High-performance liquid chromatographic analysis of fatty acid compositions of platelet phospholipids as their 2-nitrophenylhydrazides.

A novel high-performance liquid chromatographic method for biologically important fatty acids incorporated into platelet phospholipids in esterified form has been developed. 2-Nitrophenylhydrazine hydrochloride was used as a pre-column labelling agent to convert the saponified platelet phospholipids directly into corresponding fatty acid hydrazides, without a complicated isolation procedure. Isocratic separation was achieved within only 36 min for twenty-five saturated and mono- and polyunsaturated fatty acids (C8:0-C22:6), including cis and trans isomers, on a YMC-FA column. The analytical results showed good quantitative accuracy. Fatty acid compositions were determined in platelet phospholipids obtained from normal subjects and patients with diabetes mellitus. The method is simple, rapid and adequate for labelling esterified fatty acids in biological materials, and has several advantages with regard to resolution, analysis time and sensitivity over previously published methods.