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.

Microanalysis, Pharmacokinetics and Tissue Distribution of Polysaccharide-Protein Complexes from Longan Pulp in Mice

“…Intermediate precision was analyzed by the same experiment additionally repeated on three consecutive days. Precision and accuracy were calculated using the formulas described by Skidana et al [ 35 ].…”

Micellar formulations of pro-apoptotic DM-PIT-1 analogs and TRAILin vitroandin vivo

“…Drug incorporation efficiency was measured by reverse phase high performance liquid chromatography (HPLC) using an Xbridge C 18 (2.1 cm  250 cm) column (Waters Corporation, Milford, MA) on a Hitachi Elite LaChrom HPLC with autosampler (Pleasanton, CA). The HPLC method was modified from previously published method (Skidan et al, 2011). The mobile phase consisted of either 70:30 acetonitrile:water (DM-PIT-1) or 60:40 acetonitrile:water (NCL-176, NCL-179, NCL-198, NCL-240) with a flow rate of 1 ml/min.…”