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Pre-column derivatization method and method for detecting histamine in fermented milk product

A technology of pre-column derivatization and detection method, which is applied in the field of pre-column derivatization and the detection of histamine in fermented dairy products, can solve problems such as the absence of factor optimization research and the inability to find the optimal combination and optimal value of factors. , to achieve the effect of simple derivatization conditions, high accuracy and precision

Active Publication Date: 2017-05-31
BRIGHT DAIRY & FOOD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to overcome the fact that when the prior art detects the content of histamine in fermented milk products, there is no optimization research on many factors in the pre-column derivatization process, and there are many factors that affect the detection results. Deficiency in finding the optimal combination and optimal value of factors, providing a method for pre-column derivatization and detection of histamine in fermented dairy products

Method used

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  • Pre-column derivatization method and method for detecting histamine in fermented milk product
  • Pre-column derivatization method and method for detecting histamine in fermented milk product
  • Pre-column derivatization method and method for detecting histamine in fermented milk product

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Experimental program
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Effect test

Embodiment 1

[0059] The derivatization conditions optimized by response surface methodology were obtained through the following steps:

[0060] 1. Prepare the solution

[0061] (1) Histamine standard solution

[0062] Accurately weigh an appropriate amount of histamine standard substance (chromatographic grade) and dissolve it in 0.1 mol / L hydrochloric acid to prepare a 1000 mg / L mother solution, and store it in a refrigerator at 4°C. Use 0.1mol / L hydrochloric acid to dilute the mother liquor into a 100mg / L standard solution for later use.

[0063] (2) Internal standard solution

[0064] Accurately weigh an appropriate amount of 1,7-heptanediamine (chromatographic grade) standard substance and dissolve it in 0.1mol / L hydrochloric acid to prepare a 1000mg / L mother liquor, and store it in a refrigerator at 4°C. Use 0.1mol / L hydrochloric acid to dilute the mother liquor into a 100mg / L standard solution for use.

[0065] 2. Pretreatment of samples to be tested

[0066] Take weighing 5g gr...

Embodiment 2

[0103] (1) Sample pretreatment

[0104] Take fermented milk sample A (commercially available product, Inner Mongolia old yoghurt, manufactured by Mengniu Company) weighing 5g that has been stirred evenly, add 15mL, 0.6mol / L perchloric acid aqueous solution, mix well, shake for 30min, centrifuge at 2000G for 10min, and take supernatant. The samples were extracted 3 times with perchloric acid according to the above steps, and the supernatants were combined and filtered into a volumetric flask. Dilute the filtrate to 50 mL with 0.6 mol / L perchloric acid aqueous solution, shake well and set aside. The concentration of the sample to be tested after pretreatment was 100.10g / L.

[0105] (2) Derivatization reaction

[0106] Get above-mentioned sample solution 0.5mL after adding the 10 μ L internal standard solution that embodiment 1 makes, then add 1.5mL, the carbonate buffer of pH 10.0 and 2mL, 7.25g / L dansyl chloride solution (solvent is acetone), in React at 49.0°C for 60 min, ...

Embodiment 3

[0113] (1) Sample pretreatment

[0114] Take 5g of grated natural cheese B (commercially available product, cheddar cheese, manufactured by Castle Combe, UK), add 15mL, 0.6mol / L perchloric acid aqueous solution, mix well, shake for 30min, then centrifuge at 2000G for 10min , take the supernatant. The samples were extracted 3 times with perchloric acid according to the above steps, and the supernatants were combined and filtered into a volumetric flask. Dilute the filtrate to 50 mL with 0.6 mol / L perchloric acid aqueous solution, shake well and set aside. The concentration of the sample to be tested after pretreatment was 104.77g / L.

[0115] (2) Derivatization reaction

[0116] Get above-mentioned sample solution 0.5mL after adding the 10 μ L internal standard solution that embodiment 1 makes, then add 1.5mL, the carbonate buffer of pH 10.0 and 2mL, 7.25g / L dansyl chloride solution (solvent is acetone), in React at 49.0°C for 60 min, remove acetone by nitrogen blowing at 40...

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Abstract

The present invention discloses a pre-column derivatization method and a method for detecting histamine in a fermented milk product. The pre-column derivatization method comprises: (1) pre-treating a sample to be detected, wherein a sample to be detected is subjected to a mixed reaction in a perchloric acid aqueous solution, centrifugation is performed, the supernatant is taken, and the pre-treated sample having a concentration of 50-250 g / L is prepared with the perchloric acid aqueous solution; and (2) adding an internal standard solution to the pre-treated sample, carrying out a derivatization reaction under a derivatization condition, removing acetone in the derivatization reaction process, extracting with a solvent, and taking the extraction phase. According to the present invention, the pre-column derivatization conditions are simple and efficient, have high accuracy and high precision, and can meet the accurate detection requirements of laboratories, factories and the like on the histamine content in milk products during the subsequent histamine detection process.

Description

technical field [0001] The invention relates to a pre-column derivatization method and a detection method for histamine in fermented milk products. Background technique [0002] Histamine is a kind of biogenic amine with the most content in food and the greatest impact on human health. It is formed by decarboxylation of histidine by decarboxylase. Many modern food ingredients contain histamine, which is formed by the decarboxylation reaction of histidine rich in raw materials and corresponding decarboxylase under suitable process conditions. Among them, the content of histamine in fish food is the highest, and the content of histamine in dairy products (especially fermented milk products) is second only to fish food. [0003] An appropriate amount of histamine has physiological effects such as releasing hormones and stimulating nerves in the human body. It is only toxic when the accumulation in the body reaches a high level or the intake is too high. Toxicological studies ...

Claims

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Application Information

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IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/884
Inventor 刘景刘振民游春苹王渊龙
Owner BRIGHT DAIRY & FOOD
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