Method for detecting biogenic amine through needle sampling-microextraction-catalytic color development

A technology of biogenic amines and needles, which is applied in the field of food analysis, can solve the problems of sample waste and achieve the effects of low cost, high detection efficiency and simple operation

Active Publication Date: 2021-09-03
JIANGSU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, researchers have developed a variety of methods that can be used for BAs detection, such as chromatography, capillary electrophoresis, chemiluminescence, colorimetric analysis, and enzyme-linked immunosorbent assay, etc., but these methods are difficult to avoid complex and expensive instruments and complicated pretreatment process, and it is easy to cause waste of samples, so a more economical, environmentally friendly, and easy-to-operate BAs analysis method needs to be developed urgently

Method used

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  • Method for detecting biogenic amine through needle sampling-microextraction-catalytic color development
  • Method for detecting biogenic amine through needle sampling-microextraction-catalytic color development
  • Method for detecting biogenic amine through needle sampling-microextraction-catalytic color development

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Specific detection method:

[0050] (1) First add 100 μL DAO solution to the centrifuge tube, then add 5 μL BAs solution, incubate in a water bath at 37°C for 20-35 minutes, add 45 μL mixed chromogenic solution after cooling, react for 10-25 minutes, and pipette 2 μL , followed by detection at 650nm with an ultra-micro UV-Vis spectrophotometer, and drawing a standard curve related to BAs and UV intensity;

[0051] (2) Use a syringe pump to absorb 100 μL of DAO solution, insert the needle into the meat sample, rotate clockwise and counterclockwise once, press the inclined surface of the needle on the meat sample to take it out, assemble the syringe pump with the needle, and push the syringe pump piston, Add a drop of DAO solution, soak the meat sample in it and react for 25-40min;

[0052] (3) Pull the plunger of the syringe to withdraw the BAs-extracted droplet, take out the meat sample with a clean needle, draw 45 μL of the mixed chromogenic solution with the syringe,...

Embodiment 2

[0059] Detection of BAs in fish samples:

[0060] In order to compare with the conventional method (i.e. HPLC-UV), the BAs content in the fish sample was measured by the method of Example 1, and the results were compared with the results from the HPLC-UV analysis; 20.51 The Bas of μ M; HPLC-UV measuring value is 20.01 μ M; In order to evaluate matrix effect, these samples have also carried out standard addition detection; As shown in Table 1, obtained good rate of recovery (96.76-97.76%); Above-mentioned result and The results of HPLC-UV detection were comparable.

[0061] Table 1 Comparison of detection results between HPLC-UV and this method

[0062]

[0063] a Relative recovery = (total concentration - blank concentration) / spiked concentration

Embodiment 3

[0065] Detection of changes in BAs of fish and pork under different storage conditions and different parts of fish over time:

[0066] (1) Divide the pork (purchased in the supermarket) into 6 portions, which are sealed at room temperature, exposed at room temperature, sealed at 4°C, exposed at 4°C, frozen at -20°C and frozen at -20°C. mouth packaging. Take the abdomen of a fish (purchased in a supermarket) and divide it into 4 parts, which are sealed at room temperature, exposed at room temperature, sealed at 4°C and refrigerated at 4°C; ) is divided into four parts: fish head, fish back, fish belly and fish tail, packed in separate bags, and stored in a refrigerator at 4°C;

[0067] (2), adopt embodiment 1 method, the situation that pork and fish meat BAs content changes with time under different storage conditions is detected; Figure 4 As shown in a, under the four storage states, the total amount of the three amines in the samples accumulated continuously over time, and...

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Abstract

The invention discloses a method for detecting biogenic amine through needle sampling-microextraction-catalytic color development and belongs to the technical field of food analysis. According to the basic principle of the method, BAs is oxidized under the catalytic action of diamine oxidase (DAO) to generate H2O2; ATO catalyzes H2O2 generated by the BAs to oxidize 3, 3, 5, 5-tetramethyl benzidine (TMB) to develop color, and the content of the BAs is indirectly detected by detecting the amount of the H2O2. The method has good selectivity on histamine, putrescine and cadaverine; the linear range of a standard curve is 5-50 [mu] M and 50-1000 [mu] M; and the detection limit is 1.52 [mu] M. The method provided by the invention successfully detects the content of BAs (the total amount of histamine, putrescine and cadaverine in a meat sample is taken as a measurement standard) in order to indicate the putrefaction degree of the meat.

Description

technical field [0001] The invention belongs to the technical field of food analysis, and relates to a needle sampling-microextraction-catalyzed color development method for detecting biogenic amines, in particular to a needle sampling method combined with liquid phase microextraction and peroxidase-like catalyzed color reaction Method for the detection of biogenic amines in meat samples. Background technique [0002] Biogenic amines (BAs) exist in various protein-rich foods, such as meat, dairy products, and aquatic products. They are a class of low-molecular-weight nitrogen-containing compounds, including putrescine, cadaverine, and histamine. Certain proteins in food may be broken down into free amino acids that exist in food. If the food is contaminated with decarboxylase-containing bacteria, these free amino acids undergo decarboxylation to release BAs. The BA content in food is closely related to its spoilage and quality safety, so BAs are considered to be a marker o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/78
CPCG01N21/31G01N21/78
Inventor 唐盛许孟婵宋畅姜依依刘畅鞠嘉和陈一桐
Owner JIANGSU UNIV OF SCI & TECH
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