Lipid peroxidation and the thiobarbituric acid assay: standardization of the assay when using saturated and unsaturated fatty acids.
Saturated fatty acids are less vulnerable to lipid peroxidation than their unsaturated counterparts. In this investigation, individual fatty acids of the C(16), C(18) and (20) families were subjected to the thiobarbituric (TBA) assay. These fatty acids were chosen based on their degree of saturation and configuration of double bonds. Interestingly, an assay threshold was reached where increasing the fatty acid concentration resulted in no additional decrease in the TBARS concentrations. Therefore, the linear range of TBARS inhibition was determined for fatty acids in the C(16) and C(20) families. The rate of TBARS inhibition was greater for the saturated than for unsaturated fatty acids, as measured from the slope of the linear range. These findings demonstrate the need to standardize the TBARS assay using multiple fatty acid concentrations when using this assay for measuring in vitro lipid peroxidation.
Rael
Leonard T
LT
Department of Trauma Research and Trauma Services, Swedish Medical Center, 501 E. Hampden Avenue, Englewood, CO 80113, USA.
Thomas
Gregory W
GW
Craun
Michael L
ML
Curtis
C Gerald
CG
Bar-Or
Raphael
R
Bar-Or
David
D
D000818:Animals; D001681:Biological Assay / Q000379:methods / Q000592:standards; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000737:chemistry / Q000378:metabolism*; D015227:Lipid Peroxidation; D012015:Reference Standards; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism*
D005227:Fatty Acids; D005231:Fatty Acids, Unsaturated; D017392:Thiobarbituric Acid Reactive Substances