Development and Validation of a High-Performance Liquid Chromatography Method With UV Detection for Determination of 1-(2-phenylethyl)-5- (quinaldin-4-yl) Biuret in Rat Plasma
Jundishapur Journal of Natural Pharmaceutical Products: 8 (2); 81-85
May 3, 2013
Article Type: Research Article
October 27, 2012
December 18, 2012
M J, Rezaee
S. Development and Validation of a High-Performance Liquid Chromatography Method With UV Detection for Determination of 1-(2-phenylethyl)-5- (quinaldin-4-yl) Biuret in Rat Plasma,
Jundishapur J Nat Pharm Prod.
Online ahead of Print
Recently, biuret derivatives have been reported as showing moderate to good cytotoxic effect against certain cancer cell lines. In this study, a high-performance liquid chromatography method was developed for determination of 1-(2-phenylethyl)-5-(quinaldin-4-yl) biuret (PEQB) in rat plasma to use in future studies on this compound and related derivatives.
In this study, we describe a simple and sensitive high-performance liquid chromatography method with UV detection for determination of 1-(2-phenylethyl)-6-(quinaldin-4-yl) biuret (PEQB) in rat plasma.
Materials and Methods:
Separations were performed on a Nucleosil-100 CN HPLC column (125 4.0 mm) (5 m), using a mixture of acetonitrile: methanol: potassium dihydrogen phosphate buffer (0.05 M, pH 3.5) (10:10:80) as mobile phase delivered at a flow rate of 1 mL/minute. Detection of PEQB and internal standard (1-([[3-(1,3-benzothiazol-2-ylsulfanyl)propyl]carbamoyl]amino)-N-phenylformamide) was performed at 235 nm and ambient temperature. Plasma samples (200 L) were prepared by addition of 40 L internal standard (100 g/mL), and 400 L acetonitrile. After vortex mixing and centrifugation at 10000 g, 50 L of the clear supernatant was directly injected onto the chromatography column. Calibration curves were constructed by fitting the peak area ratio of the biuret to internal standard against concentration of biuret to a power model using generalized least squares nonlinear regression method.
Under the above chromatography condition, biuret compound (PEQB) and the internal standard were detected at 4.5 and 13.5 minutes, respectively. Limit of quantitation of the PEQB was 0.1 g/mL. Accuracy of the method over the concentration range of 0.1-100 g/mL was between 88-109%. Inter- and intraday precisions were 4-19% and 6-8%, respectively. A good relationship in the form of a power model was found for two separate concentration ranges of 0.1-1 and 2.5-100 g/mL (R 2> 0.99).
The presented simple HPLC method is sufficiently accurate, precise and sensitive for the quantitation of 1-(2-phenylethyl)-5-(quinaldin-4-yl) biuret in rat plasma.
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