Determination of glycerol turnover by [1,1,2,3,3-2H5]glycerol in humans: evaluation of a new derivative for mass spectrometry analysis

Schricker, Th.; Albuszies, Gerd; Kugler, Benjamin A.; Wagner, Deborah; Georgieff, Michael

doi:10.1016/0261-5614(93)90213-n

Cited by 3 publications

(3 citation statements)

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“…Breath 13 CO 2 isotopic enrichment was determined by isotope ratio MS on a Tracermass C/N (SerCon Limited, Crewe, Cheshire, UK). Isotopic enrichments of plasma 6,6- 2 H 2 -glucose and 2 H 5 -glycerol were measured by GC–MS (Agilent Technologies) as described previously ( 14 , 15 ) . For the analysis of plasma 13 C glucose, plasma glucose was purified by sequential anion/cation exchange chromatography using resins (AG 50 W-X8 and AG 1-X8; Bio-Rad, Richmond, CA, USA) followed by HPLC ( 16 ) .…”

Section: Methodsmentioning

confidence: 99%

Sex differences in lipid and glucose kinetics after ingestion of an acute oral fructose load

et al. 2010

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The increase in VLDL TAG concentration after ingestion of a high-fructose diet is more pronounced in men than in pre-menopausal women. We hypothesised that this may be due to a lower fructose-induced stimulation of de novo lipogenesis (DNL) in pre-menopausal women. To evaluate this hypothesis, nine healthy male and nine healthy female subjects were studied after ingestion of oral loads of fructose enriched with 13 C 6 fructose. Incorporation of 13 C into breath CO 2 , plasma glucose and plasma VLDL palmitate was monitored to evaluate total fructose oxidation, gluconeogenesis and hepatic DNL, respectively. Substrate oxidation was assessed by indirect calorimetry. After 13 C fructose ingestion, 44·0 (SD 3·2) % of labelled carbons were recovered in plasma glucose in males v. 41·9 (SD 2·3) % in females (NS), and 42·9 (SD 3·7) % of labelled carbons were recovered in breath CO 2 in males v. 43·0 (SD 4·5) % in females (NS), indicating similar gluconeogenesis from fructose and total fructose oxidation in males and females. The area under the curve for 13 C VLDL palmitate tracer-to-tracee ratio was four times lower in females (P,0·05), indicating a lower DNL. Furthermore, lipid oxidation was significantly suppressed in males (by 16·4 (SD 5·2), P, 0·05), but it was not suppressed in females (2 1·3 (SD 4·7) %). These results support the hypothesis that females may be protected against fructose-induced hypertriglyceridaemia because of a lower stimulation of DNL and a lower suppression of lipid oxidation. De novo lipogenesis: VLDL TAG: Endogenous glucose productionThe metabolic effects of a high-fructose diet have been widely studied in both animals and human subjects (1) . High-fructose diet is associated with the development of at least two features of the metabolic syndrome, i.e. insulin resistance and increased plasma VLDL TAG. The latter may in turn be associated with an increased production of small, dense LDL particle with high atherogenic potential (2,3) . It has been reported by several authors that a high-fructose diet increases plasma TAG less and does not reduce insulin sensitivity in both pre-menopausal women and female rodents compared with males (4 -11) . This may be due to a sex-specific difference in either fructose metabolism or adaptations to chronic fructose overfeeding. To our knowledge, whether the metabolism of an acute fructose load differs in females and males has not been specifically assessed. Fructose disposal relies mainly on the stimulation of carbohydrate oxidation (fructose oxidation in splanchnic organs and indirect oxidation of glucose and lactate synthesised from the fructose), gluconeogenesis, hepatic glycogen storage and hepatic de novo lipogenesis (DNL). The latter, although a quantitatively minor pathway for fructose disposal, may be closely linked to hepatic VLDL TAG production and synthesis of intra-hepatic lipids. Therefore, we hypothesised that fructose-induced stimulation of DNL may be blunted in pre-menopausal women.To evaluate this hypothesis, we compared the metabolic fate of i...

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Section: Methodsmentioning

confidence: 99%

Sex differences in lipid and glucose kinetics after ingestion of an acute oral fructose load

et al. 2010

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“…Several software tools for automated analysis of GC-EI MS data are available, − including tools for stable isotope-labeling data analysis, − which must address the inherent difficulty of deconvoluting the intensity of overlapping isotopologues from EI mass spectral fragmentation. Interestingly, the use of GC-CI MS for stable isotope tracing is largely unexplored, − possibly prompted by suboptimal configurations that favored CI source contamination, lower sensitivity, and reproducibility in comparison with GC-EI MS. Here, we have studied the suitability of GC-CI-MS for stable isotope tracing using multiple analytical configurations based on low-resolution (LRMS) and high-resolution (HRMS) mass spectrometry.…”

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confidence: 91%

Exploring the Use of Gas Chromatography Coupled to Chemical Ionization Mass Spectrometry (GC-CI-MS) for Stable Isotope Labeling in Metabolomics

et al. 2020

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Isotopic-labeling experiments have been valuable to monitor the flux of metabolic reactions in biological systems, which is crucial to understand homeostatic alterations with disease. Experimental determination of metabolic fluxes can be inferred from a characteristic rearrangement of stable isotope tracers (e.g., 13C or 15N) that can be detected by mass spectrometry (MS). Metabolites measured are generally members of well-known metabolic pathways, and most of them can be detected using both gas chromatography (GC)-MS and liquid chromatography (LC)-MS. In here, we show that GC methods coupled to chemical ionization (CI) MS have a clear advantage over alternative methodologies due to GC’s superior chromatography separation efficiency and the fact that CI is a soft ionization technique that yields identifiable protonated molecular ion peaks. We tested diverse GC-CI-MS setups, including methane and isobutane reagent gases, triple quadrupole (QqQ) MS in SIM mode, or selected ion clusters using optimized narrow windows (∼10 Da) in scan mode, and standard full scan methods using high resolution GC-(q)TOF and GC-Orbitrap systems. Isobutane as a reagent gas in combination with both low-resolution (LR) and high-resolution (HR) MS showed the best performance, enabling precise detection of isotopologues in most metabolic intermediates of central carbon metabolism. Finally, with the aim of overcoming manual operations, we developed an R-based tool called isoSCAN that automatically quantifies all isotopologues of intermediate metabolites of glycolysis, TCA cycle, amino acids, pentose phosphate pathway, and urea cycle, from LRMS and HRMS data.

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“…[ 2 H]glycerol enrichment and concentration and [ 2 H]glucose enrichment were measured simultaneously: 2á63 nmol of [1,2,[3][4][5][6][7][8][9][10][11][12][13] C]glycerol as internal standard was added to 500 ll plasma or 20 ll dialysate samples. After evaporation to dryness, the samples were acetylated to glucose pentaacetylated and glycerol triacetylated and analysed in chemical ionization mode with methane as reagent gas and with selective monitoring of m/z 159, 162 and 164 for glycerol derivatives (Schrieker et al, 1994), and of m/z 331 and 333 for glucose derivatives (Wolfe, 1992). Glycerol enrichment was calculated from the ratio of signals at m/z 164 and m/z 159, while glycerol concentration was calculated from the ratio of signals at m/z 159 and m/ z 162.…”

Section: Mass Spectrometry Analysismentioning

confidence: 99%

Effects of a sympathetic activation by lower body negative pressure on glucose and lipid metabolism

Henry

Trueb

Sartori³

et al. 1998

Clinical Physiology

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The effects of a sympathetic activation elicited by a lower body negative pressure (LBNP) (at -15 mmHg for 75 min) were assessed in 7 healthy subjects on two occasions: (i) in post-absorptive conditions, and (ii) during glucose infusion (22.2 mumol kg-1 min-1). LBNP increased plasma norepinephrine concentration and heart rate. It did not alter whole-body glucose metabolism (measured with [6,6-2H]glucose) and glycerol turnover (measured with [1,1,2,3,3-2H]glycerol). Interstitial glycerol concentrations were monitored with microdialysis in subcutaneous adipose tissue and in skeletal muscle. LBNP increased dialysate glycerol concentrations in muscle by 16% (P < 0.03) but not in adipose tissue in post-absorptive conditions, and by 37% in adipose tissue (P < 0.05) but not in muscle during glucose infusion. These results indicate that an LBNP-induced sympathetic activation (i) does not increase endogenous glucose production, and (ii) induces only a slight stimulation of lipolysis in adipose tissue during glucose infusion.

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Determination of glycerol turnover by [1,1,2,3,3-2H5]glycerol in humans: evaluation of a new derivative for mass spectrometry analysis

Cited by 3 publications

References 0 publications

Sex differences in lipid and glucose kinetics after ingestion of an acute oral fructose load

Sex differences in lipid and glucose kinetics after ingestion of an acute oral fructose load

Exploring the Use of Gas Chromatography Coupled to Chemical Ionization Mass Spectrometry (GC-CI-MS) for Stable Isotope Labeling in Metabolomics

Effects of a sympathetic activation by lower body negative pressure on glucose and lipid metabolism

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