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: ma786_786@yahoo.com
Abstract
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.