A HPLC method for the quantitative determination of N-(2-hydroxy-5-nitrophenylcarbamothioyl)-3,5-dimethylbenzamide in biological samples.

Abstract

A sensitive and simple HPLC method was developed for the determination of a novel compound, a potential anti-cancer drug, N-(2-hydroxy-5-nitrophenylcarbamothioyl)-3,5-dimethylbenzamide (DM-PIT-1), a member of the new structural class of non-phosphoinositide small molecule antagonist of phosphatidylinositol-3,4,5-trisphosphate-pleckstrin-homology domain interactions, in mouse plasma and tumor tissue homogenates. The chromatographic separation of DM-PIT-1 was achieved on C18 column using isocratic elution with acetonitrile-water (70:30) containing 0.1% formic acid (v/v). DM-PIT-1 was detected by UV absorbance at 320 nm and confirmed by LC-MS. The extraction of the DM-PIT-1 from the plasma and tumor tissue with methylene chloride resulted in its high recovery (70-80%). HPLC calibration curves for DM-PIT-1 based on the extracts from the mouse plasma and tumor tissue samples were linear over a broad concentration range of 0.25-20 μg/ml/g, with intra/inter-day accuracy of 95% and the precision of variation below 10%. The limits of detection and quantification were 0.1 ng and 0.2 ng, respectively. The described method was successfully applied to study the pharmacokinetics of the DM-PIT-1 following the parenteral injections of DM-PIT-1 entrapped in 1,2-disteratoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene-glycol)-2000] (PEG-PE) micelles.

• Authors: Igor Skidan, Jacob Grunwald, Ritesh Thekkedath, Alexei Degterev, Vladimir Torchilin
• Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (J Chromatogr B Analyt Technol Biomed Life Sci) Vol. 879 Issue 19 Pg. 1610-6 (Jun 01 2011) ISSN: 1873-376X [Electronic] Netherlands
• PMID: 21514904 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: Copyright © 2011 Elsevier B.V. All rights reserved.

Chemical References

• Benzamides
• Micelles
• N-(((2-hydroxy-5-nitrophenyl)amino)carbonothioyl)-3,5-dimethylbenzamide
• Phosphatidylethanolamines
• dioleoyl-N-(monomethoxypolyethylene glycol succinyl)phosphatidylethanolamine
• Polyethylene Glycols

Topics

• Animals
• Area Under Curve
• Benzamides (analysis, blood, pharmacokinetics)
• Cell Line, Tumor
• Chromatography, High Pressure Liquid (methods)
• Drug Stability
• Female
• Mice
• Mice, Inbred BALB C
• Micelles
• Neoplasms (chemistry)
• Phosphatidylethanolamines (chemistry)
• Polyethylene Glycols (chemistry)
• Reproducibility of Results
• Sensitivity and Specificity