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Method for simultaneously rapidly determining total amount of three kinds of ochratoxin in samples

A technology for rapid determination of ochratoxin, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of not being able to detect three kinds of ochratoxin at the same time, not suitable for wine detection, target detection interference, etc., and achieve low detection. The effect of limit value, low cost, accurate data

Inactive Publication Date: 2018-06-05
INSPECTION & QUARANTINE TECH CENT OF GUANGDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In view of the complex composition of wine, the content of ochratoxin in it is not high, which may interfere with the target detection. In daily detection, enrichment and purification technologies such as IAC or SPE are usually used to pre-treat the sample, and the enrichment degree of ochratoxin is high after treatment. , The background is pure, but the cost is high, so it is not suitable for the detection of large quantities of wine and other samples
[0007] At present, the extraction and purification conditions of OTB and OTC, the optimization of liquid chromatography and liquid chromatography-mass spectrometry conditions are all blank in China; there is no technical report on the simultaneous detection of three ochratoxins

Method used

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  • Method for simultaneously rapidly determining total amount of three kinds of ochratoxin in samples
  • Method for simultaneously rapidly determining total amount of three kinds of ochratoxin in samples
  • Method for simultaneously rapidly determining total amount of three kinds of ochratoxin in samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] The high-performance liquid chromatography detection of three kinds of ochratoxins in wine of embodiment 1

[0059] 1. If figure 1 , a method for simultaneous rapid determination of three ochratoxins in wine, the steps are as follows:

[0060] (1) Sample pretreatment and QuEChERS purification

[0061] Accurately measure 10.0ml of the white wine sample, and then follow the steps in A) below to obtain the liquid to be tested;

[0062] A) Add 10ml of a mixed solvent of acetonitrile and glacial acetic acid (the volume ratio of acetonitrile and glacial acetic acid is 90:10) to the sample, shake or vortex to mix (vortex for about 10s). Then add QuEChERS adsorbent 1 (anhydrous magnesium sulfate, sodium chloride, sodium citrate dihydrate and disodium hydrogen citrate sesquihydrate, the weight ratio is 4:1:1:0.5) 2.6g, shake or Vortex for 10s, centrifuge for the first time (4500rpm for 10min); pipette 1ml of the supernatant into a 1.5ml PE tube, add QuEChERS purification agen...

Embodiment 2

[0076] Example 2 Detection of three kinds of ochratoxins in wine by liquid chromatography-tandem mass spectrometry

[0077] 1. Method

[0078] (1) Sample pretreatment and QuEChERS purification are the same as in Example 1.

[0079] (2) Detection conditions of liquid chromatography-tandem mass spectrometry

[0080] 1) Chromatographic working conditions:

[0081] HPLC column: Agilent ZORBA-LC-C18 column (3.0mm×150mm, 3.5μm) column; column temperature: 40°C; flow rate: 0.3ml / min; injection volume: 20μl;

[0082] HPLC elution conditions: mobile phase: A is 0.1% formic acid, B is acetonitrile;

[0083] Gradient elution program: the elution program of the gradient elution is: the elution program of the gradient elution is: 0.5min: 90% solution A, 2.0min: 10% solution A, 5.0min: 10% Solution A, 5.10min: 90% solution A, 10.0min: 90% solution A;

[0084] 2) Working conditions of mass spectrometry (triple quadrupole tandem mass spectrometer):

[0085] Ion source: Ion source: elect...

Embodiment 3

[0097] Embodiment 3 The optimization of the process condition of the present invention's method

[0098] 1. Optimization of sample pretreatment conditions

[0099] (1) Optimization of Adsorbent 1

[0100] Literature at home and abroad show that the adsorbents analyzed by QuEChERS are mostly anhydrous magnesium sulfate, sodium chloride, sodium acetate, sodium citrate, sodium citrate dihydrate, disodium hydrogen citrate sesquihydrate and so on. Through the dual optimization experiment screening of combination matching and proportioning, the results show that the combination of anhydrous magnesium sulfate, sodium chloride, sodium citrate dihydrate and disodium hydrogen citrate sesquihydrate has a weight ratio of 4 :1:1:0.5, the extraction effect is the best.

[0101] (2) Optimization of Purifier 2

[0102] Through the literature on the analysis of related mycotoxins QuEChERS, the commonly used purifiers are PSA, GCB, C18, neutral alumina, NH 2 , Florisil, SiO 2 etc. Due to ...

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Abstract

The invention discloses a method for simultaneously rapidly determining total amount of three kinds of ochratoxin in samples. The method comprises the following steps: adding a mixed solvent of acetonitrile and glacial acetic acid of the same volume into a to-be-determined sample, uniformly mixing, adding a QuEChERS cleaning agent 1, uniformly mixing, performing centrifugal separation, and takingthe supernatant; adding a QuEChERS cleaning agent 2, uniformly mixing, performing centrifugal separation, taking the supernatant, and passing through an organic filter membrane so as to obtain a to-be-determined solution; simultaneously preparing a standard working solution and making a standard curve; and finally, adopting high performance liquid chromatography or high performance liquid chromatography-tandem mass spectrometry, simultaneously detecting ochratoxins A, B and C in the to-be-determined solution, and performing quantitative analysis according to the established standard curve. According to the detection method, the three kinds of ochratoxin in samples can be simultaneously and rapidly determined, and the method is excellent in purification effect, simple and convenient to operate, rapid, low in cost and less in dose of organic reagents, has high sensitivity, accuracy and precision, can be applied to routine testing of the ochratoxin, and has an important application valueand practical significance.

Description

technical field [0001] The invention belongs to the technical field of food safety detection. More specifically, it relates to a method for simultaneously and rapidly determining the total amount of three ochratoxins in a sample. Background technique [0002] Ochratoxins (OT) are a group of toxic metabolites with similar structures produced by fungi such as Penicillium verrucosum, Aspergillus ochraceus and A. carbonarius. The common ones are OchratoxinA (OTA), Ochratoxin B (OTB) and Ochratoxin C (OTC), etc. Among them, OTA is the most toxic and the most serious food contamination. Studies have shown that OTA not only has immunotoxicity, nephrotoxicity, and hepatotoxicity, but also has teratogenic, mutagenic, and carcinogenic effects. The International Agency for Research on Cancer (IARC) has defined it as a group 2B carcinogen. For example, in the process of grape planting and wine brewing, due to the influence of growth conditions and production techniques, it is easy to...

Claims

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Application Information

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IPC IPC(8): G01N30/88
CPCG01N30/88
Inventor 刘青庞世琦陈文锐熊欣曾广丰何素媚潘炳珍
Owner INSPECTION & QUARANTINE TECH CENT OF GUANGDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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