Abstract
A simple, sensitive, selective and reproducible method based on a reversed-phase chromatography was developed for the determination of Praziquantel in rat plasma using internal standard as Diazepam. Praziquantel & Diazepam was separated on a C18 column Enable (250 mm × 4.6 mm, 5 µm particle size: Spinco Biotech Pvt Ltd), with retention times of 6.4 & 8.5 min. Ultraviolet detection was set at 225 nm. The mobile phase consisted of acetonitrile and water (60:40, v/v), running through the column at a flow rate of 1.0 mL min-1. The chromatographic analysis was operated at an ambient temperature. Sample preparation (200 µl plasma) was done by protein precipitation using perchloric acid. Calibration curve in plasma was plotted at different concentration level 5, 50, 100, 500, 1000, 2000, 3000 & 5000 ng mL-1 were found to be linear with correlation coefficients (r) is 0.9989. The precision of the method based on within-day repeatability and reproducibility (day-to-day variation) was within 15% (relative standard deviation: R.S.D. should be less than 15 according to CDER guidance for Bio-analytical Method Validation). Good accuracy was observed for both the intra-day or inter-day assays, as indicated by the minimal deviation of mean values. Limit of quantitation (LOQ) was accepted as 5 ng mL-1 using 200 µl samples. The mean recovery was found to be greater than 90% for praziquantel. The method appears to be robust and has been applied for pharmacokinetic study of praziquantel, in three different groups of rats following a single oral dose of 40 mg kg-1 body weight of praziquantel.
Highlights
Praziquantel is a pyrazinoisoquinoline derivative [2-(cyclohexyl-carbonyl)-1,2,3,6,7,11bhexahydro-4H-pyrazino[2,1-a]isoquinoline-4-one], which is the treatment of choice for most human trematode and cestode infections, and is widely used in schistosomiasis, as well as other fluke infections pathogenic to human [1]
Most of the high-performance liquid chromatography (HPLC) with UV detection methods described previously are based principally on the method developed by Xio et al [16]
We have described in this paper, a simple, sensitive, and selective HPLC method for determination of Praziquantel (PRQ) in plasma using Diazepam as internal standard (IS) and applying the method in pharmacokinetic study to calculate the all the necessary pharmacokinetic parameters which was not mentioned in the previous articles
Summary
Praziquantel is a pyrazinoisoquinoline derivative [2-(cyclohexyl-carbonyl)-1,2,3,6,7,11bhexahydro-4H-pyrazino[2,1-a]isoquinoline-4-one], which is the treatment of choice for most human trematode and cestode infections, and is widely used in schistosomiasis, as well as other fluke infections pathogenic to human [1]. A number of analytical methods have been reported for determination of praziquantel in human and animal biological fluids and tissue organ extracts These methods involve radiometric assay [2], fluorometric assay [3], enzyme-linked immunosorbent assay [4], thin-layer chromatography (TLC), gas chromatography [5,7] and high-performance liquid chromatography (HPLC) [6,8,9,10,11,12,13,14,15]. The protein precipitation method is highly effective in giving a clear chromatogram and does not require solvent evaporation by gentle steam of nitrogen or oxygen. More over this method uses less plasma in this study
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