title: 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.
abstract: BACKGROUND
  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.
  
  
  OBJECTIVE
  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.
  
  
  METHODS
  Separations were performed on a Nucleosil-100 CN HPLC column (125 × 4.0 mm) (5 µm), 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 µL) were prepared by addition of 40 µL internal standard (100 µg/mL), and 400 µL acetonitrile. After vortex mixing and centrifugation at 10000 g, 50 µL 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.
  
  
  RESULTS
  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 µg/mL. Accuracy of the method over the concentration range of 0.1-100 µg/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 µg/mL (R (2)> 0.99).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Medicinal Chemistry, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IR Iran.
authors.forename: Neda
authors.initials: N
authors.lastname: Adibpour
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IR Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Ahmadnasr
authors.affiliation: Department of Pharmacology and Toxicology, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IR Iran.
authors.forename: Mohammad Javad
authors.initials: MJ
authors.lastname: Khodayar
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IR Iran.
authors.forename: Saeed
authors.initials: S
authors.lastname: Rezaee
keywords: Biuret; High-pressure Liquid Chromatography; chromatography