LC–MS Lipidomics: Exploiting a Simple High-Throughput Method for the Comprehensive Extraction of Lipids in a Ruminant Fat Dose-Response Study
Typical lipidomics methods incorporate a liquid–liquid extraction with LC–MS quantitation; however, the classic sample extraction methods are not high-throughput and do not perform well at extracting the full range of lipids especially, the relatively polar species (e.g., acyl-carnitines and glycosp...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-07-01
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| Series: | Metabolites |
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| Online Access: | https://www.mdpi.com/2218-1989/10/7/296 |
| _version_ | 1797562138972127232 |
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| author | Benjamin Jenkins Martin Ronis Albert Koulman |
| author_facet | Benjamin Jenkins Martin Ronis Albert Koulman |
| author_sort | Benjamin Jenkins |
| collection | DOAJ |
| description | Typical lipidomics methods incorporate a liquid–liquid extraction with LC–MS quantitation; however, the classic sample extraction methods are not high-throughput and do not perform well at extracting the full range of lipids especially, the relatively polar species (e.g., acyl-carnitines and glycosphingolipids). In this manuscript, we present a novel sample extraction protocol, which produces a single phase supernatant suitable for high-throughput applications that offers greater performance in extracting lipids across the full spectrum of species. We applied this lipidomics pipeline to a ruminant fat dose–response study to initially compare and validate the different extraction protocols but also to investigate complex lipid biomarkers of ruminant fat intake (adjoining onto simple odd chain fatty acid correlations). We have found 100 lipids species with a strong correlation with ruminant fat intake. This novel sample extraction along with the LC–MS pipeline have shown to be sensitive, robust and hugely informative (>450 lipids species semi-quantified): with a sample preparation throughput of over 100 tissue samples per day and an estimated ~1000 biological fluid samples per day. Thus, this work facilitating both the epidemiological involvement of ruminant fat, research into odd chain lipids and also streamlining the field of lipidomics (both by sample preparation methods and data presentation). |
| first_indexed | 2024-03-10T18:25:14Z |
| format | Article |
| id | doaj.art-f8a773033b294261ac42f6b168e79ae9 |
| institution | Directory Open Access Journal |
| issn | 2218-1989 |
| language | English |
| last_indexed | 2024-03-10T18:25:14Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Metabolites |
| spelling | doaj.art-f8a773033b294261ac42f6b168e79ae92023-11-20T07:07:55ZengMDPI AGMetabolites2218-19892020-07-0110729610.3390/metabo10070296LC–MS Lipidomics: Exploiting a Simple High-Throughput Method for the Comprehensive Extraction of Lipids in a Ruminant Fat Dose-Response StudyBenjamin Jenkins0Martin Ronis1Albert Koulman2NIHR BRC Core Metabolomics and Lipidomics Laboratory, University of Cambridge, Pathology Building Level 4, Addenbrooke’s Hospital, Cambridge CB2 0QQ, UKCollege of Medicine, Department of Pharmacology & Experimental Therapeutics, Louisiana State University Health Sciences Centre, 1901 Perdido Str., New Orleans, LA 70112, USANIHR BRC Core Metabolomics and Lipidomics Laboratory, University of Cambridge, Pathology Building Level 4, Addenbrooke’s Hospital, Cambridge CB2 0QQ, UKTypical lipidomics methods incorporate a liquid–liquid extraction with LC–MS quantitation; however, the classic sample extraction methods are not high-throughput and do not perform well at extracting the full range of lipids especially, the relatively polar species (e.g., acyl-carnitines and glycosphingolipids). In this manuscript, we present a novel sample extraction protocol, which produces a single phase supernatant suitable for high-throughput applications that offers greater performance in extracting lipids across the full spectrum of species. We applied this lipidomics pipeline to a ruminant fat dose–response study to initially compare and validate the different extraction protocols but also to investigate complex lipid biomarkers of ruminant fat intake (adjoining onto simple odd chain fatty acid correlations). We have found 100 lipids species with a strong correlation with ruminant fat intake. This novel sample extraction along with the LC–MS pipeline have shown to be sensitive, robust and hugely informative (>450 lipids species semi-quantified): with a sample preparation throughput of over 100 tissue samples per day and an estimated ~1000 biological fluid samples per day. Thus, this work facilitating both the epidemiological involvement of ruminant fat, research into odd chain lipids and also streamlining the field of lipidomics (both by sample preparation methods and data presentation).https://www.mdpi.com/2218-1989/10/7/296odd chain lipidslipid profilingFolchprotein precipitationsample preparationrelative lipid composition (Mol%) |
| spellingShingle | Benjamin Jenkins Martin Ronis Albert Koulman LC–MS Lipidomics: Exploiting a Simple High-Throughput Method for the Comprehensive Extraction of Lipids in a Ruminant Fat Dose-Response Study Metabolites odd chain lipids lipid profiling Folch protein precipitation sample preparation relative lipid composition (Mol%) |
| title | LC–MS Lipidomics: Exploiting a Simple High-Throughput Method for the Comprehensive Extraction of Lipids in a Ruminant Fat Dose-Response Study |
| title_full | LC–MS Lipidomics: Exploiting a Simple High-Throughput Method for the Comprehensive Extraction of Lipids in a Ruminant Fat Dose-Response Study |
| title_fullStr | LC–MS Lipidomics: Exploiting a Simple High-Throughput Method for the Comprehensive Extraction of Lipids in a Ruminant Fat Dose-Response Study |
| title_full_unstemmed | LC–MS Lipidomics: Exploiting a Simple High-Throughput Method for the Comprehensive Extraction of Lipids in a Ruminant Fat Dose-Response Study |
| title_short | LC–MS Lipidomics: Exploiting a Simple High-Throughput Method for the Comprehensive Extraction of Lipids in a Ruminant Fat Dose-Response Study |
| title_sort | lc ms lipidomics exploiting a simple high throughput method for the comprehensive extraction of lipids in a ruminant fat dose response study |
| topic | odd chain lipids lipid profiling Folch protein precipitation sample preparation relative lipid composition (Mol%) |
| url | https://www.mdpi.com/2218-1989/10/7/296 |
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