Pretreatment kit for detecting ochratoxin a in porcine tissue and its application

A technology of ochratoxin and kit, which is applied in the field of pretreatment kits for detecting ochratoxin A in porcine tissues, which can solve the problems such as the introduction of no standard for ochratoxin A, and achieve high sensitivity, guaranteed stability, and strong specificity Effect

Inactive Publication Date: 2016-08-24
重庆市动物疫病预防控制中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] my country has successively promulgated many legal standards for mycotoxins in food (see GB2715-2005 Mycotoxin Limits in Food [s] and GB2761-2011 Mycotoxin Limits in Food [S]. The inspection is limited to grains and their products, beans and their products, and there is no standard for ochratoxin A in pig tissues

Method used

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  • Pretreatment kit for detecting ochratoxin a in porcine tissue and its application
  • Pretreatment kit for detecting ochratoxin a in porcine tissue and its application
  • Pretreatment kit for detecting ochratoxin a in porcine tissue and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The preparation of embodiment 1 pretreatment kit

[0053] Preparation steps:

[0054]Prepare 1mol / L phosphoric acid solution as buffer; use ethyl acetate as extract 1; prepare 0.5mol / L, pH8.4 sodium bicarbonate solution as extract 2; prepare 150g sodium chloride, 2.5g sodium bicarbonate Dissolve in water, add 0.1ml Tween-20 and dilute to 1L with water to prepare an eluent; prepare methanol: acetic acid solution at a volume ratio of 49:1 as the eluent; assemble the above solution together with an IAC column to form a pretreatment reagent box.

Embodiment 2

[0055] Example 2 Determination of Ochratoxin A in No. 1 Pig Tissue

[0056] step :

[0057] 1. Obtain a blank sample

[0058] Several porcine tissue samples purchased on the market were tested by mass spectrometry, and it was finally determined that the porcine tissue without OTA was used as a blank sample to obtain a standard curve, and this was added to the OTA standard solution as a matrix to obtain a standard curve solution to avoid the entire The interference of the matrix effect occurs during the detection process, ensuring the rigor and accuracy of the present invention.

[0059] 2. Obtain the standard curve:

[0060] a. Dissolve ochratoxin A standard substance in glacial acetic acid: benzene (99:1 V / V) to prepare 1 μg / mL OTA standard stock solution;

[0061] b. Dilute the OTA standard stock solution with a chromatographic mobile phase solution into a series of OTA standard solutions of different concentrations;

[0062] c. The blank sample pig tissue is homogeniz...

Embodiment 32

[0088] The determination of ochratoxin A in the pig tissue of embodiment 3.2

[0089] step :

[0090] 1. Acquisition of standard curve:

[0091] Detailed steps are the same as the 2nd step of embodiment 2.

[0092] 2. Detection of samples to be tested

[0093] a. Collect No. 2 pig tissue for homogenization;

[0094] b. same as step b of the 3rd step of embodiment 2, obtain No. 2 need testing solution;

[0095] c. No. 10 μl of need testing solution is injected into the ultra-high liquid chromatography-tandem mass spectrometer, and the chromatographic conditions and mass spectrometry conditions of the ultra-high liquid chromatography-tandem mass spectrometer when detecting are the same as those in the second step of Example 2 The same as described in step f. Substitute the peak area of ​​the measured quantifier ion into the above-mentioned standard curve formula to calculate the concentration of ochratoxin A in the No. 2 test solution.

[0096] 3. Same as step 4 of embod...

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Abstract

The invention provides a pretreatment kit for detecting ochratoxin A in pig tissue. The pretreatment kit is characterized by comprising a buffer solution, an extracting solution 1, an extracting solution 2, an IAC column, a leacheate and an elution solution. The invention also provides a method for detecting the ochratoxin A in the pig tissue by utilizing the pretreatment kit. The method is characterized in that after the pig tissue is pretreated by utilizing the kit, the ochratoxin A is quantitatively detected by utilizing liquid chromatography-mass spectrometry under a certain liquid and mass spectrometry condition. The kit and the method have characteristics of low detection limit and high sensitivity and are relatively high in stability, precision and accuracy.

Description

technical field [0001] The invention belongs to the field of food chemical detection, in particular to a pretreatment kit for detecting ochratoxin A in pig tissue and application thereof. Background technique [0002] Ochratoxin A (OTA) is a kind of toxic compound produced by Aspergillus and Penicillium, which often contaminates food and feed. OTA is the second largest mycotoxin in causing economic losses worldwide, second only to aflatoxin. Ochratoxin is teratogenic, carcinogenic, mutagenic, and immunologically toxic, and the International Agency for Research on Cancer (IARC) classifies hertoxin A as a Class 2B carcinogen. The main target organ of ochratoxin is the kidney, which can cause kidney disease. Animal experiments have shown that ochratoxin A can cause liver and kidney damage in experimental animals, induce liver and kidney tumors, and cause kidney enlargement; it can also cause diseases such as enteritis, lymphatic gangrene, and hepatomegaly. [0003] Hertoxin ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 侯亚莉李玉平朱英才周莉丁平苏亮何义刚米自由范首军
Owner 重庆市动物疫病预防控制中心
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