The Pharmacokinetics Analysis of the Phosphoryl Peptides in MCF-7/ADR Cells

Abstract

We have developed and validated a practical high-performance liquid chromatography method to determine the concentration of N-diisopropyloxyphosphoryl-valine-phenylalanine-3, 4, 5-trimethoxy aniline (DIPP-Val-Phe-TMOA) in Adriamycin-resistant human breast adenocarcinoma (MCF-7/ADR) cells. First, DIPP-Val-Phe-TMOA was extracted from MCF-7 cells with perchloric acid, followed by a cleanup procedure with centrifugal separation, and then determinated by HPLC using a Waters C18 column with a gradient elution (solvent A: 10 mM ammonium acetate at pH 7.0, B: acetonitrile). The flow rate was maintained at 1.0 ml/min at room temperature. The detection wavelength was at 254 nm. Meanwhile, this validated method was successfully applied to analyze the concentration levels and the pharmacokinetics parameters of DIPP-Val-Phe-TMOA in MCF-7/ADR cells.

Keywords:

• Determination
• high-performance liquid chromatography
• MCF-7/ADR cells
• N-diisopropyloxyphosphoryl-valine-phenylalanine-3,4,5-trimethoxyaniline

Acknowledgments

This work was supported by the National Natural Science Foundation (China, 20472043, 20572060, 20672068), the Project of Science and Technology of Guangdong Province (2005A11601008), Mega-Project of Science Research of Ministry of Science and Technology (China, 2005CCA03400), and the Natural Science Foundation of Guangdong Province (China, 06028200).