Distribution of thiobarbituric acid-reactive substances in lipoproteins and proteins in serum.
We assessed the distribution of malondialdehyde (MDA) in lipoproteins and proteins in serum after using two procedures to separate the lipoproteins: sequential ultracentrifugation or selective precipitation with a sodium phosphotungstate and magnesium chloride reagent followed by ultracentrifugation of the supernate. MDA concentrations were determined by the thiobarbituric acid reaction and quantified by fluorometry. We found that 43% of the thiobarbituric acid-reactive substances (TBARS) was bound to the lipoproteins--27% to very-low- and low-density lipoproteins (VLDL-LDL) and 16% to high-density lipoproteins (HDL)--and from 11.5% to 15.8% to proteins, depending on the separation procedure. Residual unbound TBARS were located in the ultracentrifugation layers that contained no lipoproteins or proteins. The TBARS concentration in serum lipoproteins containing apolipoprotein B (i.e., VLDL-LDL) was the same after ultracentrifugation or selective precipitation. We therefore consider the precipitation method more suitable for routine TBARS determination in these lipoproteins, because it is easier to handle and faster. However, for determination of TBARS in HDL, selective precipitation requires subsequent ultracentrifugation at a density of 1.21 kg/L.
Bonnefont
D
D
Laboratoire de Biochimie, Hôpital de la Salpêtrière, Paris, France.
Legrand
A
A
Peynet
J
J
Emerit
J
J
Delattre
J
J
Galli
A
A
D000328:Adult; D001798:Blood Proteins / Q000302:isolation & purification*; D011232:Chemical Precipitation; D006801:Humans; D007202:Indicators and Reagents; D008074:Lipoproteins / Q000302:isolation & purification*; D008075:Lipoproteins, HDL / Q000302:isolation & purification; D008077:Lipoproteins, LDL / Q000302:isolation & purification; D008079:Lipoproteins, VLDL / Q000302:isolation & purification; D008314:Malonates; D008315:Malondialdehyde / Q000032:analysis*; D013050:Spectrometry, Fluorescence; D013858:Thiobarbiturates; D014461:Ultracentrifugation
D001798:Blood Proteins; D007202:Indicators and Reagents; D008074:Lipoproteins; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D008079:Lipoproteins, VLDL; D008314:Malonates; D013858:Thiobarbiturates; D008315:Malondialdehyde; C029684:thiobarbituric acid