Mass spectrometry assay method for detection and quantitation of organic acid metabolites

A mass spectrometry, caproic acid technology, applied in the field of mass spectrometry for detecting and quantifying organic acid metabolites, can solve the problems of reduced sensitivity, difficult to detect SCFA, difficult to retain, etc.

Active Publication Date: 2019-12-13
METABOLON
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005]SCFAs in their pristine state are not easily detected by LC-MS due to their volatility
SCFAs and other organic acids, including non-volatile organic acids such as lactic acid, pyruvic acid, and intermediates of the tricarboxylic acid (TCA) cycle are often poorly retained by traditional reversed-phase liquid chromatography methods and are difficult to detect in mass spectrometry Reduced sensitivity

Method used

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  • Mass spectrometry assay method for detection and quantitation of organic acid metabolites
  • Mass spectrometry assay method for detection and quantitation of organic acid metabolites
  • Mass spectrometry assay method for detection and quantitation of organic acid metabolites

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0115] Example 1: Measuring SCFA and energy metabolites

[0116] Chromatographic method

[0117] A liquid chromatography method was developed for the purification and separation of one or more, two or more and up to a total of twelve analytes selected from the group consisting of acetic acid (C2) , propionic acid (C3), isobutyric acid (C4), butyric acid (C4), 2-methylbutyric acid (C5), isovaleric acid (C5), valeric acid (C5), 3-methylvaleric acid, 4-Methylpentanoic acid (isocaproic acid), caproic acid (caproic acid, C6), pivalic acid, lactic acid, pyruvic acid, fumaric acid, succinic acid, malic acid, alpha-ketoglutaric acid, aconitic acid , citric acid, isocitric acid and combinations thereof. An Agilent 1290 Infinity UHPLC system equipped with a binary solvent pump unit, a refrigerated autosampler (set at 18°C) and a column heater (set at 60°C) was used with a reversed-phase column (Waters ACQUIT Y C18 BEH Shield, 1.7 μm, 2.1x100 mm) liquid chromatography. Mobile phase A...

Embodiment 2

[0131] Example 2: Measurement of SCFA Analytes in Experimental Samples

[0132] SCFAs in the samples were measured using derivatization method 2 using the method described in Example 1. The method was developed for the determination of the SCFA analytes acetate (C2), propionate (C3), isobutyrate (C4), Absolute amounts of butyric acid (C4), 2-methylbutyric acid (C5), isovaleric acid (C5), valeric acid (C5), caproic acid (caproic acid, C6).

[0133] In another example, 8 SCFA analytes were measured in 59 plasma samples. Results for representative samples are presented in Table 6.

[0134] In another example, 8 SCFA analytes were measured in 120 serum samples. Results for representative samples are presented in Table 6.

[0135] In another example, 8 SCFA analytes were measured in 50 urine samples. Results for representative samples are presented in Table 6.

[0136] In another example, 8 SCFA analytes were measured in 197 stool samples. Results for representative samples ...

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Abstract

A method for determining in a sample, by mass spectrometry, the presence, absence, or amount of one or more analytes is described herein. The run time is less than six minutes. The method includes subjecting the sample to an ionization source under conditions suitable to produce one or more ions detectable by mass spectrometry from each of the one or more analytes, wherein the one or more analytesare derivatized prior to ionization; measuring, in a single injection, by mass spectrometry, the amount of the one or more ions from each of the one or more analytes; and using the measured amount ofthe one or more ions to determine the amount of each of the one or more analytes in the sample.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 487,542, filed April 20, 2017, which is hereby incorporated by reference in its entirety. Background technique [0003] The following information describing the background of the present invention is provided to aid in the understanding of the present invention and is not considered to constitute or describe prior art to the present invention. [0004] During the fermentation of dietary fiber, bacteria in the colon produce short-chain fatty acids (SCFA) as well as lactic acid. Once produced, SCFAs are transported into the blood and taken up by organs and serve as substrates and signaling molecules to regulate energy homeostasis. SCFA and their effects on energy metabolism have been associated with a variety of diseases including inflammatory bowel disease, irritable bowel syndrome, diarrhea, metabolic syndrome, and cancer (Han, J. et al. A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07B61/00C07H19/06C07H19/10
CPCG01N33/6848G01N30/7266G01N33/493G01N2030/027H01J49/165
Inventor K·P·亚当A·J·汤普森
Owner METABOLON
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