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A method to correct for errors caused by generation of interfering compounds during erythrocyte lipid peroxidation
Authors:H S Gilbert  D D Stump  E F Roth
Affiliation:Polly Annenberg Levee Hematology Center, Department of Medicine, Mount Sinai School of Medicine of City University of New York, 19 East 98th Street, New York, New York 10029 USA
Abstract:A commonly used method for quantification of lipid peroxidation depends upon measurement of a malonaldehyde-thiobarbituric acid derivative with absorbance at 532 nm. Investigation of this assay demonstrated that erythrocyte peroxidation produces compounds that react with thiobarbituric acid to interfere with the malonaldehyde assay. Interference results from carryover absorbance at 532 nm, equivalent to 20% of the intensity of the maximum absorption peak at 453 nm. These compounds are not products of lipid peroxidation but are derived from erythrocyte hemolysate and reduced glutathione. A specific HPLC assay for malonaldehyde corroborated the improved accuracy of measuring absorbance at 453 nm and correcting for the absorbance of the interfering compounds at 532 nm when assaying erythrocyte malonaldehyde production.
Keywords:peroxidation  erythrocytes  lipids  malonaldehyde  high-performance liquid chromatography
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