Characterization and quantitation of apolipoprotein B-100 by capillary electrophoresis
The interaction of low density lipoproteins (LDL) with different surfactants was studied by capillary electrophoresis (CE) and sucrose density gradient ultracentrifugation as part of developing a method for quantitation of apoB-100 in serum. A mixture of surfactants consisting of 70% sodium dodecyl...
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| Format: | Article |
| Language: | English |
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Elsevier
1998-01-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520342164 |
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| author | Ingrid D. Cruzado Steven L. Cockrill Catherine J. McNeal Ronald D. Macfarlane |
| author_facet | Ingrid D. Cruzado Steven L. Cockrill Catherine J. McNeal Ronald D. Macfarlane |
| author_sort | Ingrid D. Cruzado |
| collection | DOAJ |
| description | The interaction of low density lipoproteins (LDL) with different surfactants was studied by capillary electrophoresis (CE) and sucrose density gradient ultracentrifugation as part of developing a method for quantitation of apoB-100 in serum. A mixture of surfactants consisting of 70% sodium dodecyl sulfate (SDS), 25% sodium myristyl sulfate, and 5% sodium cetyl sulfate was found to delipidate LDL particles more effectively than pure SDS or sodium decyl sulfate. The delipidation products of LDL [apolipoprotein B-100 (apoB-100) and lipids] were resolved as two distinct peaks by CE when using a 3.5 mm 70% SDS mixture, 20% (v/v) acetonitrile, 50 mm sodium borate, pH 9.1 buffer. This CE method was also used to characterize apoB-100 derived from samples of lipoprotein [a] and very low density lipoproteins (VLDL). A CE-based quantitation method for apoB-100 was developed utilizing the observed linear relationship between apoB-100 concentration and its corrected 214 nm absorbance peak area measured on-line by CE. Concentration values of apoB-100 in LDL and VLDL samples were determined by CE and found to be accurate when compared to values obtained by immunoturbidimetric analysis and the Lowry method. Capillary electrophoresis can be used as a precise, accurate, and specific on-line method for the qualitative and quantitative analysis of the apoB-100 component of VLDL and LDL-related lipoproteins.—Cruzado, I. D., S. L. Cockrill, C. J. McNeal, and R. D. Macfarlane. Characterization and quantitation of apolipoprotein B-100 by capillary electrophoresis. J. Lipid Res. 1998. 39: 205–217. |
| first_indexed | 2024-12-17T23:23:30Z |
| format | Article |
| id | doaj.art-8c4172ded2e74384a578840673644540 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-17T23:23:30Z |
| publishDate | 1998-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-8c4172ded2e74384a5788406736445402022-12-21T21:28:50ZengElsevierJournal of Lipid Research0022-22751998-01-01391205217Characterization and quantitation of apolipoprotein B-100 by capillary electrophoresisIngrid D. Cruzado0Steven L. Cockrill1Catherine J. McNeal2Ronald D. Macfarlane3Department of Chemistry, Texas A&M University, College Station, TX 77843–3255Department of Chemistry, Texas A&M University, College Station, TX 77843–3255Department of Internal Medicine, Scott and White Hospital, Temple, TX 76503To whom correspondence should be addressed.; Department of Chemistry, Texas A&M University, College Station, TX 77843–3255The interaction of low density lipoproteins (LDL) with different surfactants was studied by capillary electrophoresis (CE) and sucrose density gradient ultracentrifugation as part of developing a method for quantitation of apoB-100 in serum. A mixture of surfactants consisting of 70% sodium dodecyl sulfate (SDS), 25% sodium myristyl sulfate, and 5% sodium cetyl sulfate was found to delipidate LDL particles more effectively than pure SDS or sodium decyl sulfate. The delipidation products of LDL [apolipoprotein B-100 (apoB-100) and lipids] were resolved as two distinct peaks by CE when using a 3.5 mm 70% SDS mixture, 20% (v/v) acetonitrile, 50 mm sodium borate, pH 9.1 buffer. This CE method was also used to characterize apoB-100 derived from samples of lipoprotein [a] and very low density lipoproteins (VLDL). A CE-based quantitation method for apoB-100 was developed utilizing the observed linear relationship between apoB-100 concentration and its corrected 214 nm absorbance peak area measured on-line by CE. Concentration values of apoB-100 in LDL and VLDL samples were determined by CE and found to be accurate when compared to values obtained by immunoturbidimetric analysis and the Lowry method. Capillary electrophoresis can be used as a precise, accurate, and specific on-line method for the qualitative and quantitative analysis of the apoB-100 component of VLDL and LDL-related lipoproteins.—Cruzado, I. D., S. L. Cockrill, C. J. McNeal, and R. D. Macfarlane. Characterization and quantitation of apolipoprotein B-100 by capillary electrophoresis. J. Lipid Res. 1998. 39: 205–217.http://www.sciencedirect.com/science/article/pii/S0022227520342164ultracentrifugationlow density lipoproteinsvery low density lipoproteinslipoprotein[a]sodium dodecyl sulfatedelipidation |
| spellingShingle | Ingrid D. Cruzado Steven L. Cockrill Catherine J. McNeal Ronald D. Macfarlane Characterization and quantitation of apolipoprotein B-100 by capillary electrophoresis Journal of Lipid Research ultracentrifugation low density lipoproteins very low density lipoproteins lipoprotein[a] sodium dodecyl sulfate delipidation |
| title | Characterization and quantitation of apolipoprotein B-100 by capillary electrophoresis |
| title_full | Characterization and quantitation of apolipoprotein B-100 by capillary electrophoresis |
| title_fullStr | Characterization and quantitation of apolipoprotein B-100 by capillary electrophoresis |
| title_full_unstemmed | Characterization and quantitation of apolipoprotein B-100 by capillary electrophoresis |
| title_short | Characterization and quantitation of apolipoprotein B-100 by capillary electrophoresis |
| title_sort | characterization and quantitation of apolipoprotein b 100 by capillary electrophoresis |
| topic | ultracentrifugation low density lipoproteins very low density lipoproteins lipoprotein[a] sodium dodecyl sulfate delipidation |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520342164 |
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