A rapid GC–MS method for quantification of positional and geometric isomers of fatty acid methyl esters

Ecker, Josef; Scherer, Max; Schmitz, Gerd; Liebisch, Gerhard

doi:10.1016/j.jchromb.2012.04.015

Cited by 153 publications

(119 citation statements)

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“…Treated mouse hepatocytes were taken up into 200 ml of PBS and lysed by freeze-thawing. After centrifugation, lysates were Lipoprotection by UDCA-LPE at ASPET Journals on May 10, 2018 molpharm.aspetjournals.org subjected to fatty acid analyses by gas chromatography coupled to mass spectrometry (GC-MS) as previously described (Ecker et al, 2012), and quantification of phospholipids by electrospray ionization tandem mass spectrometry (ESI-MS/MS) in positive mode was done using the setup described previously (Liebisch et al, 2004). …”

Section: Methodsmentioning

confidence: 99%

Ursodeoxycholyl Lysophosphatidylethanolamide Inhibits Lipoapoptosis by Shifting Fatty Acid Pools toward Monosaturated and Polyunsaturated Fatty Acids in Mouse Hepatocytes

Chamulitrat

Liebisch

et al. 2013

Mol Pharmacol

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Ursodeoxycholyl lysophosphatidylethanolamide (UDCA-LPE) is a hepatoprotectant in inhibiting apoptosis, inflammation, and hyperlipidemia in mouse models of nonalcoholic steatohepatitis (NASH). We studied the ability of UDCA-LPE to inhibit palmitate (Pal)-induced apoptosis in primary hepatocytes and delineate cytoprotective mechanisms. We showed that lipoprotection by UDCA-LPE was mediated by cAMP and was associated with increases in triglycerides (TGs) and phospholipids (PLs). An inhibitor of cAMP-effector protein kinase A partially reversed the protective effects of UDCA-LPE. Lipidomic analyses of fatty acids and PL composition revealed a shift of lipid metabolism from saturated Pal to monounsaturated and polyunsaturated fatty acids, mainly, oleate, docosapentaenoate, and docosahexaenoate. The latter two v-3 fatty acids were particularly found in phosphatidylcholine and phosphatidylserine pools. The catalysis of Pal by stearoylCoA desaturase-1 (SCD-1) is a known mechanism for the channeling of Pal away from apoptosis. SCD-1 protein was upregulated during UDCA-LPE lipoprotection. SCD-1 knockdown of Pal-treated cells showed further increased apoptosis, and the extent of UDCA-LPE protection was reduced. Thus, the major mechanism of UDCA-LPE lipoprotection involved a metabolic shift from toxic saturated toward cytoprotective unsaturated fatty acids in part via SCD-1. UDCA-LPE may thus be a therapeutic agent for treatment of NASH by altering distinct pools of fatty acids for storage into TGs and PLs, and the latter may protect lipotoxicity at the membrane levels.

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Section: Methodsmentioning

confidence: 99%

Ursodeoxycholyl Lysophosphatidylethanolamide Inhibits Lipoapoptosis by Shifting Fatty Acid Pools toward Monosaturated and Polyunsaturated Fatty Acids in Mouse Hepatocytes

Chamulitrat

Liebisch

et al. 2013

Mol Pharmacol

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“…For this reason, we used the method described by Ecker et al [30] with minor modifications to determine plasma fatty acids as methyl esters. Using a GC single quadrupole instrument (Shimadzu GCMS QP2010 Ultra) and a BPX90 column (Trajan Scientific), 48 fatty acids could be determined in plasma.…”

Section: Targeted Gc-ms Analysis Of Fatty Acids In Plasmamentioning

confidence: 99%

Metabolite profiles evaluated, according to sex, do not predict resting energy expenditure and lean body mass in healthy non-obese subjects

et al. 2018

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Purpose Differences in resting energy expenditure (REE) between men and women mainly result from sex-related differences in lean body mass (LBM). So far, a little is known about whether REE and LBM are reflected by a distinct human metabolite profile. Therefore, we aimed to identify plasma and urine metabolite patterns that are associated with REE and LBM of healthy subjects. Methods We investigated 301 healthy male and female subjects (18-80 years) under standardized conditions in the crosssectional KarMeN (Karlsruhe Metabolomics and Nutrition) study. REE was determined by indirect calorimetry and LBM by dual X-ray absorptiometry. Fasting blood and 24 h urine samples were analyzed by targeted and non-targeted metabolomics methods using GC × GC-MS, GC-MS, LC-MS, and NMR. Data were evaluated by predictive modeling of combined data using different machine learning algorithms, namely SVM, glmnet, and PLS. Results When evaluating data of men and women combined, we were able to predict REE and LBM with high accuracy (> 90%). This, however, was a clear effect of sex, which is supported by the high degree of overlap in identified important metabolites for LBM, REE, and sex, respectively. The applied machine learning algorithms did not reveal a metabolite pattern predictive of REE or LBM, when analyzing data for men and women, separately. Conclusions We could not identify a sex independent predictive metabolite pattern for REE or LBM. REE and LBM have no impact on plasma and urine metabolite profiles in the KarMeN Study participants. Studies applying metabolomics in healthy humans need to consider sex specific data evaluation.

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“…In the majority of cases, the solvents used for the extraction of fatty acids from biological samples can be either hexane (5,6,8,11,13,18,21,22) or CHCl3/methanol (7,17,(18)(19)(20). Before the derivatization of FA to their methyl esters (FAME), a saponification can be done, which separates FA from other lipids.…”

Section: Sample Preparation -Extraction Of Fa and Derivatizationmentioning

confidence: 99%

“…However, acid catalyzed reactions are used and H2SO4 has also been considered as an option for the transesterification of FA to FAME (5,19). The research group of Schmitz (21,22) have used an unusual derivatization agent, which is acetyl-chloride, the derivatization being done at 20°C, at 150 rpm, overnight. At the same time, for FA dosing, C17:0 can be added to the sample solution as an internal standard, before the derivatization process.…”

Section: Sample Preparation -Extraction Of Fa and Derivatizationmentioning

confidence: 99%

“…The methods described in the literature for the separation and/or quantification of fatty acids in different samples are mostly gas-chromatographic (1)(2)(3)(5)(6)(7)(8)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22), among which most developed methods use a mass-spectrometer detector. However, liquid-chromatography (4,17,(23)(24)(25)(26)(27)(28)(29)(30)(31) has been presented by several authors and also, there are few articles which deal with other techniques such as capillary electrophoresis or infrared (IR) and Raman spectroscopy (16,31,32).…”

Section: Introductionmentioning

confidence: 99%

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Recent developments in fatty acids profile determination in biological samples - a review

Tiuca

Nagy

Oprean

2015

Revista Romana De Medicina De Laborator

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A rapid GC–MS method for quantification of positional and geometric isomers of fatty acid methyl esters

Cited by 153 publications

References 20 publications

Ursodeoxycholyl Lysophosphatidylethanolamide Inhibits Lipoapoptosis by Shifting Fatty Acid Pools toward Monosaturated and Polyunsaturated Fatty Acids in Mouse Hepatocytes

Ursodeoxycholyl Lysophosphatidylethanolamide Inhibits Lipoapoptosis by Shifting Fatty Acid Pools toward Monosaturated and Polyunsaturated Fatty Acids in Mouse Hepatocytes

Metabolite profiles evaluated, according to sex, do not predict resting energy expenditure and lean body mass in healthy non-obese subjects

Recent developments in fatty acids profile determination in biological samples - a review

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