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Development and Optimization of Fast and New Reversed-Phase HPLC Method for Analysis of 5-Fluorouracil in Human and Rabbit Plasma
Muhammad Usman Minhas, Mahmood Ahmad*, Muhammad Sohail and Faryal Siddique
Faculty of Pharmacy and Alternative Medicine, The Islamia University of Bahawalpur, Punjab, Pakistan
*Corresponding author:


The objective was to develop and compare an accurate and reproducible HPLC method for determination of 5-fluouracil (5-FU) in rabbit and human plasma. A simple, rapid, accurate, precise and sensitive high pressure liquid chromatography method has been developed and subsequently validated for determination of 5-FU. The current method has been developed with simple mobile phase of de-ionized distilled water, pH adjusted at 3.2 with perchloric acid. An isocratic HPLC system (Agilent Technologies, 1200 series, USA) was used with variable wavelength detector and data processing software, ChemStation. Flow rate was 0.8 ml/min and 20 µl sample was injected and eluate was monitored at 260 nm wavelength. The developed method showed excellent linearity, reproducibility and sensitivity. The studied method was found robust for analysis of 5-FU in both, rabbit and human plasma samples. Run time was short (10 min) and simple extraction procedure with good response even at low drug concentration made this method suitable for pharmacokinetic and other applications.  A rapid, accurate and precise high pressure liquid chromatography method was developed and validated. It is concluded that developed method is simple, cost effective, fast and reproducible for the analysis of drug concentrations, in rabbit and human plasma.

Key words: 5-Fluorouracil, Anticancer, HPLC, Pharmacokinetics, Validation


ISSN 0253-8318 (Print)
ISSN 2074-7764 (Online)