Pitfalls in the sample preparation and analysis of N-acylethanolamines[S]
N-acylethanolamines (NAEs) are a group of lipid mediators synthesized in response to a number of physiological and pathological stimuli. Because of the low tissue concentrations of NAEs, analyses often include liquid extraction followed by solid-phase extraction and subsequent quantitation by LC/MS...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2010-10-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520409964 |
| _version_ | 1819127482061684736 |
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| author | Christian Skonberg Andreas Artmann Claus Cornett Steen Honoré Hansen Harald S. Hansen |
| author_facet | Christian Skonberg Andreas Artmann Claus Cornett Steen Honoré Hansen Harald S. Hansen |
| author_sort | Christian Skonberg |
| collection | DOAJ |
| description | N-acylethanolamines (NAEs) are a group of lipid mediators synthesized in response to a number of physiological and pathological stimuli. Because of the low tissue concentrations of NAEs, analyses often include liquid extraction followed by solid-phase extraction and subsequent quantitation by LC/MS or GC/MS. Reported levels of NAEs vary considerably, however, and often no explanation is given for these discrepancies. Brought on by difficulties encountered during method development, the effects of using four different brands of silica-containing solid phase extraction (SPE) columns and five different brands of chloroform for sample preparation were investigated. Considerable variation in the retention and recoveries of seven NAEs and 2-arachidonoylglycerol existed between the SPE columns. Furthermore, it was found that some chloroforms contained quantifiable amounts of N-palmitoylethanolamine and N-stearoylethanolamine. Finally, it was found that use of one of the chloroforms resulted in a loss of N-oleoylethanolamine from solution due to addition of chlorine to the ω-9 bond. The identity of this reaction product was confirmed by LC-MS/MS and NMR. It is recommended that these aspects of sample preparation and analysis should be thoroughly validated during method development and the relevant information on specific brands used be reported in future communications in order to better estimate the validity of reported quantitative data. |
| first_indexed | 2024-12-22T08:12:37Z |
| format | Article |
| id | doaj.art-79c0760bfac040c489ce044c68f18db1 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-22T08:12:37Z |
| publishDate | 2010-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-79c0760bfac040c489ce044c68f18db12022-12-21T18:32:59ZengElsevierJournal of Lipid Research0022-22752010-10-01511030623073Pitfalls in the sample preparation and analysis of N-acylethanolamines[S]Christian Skonberg0Andreas Artmann1Claus Cornett2Steen Honoré Hansen3Harald S. Hansen4To whom correspondence should be addressed. cs@farma.ku.dk; Department of Pharmaceutics and Analytical Chemistry, Faculty of Pharmaceutical Sciences, and University of Copenhagen, Copenhagen, DenmarkDepartment of Pharmacology and Pharmacotherapy, Faculty of Pharmaceutical Sciences, University of Copenhagen, Copenhagen, DenmarkDepartment of Pharmaceutics and Analytical Chemistry, Faculty of Pharmaceutical Sciences, and University of Copenhagen, Copenhagen, DenmarkDepartment of Pharmaceutics and Analytical Chemistry, Faculty of Pharmaceutical Sciences, and University of Copenhagen, Copenhagen, DenmarkDepartment of Pharmacology and Pharmacotherapy, Faculty of Pharmaceutical Sciences, University of Copenhagen, Copenhagen, DenmarkN-acylethanolamines (NAEs) are a group of lipid mediators synthesized in response to a number of physiological and pathological stimuli. Because of the low tissue concentrations of NAEs, analyses often include liquid extraction followed by solid-phase extraction and subsequent quantitation by LC/MS or GC/MS. Reported levels of NAEs vary considerably, however, and often no explanation is given for these discrepancies. Brought on by difficulties encountered during method development, the effects of using four different brands of silica-containing solid phase extraction (SPE) columns and five different brands of chloroform for sample preparation were investigated. Considerable variation in the retention and recoveries of seven NAEs and 2-arachidonoylglycerol existed between the SPE columns. Furthermore, it was found that some chloroforms contained quantifiable amounts of N-palmitoylethanolamine and N-stearoylethanolamine. Finally, it was found that use of one of the chloroforms resulted in a loss of N-oleoylethanolamine from solution due to addition of chlorine to the ω-9 bond. The identity of this reaction product was confirmed by LC-MS/MS and NMR. It is recommended that these aspects of sample preparation and analysis should be thoroughly validated during method development and the relevant information on specific brands used be reported in future communications in order to better estimate the validity of reported quantitative data.http://www.sciencedirect.com/science/article/pii/S0022227520409964anandamideartefactchloroformcontaminationextractionoleoylethanolamide |
| spellingShingle | Christian Skonberg Andreas Artmann Claus Cornett Steen Honoré Hansen Harald S. Hansen Pitfalls in the sample preparation and analysis of N-acylethanolamines[S] Journal of Lipid Research anandamide artefact chloroform contamination extraction oleoylethanolamide |
| title | Pitfalls in the sample preparation and analysis of N-acylethanolamines[S] |
| title_full | Pitfalls in the sample preparation and analysis of N-acylethanolamines[S] |
| title_fullStr | Pitfalls in the sample preparation and analysis of N-acylethanolamines[S] |
| title_full_unstemmed | Pitfalls in the sample preparation and analysis of N-acylethanolamines[S] |
| title_short | Pitfalls in the sample preparation and analysis of N-acylethanolamines[S] |
| title_sort | pitfalls in the sample preparation and analysis of n acylethanolamines s |
| topic | anandamide artefact chloroform contamination extraction oleoylethanolamide |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520409964 |
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