A RAPID AND SENSITIVE HPLC METHOD FOR THE ANALYSIS OF PROGUNAIL AND CYCLOGUANIL IN PLASMA: APPLICATION TO SINGLE DOSE PHARMACOKINETIC STUDIES
A simple, sensitive cost-effective and reproducible reverse phase high performance liquid
chromatographic (HPLC) method was developed to quantitate plasma levels of proguanil
(PGN) and its active metabolite, cycloguanil (CGN) in order to conduct single dose
pharmacokinetic studies. The drug and the internal standard were added to plasma samples,
vortexed and rendered alkaline with 2 M NaoH and the samples extracted with ether,
evaporated to dryness and the residue was reconstituted in methanol, whirlmixed before
injecting an aliquot onto the HPLC system. The calibration plots were linear over the
concentration range up to 4.0 μg /ml. The correlation coefficients (r) were of the order of 0.99
and above for both PGN and CGN. The ion pair method was carried out on a 5 μ reverse
phase C-18 column, using perchlorate ion as the counter ion and ultra violet detection at 254
nm. The method was reproducible with coefficient of variation for PGN and CGN, being less
than 4.0 %. PGN was well resolved from its active metabolite, CGN, and the internal
standard, pyrimethamine. The limit of detection of PGN was 10 ng /ml and the recovery was
greater than 95% in plasma. The analytical method therefore, exhibits good precision and
sensitivity in detecting and quantifying PGN and CGN and has been demonstrated to be
suitable for the pharmacokinetic studies of proguanil. The clinical applicability of the method
was assessed by the preliminary pharmacokinetic study of PGN and CGN, in fifteen healthyvolunteers. The in vivo study was carried out according to a single dose randomized design.
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