Optimized determination of malondialdehyde in plasma lipid extracts using 1,3-diethyl-2-thiobarbituric acid: influence of detection method and relations with lipids and fatty acids in plasma from healthy adults.
We investigated the influence of different concentrations of Fe3+, phosphoric acid, butylated hydroxytoluene and glutathione on the production of the malondialdehyde-1,3-diethyl-2-thiobarbituric acid adduct in plasma lipid extracts. Following organic solvent extraction the stable product was analyzed by spectrophotometry (537 nm), fluorometry (547 nm) and high-performance liquid chromatography with fluorometric detection. Using optimized reaction conditions there was good agreement between the three methods, with slightly higher values for the spectrophotometric method. Plasma total lipid malondialdehyde reference values for 24 healthy adults amounted to 1.30 +/- 0.23 mumol/l (spectrophotometric method) and 1.11 +/- 0.31 mumol/l (fluorometric method). Plasma lipid malondialdehyde concentrations correlated significantly with plasma triglycerides (r = 0.527), total cholesterol (r = 0.612) and total fatty acids (r = 0.810) and with the total number of double bonds present in plasma fatty acids with three or more double bonds (r = 0.923).
Hoving
E B
EB
Central Laboratory for Clinical Chemistry, University Hospital, Groningen, The Netherlands.
Laing
C
C
Rutgers
H M
HM
Teggeler
M
M
van Doormaal
J J
JJ
Muskiet
F A
FA
D000328:Adult; D002084:Butylated Hydroxytoluene; D002712:Chlorides; D002851:Chromatography, High Pressure Liquid; D005260:Female; D005290:Ferric Compounds; D005978:Glutathione; D006801:Humans; D015227:Lipid Peroxidation; D008055:Lipids / Q000097:blood / Q000737:chemistry*; D008297:Male; D008315:Malondialdehyde / Q000097:blood* / Q000378:metabolism; D008875:Middle Aged; D010756:Phosphoric Acids; D013050:Spectrometry, Fluorescence; D013054:Spectrophotometry, Atomic; D013858:Thiobarbiturates / Q000097:blood* / Q000378:metabolism
D002712:Chlorides; D005290:Ferric Compounds; D008055:Lipids; D010756:Phosphoric Acids; D013858:Thiobarbiturates; D002084:Butylated Hydroxytoluene; D008315:Malondialdehyde; C046058:1,3-diethyl-2-thiobarbituric acid; C030242:phosphoric acid; D005978:Glutathione; C029684:thiobarbituric acid; C024555:ferric chloride