Method for determining content of biogenic amines in foods by adopting ultra-efficient bonded phase chromatography

An ultra-efficient phase-convergence and biogenic amine technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of high cost, long analysis time, and large organic solvent load, and achieve low environmental pollution and good mass transfer performance , the effect of less solvent loading

Active Publication Date: 2014-06-04
广东广垦农产品质量安全检测中心有限公司
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are many methods for the determination of biogenic amines in food, each with advantages and disadvantages in terms of sensitivity, precision, specificity, detection cost and time-consuming
Among them, capillary electrophoresis (CZE) has the advantages of direct detection without derivatization, high sensitivity, fast separation speed, and simple instrument, but there are certain limitations in sample pretreatment, and too much organic solvent cannot be introduced, which limits liquid The application of liquid extraction method (LLE); enzyme biosensor method, which is simple and fast, with low detection limit and strong specificity, but the target enzyme is difficult to screen and easy to inactivate, resulting in high cost and poor reproducibility; thin layer chromatography ( TLC) is low in cost, simple and fast in operation, but has a slightly lower sensitivity and can only be used for qualitative and semi-quantitative determination of biogenic amines; high performance liquid chromatography (HPLC) or gas chromatography (GC) has good accuracy, high resolution and sensitivity High, but the analysis time is long, the organic solvent load is large, and the cost is high

Method used

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  • Method for determining content of biogenic amines in foods by adopting ultra-efficient bonded phase chromatography
  • Method for determining content of biogenic amines in foods by adopting ultra-efficient bonded phase chromatography
  • Method for determining content of biogenic amines in foods by adopting ultra-efficient bonded phase chromatography

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Experimental program
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Effect test

Embodiment 1

[0049] (a) Sample pretreatment: Accurately weigh 3.0g of the sample to be tested, place it in a 50ml centrifuge tube, add 10ml HClO 4 Solution (0.4mol / L), vortexed for 5min, centrifuged at 3040×g for 10min at 4°C, filtered through filter paper, and collected the supernatant. Precipitate with 10ml HClO 4 (0.4mol / L) solution was repeatedly extracted once, and the supernatant was combined.

[0050] (b) Purification treatment: Take the supernatant of the above sample, add an equal volume of n-hexane solution, vortex for 5 minutes, discard the upper organic phase, and collect the HClO-containing 4 part, repeated twice, transferred to a 25ml brown volumetric flask, and filled with 0.4mol / L HClO 4 The solution is brought up to volume.

[0051] (c) Pre-column derivatization: Take 1.0ml of the purified extract, 200μl of 2mol / L sodium hydroxide solution, 300μl of saturated sodium bicarbonate solution and 1ml of acetone solution of 10mg / ml dansyl chloride. ℃, protected from light for...

Embodiment 2

[0056] (a) Sample pretreatment: Accurately weigh 4.5g of the sample to be tested, place it in a 50ml centrifuge tube, add 20ml HClO 4 Solution (0.4mol / L), vortexed for 7min, centrifuged at 3040×g for 10min at 4°C, filtered through filter paper, and collected the supernatant.

[0057] (b) Purification treatment: take the supernatant, add an equal volume of n-hexane solution, vortex for 3 minutes, discard the upper organic phase, and collect the HClO-containing 4 part, transferred to a 25ml brown volumetric flask, with 0.4mol / L HClO 4 The solution is brought up to volume.

[0058] (c) Pre-column derivatization: take 1.0ml of the purified extract, 200μl of 2mol / L sodium hydroxide solution, 300μl of saturated sodium bicarbonate solution and 1ml of 10mg / ml dansyl chloride acetone solution, shake and mix well, and place at 40 ℃, protected from light for 45 minutes, shaking once every 9 minutes; adding 100 μl of ammonia water to terminate the reaction, standing in the dark at room ...

Embodiment 3

[0063] (a) Sample pretreatment: Accurately weigh 6.0g of the sample to be tested, place it in a 50ml centrifuge tube, add 10ml HClO 4 Solution (0.4mol / L), vortexed for 10min, centrifuged at 3040×g for 10min at 4°C, filtered through filter paper, and collected the supernatant. Precipitate with 10ml HClO 4 (0.4mol / L) solution was repeatedly extracted once, and the supernatant was combined.

[0064] (b) Purification treatment: Take the supernatant of the above sample, add an equal volume of n-hexane solution, vortex for 8 minutes, discard the upper organic phase, and collect the HClO-containing 4 part, repeated twice, transferred to a 25ml brown volumetric flask, and filled with 0.4mol / L HClO 4 The solution is brought up to volume.

[0065] (c) Pre-column derivatization: take 1.0ml of the purified extract, 200μl of 2mol / L sodium hydroxide solution, 300μl of saturated sodium bicarbonate solution and 1ml of 10mg / ml dansyl chloride acetone solution, shake and mix well, and place ...

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Abstract

The invention relates to the field of detection of harmful components in foods and particularly relates to a method for determining the content of biogenic amines in foods by adopting an ultra-efficient bonded phase chromatography. The method comprises the following steps: carrying out biogenic amines extraction treatment on foods to be detected with the mass of m by using an HClO4 solution to obtain liquid supernatant with the volume of V; adding an n-hexane solution with the equal volume into the liquid supernatant, extracting, and repeating for 1-2 times; removing an upper-layer organic phase part to obtain a purified extracting solution; carrying out pre-column derivatization on the purified extracting solution to obtain a derivatized extracting solution; and detecting the content of the biogenic amines in the derivatized extracting solution by using an ultra-efficient bonded phase chromatographic instrument to obtain the mass concentration of the biogenic amines in the extracting solution. According to the method, the content of the biogenic amines in the foods can be determined by adopting the ultra-efficient bonded phase chromatography and a mixed system of supercritical carbon dioxide and n-hexane/isopropanol/an ammonia water solution is used as a flowing phase; the viscosity is low, the mass transfer performance is good, the analyzing time is rapid, the solvent loading amount is samll and the environmental pollution is small.

Description

technical field [0001] The invention relates to the field of detection of harmful components in food, in particular to a method for determining the content of biogenic amine in food by ultra-high performance convergence chromatography. Background technique [0002] Biogenic amines are a general term for a class of biologically active nitrogen-containing organic compounds, which can be regarded as low-molecular-weight alkaloids formed after 1-3 hydrogen atoms in ammonia molecules are replaced by alkyl or aryl groups. According to the structure, biogenic amines can be divided into aliphatic (putrescine, cadaverine, spermine, spermidine, etc.), aromatic (tyramine, phenethylamine, etc.) and heterocyclic (histamine, tryptamine, etc.) three category. Biogenic amines participate in the normal metabolism of the body and are important precursors for the synthesis of hormones, alkaloids, nucleic acids, and proteins. Most of them have strong physiological activities, such as promoting...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
Inventor 龚霄齐宁利陈珂林丽静李积华
Owner 广东广垦农产品质量安全检测中心有限公司
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