High-performance liquid chromatography as a tool for identification of linolenic acid hydroperoxide prepared with soybean lipoxygenase.

Linolenic acid hydroperoxide, enzymatically produced by soybean lipoxygenase from linolenic acid, was purified by high-performance liquid chromatography (HPLC) using a Supelco LC-8 (5 microns), 22 cm X 4.6 mm I.D. column with a solvent system of acetonitrile-tetrahydrofuran-0.1% phosphoric acid (50.4:21.6:28, v/v/v). The hydroperoxide was converted to several derivatives, the hydroxy, methyl, and stearate which were analyzed by HPLC and identified by gas chromatography-mass spectrometry. The 13-hydroperoxy linolenic acid was the major hydroperoxide produced by soybean lipoxygenase.