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Quantitative determination of DRF‐1042 in human plasma by HPLC: validation and application in clinical pharmacokinetics
Author(s) -
Upreti Vijay V.,
Mamidi Rao N.V.S.,
Katneni Kasiram,
Srinivas Nuggehally R.
Publication year - 2003
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.253
Subject(s) - chromatography , chemistry , high performance liquid chromatography , calibration curve , triethylamine , pharmacokinetics , detection limit , standard curve , acetonitrile , quantitative analysis (chemistry) , serial dilution , analytical chemistry (journal) , pharmacology , medicine , alternative medicine , organic chemistry , pathology
Abstract 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 acidied 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 ow rate of 1.0 mL/min. The eluate was monitored with a uorescence detector set at excitation and emission wavelengths of 370 and 430 nm, respectively. The standard curves were linear ( r 2 > 0.999) in the concentration ranges 5.0–2004 ng/mL. The lower limit of quantication (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 conrmed 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.