration of 8 g/mL. The parallelism tests were also performed to evaluate whether overrange sample can be analyzed after dilution, without compromising the analytical performances of the validated method. The intra- and inter-day precision (RSD%) values were found ≤13.1% and ≤13.9%, respectively. The intra- and inter-day trueness (bias%) values were found in the range from -12.1% to 10.5%. The performances of the validated FPSE-HPLC-PDA were further tested on real samples collected from healthy volunteers after a single dose administration of itraconazole and miconazole showing the adoptability of herein reported procedure as a rapid and robust green analytical tool for clinical and pharmaceutical applications