Lipid profiling and lipidomics

Reported lipid biomarkers

Many studies have determined the association between specific lipids and specific pathologies, nutrients or diets. This does not mean that these lipids are biomarkers.

Table 1 gives an overview of associations between lipids and nutrition that have been reproduced by different studies, either using independent methods or independent cohorts.

Legend: PC(36:2): phosphatidylcholine(36:2); SM(39:1): sphingomyelin(39:1); FA(22:6n3): fatty acid (22:6n3) or DHA; FA(20:5n3) fatty acid (20:5n3) or EPA; CE(22:6):  Cholesteryl ester (22:6), CE(20:5): Cholesteryl ester (20:5); PC(36:5): phosphatidylcholine (36:5); PC(38:6): phosphatidylcholine(36:5); FA(15:0): fatty acid (15:0); FA(23:0): fatty acid (23:0); PC(35:1): phosphatidylcholine(35:1); SM(39:1): sphingomyelin(39:1); LCMS: Liquid chromatography hyphenated with mass spectrometry; DIMS: direct infusion mass spectrometry; GC: Gas chromatography.

Methods of analysis

Direct infusion mass spectrometry (DIMS)
Organic extracts are infused into a high-resolution mass spectrometer using chip-based nanospray. Data can be collected in positive mode or in negative mode or both. Data processing protocols are necessary to deconvolute the spectra and obtain measurements per lipid signal. Internal standards can be used to quantify the results.

Liquid chromatography hyphenated with mass spectrometry (LCMS)
Instead of direct infusion, it is possible to separate the lipids first by chromatography, most approaches will use a reversed phase column material, that will retain lipids by their polarity (retaining more apolar lipids). The mass spectrometer can by based on high resolution or on specific fragmentations (triple quadrupole). Each lipid will have a specific retention time and mass spectral feature that will be detected. Again, internal standards can be used to enable quantification.

Gas Chromatography
It is possible to measure the individual fatty acids which are with a lipid fraction (e.g. phospholipids) or within the whole sample. The standard method is to hydrolyse the extracts and derivatise the fatty acids into their methyl esters (FAMEs). These can be separated by Gas chromatography and both a flame ionisation detector or an electron impact mass spectrometer can be used to for identification and quantification.

Quality Control

The National Institute of Standards and Technology produces Standard Reference Material (SRM) 1950 for which quantified lipid concentration are available.

Technical assistance

For technical assistance and questions on lipids or lipidomics, contact sophie.moore@kcl.ac.uk, or write to Dr Albert Koulman, (Ak675@medschl.cam.ac.uk) PhD Head of NIHR BRC core Metabolomics and Lipidomics Laboratory, University of Cambridge, UK.

Useful links

cMaLL combined lipidomics training and sample analysis course: for more information, see the OpeN-Global page on training opportunities, or link directly to the course information.

Further reading 

Zheng, Sharp, Imamura, Koulman, et al. Association between plasma phospholipid saturated fatty acids and metabolic markers of lipid, hepatic, inflammation and glycaemic pathways in eight European countries: a cross-sectional analysis in the EPIC-InterAct study. BMC Med. 2017 Nov 17;15(1):203. doi: 10.1186/s12916-017-0968-4.

Stanley, Jenkins, Walker, Koulman, et al. Lipidomics Profiling of Human Adipose Tissue Identifies a Pattern of Lipids Associated with Fish Oil Supplementation. J Proteome Res. 2017 Sep 1;16(9):3168-3179. doi: 10.1021/acs.jproteome.7b00161.

Markey, Vasilopoulou, Kliem, Koulman, et al. Plasma phospholipid fatty acid profile confirms compliance to a novel saturated fat-reduced, monounsaturated fat-enriched dairy product intervention in adults at moderate cardiovascular risk: a randomized controlled trial. Nutr J. 2017 May 23;16(1):33. doi: 10.1186/s12937-017-0249-2.

Jenkins, Seyssel, Chiu,…Koulman. Odd Chain Fatty Acids; New Insights of the Relationship Between the Gut Microbiota, Dietary Intake, Biosynthesis and Glucose Intolerance. Sci Rep. 2017 Mar 23;7:44845. doi:10.1038/srep44845.

Acharjee, Prentice, Acerini, …Koulman. The translation of lipid profiles to nutritional biomarkers in the study of infant metabolism. Metabolomics. 2017;13(3):25. doi: 10.1007/s11306-017-1166-2

References

1.    Roberts LD, Koulman A, Griffin JL. Towards metabolic biomarkers of insulin resistance and type 2 diabetes: progress from the metabolome. Lancet Diabetes Endocrinol. 2014 Jan;2(1):65-75.

2.    Koulman A, Prentice P, Wong MCY, Matthews L, Bond NJ, Eiden M, Griffin JL, Dunger DB. The development and validation of a fast and robust dried blood spot based lipid profiling method to study infant metabolism. Metabolomics. 2014;10(5):1018-1025.

3.     Bowden JA, Heckert A, Ulmer CZ, Jones CM, Koelmel JP, Abdullah L, Ahonen L, Alnouti Y, Armando AM, Asara JM, Bamba T, Barr JR, Bergquist J, Borchers CH, Brandsma J, Breitkopf SB, Cajka T, Cazenave-Gassiot A, Checa A, Cinel MA, Colas RA, Cremers S, Dennis EA, Evans JE, Fauland A, Fiehn O, Gardner MS, Garrett TJ, Gotlinger KH, Han J, Huang Y, Neo AH, Hyötyläinen T, Izumi Y, Jiang H, Jiang H, Jiang J, Kachman M, Kiyonami R, Klavins K, Klose C, Köfeler HC, Kolmert J, Koal T, Koster G, Kuklenyik Z, Kurland IJ, Leadley M, Lin K, Maddipati KR, McDougall D, Meikle PJ, Mellett NA, Monnin C, Moseley MA, Nandakumar R, Oresic M, Patterson R, Peake D, Pierce JS, Post M, Postle AD, Pugh R, Qiu Y, Quehenberger O, Ramrup P, Rees J, Rembiesa B, Reynaud D, Roth MR, Sales S, Schuhmann K, Schwartzman ML, Serhan CN, Shevchenko A, Somerville SE, St John-Williams L, Surma MA, Takeda H, Thakare R, Thompson JW, Torta F, Triebl A, Trötzmüller M, Ubhayasekera SJK, Vuckovic D, Weir JM, Welti R, Wenk MR, Wheelock CE, Yao L, Yuan M, Zhao XH, Zhou S. Harmonizing lipidomics: NIST interlaboratory comparison exercise for lipidomics using SRM 1950-Metabolites in Frozen Human Plasma. J Lipid Res. 2017 Dec;58(12):2275-2288.

4.    Eiden M, Koulman A, Hatunic M, West JA, Murfitt S, Osei M, Adams C, Wang X, Chu Y, Marney L, Roberts LD, O’Rahilly S, Semple RK, Savage DB, Griffin JL. Mechanistic insights revealed by lipid profiling in monogenic insulin resistance syndromes. Genome Med. 2015 Jun 28;7:63.

5.    Stanley EG, Jenkins BJ, Walker CG, Koulman A, Browning L, West AL, Calder PC, Jebb SA, Griffin JL. Lipidomics Profiling of Human Adipose Tissue Identifies a Pattern of Lipids Associated with Fish Oil Supplementation. J Proteome Res. 2017 Sep 1;16(9):3168-3179.

6.    Jacobs S, Schiller K, Jansen E, Fritsche A, Weikert C, di Giuseppe R, Boeing H, Schulze MB, Kröger J. Association between erythrocyte membrane fatty acids and biomarkers of dyslipidemia in the EPIC-Potsdam study. Eur J Clin Nutr. 2014 Apr;68(4):517-25.

7.    Acharjee A, Prentice P, Acerini C, Smith J, Hughes IA, Ong K, Griffin JL, Dunger D, Koulman A. The translation of lipid profiles to nutritional biomarkers in the study of infant metabolism. Metabolomics. 2017;13(3):25.

8.    Lu L, Koulman A, Petry CJ, Jenkins B, Matthews L, Hughes IA, Acerini CL, Ong KK, Dunger DB. An Unbiased Lipidomics Approach Identifies Early Second Trimester Lipids Predictive of Maternal Glycemic Traits and Gestational Diabetes Mellitus. Diabetes Care. 2016 Dec;39(12):2232-2239.

9.    Sanders FWB, Acharjee A, Walker C, Marney L, Roberts LD, Imamura F, Jenkins B, Case J, Ray S, Virtue S, Vidal-Puig A, Kuh D, Hardy R, Allison M, Forouhi N, Murray AJ, Wareham N, Vacca M, Koulman A, Griffin JL. Hepatic steatosis risk is partly driven by increased de novo lipogenesis following carbohydrate consumption. Genome Biol. 2018 Jun 20;19(1):79.

10.  Proitsi P, Kim M, Whiley L, Simmons A, Sattlecker M, Velayudhan L, Lupton MK, Soininen H, Kloszewska I, Mecocci P, Tsolaki M, Vellas B, Lovestone S, Powell JF, Dobson RJ, Legido-Quigley C. Association of blood lipids with Alzheimer’s disease: A comprehensive lipidomics analysis. Alzheimers Dement. 2017 Feb;13(2):140-151.

11.  Browning LM, Walker CG, Mander AP, West AL, Madden J, Gambell JM, Young S, Wang L, Jebb SA, Calder PC. Incorporation of eicosapentaenoic and docosahexaenoic acids into lipid pools when given as supplements providing doses equivalent to typical intakes of oily fish. Am J Clin Nutr. 2012 Oct;96(4):748-58.

12.  Jenkins BJ, Seyssel K, Chiu S, Pan PH, Lin SY, Stanley E, Ament Z, West JA, Summerhill K, Griffin JL, Vetter W, Autio KJ, Hiltunen K, Hazebrouck S, Stepankova R, Chen CJ, Alligier M, Laville M, Moore M, Kraft G, Cherrington A, King S, Krauss RM, de Schryver E, Van Veldhoven PP, Ronis M, Koulman A. Odd Chain Fatty Acids; New Insights of the Relationship Between the Gut Microbiota, Dietary Intake, Biosynthesis and Glucose Intolerance. Sci Rep. 2017 Mar 23;7:44845.

Page 1: Lipid profiling and lipidomics: home // Page 2: Importance of lipids for health // Page 3: Background to lipid profiling and lipidomics // Page 4: Methods of analysis // Page 5: Quality control and technical assistance // Page 6: Further reading // Page 7: References