Simultaneous Determination of Prostaglandin E1 and Prostaglandin E1 Ethyl Ester in Hairless Mouse Skin Homogenate by High-Performance Liquid Chromatography

Abstract

A rapid and specific high-performance liquid chromatographic method was developed and validated for the simultaneous determination of prostaglandin E1 (PGE1) and prostaglandin E1 ethyl ester (PGE1-EE) in hairless mouse skin homogenate. The sample treatment procedure involved deproteination and precipitation by acetonitrile. PGE1 and PGE1-EE in supernatant were separated in a reversed-phase C18 column without being interfered by other components present in hairless mouse skin homogenate. 9-Anthracenecarboxylic acid was used as an internal standard. The retention times of PGE1, 9-anthracenecarboxylic acid and PGE1-EE were, 4.5, 9.5 and 18.0 min, respectively. The assay showed linearity from 1 to 40 g/ml for both PGE1 and PGE1-EE. Precision expressed as RSD ranged from 2.3 to 14.1% for PGE1 and 1.6 to 11.0% for PGE1-EE. Accuracy ranged from 100.5 to 119.6 % for PGE1 and from 98.0 to 103.7% for PGE1-EE. This method was employed successfully to follow the time course of concentrations of PGE1 and PGE1-EE in hairless mouse skin homogenate for stability study.