Femtomole analysis of 9-oxononanoyl cholesterol by high performance liquid chromatography1
9-Oxononanoyl cholesterol, a cholesterol core-aldehyde formed during lipoprotein oxidation, was recently identified in advanced human atherosclerotic lesions. Here we present a rapid and sensitive HPLC method for 9-oxononanoyl cholesterol analysis. 9-Oxononanoyl cholesterol was converted to the corr...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
1998-07-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520325335 |
| _version_ | 1819076881949917184 |
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| author | Barbara Karten Herbert Boechzelt Peter M. Abuja Martin Mittelbach Karl Oettl Wolfgang Sattler |
| author_facet | Barbara Karten Herbert Boechzelt Peter M. Abuja Martin Mittelbach Karl Oettl Wolfgang Sattler |
| author_sort | Barbara Karten |
| collection | DOAJ |
| description | 9-Oxononanoyl cholesterol, a cholesterol core-aldehyde formed during lipoprotein oxidation, was recently identified in advanced human atherosclerotic lesions. Here we present a rapid and sensitive HPLC method for 9-oxononanoyl cholesterol analysis. 9-Oxononanoyl cholesterol was converted to the corresponding fluorescent decahydroacridine derivative by reaction with 1,3-cyclohexanedione. The derivatives formed were purified by solid-phase extraction on C-18 columns, separated by reversed phase HPLC with isocratic elution, and detected by their fluorescence. Decahydroacridine derivatives of 9-oxononanoyl cholesterol were stable for at least 160 h. The limit of quantitation of the method presented is at the low (≈50) femtomole level, with an absolute limit of detection (signal: noise = 6) of 15 fmol. Intra-assay variation was ≤5%, while inter-assay variations were between 5 and 15%, depending on the concentration of the analyte. Standard curves were linear over nearly three orders of magnitude (50 fmol–12.5 pmol). 9-Oxononanoyl cholesterol proved to be the major cholesterol core-aldehyde formed during t-BuOOH/FeSO4 oxidation of cholesteryl linoleate and Cu2+-induced LDL oxidation, findings confirmed by atmospheric pressure chemical ionization–mass spectrometry. Analysis of lipid extracts obtained from advanced human atherosclerotic lesions revealed the presence of 9-oxononanoyl cholesterol in all tissue samples analyzed (28 ± 14 μmol/mol cholesterol, n = 9) despite the presence of α-tocopherol (4 ± 1.2 mmol/mol cholesterol, n = 9).—Karten, B., H. Boechzelt, P. M. Abuja, M. Mittelbach, K. Oettl, and W. Sattler. Femtomole analysis of 9-oxononanoyl by high performance liquid chromatography. J. Lipid Res. 1998. 39: 1508–1519. |
| first_indexed | 2024-12-21T18:48:21Z |
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| id | doaj.art-a989ca9936e54e7cb81a932fea675946 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-21T18:48:21Z |
| publishDate | 1998-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-a989ca9936e54e7cb81a932fea6759462022-12-21T18:53:50ZengElsevierJournal of Lipid Research0022-22751998-07-0139715081519Femtomole analysis of 9-oxononanoyl cholesterol by high performance liquid chromatography1Barbara Karten0Herbert Boechzelt1Peter M. Abuja2Martin Mittelbach3Karl Oettl4Wolfgang Sattler5Institute of Biochemistry, SFB Biomembranes Research Centre; University of Graz; 8010 Graz, AustriaInstitute of Organic Chemistry, SFB Biomembranes Research Centre; University of Graz; 8010 Graz, AustriaInstitute of Biochemistry, SFB Biomembranes Research Centre; University of Graz; 8010 Graz, AustriaInstitute of Organic Chemistry, SFB Biomembranes Research Centre; University of Graz; 8010 Graz, AustriaInstitute of Medical Chemistry, SFB Biomembranes Research Centre; University of Graz; 8010 Graz, AustriaTo whom correspondence should be addressed.; Institute of Medical Biochemistry, SFB Biomembranes Research Centre; University of Graz; 8010 Graz, Austria9-Oxononanoyl cholesterol, a cholesterol core-aldehyde formed during lipoprotein oxidation, was recently identified in advanced human atherosclerotic lesions. Here we present a rapid and sensitive HPLC method for 9-oxononanoyl cholesterol analysis. 9-Oxononanoyl cholesterol was converted to the corresponding fluorescent decahydroacridine derivative by reaction with 1,3-cyclohexanedione. The derivatives formed were purified by solid-phase extraction on C-18 columns, separated by reversed phase HPLC with isocratic elution, and detected by their fluorescence. Decahydroacridine derivatives of 9-oxononanoyl cholesterol were stable for at least 160 h. The limit of quantitation of the method presented is at the low (≈50) femtomole level, with an absolute limit of detection (signal: noise = 6) of 15 fmol. Intra-assay variation was ≤5%, while inter-assay variations were between 5 and 15%, depending on the concentration of the analyte. Standard curves were linear over nearly three orders of magnitude (50 fmol–12.5 pmol). 9-Oxononanoyl cholesterol proved to be the major cholesterol core-aldehyde formed during t-BuOOH/FeSO4 oxidation of cholesteryl linoleate and Cu2+-induced LDL oxidation, findings confirmed by atmospheric pressure chemical ionization–mass spectrometry. Analysis of lipid extracts obtained from advanced human atherosclerotic lesions revealed the presence of 9-oxononanoyl cholesterol in all tissue samples analyzed (28 ± 14 μmol/mol cholesterol, n = 9) despite the presence of α-tocopherol (4 ± 1.2 mmol/mol cholesterol, n = 9).—Karten, B., H. Boechzelt, P. M. Abuja, M. Mittelbach, K. Oettl, and W. Sattler. Femtomole analysis of 9-oxononanoyl by high performance liquid chromatography. J. Lipid Res. 1998. 39: 1508–1519.http://www.sciencedirect.com/science/article/pii/S0022227520325335LDLatherosclerosisplaque lipidsreactive aldehydeslipid peroxidationcore-aldehydes |
| spellingShingle | Barbara Karten Herbert Boechzelt Peter M. Abuja Martin Mittelbach Karl Oettl Wolfgang Sattler Femtomole analysis of 9-oxononanoyl cholesterol by high performance liquid chromatography1 Journal of Lipid Research LDL atherosclerosis plaque lipids reactive aldehydes lipid peroxidation core-aldehydes |
| title | Femtomole analysis of 9-oxononanoyl cholesterol by high performance liquid chromatography1 |
| title_full | Femtomole analysis of 9-oxononanoyl cholesterol by high performance liquid chromatography1 |
| title_fullStr | Femtomole analysis of 9-oxononanoyl cholesterol by high performance liquid chromatography1 |
| title_full_unstemmed | Femtomole analysis of 9-oxononanoyl cholesterol by high performance liquid chromatography1 |
| title_short | Femtomole analysis of 9-oxononanoyl cholesterol by high performance liquid chromatography1 |
| title_sort | femtomole analysis of 9 oxononanoyl cholesterol by high performance liquid chromatography1 |
| topic | LDL atherosclerosis plaque lipids reactive aldehydes lipid peroxidation core-aldehydes |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520325335 |
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