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Titolo:
RAPID ISOLATION OF LOW-DENSITY LIPOPROTEINS IN A CONCENTRATED FRACTION FREE FROM WATER-SOLUBLE PLASMA ANTIOXIDANTS
Autore:
VIEIRA OV; LARANJINHA JAN; MADEIRA VMC; ALMEIDA LM;
Indirizzi:
UNIV COIMBRA,FAC FARM,LAB BIOQUIM P-3000 COIMBRA PORTUGAL UNIV COIMBRA,FAC FARM,LAB BIOQUIM P-3000 COIMBRA PORTUGAL UNIV COIMBRA,CTR NEUROCIENCIAS P-3049 COIMBRA PORTUGAL
Titolo Testata:
Journal of lipid research
fascicolo: 12, volume: 37, anno: 1996,
pagine: 2715 - 2721
SICI:
0022-2275(1996)37:12<2715:RIOLLI>2.0.ZU;2-C
Fonte:
ISI
Lingua:
ENG
Soggetto:
VITAMIN-E; LIPID-PEROXIDATION; INDUCED OXIDATION; ULTRACENTRIFUGATION; ATHEROSCLEROSIS; CHOLESTEROL; INHIBITION; SEPARATION; BLOOD; LDL;
Keywords:
ULTRACENTRIFUGATION; ALPHA-TOCOPHEROL; OXIDATION STUDIES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
28
Recensione:
Indirizzi per estratti:
Citazione:
O.V. Vieira et al., "RAPID ISOLATION OF LOW-DENSITY LIPOPROTEINS IN A CONCENTRATED FRACTION FREE FROM WATER-SOLUBLE PLASMA ANTIOXIDANTS", Journal of lipid research, 37(12), 1996, pp. 2715-2721

Abstract

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

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 11/07/20 alle ore 14:07:35