An improved assay for platelet-activating factor using HPLC-tandem mass spectrometry
We describe an improved assay for platelet-activating factor (PAF; 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) using HPLC-tandem mass spectrometry (LC-MS/MS). The present method can readily detect as little as 1 pg (1.9 fmol) of PAF, a significant improvement over previously described LC-MS/MS m...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2005-02-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520340712 |
| _version_ | 1818602518501916672 |
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| author | John S. Owen Robert L. Wykle Michael P. Samuel Michael J. Thomas |
| author_facet | John S. Owen Robert L. Wykle Michael P. Samuel Michael J. Thomas |
| author_sort | John S. Owen |
| collection | DOAJ |
| description | We describe an improved assay for platelet-activating factor (PAF; 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) using HPLC-tandem mass spectrometry (LC-MS/MS). The present method can readily detect as little as 1 pg (1.9 fmol) of PAF, a significant improvement over previously described LC-MS/MS methods, and gives a linear response up to 1,000 pg of PAF. Our method also overcomes the artifacts from isobaric lipids that have limited the usefulness of certain existing LC-MS/MS assays for PAF. In the course of these studies, we detected three novel lipid species in human neutrophils. One of the novel lipids appears to be a new molecular species of PAF, and the other two have chromatographic and mass spectrometric properties consistent with stearoyl-formyl-glycerophosphocholine and oleoyl-formyl-glycerophosphocholine.These observations identify previously unknown potential interferences in the measurement of PAF by LC-MS/MS. Moreover, our data suggest that the previously described palmitoyl-formyl-glycerophosphocholine is not unique but rather is a member of a new and poorly understood family of formylated lipids. |
| first_indexed | 2024-12-16T13:08:33Z |
| format | Article |
| id | doaj.art-ebbec803eb684cbeb4555e518d332ccc |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-16T13:08:33Z |
| publishDate | 2005-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-ebbec803eb684cbeb4555e518d332ccc2022-12-21T22:30:41ZengElsevierJournal of Lipid Research0022-22752005-02-01462373382An improved assay for platelet-activating factor using HPLC-tandem mass spectrometryJohn S. Owen0Robert L. Wykle1Michael P. Samuel2Michael J. Thomas3Department of Biochemistry, Wake Forest University School of Medicine, Medical Center Boulevard, Winston-Salem, NCDepartment of Biochemistry, Wake Forest University School of Medicine, Medical Center Boulevard, Winston-Salem, NCDepartment of Biochemistry, Wake Forest University School of Medicine, Medical Center Boulevard, Winston-Salem, NCDepartment of Biochemistry, Wake Forest University School of Medicine, Medical Center Boulevard, Winston-Salem, NCWe describe an improved assay for platelet-activating factor (PAF; 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) using HPLC-tandem mass spectrometry (LC-MS/MS). The present method can readily detect as little as 1 pg (1.9 fmol) of PAF, a significant improvement over previously described LC-MS/MS methods, and gives a linear response up to 1,000 pg of PAF. Our method also overcomes the artifacts from isobaric lipids that have limited the usefulness of certain existing LC-MS/MS assays for PAF. In the course of these studies, we detected three novel lipid species in human neutrophils. One of the novel lipids appears to be a new molecular species of PAF, and the other two have chromatographic and mass spectrometric properties consistent with stearoyl-formyl-glycerophosphocholine and oleoyl-formyl-glycerophosphocholine.These observations identify previously unknown potential interferences in the measurement of PAF by LC-MS/MS. Moreover, our data suggest that the previously described palmitoyl-formyl-glycerophosphocholine is not unique but rather is a member of a new and poorly understood family of formylated lipids.http://www.sciencedirect.com/science/article/pii/S0022227520340712high-performance liquid chromatography tandem mass spectrometrytrace analysisneutrophil |
| spellingShingle | John S. Owen Robert L. Wykle Michael P. Samuel Michael J. Thomas An improved assay for platelet-activating factor using HPLC-tandem mass spectrometry Journal of Lipid Research high-performance liquid chromatography tandem mass spectrometry trace analysis neutrophil |
| title | An improved assay for platelet-activating factor using HPLC-tandem mass spectrometry |
| title_full | An improved assay for platelet-activating factor using HPLC-tandem mass spectrometry |
| title_fullStr | An improved assay for platelet-activating factor using HPLC-tandem mass spectrometry |
| title_full_unstemmed | An improved assay for platelet-activating factor using HPLC-tandem mass spectrometry |
| title_short | An improved assay for platelet-activating factor using HPLC-tandem mass spectrometry |
| title_sort | improved assay for platelet activating factor using hplc tandem mass spectrometry |
| topic | high-performance liquid chromatography tandem mass spectrometry trace analysis neutrophil |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520340712 |
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