A Simple and Sensitive High Performance Liquid Chromatography-Electrospray Ionization/Mass Spectrometry Method for the Quantification of Ethyl Pyruvate in Rat Plasma

Ethyl pyruvate (EP) is known as a scavenger of reactive oxygen species (ROS) in the body through its role in the donation of diketone groups to metals to form an EP-metal complex. In order to develop a method for the quantification of EP in biological media, a sensitive and specific, high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI/MS) method is used to determine the EP-alkali metal ion binding species. The analyte was separated on a ZORBOX SB-C8 (3.5 mu m, 30 mm x 2.1 mm I.D.) column and analyzed in selected ion monitoring (SIM) mode with a positive ESI interface using the m/z 255 [2M + Na](+) ion. The method was validated over the concentration range of 0.5-60.0 mu g/mL under 1/9 (v/v) of acetonitrile/methanol solvent system with flow rate 0.05 mL/min. The limit of quantification (LOQ) was 0.5 mu g/mL.