An optimized analytical method for cellular targeted quantification of primary metabolites in tricarboxylic acid cycle and glycolysis using gas chromatography-tandem mass spectrometry and its application in three kinds of hepatic cell lines

Acetate, 3-hydroxybutyrate, pyruvate, lactate, citrate, 2-oxoglutarate, succinate, fumarate and malate were analysed in rat bile by gas chromatography and gas chromatography/mass spectrometry of their O-melthyloxime-t-butyldimethylsilyl derivatives. The concentration of acetate increased to about 1.8 mmol/l after administration of [2,2,2-2H3]ethanol. Acetate was formed from ethanol to an extent of about 82% and retained all of the 2H at C-2, whereas 15% of the 2H had been lost in the tricarboxylic acid cycle intermediates and 24% in 3-hydroxybutyrate. Thus the exchange of 2H for 1H takes place after formation of acetyl CoA. For citrate and 3-hydroxybutyrate, 41% and 11% respectively was formed from [2,2,2-2H3]ethanol. These results indicate that different pools of acetyl CoA are used for the synthesis of ketone bodies and citrate, with the latter being derived from ethanol to a much larger extent. Smaller fractions of 2-oxoglutarate (16%) and succinate (5%) were derived from [2,2,2--2H3]ethanol, indicating significant contributions from amino acids.

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Analysis of Metabolites from the Tricarboxylic Acid Cycle for Yeast and Bacteria Samples Using Gas Chromatography Mass Spectrometry

Methods in Molecular Biology - Clinical Metabolomics ◽

10.1007/978-1-4939-7592-1_20 ◽

2018 ◽

pp. 277-282

Author(s):

Reza Maleki Seifar ◽

Angela ten Pierick ◽

Patricia T. N. van Dam

Keyword(s):

Mass Spectrometry ◽

Gas Chromatography ◽

Tricarboxylic Acid Cycle ◽

Tricarboxylic Acid ◽

Gas Chromatography Mass Spectrometry ◽

Acid Cycle ◽

Chromatography Mass Spectrometry

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ANALYsis oF RYRRoLIDiNopHENoNE DERivATivEs iN BioLoGiOAL FLuiDs

Aspirantskiy Vestnik Povolzhiya ◽

10.17816/2072-2354.2019.19.1.33-40 ◽

2019 ◽

Vol 19 (1-2) ◽

pp. 33-40 ◽

Cited By ~ 1

Author(s):

IV. V Synbulatov ◽

A. V Voronin ◽

T. V Voronina

Keyword(s):

Mass Spectrometry ◽

Gas Chromatography ◽

Tandem Mass Spectrometry ◽

High Performance ◽

Biological Fluids ◽

Gas Chromatography Mass Spectrometry ◽

Tandem Mass ◽

Primary Metabolites ◽

Chromatography Tandem Mass Spectrometry ◽

Chromatography Mass Spectrometry

Pyrrolidinophenone derivatives are the group of narcotic drugs controlled in the Russian Federation. The review presents the trends of biotransformation of а-pyrrolidinovalerophenone and 3,4-methylenedioxypyrovalerone, the data about their primary metabolites is provided. Various techniques of the sample preparation of biological fluids for analytical toxicology studies for substances of the pyrrolidinophenone derivative group are discussed. The use of enzymatic hydrolysis followed by solid-phase extraction (sorption) provides low detection limits for native sub- PHARMACY ФАРМАЦИЯ stances of this group and primary metabolites using small volumes of biological fluids (0.5 and 1.0 ng/ml blood for a-pyrrolidinovalerophenone and 3,4-methylenedioxypirovalonone respectively). The main characteristics of pyrro-lidinophenone derivatives (Kovac’s retention indices in nonpolar stationary liquid phases and the main characteristic ions in the mass spectra of electron impact) are presented. They allow to identify pyrrolidinophenone derivatives and their primary metabolites in biological fluids during chromatographic-mass spectrometric screening. Analytical possibilities of an alternative variant of screening for biological fluids i.e. analysis by using current immunochemical test systems, including “biochips” are discussed. The main methods of reliable identification and quantitative determination of pyrrolidinophenone derivatives are chromatography-mass spectrometry and high-performance liquid chromatography-tandem mass spectrometry. The detection limit of 3,4-methylenedioxypyralovalone in blood by gas chromatography-mass spectrometry is 1.0 ng/ml. The ranges of the determined concentrations of the method of quantitation by gas chromatography-mass spectrometry are 2.0-2000.0 ng/ml for blood and 0.05-50.0 ng/10 mm for hair. The high-performance chromatography-tandem mass spectrometry method with a triple quadrupole in the monitoring mode of multiple molecular reactions makes it possible to achieve a nearly complete suppression of analytical background “noise” for a sample, and to obtain detection and quantification limits for 3,4-methylenedioxypy-raloperone in cadaveric blood at a level of 10.0-100.0 pg/ml and 1.0-10.0 ng/ml, respectively. One of the advantages of the high-performance chromatography-tandem mass spectrometry system is the screening possibility.

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