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Method for testing biogenic amine in kudzu vine root

A technology of biogenic amines and pueraria, applied in the field of detection, can solve the problems of harmful health of experimenters, cumbersome pretreatment, complicated treatment methods, etc., and achieves the effect of good reproducibility and simple method.

Inactive Publication Date: 2019-03-29
南宁海关技术中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Existing national standards [19] It is aimed at the determination of biogenic amines in food, but the pretreatment method is very complicated
Sample pretreatment is relatively cumbersome, including n-hexane degreasing, liquid-liquid extraction, pH adjustment, nitrogen blowing and other steps, requiring a large amount of organic solvents, which is harmful to the health of experimenters

Method used

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  • Method for testing biogenic amine in kudzu vine root
  • Method for testing biogenic amine in kudzu vine root
  • Method for testing biogenic amine in kudzu vine root

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] The contrast of embodiment 1 chromatographic analysis method

[0063] In order to compare the LC separation performance, two systems were used for analysis in this study.

[0064] One is a Waters Acquity UPLC system (Waters, USA) including a binary solvent manager pump, a sample manager and a PDA (photodiode array) detector. Nine biogenic amines were separated by UPLC BEH C18 (1.7 μm, 2.1 mm×100 mm) (WATES, USA) column. The temperature of the UPLC column was set at 35°C. The mobile phase was pumped at a rate of 0.4 mL / min, and the gradient elution was shown in Table 1. Mobile phase A was 5 mmol / L ammonium acetate, and mobile phase B was acetonitrile. The injection volume was 5 μL, and nine biogenic amines were determined at a wavelength of 254 nm.

[0065] Table 1 Gradient elution program

[0066]

[0067] Another analytical system is an Agilent HPLC 1260 system (Agilent Corporation, USA), including a G1311C1260Quat pump VL, a G 13298 1260ALS autosampler and a G 13...

Embodiment 2

[0070] The selection of embodiment 2 chromatographic columns

[0071] On the basis of the UPLC parameters in Example 1, a comparative test was carried out on the chromatographic column, the difference being that: Acquity UPLC BEH C18 (1.7 μm, 2.1×100mm) column and Acquity UPLC BEH shielding RP18 (1.7 μm, 2.1×100mm) were studied )column. When Acquity UPLC BEH Shield RP18 column was used, the peak completion time of 9 biogenic amines was shorter than that of Acquity UPLC BEH C18 column, but cadaverine and histamine could not be completely separated. When using the Acquity UPLC BEH C18 column, the peak response values ​​of the 9 biogenic amines were higher than the Acquity UPLC BEH Shield RP18 column. For this, a UPLCBEH C18 column was chosen.

Embodiment 3

[0072] The selection of embodiment 3 mobile phases

[0073] On the basis of the UPLC parameters of Example 1, a comparative test was carried out on the mobile phase, the difference being: compared acetonitrile-water (with Example 1), acetonitrile-10mmol / L ammonium acetate buffer solution (volume ratio 1:1) and methanol-water (volume ratio 1:1) are the three most widely used mobile phase systems. It was found that the chromatographic peaks of phenethylamine and putrescine, cadaverine and tyramine could not be completely separated when methanol-water was used as the mobile phase. When acetonitrile-water was used as the mobile phase, the retention time of cadaverine and histamine was not stable enough, and due to the drift of retention time, the two components could not be completely separated, which affected the accuracy of quantification. When the acetonitrile-ammonium acetate buffered saline mobile phase system is used, the target peaks have a good shape, symmetry, no tail ph...

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Abstract

The invention firstly establishes an ultrahigh-efficiency liquid chromatography method which is used for quickly extracting and determining nine medicinal biogenic amines in the kudzu vine root whichis a medicine-and-food homologous product. The nine biogenic amines comprise tryptamine, phenylethylamine, putrescine, cadaverine, histamine, octopamine, tyramine, spermidine and spermine. After breaking a kudzu vine root sample, extraction by 0.4mol / L hydrochloric acid is utilized. Derivatization is performed in front of a dansyl chloride column. Ammoniacal liquor is added for terminating reaction, and acetonitrile is added for realizing a constant volume. A UPLC chromatographic column is utilized. Ammonium acetate and acetonitrile are used as mobile phase for performing gradient elution. A diode array detector is utilized for detecting in a wavelength of 254nm. The detecting method has an excellent linear relation. The sample recovery rate is 75.6-97.2%, and the relative standard deviation is 0.12-5.32%. The method limit of quantitation of the nine biogenic amines are respectively 10mg / kg. The method has advantages of simple operation, high speed and high sensitivity. The method canbe used for simultaneously measuring the residual amount of nine biogenic amines in the animal and botanical food such as kudzu vine root.

Description

technical field [0001] The invention relates to a method for detecting biogenic amine in kudzu root, belonging to the technical field of detection. Background technique [0002] Biogenic amines (BAS) are a class of low-molecular-weight basic compounds with biological activity and nitrogen. They are mainly formed from the corresponding amino acids under microbial decarboxylation, or from aldehydes and ketones under the action of amino acid transaminases [1] . Common biogenic amines include tryptamine, putrescine, tyramine, histamine, spermine, spermidine, phenethylamine, octopamine, cadaverine, etc. [2] . In general, biogenic amines are toxic, and when they accumulate in the human body to a certain extent, they can cause poisoning to the human body. They can cause headaches, nausea, vomiting, diarrhea, heart palpitations, blood pressure changes, breathing disturbances, and other allergic reactions that can be life-threatening in severe cases [3] . Among biogenic amines,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/04G01N30/06
CPCG01N30/02G01N30/04G01N30/06G01N2030/045G01N2030/067
Inventor 吴玉杰蔡翔宇江思华
Owner 南宁海关技术中心
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