Spectrophotometric and high-performance chromatographic assays of hydroperoxides by the iodometric technique.
Samples containing between 1 and 50 nmol of hydroperoxides in oxygen-free methanol-acetic acid containing I- were incubated at 50 degrees C. This ensured reduction of all hydroperoxides tested in 15 min. Addition of sufficient Cd acetate to combine with the remaining I- allowed reading of absorbance of the I3- produced in open cuvettes. The absorbance was stable for several hours. It had a maximum at 358 nm with molar extinction coefficient of 2.97 X 10(4) 1 mol-1 cm-1. Sensitivity of the assay could be improved by injecting the I3- into a C-18 HPLC column eluted with methanol-water-acetic acid solution. Both methods are potentially suitable for assay of hydroperoxides in a wide range of biological materials.
Gebicki
J M
JM
School of Biological Sciences, Macquarie University, Sydney, New South Wales, Australia.
Guille
J
J
D000085:Acetates; D002104:Cadmium / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006861:Hydrogen Peroxide / Q000032:analysis*; D007454:Iodides / Q000032:analysis*; D008054:Lipid Peroxides / Q000032:analysis; D008055:Lipids / Q000302:isolation & purification; D010084:Oxidation-Reduction; D012996:Solutions; D013025:Glycine max; D013053:Spectrophotometry / Q000379:methods
D000085:Acetates; D007454:Iodides; D008054:Lipid Peroxides; D008055:Lipids; D012996:Solutions; D002104:Cadmium; C028031:cadmium acetate; D006861:Hydrogen Peroxide