Quantitation of fatty acyl-coenzyme As in mammalian cells by liquid chromatography-electrospray ionization tandem mass spectrometry*
Fatty acyl-CoAs participate in numerous cellular processes. This article describes a method for the quantitation of subpicomole amounts of long-chain and very-long-chain fatty acyl-CoAs by reverse-phase LC combined with electrospray ionization tandem mass spectrometry in positive ion mode with odd-c...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2008-05-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520425982 |
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| author | Christopher A. Haynes Jeremy C. Allegood Kacee Sims Elaine W. Wang M. Cameron Sullards Alfred H. Merrill, Jr. |
| author_facet | Christopher A. Haynes Jeremy C. Allegood Kacee Sims Elaine W. Wang M. Cameron Sullards Alfred H. Merrill, Jr. |
| author_sort | Christopher A. Haynes |
| collection | DOAJ |
| description | Fatty acyl-CoAs participate in numerous cellular processes. This article describes a method for the quantitation of subpicomole amounts of long-chain and very-long-chain fatty acyl-CoAs by reverse-phase LC combined with electrospray ionization tandem mass spectrometry in positive ion mode with odd-chain-length fatty acyl-CoAs as internal standards. This method is applicable to a wide range of species [at least myristoyl- (C14:0-) to cerotoyl- (C26:0-) CoA] in modest numbers of cells in culture (∼106–107), with analyses of RAW264.7 cells and MCF7 cells given as examples. Analysis of these cells revealed large differences in fatty acyl-CoA amounts (12 ± 1.0 pmol/106 RAW264.7 cells vs. 80.4 ± 6.1 pmol/106 MCF7 cells) and subspecies distribution. Very-long-chain fatty acyl-CoAs with alkyl chain lengths > C20 constitute <10% of the total fatty acyl-CoAs of RAW264.7 cells versus >50% for MCF7 cells, which somewhat astonishingly contain approximately as much C24:0- and C26:0-CoAs as C16:0- and C18:0-CoAs and essentially equal amounts of C26:1- and C18:1-CoAs. This simple and robust method should facilitate the inclusion of this family of compounds in “lipidomics” and “metabolomics” studies. |
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| format | Article |
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| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-17T23:23:42Z |
| publishDate | 2008-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-b42f584bd718495c92d74e94e01dfa002022-12-21T21:28:50ZengElsevierJournal of Lipid Research0022-22752008-05-0149511131125Quantitation of fatty acyl-coenzyme As in mammalian cells by liquid chromatography-electrospray ionization tandem mass spectrometry*Christopher A. Haynes0Jeremy C. Allegood1Kacee Sims2Elaine W. Wang3M. Cameron Sullards4Alfred H. Merrill, Jr.5Schools of Biology, Georgia Institute of Technology, Atlanta, GA 30332-0230Chemistry and Biochemistry and the Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332-0230Chemistry and Biochemistry and the Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332-0230Schools of Biology, Georgia Institute of Technology, Atlanta, GA 30332-0230Schools of Biology, Georgia Institute of Technology, Atlanta, GA 30332-0230; Chemistry and Biochemistry and the Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332-0230Schools of Biology, Georgia Institute of Technology, Atlanta, GA 30332-0230; Chemistry and Biochemistry and the Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332-0230Fatty acyl-CoAs participate in numerous cellular processes. This article describes a method for the quantitation of subpicomole amounts of long-chain and very-long-chain fatty acyl-CoAs by reverse-phase LC combined with electrospray ionization tandem mass spectrometry in positive ion mode with odd-chain-length fatty acyl-CoAs as internal standards. This method is applicable to a wide range of species [at least myristoyl- (C14:0-) to cerotoyl- (C26:0-) CoA] in modest numbers of cells in culture (∼106–107), with analyses of RAW264.7 cells and MCF7 cells given as examples. Analysis of these cells revealed large differences in fatty acyl-CoA amounts (12 ± 1.0 pmol/106 RAW264.7 cells vs. 80.4 ± 6.1 pmol/106 MCF7 cells) and subspecies distribution. Very-long-chain fatty acyl-CoAs with alkyl chain lengths > C20 constitute <10% of the total fatty acyl-CoAs of RAW264.7 cells versus >50% for MCF7 cells, which somewhat astonishingly contain approximately as much C24:0- and C26:0-CoAs as C16:0- and C18:0-CoAs and essentially equal amounts of C26:1- and C18:1-CoAs. This simple and robust method should facilitate the inclusion of this family of compounds in “lipidomics” and “metabolomics” studies.http://www.sciencedirect.com/science/article/pii/S0022227520425982lipidomicsmetabolomicsRAW264.7MCF7lignoceroyl-CoAnervonoyl-CoA |
| spellingShingle | Christopher A. Haynes Jeremy C. Allegood Kacee Sims Elaine W. Wang M. Cameron Sullards Alfred H. Merrill, Jr. Quantitation of fatty acyl-coenzyme As in mammalian cells by liquid chromatography-electrospray ionization tandem mass spectrometry* Journal of Lipid Research lipidomics metabolomics RAW264.7 MCF7 lignoceroyl-CoA nervonoyl-CoA |
| title | Quantitation of fatty acyl-coenzyme As in mammalian cells by liquid chromatography-electrospray ionization tandem mass spectrometry* |
| title_full | Quantitation of fatty acyl-coenzyme As in mammalian cells by liquid chromatography-electrospray ionization tandem mass spectrometry* |
| title_fullStr | Quantitation of fatty acyl-coenzyme As in mammalian cells by liquid chromatography-electrospray ionization tandem mass spectrometry* |
| title_full_unstemmed | Quantitation of fatty acyl-coenzyme As in mammalian cells by liquid chromatography-electrospray ionization tandem mass spectrometry* |
| title_short | Quantitation of fatty acyl-coenzyme As in mammalian cells by liquid chromatography-electrospray ionization tandem mass spectrometry* |
| title_sort | quantitation of fatty acyl coenzyme as in mammalian cells by liquid chromatography electrospray ionization tandem mass spectrometry |
| topic | lipidomics metabolomics RAW264.7 MCF7 lignoceroyl-CoA nervonoyl-CoA |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520425982 |
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