15-hydroperoxy-5-8-11-13-eicosatetraenoic-acid and acetonitrile

A complex mixture of different lipid compounds, including phosphatidylcholine, phosphatidylserine, all trans-retinol, 15(S)-hydroperoxyeicosatetraenoic acid, D-alpha-tocopherol, saturated and unsaturated fatty acids can be separated by reversed phase HPLC by using a C-18, 120 mm x 4 mm, 3 microns particle size column and a step gradient from acetonitrile/water (1:1; v:v) to 100% acetonitrile at a flow rate of 0.8 ml/min. By applying this elution condition, separation of various groups of lipid hydroperoxides and lipid derivatives, each one originating from a different in vitro peroxidized polyunsaturated fatty acid, can be obtained. Simultaneous detection is carried out by a diode array detector at a wavelength accumulation range set up between 195 and 400 nm. The possibility of simultaneously having such a large number of measurements renders this chromatographic method particularly suitable in studies concerning lipid peroxidation where, in addition to the detection of free radical-induced lipid hydroperoxides, data on some key antioxidant molecules, i.e. vitamin A and E, as well as that of structural compounds of biological membranes, i.e. phosphatidylcholine and phosphatidylserine, can be achieved.

Topics: Acetonitriles; Arachidonic Acid; Chromatography, High Pressure Liquid; Fatty Acids, Unsaturated; gamma-Linolenic Acid; Leukotrienes; Linoleic Acid; Linoleic Acids; Lipid Peroxidation; Lipid Peroxides; Lipids; Membrane Lipids; Oleic Acid; Palmitic Acid; Phosphatidylcholines; Phosphatidylserines; Stearic Acids; Vitamin A; Vitamin E