The Analysis of Prostaglandins by High Performance Liquid Chromatography Using Pre Column Derivatization with 9-Anthryldiazomethane

Abstract

The analysis of prostaglandins (PGs) were investigated by high performance liquid chromatography (HPLC) using 9-anthryldiazomethane (ADAM).
ADAM, which is the labeling reagent of carboxylic group, can react with PGs rapidly. The derivatives were separated in reversed phase chromatography using Zorbax ODS column (4.6mm i. d.×25cm). Methanol-water was found to be suitable for complete separation of ADAM esters of PGE₁, PGE₂, PGF_1α and PGF_2α within 30 minutes.
The PGs-ADAM esters can be detected by fluorophotometer at Ex. 365nm Em. 412nm.
In this method, the fluorescence intensity of the derivatives have wide linearity over 100ng and detection limit was 100pg.