High sensitivity tracer detection using high-precision gas chromatography-combustion isotope ratio mass spectrometry and highly enriched uniformly carbon-13 labeled precursors

Goodman, Keith J.; Brenna, J. Thomas

doi:10.1021/ac00034a004

Cited by 156 publications

(90 citation statements)

References 27 publications

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“…Tracer analysis for 13 C-DHA was performed on the FAME mixtures using similar GC column conditions as for the quantitative analysis, as has been described in detail previously ( 27 ). Instrumentation for tracer analysis was an Agilent 6890 gas chromatograph coupled to a combustion furnace interface, and to a Thermo Scientifi c 253 isotope ratio mass spectrometer.…”

Section: C-dha Analysismentioning

confidence: 99%

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Higher efficacy of dietary DHA provided as a phospholipid than as a triglyceride for brain DHA accretion in neonatal piglets

Liu

Bartke

Daele

et al. 2014

Journal of Lipid Research

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Section: C-dha Analysismentioning

confidence: 99%

“…Data processing was as described previously ( 21 ). Isotope ratios in the conventional high precision notation, ␦ 13 C [defi ned previously ( 27 )], were converted to fraction of 13 C. For each FA, the mean by guest, on May 11, 2018 www.jlr.org…”

Section: C-dha Analysismentioning

confidence: 99%

Higher efficacy of dietary DHA provided as a phospholipid than as a triglyceride for brain DHA accretion in neonatal piglets

Liu

Bartke

Daele

et al. 2014

Journal of Lipid Research

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“…Gas chromatography-combustion-isotope ratio mass spectrometry was used to analyze 13 C-fatty acids ( 17 ). After 48 h, mice were euthanized by CO 2 inhalation.…”

Section: In Vivo D6d Activity Analysismentioning

confidence: 99%

Disruption of FADS2 gene in mice impairs male reproduction and causes dermal and intestinal ulceration

Stroud

Nara

Roqueta‐Rivera

et al. 2009

Journal of Lipid Research

158

135

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This article is available online at http://www.jlr.org identifi ed as essential fatty acids that must be consumed in the diet. Once consumed, however, LA and ALA can both be desaturated and elongated into more highly unsaturated fatty acids (HUFA) such as arachidonic (AA), docosapentaenoic (DPA n-6), and docosahexaenoic (DHA) acids via the pathway shown in Fig. 1A . Delta-6 desaturase (D6D) performs the fi rst and rate-limiting step in this process, as well as the last step of desaturation for DHA and DPA n-6 synthesis. The D6D gene FADS2 was cloned in 1999 ( 2 ), and subsequently, a human case of D6D deficiency was identifi ed ( 3 ). The patient exhibited growth retardation accompanied by skin abnormalities, corneal ulceration, and feeding intolerance. Treatment with dietary AA and DHA restored normal growth and eliminated most other symptoms, underscoring the importance of the endogenous synthesis of these HUFAs.AA is a precursor to a host of signaling molecules known as eicosanoids, which include thromboxanes, leukotrienes, prostacyclins, and prostaglandins produced from the oxygenation of AA by cyclooxygenase and lipoxygenase enzymes. However, the symptoms of classic essential fatty acid defi ciency, growth retardation and dermatitis ( 1 ), are attributed to a loss of LA, not AA or eicosanoids. Because LA is an essential component of skin ceramides, LA defi ciency results in the disruption of the skin's water barrier function ( 4 ) and heat loss from skin ( 5 ). These side effects make investigation of AA defi ciency impossible by dietary manipulation without complications from LA defi ciency.DHA is found in large amounts in the retina, brain, and testes ( 6, 7 ). The role of DHA has been largely thought to be structural, increasing the fl uidity of cellular memAbstract Delta-6 desaturase (D6D) catalyzes the fi rst step in the synthesis of highly unsaturated fatty acids (HUFA) such as arachidonic (AA), docosapentaenoic (DPAn-6), and docosahexaenoic (DHA) acids, as well as the last desaturation of DPAn-6 and DHA. We created D6D-null mice ( ؊ / ؊ ), which enabled us to study HUFA defi ciency without depleting their precursors. In ؊ / ؊ , no in vivo AA synthesis was detected after administration of [U-

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“…5). By simultaneously opening both valves, helium is added such that flow through the reactor is reversed and GC column effluent is vented to the atmosphere [25,73]. Such a system is commonly called "backflushing," and is operated under computer control.…”

Section: Solvent Diversionmentioning

confidence: 99%

Isotope‐ratio detection for gas chromatography

Sessions¹

2006

J of Separation Science

244

270

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Isotope-ratio detection for gas chromatographyInstrumentation and methods exist for highly precise analyses of the stable-isotopic composition of organic compounds separated by GC. The general approach combines a conventional GC, a chemical reaction interface, and a specialized isotoperatio mass spectrometer (IRMS). Most existing GC hardware and methods are amenable to isotope-ratio detection. The interface continuously and quantitatively converts all organic matter, including column bleed, to a common molecular form for isotopic measurement. C and N are analyzed as CO 2 and N 2 , respectively, derived from combustion of analytes. H and O are analyzed as H 2 and CO produced by pyrolysis/reduction. IRMS instruments are optimized to provide intense, highly stable ion beams, with extremely high precision realized via a system of differential measurements in which ion currents for all major isotopologs are simultaneously monitored. Calibration to an internationally recognized scale is achieved through comparison of closely spaced sample and standard peaks. Such systems are capable of measuring 13 C/ 12 C ratios with a precision approaching 0.1? (for values reported in the standard delta notation), four orders of magnitude better than that typically achieved by conventional "organic" mass spectrometers. Detection limits to achieve this level of precision are typically a1 nmol C (roughly 10 ng of a typical hydrocarbon) injected on-column. Achievable precision and detection limits are correspondingly higher for N, O, and H, in that order.

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High sensitivity tracer detection using high-precision gas chromatography-combustion isotope ratio mass spectrometry and highly enriched uniformly carbon-13 labeled precursors

Cited by 156 publications

References 27 publications

Higher efficacy of dietary DHA provided as a phospholipid than as a triglyceride for brain DHA accretion in neonatal piglets

Higher efficacy of dietary DHA provided as a phospholipid than as a triglyceride for brain DHA accretion in neonatal piglets

Disruption of FADS2 gene in mice impairs male reproduction and causes dermal and intestinal ulceration

Isotope‐ratio detection for gas chromatography

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