Rapid isolation of low density lipoproteins in a concentrated fraction free from water-soluble plasma antioxidants.

摘要: A rapid method is described for isolation and concentration of plasma low density lipoproteins (LDL) using a Beckman L80 ultracentrifuge equipped with 70.1 Ti fixed angle rotor. The LDL achieved by discontinuous gradient step (180 min) was followed simultaneous purification (45 ultrafiltration through collodium bag under nitrogen. This dialysis/concentration step, in contrast to the standard dialysis techniques batch or filtration short gel columns, prevents oxidation dilution sample. Electrophoresis agarose sodium dodecylsulfate-polyacrylamide (SDS-PAGE) gels were used monitor surface charge, purity, contamination proteins. artifactual during subsequent handling, thus ability preparation oxidation/antioxidation studies, assessed determination endogenous hydroperoxides thiobarbituric acid reactive substances. that allows obtention concentrated purified validated absence ascorbate urate, as measured HPLC. led preparations free water-soluble antioxidants minimally oxidized suitable reliable vitro inhibition studies. applicability this methodology tested studying alpha-tocopherol content Portuguese population university students.