Quantitative determination of DRF-1042 in human plasma by HPLC: validation and application in clinical pharmacokinetics

• Published in: Biomedical chromatography Vol. 17; no. 6; pp. 385 - 390
• Main Authors: Upreti, Vijay V., Mamidi, Rao N.V.S., Katneni, Kasiram, Srinivas, Nuggehally R.
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01.09.2003
• Subjects: anticancer agent; Antineoplastic Agents - blood; Antineoplastic Agents - pharmacokinetics; Antineoplastic Agents - therapeutic use; Calibration; Camptothecin - analogs & derivatives; Camptothecin - blood; Camptothecin - pharmacokinetics; Camptothecin - therapeutic use; camptothecin analog; Chromatography, High Pressure Liquid - methods; Humans; Neoplasms - blood; Neoplasms - drug therapy; phase I clinical trial; Reproducibility of Results; Sensitivity and Specificity; fluorescence detection
• ISSN: 0269-3879; 1099-0801
• DOI: 10.1002/bmc.253

A simple and sensitive high‐performance liquid chromatography (HPLC) method has been developed and validated for the determination of DRF‐1042, a novel orally active camptothecin (CPT) analog, in human plasma. The sample preparation was a simple deproteinization with acidified methanol yielding almost 100% recovery of DRF‐1042. An isocratic reverse‐phase HPLC separation was developed on a Supelcosil‐LC318 column (250 × 4.6 mm, 5 µm) with mobile phase consisting of 1% v/v triethylamine acetate, pH 5.5 and acetonitrile (80:20, v/v) at a flow rate of 1.0 mL/min. The eluate was monitored with a fluorescence detector set at excitation and emission wavelengths of 370 and 430 nm, respectively. The standard curves were linear (r2 > 0.999) in the concentration ranges 5.0–2004 ng/mL. The lower limit of quantification (LLQ) of the assay was 5 ng/mL. The mean measured quality control (QC) concentrations (range 5 ng/mL to 40 µg/mL) deviated from the nominal concentrations in the range of −10.5–0.08 and −14.5–7.97%, inter‐ and intra‐day, respectively. The inter‐ and intra‐day precisions in the measurement of QC samples at four tested concentrations, were in the range 0.64–5.89% relative standard deviation (RSD) and 0.33–14.7% RSD, respectively. The method was found to be suitable for measurement of plasma concentrations above the calibration curve after serial dilutions. Stability of DRF‐1042 was confirmed in a battery of studies, viz., on bench‐top, in the auto‐sampler, in the stock solutions, after four quick freeze–thaw cycles, up to one month at −20C in human plasma and up to 2 months in the ex vivo samples. The method is simple, sensitive and reliable and has been successfully implemented to investigate the clinical pharmacokinetics of DRF‐1042 in cancer patients in a phase I clinical trial. Copyright © 2003 John Wiley & Sons, Ltd.