Sample pretreatment extraction kit for ochratoxin A

A technology of ochratoxin and kit, which is applied in the field of sample pretreatment and extraction kit of ochratoxin A, which can solve the problems of low separation efficiency and complex purification and separation operation, so as to improve accuracy and precision and reduce background substances The effect of interference, easy operation

Pending Publication Date: 2019-07-12
沭阳康源泰博生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to make up for the deficiencies in the prior art, and to provide a sample pretreatment and extraction kit for ochratoxin A, which is used for the enrichment and purification of ochratoxin A in samples, so as to solve the problem of ochratoxin A sample Technical problems such as complex purification and separation operations and low separation efficiency

Method used

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  • Sample pretreatment extraction kit for ochratoxin A
  • Sample pretreatment extraction kit for ochratoxin A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1. Preparation of anti-ochratoxin A antibody immunomagnetic beads:

[0033] 1. Put the magnetic beads into a centrifuge tube, add activation buffer to wash the magnetic beads, repeat three times, each time with an interval of 3 minutes, remove the activation buffer, add 200 μL of activator EDC and NHS each, mix well, and incubate at 37°C for 2 hours , to activate the magnetic beads;

[0034] 2. Wash the magnetic beads with coupling buffer three times, with an interval of 3 minutes each time, add ochratoxin A antibody, mix well, and incubate at 37°C for 3 hours to prepare anti-ochratoxin A antibody immunomagnetic beads;

[0035] 3. Wash the immunomagnetic beads three times with wash buffer at 3 min intervals. Incubate with blocking solution at 37°C for 2 hours, store in storage buffer at 4°C until use;

[0036] 2. Application of anti-ochratoxin A immunomagnetic beads for the enrichment of ochratoxin A in wheat samples:

[0037] 1. Weigh 5.0 g of wheat sample and mi...

Embodiment 2

[0040] 1. Preparation of anti-ochratoxin A antibody immunomagnetic beads:

[0041] 1. Put the magnetic beads into a centrifuge tube, add activation buffer to wash the magnetic beads, repeat three times with an interval of 5 minutes each time, remove the activation buffer, add 180 μL of activator EDC and NHS each, mix well, and incubate at 37°C for 2 hours , to activate the magnetic beads;

[0042] 2. Wash the magnetic beads with coupling buffer three times, with an interval of 3 minutes each time, add ochratoxin A antibody, mix well, and incubate at 37°C for 3 hours to prepare ochratoxin A antibody immunomagnetic beads;

[0043]3. Wash the immunomagnetic beads with washing buffer three times, with an interval of 3 minutes between each time. Incubate with blocking solution at 37°C for 2 hours, store in storage buffer at 4°C until use;

[0044] 2. Application of anti-ochratoxin A immunomagnetic beads for the enrichment of ochratoxin A in feed samples:

[0045] 1. Weigh 2.0...

Embodiment 3

[0048] 1. Preparation of anti-ochratoxin A antibody immunomagnetic beads:

[0049] 1. Put the magnetic beads into a centrifuge tube, add activation buffer to wash the magnetic beads, repeat three times with an interval of 4 minutes each time, remove the activation buffer, add 300 μL of activator EDC and NHS each, mix well, and incubate at 37°C for 4 hours , to activate the magnetic beads;

[0050] 2. Wash the magnetic beads with coupling buffer three times, with an interval of 4 minutes each time, add ochratoxin A antibody, mix well, and incubate at 37°C for 4 hours to prepare ochratoxin A antibody immunomagnetic beads;

[0051] 3. Wash the immunomagnetic beads with washing buffer three times, with an interval of 5 minutes between each time. Incubate with blocking solution at 37°C for 4 hours, store in storage buffer at 4°C until use;

[0052] 2. Application of anti-ochratoxin A immunomagnetic beads for the enrichment of ochratoxin A in wheat samples:

[0053] 1. Crush t...

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Abstract

The invention provides a sample pretreatment kit for ochratoxin A. The pretreatment kit is composed of a sample cavity, a reagent cavity, a cleaning cavity and an elution cavity. Through the enrichment and purification effects of the immunomagnetic beads, the immunomagnetic beads are fully diffused in liquid, so that the binding surface area is enlarged, the immunoreaction is more thorough, the kit can be widely applied to enrichment and separation of ochratoxin A in products such as feeds and grain and oil products, the sensitivity of detection and analysis is improved, and the phenomenon ofmissing detection is avoided; and meanwhile, the use of a large amount of organic solvents is reduced. Meanwhile, the kit is compact in structure arrangement and reasonable in design.

Description

technical field [0001] The invention belongs to the field of food safety detection and analysis, in particular to a sample pretreatment and extraction kit for ochratoxin A. Background technique [0002] Ochratoxin is a kind of mycotoxin produced by Aspergillus and Penicillium, among which ochratoxin A (ochratoxin A, OTA) is the most toxic and widely distributed. OTA not only has liver / renal toxicity and immunotoxicity, but also has carcinogenic, teratogenic and mutagenic properties. All countries in the world have made strict regulations on the limit of OTA. For example, Japan stipulates that the OTA detection limit in rice, corn and beans is ≤5 µg / kg; In beans and their products, the detection limit of OTA is ≤5 µg / kg. [0003] Due to the complexity of biological components, low concentration of analytes, complex pretreatment process and other factors, the existing extraction process of ochratoxin A is not stable, resulting in low enrichment efficiency of extraction of oc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56961
Inventor 张波张学记郗日沫王鹏刘向阳雷达
Owner 沭阳康源泰博生物科技有限公司
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