Summary of Study ST000916

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000633. The data can be accessed directly via it's Project DOI: 10.21228/M8V961 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

Study ID	ST000916
Study Title	Biomarkers of NAFLD progression: a lipidomics approach to an epidemic. Part 2:Plasma
Study Type	Lipidomics Study
Study Summary	The spectrum of nonalcoholic fatty liver disease (NAFLD) includes steatosis, nonalcoholic steatohepatitis (NASH), and cirrhosis. Recognition and timely diagnosis of these different stages, particularly NASH, is important for both potential reversibility and limitation of complications. Liver biopsy remains the clinical standard for definitive diagnosis. Diagnostic tools minimizing the need for invasive procedures or that add information to histologic data are important in novel management strategies for the growing epidemic of NAFLD. We describe an 'omics' approach to detecting a reproducible signature of lipid metabolites, aqueous intracellular metabolites, SNPs, and mRNA transcripts in a double-blinded study of patients with different stages of NAFLD that involves profiling liver biopsies, plasma, and urine samples. Using linear discriminant analysis, a panel of 20 plasma metabolites that includes glycerophospholipids, sphingolipids, sterols, and various aqueous small molecular weight components involved in cellular metabolic pathways, can be used to differentiate between NASH and steatosis. This identification of differential biomolecular signatures has the potential to improve clinical diagnosis and facilitate therapeutic intervention of NAFLD.
Institute	LIPID MAPS
Department	Bioengineering
Last Name	Fahy
First Name	Eoin
Address	9500 Gilman, La Jolla, CA, 92093, USA
Email	efahy@ucsd.edu
Phone	858-534-4076
Submit Date	2018-01-14
Publications	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4340319/
Raw Data Available	Yes
Analysis Type Detail	GC-MS/LC-MS
Release Date	2018-04-05
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR000633
Project DOI:	doi: 10.21228/M8V961
Project Title:	Biomarkers of NAFLD progression: a lipidomics approach to an epidemic
Project Type:	Lipidomics Study
Project Summary:	The spectrum of nonalcoholic fatty liver disease (NAFLD) includes steatosis, nonalcoholic steatohepatitis (NASH), and cirrhosis. Recognition and timely diagnosis of these different stages, particularly NASH, is important for both potential reversibility and limitation of complications. Liver biopsy remains the clinical standard for definitive diagnosis. Diagnostic tools minimizing the need for invasive procedures or that add information to histologic data are important in novel management strategies for the growing epidemic of NAFLD. We describe an 'omics' approach to detecting a reproducible signature of lipid metabolites, aqueous intracellular metabolites, SNPs, and mRNA transcripts in a double-blinded study of patients with different stages of NAFLD that involves profiling liver biopsies, plasma, and urine samples. Using linear discriminant analysis, a panel of 20 plasma metabolites that includes glycerophospholipids, sphingolipids, sterols, and various aqueous small molecular weight components involved in cellular metabolic pathways, can be used to differentiate between NASH and steatosis. This identification of differential biomolecular signatures has the potential to improve clinical diagnosis and facilitate therapeutic intervention of NAFLD.
Institute:	University of California, San Diego
Department:	Bioengineering
Last Name:	Fahy
First Name:	Eoin
Address:	9500 Gilman, La Jolla, CA, 92093, USA
Email:	efahy@ucsd.edu
Phone:	858-534-4076
Funding Source:	NIGMS Grant GM U54069338
Publications:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4340319/
Contributors:	LIPID MAPS Consortium

Subject:

Subject ID:	SU000954
Subject Type:	Human clinical study
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Age Or Age Range:	23-83
Gender:	Male and Female
Human Ethnicity:	Mixed

Factors:

Subject type: Human clinical study; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id	local_sample_id	Diagnosis
SA053890	NASH055	Cirrhosis
SA053891	NASH049	Cirrhosis
SA053892	NASH048	Cirrhosis
SA053893	NASH064	Cirrhosis
SA053894	NASH068	Cirrhosis
SA053895	NASH005	Cirrhosis
SA053896	NASH069	Cirrhosis
SA053897	NASH047	Cirrhosis
SA053898	NASH065	Cirrhosis
SA053899	NASH052	Cirrhosis
SA053900	NASH016	Cirrhosis
SA053901	NASH013	Cirrhosis
SA053902	NASH040	Cirrhosis
SA053903	NASH007	Cirrhosis
SA053904	NASH022	Cirrhosis
SA053905	NASH009	Cirrhosis
SA053906	NASH026	Cirrhosis
SA053907	NASH029	Cirrhosis
SA053908	NASH028	Cirrhosis
SA053909	NASH027	Cirrhosis
SA053910	NASH072	NASH
SA053911	NASH057	NASH
SA053912	NASH044	NASH
SA053913	NASH074	NASH
SA053914	NASH088	NASH
SA053915	NASH039	NASH
SA053916	NASH090	NASH
SA053917	NASH087	NASH
SA053918	NASH084	NASH
SA053919	NASH031	NASH
SA053920	NASH018	NASH
SA053921	NASH012	NASH
SA053922	NASH038	NASH
SA053923	NASH010	NASH
SA053924	NASH019	NASH
SA053925	NASH015	NASH
SA053926	NASH037	NASH
SA053927	NASH035	NASH
SA053928	NASH030	NASH
SA053929	NASH021	NASH
SA053930	NASH067	Normal
SA053931	NASH070	Normal
SA053932	NASH066	Normal
SA053933	NASH060	Normal
SA053934	NASH077	Normal
SA053935	NASH053	Normal
SA053936	NASH054	Normal
SA053937	NASH086	Normal
SA053938	NASH091	Normal
SA053939	NASH051	Normal
SA053940	NASH089	Normal
SA053941	NASH085	Normal
SA053942	NASH080	Normal
SA053943	NASH082	Normal
SA053944	NASH078	Normal
SA053945	NASH014	Normal
SA053946	NASH017	Normal
SA053947	NASH020	Normal
SA053948	NASH006	Normal
SA053949	NASH004	Normal
SA053950	NASH050	Normal
SA053951	NASH003	Normal
SA053952	NASH024	Normal
SA053953	NASH011	Normal
SA053954	NASH043	Normal
SA053955	NASH045	Normal
SA053956	NASH034	Normal
SA053957	NASH042	Normal
SA053958	NASH046	Normal
SA053959	NASH041	Normal
SA053960	NASH036	Normal
SA053961	NASH075	Steatosis
SA053962	NASH073	Steatosis
SA053963	NASH071	Steatosis
SA053964	NASH076	Steatosis
SA053965	NASH081	Steatosis
SA053966	NASH083	Steatosis
SA053967	NASH062	Steatosis
SA053968	NASH079	Steatosis
SA053969	NASH002	Steatosis
SA053970	NASH032	Steatosis
SA053971	NASH023	Steatosis
SA053972	NASH001	Steatosis
SA053973	NASH033	Steatosis
SA053974	NASH056	Steatosis
SA053975	NASH059	Steatosis
SA053976	NASH058	Steatosis
SA053977	NASH061	Steatosis

Showing results 1 to 88 of 88

Showing results 1 to 88 of 88

Collection:

Collection ID:	CO000948
Collection Summary:	Human samples were collected according to a protocol approved by Vanderbilt University Medical Center's Internal Review Board (#120829) and under informed written patients' consent prior to inclusion in this study. Sample sizes were selected to minimize the invasive procedures. Plasma samples were obtained from patients' blood collected during standard of care surgical procedures. Urine samples were collected from patients' Foley catheters placed for standard of care procedure. Liver samples were obtained from the excess tissue collected as part of the standard of care liver biopsies performed at the time of surgery that would otherwise be discarded. Subsequently, studies at University of California, San Diego were conducted under further auspices of University of California, San Diego Internal Review Board #121220.
Sample Type:	Plasma

Treatment:

Treatment ID:	TR000968
Treatment Summary:	-

Sample Preparation:

Sampleprep ID:	SP000961
Sampleprep Summary:	Plasma sample extraction. One hundred microliters of thawed plasma containing 4 µl of internal standard mix were extracted with 500 µl cold (-20°C) 70% CH3OH by incubation in an ice bath for 30 min. After mixing by vortex at 4°C for 1 min and centrifugation (4°C, 18,000 g, 10 min), the solvent was evaporated from the supernatant. The residue was dissolved in 200 µl of resuspension solvent, vortexed to mix (1 min at 4°C), and centrifuged (4°C, 18,000 g, 10 min) to remove any insoluble material. GPLs: GPLs from liver samples were extracted and analyzed by MS essentially as described in (20, 21). Extraction and analysis of plasma samples was according to previously published procedures (22). Cardiolipin, coenzyme Q, and dolichol: Lipid extractions were performed based on the Bligh and Dyer method with minor modifications (23-25). FAs and eicosanoids: FFAs were extracted essentially as previously described after supplementation with deuterated internal standards (Cayman Chemicals) (26, 27). Eicosanoids were isolated via solid phase extraction, utilizing 25 deuterated internal standards (28, 29). Sterols and oxysterols: Sterols and oxysterols were extracted using previously described methods (30). Neutral lipids: Cholesteryl esters (CEs), TAGs, and DAGs were extracted from weighed liver tissue (0.5-1 mg) suspended in 0.5 ml PBS that had been homogenized by sonication. Extractions of plasma (0.05 ml diluted to 0.1 ml with PBS), urine (1 ml), and tissue sonicates were carried out using 1 ml hexane:methyl t-butyl ether (1:1, v/v), essentially as previously described (31). Sphingolipids: Sphingolipids from liver, plasma, and urine were extracted following previously published procedures (32, 33), with the exception that methylene chloride was substituted for chloroform for the single-phase extraction of sphingoid bases. 20. Ivanova P. T., Milne S. B., Byrne M. O., Xiang Y., Brown H. A. 2007. Glycerophospholipid identification and quantitation by electrospray ionization mass spectrometry. Methods Enzymol. 432: 21-57. 21. Myers D. S., Ivanova P. T., Milne S. B., Brown H. A. 2011. Quantitative analysis of glycerophospholipids by LC-MS: acquisition, data handling, and interpretation. Biochim. Biophys. Acta. 1811: 748-757. 22. Quehenberger O., Armando A. M., Brown H. A., Milne S. B., Myers D. S., Merrill A. H., Jr, Bandyopadhyay S., Jones K. N., Kelly S., Shaner R. L., et al. 2010. Lipidomics reveals a remarkable diversity of lipids in human plasma. J. Lipid Res. 51: 3299-3305. 23. Guan Z., Li S., Smith D., Shaw W., Raetz C. 2007. Identification of N-acylphosphatidylserine molecules in eukaryotic cells. Biochemistry. 46: 14500-14513. 24. Tan B. K., Bogdanov M., Zhao J., Dowhan W., Raetz C. R., Guan Z. 2012. Discovery of cardiolipin synthase utilizing phosphatidylethanolamine and phosphatidylglycerol as substrates. Proc. Natl. Acad. Sci. USA. 109: 16504-16509. 25. Wen R., Lam B., Guan Z. 2013. Aberrant dolichol chain lengths as biomarkers for retinitis pigmentosa caused by impaired dolichol biosynthesis. J. Lipid Res. 54: 3516-3522. 26. Quehenberger O., Armando A., Dumlao D., Stephens D. L., Dennis E. A. 2008. Lipidomics analysis of essential fatty acids in macrophages. Prostaglandins Leukot. Essent. Fatty Acids. 79: 123-129. 27. Quehenberger O., Armando A. M., Dennis E. A. 2011. High sensitivity quantitative lipidomics analysis of fatty acids in biological samples by gas chromatography-mass spectrometry. Biochim. Biophys. Acta. 1811: 648-656. 28. Deems R., Buczynski M. W., Bowers-Gentry R., Harkewicz R., Dennis E. A. 2007. Detection and quantitation of eicosanoids via high performance liquid chromatography-electrospray ionization-mass spectrometry. Methods Enzymol. 432: 59-82. 29. Dumlao D. S., Buczynski M. W., Norris P. C., Harkewicz R., Dennis E. A. 2011. High-throughput lipidomic analysis of fatty acid derived eicosanoids and N-acylethanolamines. Biochim. Biophys. Acta. 1811: 724-736. 30. McDonald J. G., Smith D. D., Stiles A. R., Russell D. W. 2012. A comprehensive method for extraction and quantitative analysis of sterols and secosteroids from human plasma. J. Lipid Res. 53: 1399-1409. 31. Hutchins P. M., Barkley R. M., Murphy R. C. 2008. Separation of cellular nonpolar neutral lipids by normal-phase chromatography and analysis by electrospray ionization mass spectrometry. J. Lipid Res. 49: 804-813. 32. Shaner R. L., Allegood J. C., Park H., Wang E., Kelly S., Haynes C. A., Sullards M. C., Merrill A. H., Jr 2009. Quantitative analysis of sphingolipids for lipidomics using triple quadrupole and quadrupole linear ion trap mass spectrometers. J. Lipid Res. 50: 1692-1707. 33. Sullards M. C., Liu Y., Chen Y., Merrill A. H., Jr 2011. Analysis of mammalian sphingolipids by liquid chromatography tandem mass spectrometry (LC-MS/MS) and tissue imaging mass spectrometry (TIMS). Biochim. Biophys. Acta. 1811: 838-853.

Sampleprep ID:

SP000961

Sampleprep Summary:

Plasma sample extraction. One hundred microliters of thawed plasma containing 4 µl of internal standard mix were extracted with 500 µl cold (-20°C) 70% CH3OH by incubation in an ice bath for 30 min. After mixing by vortex at 4°C for 1 min and centrifugation (4°C, 18,000 g, 10 min), the solvent was evaporated from the supernatant. The residue was dissolved in 200 µl of resuspension solvent, vortexed to mix (1 min at 4°C), and centrifuged (4°C, 18,000 g, 10 min) to remove any insoluble material. GPLs: GPLs from liver samples were extracted and analyzed by MS essentially as described in (20, 21). Extraction and analysis of plasma samples was according to previously published procedures (22). Cardiolipin, coenzyme Q, and dolichol: Lipid extractions were performed based on the Bligh and Dyer method with minor modifications (23-25). FAs and eicosanoids: FFAs were extracted essentially as previously described after supplementation with deuterated internal standards (Cayman Chemicals) (26, 27). Eicosanoids were isolated via solid phase extraction, utilizing 25 deuterated internal standards (28, 29). Sterols and oxysterols: Sterols and oxysterols were extracted using previously described methods (30). Neutral lipids: Cholesteryl esters (CEs), TAGs, and DAGs were extracted from weighed liver tissue (0.5-1 mg) suspended in 0.5 ml PBS that had been homogenized by sonication. Extractions of plasma (0.05 ml diluted to 0.1 ml with PBS), urine (1 ml), and tissue sonicates were carried out using 1 ml hexane:methyl t-butyl ether (1:1, v/v), essentially as previously described (31). Sphingolipids: Sphingolipids from liver, plasma, and urine were extracted following previously published procedures (32, 33), with the exception that methylene chloride was substituted for chloroform for the single-phase extraction of sphingoid bases.
20. Ivanova P. T., Milne S. B., Byrne M. O., Xiang Y., Brown H. A. 2007. Glycerophospholipid identification and quantitation by electrospray ionization mass spectrometry. Methods Enzymol. 432: 21-57.
21. Myers D. S., Ivanova P. T., Milne S. B., Brown H. A. 2011. Quantitative analysis of glycerophospholipids by LC-MS: acquisition, data handling, and interpretation. Biochim. Biophys. Acta. 1811: 748-757.
22. Quehenberger O., Armando A. M., Brown H. A., Milne S. B., Myers D. S., Merrill A. H., Jr, Bandyopadhyay S., Jones K. N., Kelly S., Shaner R. L., et al. 2010. Lipidomics reveals a remarkable diversity of lipids in human plasma. J. Lipid Res. 51: 3299-3305.
23. Guan Z., Li S., Smith D., Shaw W., Raetz C. 2007. Identification of N-acylphosphatidylserine molecules in eukaryotic cells. Biochemistry. 46: 14500-14513.
24. Tan B. K., Bogdanov M., Zhao J., Dowhan W., Raetz C. R., Guan Z. 2012. Discovery of cardiolipin synthase utilizing phosphatidylethanolamine and phosphatidylglycerol as substrates. Proc. Natl. Acad. Sci. USA. 109: 16504-16509.
25. Wen R., Lam B., Guan Z. 2013. Aberrant dolichol chain lengths as biomarkers for retinitis pigmentosa caused by impaired dolichol biosynthesis. J. Lipid Res. 54: 3516-3522.
26. Quehenberger O., Armando A., Dumlao D., Stephens D. L., Dennis E. A. 2008. Lipidomics analysis of essential fatty acids in macrophages. Prostaglandins Leukot. Essent. Fatty Acids. 79: 123-129.
27. Quehenberger O., Armando A. M., Dennis E. A. 2011. High sensitivity quantitative lipidomics analysis of fatty acids in biological samples by gas chromatography-mass spectrometry. Biochim. Biophys. Acta. 1811: 648-656.
28. Deems R., Buczynski M. W., Bowers-Gentry R., Harkewicz R., Dennis E. A. 2007. Detection and quantitation of eicosanoids via high performance liquid chromatography-electrospray ionization-mass spectrometry. Methods Enzymol. 432: 59-82.
29. Dumlao D. S., Buczynski M. W., Norris P. C., Harkewicz R., Dennis E. A. 2011. High-throughput lipidomic analysis of fatty acid derived eicosanoids and N-acylethanolamines. Biochim. Biophys. Acta. 1811: 724-736.
30. McDonald J. G., Smith D. D., Stiles A. R., Russell D. W. 2012. A comprehensive method for extraction and quantitative analysis of sterols and secosteroids from human plasma. J. Lipid Res. 53: 1399-1409.
31. Hutchins P. M., Barkley R. M., Murphy R. C. 2008. Separation of cellular nonpolar neutral lipids by normal-phase chromatography and analysis by electrospray ionization mass spectrometry. J. Lipid Res. 49: 804-813.
32. Shaner R. L., Allegood J. C., Park H., Wang E., Kelly S., Haynes C. A., Sullards M. C., Merrill A. H., Jr 2009. Quantitative analysis of sphingolipids for lipidomics using triple quadrupole and quadrupole linear ion trap mass spectrometers. J. Lipid Res. 50: 1692-1707.
33. Sullards M. C., Liu Y., Chen Y., Merrill A. H., Jr 2011. Analysis of mammalian sphingolipids by liquid chromatography tandem mass spectrometry (LC-MS/MS) and tissue imaging mass spectrometry (TIMS). Biochim. Biophys. Acta. 1811: 838-853.

Combined analysis:

Analysis ID	AN001491	AN001492	AN001493	AN001494	AN001495	AN001496
Analysis type	MS	MS	MS	MS	MS	MS
Chromatography type	Unspecified	Unspecified	Unspecified	Unspecified	Unspecified	Unspecified
Chromatography system	Multiple	Multiple	Multiple	Multiple	Multiple	Multiple
Column	Multiple	Multiple	Multiple	Multiple	Multiple	Multiple
MS Type	Other	Other	Other	Other	Other	Other
MS instrument type	-	-	-	-	-	-
MS instrument name	-	-	-	-	-	-
Ion Mode	UNSPECIFIED	UNSPECIFIED	UNSPECIFIED	UNSPECIFIED	UNSPECIFIED	UNSPECIFIED
Units	pmol/ml	nmol/ml	pmol/ml	nmol/ml	pmol/ml	pmol/ml

Chromatography:

Chromatography ID:	CH001048
Chromatography Summary:	FFAs were analyzed by stable isotope dilution GC-MS after derivatization, essentially as described previously (26, 27). This method quantifies 33 FAs including all major and minor saturated FAs, monounsaturated FAs, and PUFAs containing 12 to 26 carbons. Eicosanoids were analyzed by a stable isotope dilution LC/MS method utilizing 26 deuterated internal standards (28, 29). The metabolites were quantified after separation by reverse phase chromatography on a 2.1 × 100 mm BEH Shield column, 1.7 µM (Waters, Milford, MA) employing an Acquity UPLC system (Waters). Detection and quantification were performed on an AB SCIEX 6500 QTrap mass spectrometer equipped with an IonDrive Turbo V source (AB SCIEX, Framingham, MA), operated in negative ionization mode via MRM, using standard curves generated from 145 authentic quantification standards (34). The method analyzes an additional 13 metabolites based on authentic primary standards, but which cannot be quantified due to the lack of appropriate internal standards. Data analysis was performed using MultiQuant 2.1 software (AB SCIEX). 26. Quehenberger O., Armando A., Dumlao D., Stephens D. L., Dennis E. A. 2008. Lipidomics analysis of essential fatty acids in macrophages. Prostaglandins Leukot. Essent. Fatty Acids. 79: 123-129. 27. Quehenberger O., Armando A. M., Dennis E. A. 2011. High sensitivity quantitative lipidomics analysis of fatty acids in biological samples by gas chromatography-mass spectrometry. Biochim. Biophys. Acta. 1811: 648-656.
Instrument Name:	Multiple
Column Name:	Multiple
Chromatography Type:	Unspecified

Chromatography ID:	CH001049
Chromatography Summary:	Sterols and oxysterols were measured using methods previously described (30). Plasma total cholesterol was measured using a Vitros 250 chemistry system (Ortho-Clinical Diagnostics, Rochester, NY). Plasma free cholesterol and liver free and total cholesterol were measured using methods adapted from (30). 30. McDonald J. G., Smith D. D., Stiles A. R., Russell D. W. 2012. A comprehensive method for extraction and quantitative analysis of sterols and secosteroids from human plasma. J. Lipid Res. 53: 1399-1409.
Instrument Name:	Multiple
Column Name:	Multiple
Chromatography Type:	Unspecified

Chromatography ID:	CH001050
Chromatography Summary:	Cardiolipin analysis was achieved with normal phase LC coupled with high-resolution MS performed on a TripleTOF 5600 system (AB SCIEX, Foster City, CA) (24). Dolichol and coenzyme Q were analyzed by reverse phase LC coupled with multiple reaction monitoring (MRM) MS utilizing a 4000 Q-Trap hybrid triple quadrupole linear ion-trap mass spectrometer (AB SCIEX) (22, 23). For dolichol analysis, MRM was performed in negative ion mode, with the precursor ions being the (M+acetate)- adduct ions and the product ions being the acetate ions (m/z 59). For coenzyme Q analysis, MRM was carried out in positive ion mode, with ammonium adducts (M+NH4)+ as precursor ions and the proton adducts of the quinone ring of coenzyme Q (m/z 197) as product ions. For quantitation, an internal standard mixture composed of a cardiolipin mix (Avanti Polar Lipids, Inc.), nor-dolichol (13-22) (Avanti Polar Lipids, Inc.), and yeast coenzyme Q6 (Sigma) was added during the first step of lipid extraction (22). 22. Quehenberger O., Armando A. M., Brown H. A., Milne S. B., Myers D. S., Merrill A. H., Jr, Bandyopadhyay S., Jones K. N., Kelly S., Shaner R. L., et al. 2010. Lipidomics reveals a remarkable diversity of lipids in human plasma. J. Lipid Res. 51: 3299-3305. 23. Guan Z., Li S., Smith D., Shaw W., Raetz C. 2007. Identification of N-acylphosphatidylserine molecules in eukaryotic cells. Biochemistry. 46: 14500-14513. 24. Tan B. K., Bogdanov M., Zhao J., Dowhan W., Raetz C. R., Guan Z. 2012. Discovery of cardiolipin synthase utilizing phosphatidylethanolamine and phosphatidylglycerol as substrates. Proc. Natl. Acad. Sci. USA. 109: 16504-16509.
Instrument Name:	Multiple
Column Name:	Multiple
Chromatography Type:	Unspecified

Chromatography ID:	CH001051
Chromatography Summary:	The organic solvent extraction layer containing CEs, TAGs, and DAGs was taken to dryness, then derivatized with 2,5-difluorophenylisocyanate to convert DAGs to urethane derivatives (35). The derivatized extract was separated by normal phase LC as previously described (35). The CEs eluting first from the LC column were detected by 20 specific MRM transitions corresponding to each [M+NH4]+ ion being collisionally activated to m/z 369.3. During elution of TAGs, the mass spectrometer was set to carry out full mass scanning from m/z 400-1,000. The DAGs were detected by neutral loss scanning of 190 Da. 35. Leiker T. J., Barkley R. M., Murphy R. C. 2011. Analysis of diacylglycerol molecular species in cellular lipid extracts by normal-phase LC-electrospray mass spectrometry. Int. J. Mass Spectrom. 305: 103-109
Instrument Name:	Multiple
Column Name:	Multiple
Chromatography Type:	Unspecified

Chromatography ID:	CH001052
Chromatography Summary:	Sphingolipids were analyzed by LC-MS/MS essentially as described in (32, 33) with minor modifications to include the 1-deoxy-sphingolipids as generally described in (36). 32. Shaner R. L., Allegood J. C., Park H., Wang E., Kelly S., Haynes C. A., Sullards M. C., Merrill A. H., Jr 2009. Quantitative analysis of sphingolipids for lipidomics using triple quadrupole and quadrupole linear ion trap mass spectrometers. J. Lipid Res. 50: 1692-1707. 33. Sullards M. C., Liu Y., Chen Y., Merrill A. H., Jr 2011. Analysis of mammalian sphingolipids by liquid chromatography tandem mass spectrometry (LC-MS/MS) and tissue imaging mass spectrometry (TIMS). Biochim. Biophys. Acta. 1811: 838-853. 36. Zitomer N. C., Mitchell T., Voss K. A., Bondy G. S., Pruett S. T., Garnier-Amblard E. C., Liebeskind L. S., Park H., Wang E., Sullards M. C., et al. 2009. Ceramide synthase inhibition by fumonisin B1 causes accumulation of 1-deoxysphinganine: a novel category of bioactive 1-deoxysphingoid bases and 1-deoxydihydroceramides biosynthesized by mammalian cell lines and animals. J. Biol. Chem. 284: 4786-4795.
Instrument Name:	Multiple
Column Name:	Multiple
Chromatography Type:	Unspecified

Chromatography ID:	CH001053
Chromatography Summary:	Extracted GPLs from liver and plasma were analyzed by MS, as described elsewhere (20, 21). 20. Ivanova P. T., Milne S. B., Byrne M. O., Xiang Y., Brown H. A. 2007. Glycerophospholipid identification and quantitation by electrospray ionization mass spectrometry. Methods Enzymol. 432: 21-57. 21. Myers D. S., Ivanova P. T., Milne S. B., Brown H. A. 2011. Quantitative analysis of glycerophospholipids by LC-MS: acquisition, data handling, and interpretation. Biochim. Biophys. Acta. 1811: 748-757.
Instrument Name:	Multiple
Column Name:	Multiple
Chromatography Type:	Unspecified

MS:

MS ID:	MS001374
Analysis ID:	AN001491
Instrument Name:	-
Instrument Type:	-
MS Type:	Other
Ion Mode:	UNSPECIFIED

MS ID:	MS001375
Analysis ID:	AN001492
Instrument Name:	-
Instrument Type:	-
MS Type:	Other
Ion Mode:	UNSPECIFIED

MS ID:	MS001376
Analysis ID:	AN001493
Instrument Name:	-
Instrument Type:	-
MS Type:	Other
Ion Mode:	UNSPECIFIED

MS ID:	MS001377
Analysis ID:	AN001494
Instrument Name:	-
Instrument Type:	-
MS Type:	Other
Ion Mode:	UNSPECIFIED

MS ID:	MS001378
Analysis ID:	AN001495
Instrument Name:	-
Instrument Type:	-
MS Type:	Other
Ion Mode:	UNSPECIFIED

MS ID:	MS001379
Analysis ID:	AN001496
Instrument Name:	-
Instrument Type:	-
MS Type:	Other
Ion Mode:	UNSPECIFIED