Analysis method for simultaneous detection of amino acids and biogenic amines in foods

An analysis method, amino acid technology, applied in the direction of analysis materials, measurement devices, material separation, etc., can solve environmental problems such as unfriendly, and achieve the effect of improving sensitivity, good sensitivity, and good recovery rate results

Active Publication Date: 2016-08-17
食药环检验研究院(山东)集团有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For the first time, 4'-carbonyl chloride-rhodamine was used to derivatize the active groups of amino acids and biogenic amines. At the same time, bro

Method used

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  • Analysis method for simultaneous detection of amino acids and biogenic amines in foods
  • Analysis method for simultaneous detection of amino acids and biogenic amines in foods
  • Analysis method for simultaneous detection of amino acids and biogenic amines in foods

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Experimental program
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Embodiment 1

[0038] Chromatographic separation and mass spectrometry qualitative and quantitative analysis of amino acids and biogenic amines:

[0039] Amino acid and biogenic amine standards (purchased from Sigma Reagent Company) were prepared with 50% acetonitrile / water solution to obtain a concentration of 1×10 -5 mol / L amino acid and biogenic amine mixed standard solution. Take 10 µL amino acid and biogenic amine mixed standard solution, put it in a 1.5 mL centrifuge tube, add 10 µL 1×10 -5 mol / L 1,7-diaminoheptane-acetonitrile solution, add 800 µL pH 9.2 sodium bicarbonate-sodium carbonate buffer solution, then immediately add 200 µL 1×10 -2 mol / L 4’-carbonyl chloride-rhodamine acetonitrile solution and 90 μL bromobenzene, vortexed for 5 s to mix the solution evenly. The mixture was put into an ultrasonic oscillator for ultrasonic-assisted-in-situ derivatization-dispersion liquid-liquid microextraction, and it was taken out after 1 min (25 °C, ultrasonic frequency 40 KHz). Centri...

Embodiment 2

[0044] The extraction of amino acids and biogenic amines in red wine / white wine / beer includes the following steps:

[0045] Take 100 µL of each of the above three wines into three centrifuge tubes, sonicate for 3 min and then filter with an organic filter membrane. For in situ derivatization; 90 µL were placed in 1.5 mL centrifuge tubes, and 10 µL 1×10 -5 mol / L 1,7-diaminoheptane acetonitrile solution, add 500 µL pH 10 sodium bicarbonate-sodium carbonate buffer solution, then immediately add 200 µL 1×10 -2 mol / L 4'-carbonyl chloride-rhodamine-dispersant solution (red wine dispersant is methanol, white wine is acetone, beer is acetonitrile) and 120 µL extractant (red wine extractant is bromocyclohexane, white wine is 1- Bromooctane, beer is bromobenzene), vortexed and shaken for 5 s, put the mixture into an ultrasonic oscillator for ultrasonic-assisted-in-situ derivatization-dispersion liquid-liquid microextraction, 5 min (10 ℃, 40 KHz ) and taken out; high-speed centrifuga...

Embodiment 3

[0047] The extraction of amino acids and biogenic amines in cheese includes the following steps:

[0048] Accurately weigh 2.0 g of cheese, homogenize it and transfer it to a 50 mL centrifuge tube, inject 20 mL of hydrochloric acid solution with a concentration of 0.1 mol / L quickly for extraction, ultrasonicate it for 3 min at room temperature, and then centrifuge for 5 min. min, the speed is 10000 r / min, which makes the dissolution of amino acids and biogenic amines more thorough, and reduces the loss of amino acids and biogenic amines in pretreatment. Subsequently, the supernatants were combined; 100 µL were placed in 1.5 mL centrifuge tubes, and 30 µL 1×10 -5 mol / L 1,7-diaminoheptane acetonitrile solution, add 800 µL pH 8 sodium bicarbonate-sodium carbonate buffer solution, then immediately add 300 µL 1×10 -2 mol / L 4'-carbonyl chloride-rhodamine-acetonitrile solution and 200 µL 1-bromo-3-methylbutane, vortex and shake for 5 s, put the mixture into an ultrasonic oscillato...

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Abstract

The invention belongs to the field of analytical chemistry and particularly relates to an analysis method for simultaneous detection of amino acids and biogenic amines in foods. The method is characterized in that for the first time, 4'-carbonyl chloride-rhodamine is used as a derivatization reagent to realize simultaneous derivatization of the amino acids and the biogenic amines in the food by means of ultrasonic assistance, in-situ derivation and dispersive liquid-liquid microextraction, and qualitative and quantitative detection and analysis are performed by means of ultra-high performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry in multi-reaction monitoring mode. The method can be used for simultaneous analysis and detection of eight amino acids and nine biogenic amines; by adoption of 1,7-diamino heptane as an internal standard substance, contents of the amino acids and the biogenic amines in different foods can be obtained accurately according to an internal standard method for quantification. The method has advantages of simplicity, quickness, high sensitivity, high selectivity and the like, and high recovery rate is achieved. In addition, by the analysis method, accurate and reliable technical means can be provided for food quality assessment and supervision.

Description

technical field [0001] The present invention relates to the field of analytical chemistry, in particular to an analysis method for simultaneous detection of amino acids and biogenic amines in food, especially to a method using 4'-carbonyl chloride-rhodamine as a derivatization reagent, ultrasonic-assisted-in-situ derivatization-dispersion An analytical method based on liquid microextraction combined with ultra-high performance liquid chromatography triple quadrupole mass spectrometry detection. Background technique [0002] Amino acids and biogenic amines are a class of biologically active low-molecular-weight nitrogen-containing organic compounds that are widely found in various foods, such as red wine, white wine, beer, meat products, cheese, aquatic products, etc. Studies in recent years have shown that the production of biogenic amines is mainly derived from the decarboxylation of free amino acids in microorganisms, and the intake of appropriate amounts of amino acids an...

Claims

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Application Information

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IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/884
Inventor 赵先恩和永瑞王仁君颜平王玉花魏娜雷翠华朱树芸尤进茂
Owner 食药环检验研究院(山东)集团有限公司
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