Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry (UHPLC-MS/MS) Assay for Simultaneous Quantifications of CZ48, Lactone-stabilized Camptothecin, and Camptothecin and Their Pharmacokinetic and Biliary Evaluations in Rats

In this study, a UHPLC-MS/MS method was developed and validated to simultaneously quantify CZ48 and CPT in rat plasma and bile. Detection was performed using the API 3200 Q Trap triple quadrupole mass spectrometer in a positive ion mode. Chromatographic separation was achieved on Waters ACQUITY UPLC BEH Shield RP18 column with a gradient elution at a flow rate of 0.45 ml/min, using mobile phases of 0.1% acetic acid in water (A) and 0.1% acetic acid in acetonitrile (B). The method was linear at the concentration ranges of 0.98 (LLOQ) – 1,000 ng/ml of CZ48 and CPT in rat plasma and 3.9 (LLOQ) – 1,000 ng/ml in bile. Intra- and inter-day accuracy and precision values did not deviate by more than 6.57 % and 10.15 % for CZ48 and CPT, respectively, in plasma, and 12.09 and 13.48 % in bile. Extraction recoveries of CZ48 were 90.18 - 95.42 % from plasma and 86.51 - 91.66 % from bile. The recoveries of CPT were 91.56 - 97.06 % from plasma and 84.89 - 89.15 % from bile. No significant matrix effects were observed in plasma and bile within 14.00 % and 16.19 %, respectively. CZ48 and CPT in plasma were stable after extraction process and different storage conditions, including bench-top, processed sample in autosampler, three cycles of freeze and thaw, and long-term (3 month) stability at −80 °C. The application of the validated method was demonstrated by a PK study after an intravenous dose of CZ48 in rats.
Source: Journal of Pharmaceutical and Biomedical Analysis - Category: Drugs & Pharmacology Source Type: research