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Method for simultaneously detecting fatty acid, amino acid and polyfunctional group organic acid by GC-NCI-MS

A multi-functional group and amino acid technology, applied in the field of analysis and detection, can solve the problems of inability to derivatize amino acids and multi-functional organic acids, and inability to derivatize

Pending Publication Date: 2021-05-04
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of these conventional derivatization methods is that the carboxyl groups of short-chain fatty acids and short-chain organic acids and the amino groups in amino acids cannot be derivatized, so the derivatization of amino acids and polyfunctional organic acids cannot be achieved.

Method used

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  • Method for simultaneously detecting fatty acid, amino acid and polyfunctional group organic acid by GC-NCI-MS
  • Method for simultaneously detecting fatty acid, amino acid and polyfunctional group organic acid by GC-NCI-MS
  • Method for simultaneously detecting fatty acid, amino acid and polyfunctional group organic acid by GC-NCI-MS

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0108] Establishment of standard curve

[0109] Prepare a series of capric acid standard solutions with concentrations of 0.1mmol / L, 0.2mmol / L, 0.5mmol / L, 1mmol / L, 2mmol / L and 5mmol / L; at the same time, prepare the same series of fumaric acid standard solutions, series of amber Acid standard solution, series of alanine standard solution, series of valine standard solution, series of isoleucine standard solution and series of glycine standard solution;

[0110] Mix 1mL series capric acid standard solution with 0.6mL 1mol / L sodium hydroxide, 3.2mL n-propanol and 0.2mL methyl chloroformate and shake well, heat in 80°C water bath for 60min, cool to room temperature; add 1mL dichloromethane to extract, 50mmoL / L sodium bicarbonate to adjust the pH value to 12, let it stand for stratification, absorb the dichloromethane layer with a glass syringe and pass it through a 0.22μm organic phase filter membrane to obtain a series of derivative solutions;

[0111] The above-mentioned GC-NCI...

Embodiment 2

[0136] Pretreatment of fermented milk: Take 50mL of fermented milk in a 250mL Erlenmeyer flask, add 2mL of ammonia solution with a volume fraction of 25%, stopper, shake well, heat in a water bath at 60°C for 15min, and cool to room temperature; add 10mL of absolute ethanol and 25mL of ether, Add stopper and shake well, sonicate for 2 minutes; add 25 mL of petroleum ether, sonicate for 2 minutes; centrifuge at 2000 r / min for 15 minutes, repeat the above extraction process twice, combine the extracted organic layers, and distill off the solvent under reduced pressure to obtain the solution to be tested.

[0137] Pre-treatment of shiitake mushrooms: remove the roots of shiitake mushrooms, wash them with clean water, dry them in an oven at 150°C, mash them, and pass them through a 50-mesh sieve for later use; weigh 3g shiitake mushroom powder with a 1 / 10,000th balance, add 90mL distilled water for ultrasonic extraction for 10min, Centrifuge and take the supernatant as the solution...

Embodiment 3

[0155] The concentration of preparation heptanoic acid, caprylic acid, sorbic acid, fumaric acid, capric acid, benzoic acid, succinic acid, salicylic acid, leucine, glycine is respectively the mixed standard solution of 5mmol / L;

[0156] Sulfuric acid-methanol derivatization / GC-EI-MS method:

[0157] Take 1mL of the mixed standard solution, add 2mL of 20% sulfuric acid-methanol, heat in a water bath at 80°C for 60min, take 1mL of the reacted solution, add 0.5mL of distilled water and 1mL of chloroform for extraction, let it stand for layers, and absorb the chloroform with a glass syringe Layer through a 0.22 μm organic phase filter to obtain a comparative derivative solution.

[0158] Chloroformic ester derivatization / GC-NCI-MS method:

[0159] Take 1mL mixed standard solution, add 0.6mL 1mol / L sodium hydroxide, 3.2mL n-propanol, 0.2mL methyl chloroformate and shake well, heat in 80°C water bath for 60min, cool to room temperature; add 1mL dichloromethane to extract, 50mmoL / ...

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Abstract

The invention provides a method for simultaneously detecting fatty acid, amino acid and polyfunctional group organic acid by utilizing GC-NCI-MS, and belongs to the technical field of analysis and detection. According to the method, the fatty acid, the amino acid and the polyfunctional group organic acid are simultaneously derivatized by using a chloroformate derivating agent, so that the method has the advantages of rapid derivation, good reproducibility and high derivative product stability under the normal temperature condition, and can simultaneously derivatize the carboxyl of the short-chain fatty acid, the carboxyl of the short-chain polyfunctional group organic acid and the amino and carboxyl of the branched-chain amino acid. The GC-NCI-MS method is small in matrix interference, a mass spectrum is simple and clean, molecular ion information can be reserved, the obtained molecular / quasi-molecular ion information is used for artificial spectrum unfolding qualitative analysis, and accurate quantification of fatty acid, amino acid and polyfunctional group organic acid is achieved by combining a corresponding standard curve. Embodiment data show that the method provided by the invention has the advantages of strong anti-interference capability, high sensitivity, wide application range and stable detection result.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a method for simultaneous detection of fatty acids, amino acids and organic acids by using GC-NCI-MS. Background technique [0002] Gas chromatography-mass spectrometry (GC-MS) has been used as a standard detection method for detecting volatile, low boiling point, and thermally stable substances due to its high sensitivity, high resolution, and high reproducibility. At the same time, these conditions also limit the detection and analysis of polar, high boiling point and thermally unstable substances by this method. Fatty acids, amino acids, and multifunctional organic acids are common small-molecule organic substances in living organisms. Because they all contain carboxyl, amino, and hydroxyl groups in their structures, they have strong polarity and thermal instability. In order to apply the GC-MS method to the detection of fatty acids / amino acids / polyfunctional o...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 白冰刘笑男赵楠颜廷才张旋李春强程顺昌魏宝东周倩赵瑛博
Owner SHENYANG AGRI UNIV
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