Capillary electrophoresis to monitor the oxidative modification of low density lipoproteins
A procedure has been developed that uses high performance capillary electrophoresis to monitor the changes in the electrophoretic mobility of low density lipoproteins (LDL) resulting from Cu2+-catalyzed lipid peroxidation. Using uncoated fused silica capillaries, methylglucamine-Tricine, pH 9.0, as...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
1998-06-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520325566 |
| _version_ | 1819294271046418432 |
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| author | Joseph Stocks Norman E. Miller |
| author_facet | Joseph Stocks Norman E. Miller |
| author_sort | Joseph Stocks |
| collection | DOAJ |
| description | A procedure has been developed that uses high performance capillary electrophoresis to monitor the changes in the electrophoretic mobility of low density lipoproteins (LDL) resulting from Cu2+-catalyzed lipid peroxidation. Using uncoated fused silica capillaries, methylglucamine-Tricine, pH 9.0, as electrophoresis buffer and a field strength of 350 V/cm, separation of native LDL and oxidized LDL could be achieved in 8–10 min. The electrophoretic mobility of native LDL under these conditions was 1.32 × 10-4 cm2·V-1·s-1, and the migration time could be measured with a coefficient of variation of 0.44%. The increase in the electronegativity of LDLs during incubation with 10 μm Cu2+ for 0.25–2.0 h resulted in a progressive increase in migration time. Monitoring the absorbance of the migrating LDL particles at a wavelength of 234 nm showed a progressive increase in peak area, which paralleled that in diene conjugation measured spectrophotometrically. Electronegative LDL particles formed by modification with malondialdehyde could also be separated from native LDL particles under these conditions. This new procedure should be useful in studies of factors influencing low density lipoprotein oxidation in vitro and in vivo.—Stocks, J., and N. E. Miller. Capillary electrophoresis to monitor the oxidative modification of low density lipoproteins. J. Lipid Res. 1998. 39: 1305–1309. |
| first_indexed | 2024-12-24T04:23:40Z |
| format | Article |
| id | doaj.art-2167ed65bb5d4e7ea876656d446663ab |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-24T04:23:40Z |
| publishDate | 1998-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-2167ed65bb5d4e7ea876656d446663ab2022-12-21T17:15:41ZengElsevierJournal of Lipid Research0022-22751998-06-0139613051309Capillary electrophoresis to monitor the oxidative modification of low density lipoproteinsJoseph Stocks0Norman E. Miller1To whom correspondence and reprint requests should be addressed.; Department of Cardiovascular Biochemistry, St. Bartholomew's and the Royal London School of Medicine and Dentistry, Charterhouse Square, London, EC1M 6BQ, United KingdomDepartment of Cardiovascular Biochemistry, St. Bartholomew's and the Royal London School of Medicine and Dentistry, Charterhouse Square, London, EC1M 6BQ, United KingdomA procedure has been developed that uses high performance capillary electrophoresis to monitor the changes in the electrophoretic mobility of low density lipoproteins (LDL) resulting from Cu2+-catalyzed lipid peroxidation. Using uncoated fused silica capillaries, methylglucamine-Tricine, pH 9.0, as electrophoresis buffer and a field strength of 350 V/cm, separation of native LDL and oxidized LDL could be achieved in 8–10 min. The electrophoretic mobility of native LDL under these conditions was 1.32 × 10-4 cm2·V-1·s-1, and the migration time could be measured with a coefficient of variation of 0.44%. The increase in the electronegativity of LDLs during incubation with 10 μm Cu2+ for 0.25–2.0 h resulted in a progressive increase in migration time. Monitoring the absorbance of the migrating LDL particles at a wavelength of 234 nm showed a progressive increase in peak area, which paralleled that in diene conjugation measured spectrophotometrically. Electronegative LDL particles formed by modification with malondialdehyde could also be separated from native LDL particles under these conditions. This new procedure should be useful in studies of factors influencing low density lipoprotein oxidation in vitro and in vivo.—Stocks, J., and N. E. Miller. Capillary electrophoresis to monitor the oxidative modification of low density lipoproteins. J. Lipid Res. 1998. 39: 1305–1309.http://www.sciencedirect.com/science/article/pii/S0022227520325566apolipoprotein Bmalondialdehydelipid peroxideshuman |
| spellingShingle | Joseph Stocks Norman E. Miller Capillary electrophoresis to monitor the oxidative modification of low density lipoproteins Journal of Lipid Research apolipoprotein B malondialdehyde lipid peroxides human |
| title | Capillary electrophoresis to monitor the oxidative modification of low density lipoproteins |
| title_full | Capillary electrophoresis to monitor the oxidative modification of low density lipoproteins |
| title_fullStr | Capillary electrophoresis to monitor the oxidative modification of low density lipoproteins |
| title_full_unstemmed | Capillary electrophoresis to monitor the oxidative modification of low density lipoproteins |
| title_short | Capillary electrophoresis to monitor the oxidative modification of low density lipoproteins |
| title_sort | capillary electrophoresis to monitor the oxidative modification of low density lipoproteins |
| topic | apolipoprotein B malondialdehyde lipid peroxides human |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520325566 |
| work_keys_str_mv | AT josephstocks capillaryelectrophoresistomonitortheoxidativemodificationoflowdensitylipoproteins AT normanemiller capillaryelectrophoresistomonitortheoxidativemodificationoflowdensitylipoproteins |