Dialysis and gel filtration of isolated low density lipoproteins do not cause a significant loss of low density lipoprotein tocopherol and carotenoid concentration.
The resistance of isolated low density lipoprotein (LDL) to copper-initiated oxidation is often used as a measure of effectiveness of an antioxidant intervention. Prior to oxidation, excess salt and EDTA are removed via dialysis or gel filtration of the LDL sample. However, there is concern over whether the antioxidant content of dialyzed or gel-filtered LDL is truly representative of native LDL extracted from a blood sample. Previously, the experiments done after the storage of native and dialyzed LDL at -80 degrees C showed that the dialysis step can cause a loss of up to 60% in the tocopherol and carotenoid content of LDL. In the present study, a comparison of the micronutrient concentration in freshly prepared dialyzed and native LDL from 35 subjects showed that after the correction for cholesterol, only lycopene (13%, P < 0.001) and to a lesser extent alpha-carotene (8%, P < 0.02) were significantly decreased, and the absolute fall in concentration was far smaller than previously reported. Other experiments done with smaller numbers of samples suggested that there were minimal micronutrient losses following gel filtration and that it was important to include 10 micromol/L EDTA in the dialysis and elution buffer; otherwise micronutrient losses did occur. In summary, immediate dialysis of freshly isolated LDL in the presence of 10 micromol/L EDTA does not cause any major loss in the concentration of tocopherol and most carotenoids.
Chopra
M
M
NICHE, School of Biomedical Sciences, University of Ulster, Northern Ireland, United Kingdom. M.Chopra@ulst.ac.uk
Fitzsimons
P
P
Hopkins
M
M
Thurnham
D I
DI
D002338:Carotenoids / Q000097:blood*; D002850:Chromatography, Gel / Q000379:methods*; D003956:Dialysis / Q000379:methods*; D004492:Edetic Acid; D006801:Humans; D008074:Lipoproteins / Q000097:blood* / Q000302:isolation & purification*; D014810:Vitamin E / Q000097:blood
D008074:Lipoproteins; D014810:Vitamin E; D002338:Carotenoids; D004492:Edetic Acid