LS/T 6109-2014 Grain and Oil Inspection Determination of Zearalenone in Cereals by Colloidal Gold Rapid Test Card Method LS/T6109-2014 |tt||Standard compression package decompression password: www.bzxz.net
This standard specifies the principle, reagents and materials, instruments and equipment, sample preparation, sample determination, result determination, and positive sample confirmation of the colloidal gold rapid test card detection method for zearalenone. This standard is applicable to the rapid screening of zearalenone in cereals such as wheat and corn.
This standard was drafted in accordance with the rules given in GB/T1.1-2009.
This standard is under the jurisdiction of the National Technical Committee for Standardization of Cereals and Oils (SAC/TC270).
The drafting units of this standard: Beijing Cereals, Oils and Food Inspection Institute, Hubei Cereals, Oils and Food Quality Monitoring Station, Sichuan Cereals and Oils Central Monitoring Station, Liaoning Cereals and Oils Inspection and Monitoring Institute, Chongqing Cereals and Oils Quality Supervision and Inspection Station, Guangxi Cereals and Oils Quality Supervision and Inspection Station, National Grain Administration Standard Quality Center, Henan Province Shangqiu Cereals and Oils Technology Inspection Institute.
The main drafters of this standard: Guo Jian, Wang Zhengping, Liu Jian, Yang Jun, Zhu Xudong, Peng Chao, Liu Yongying, Liu Zheng, Zhang Yan, Yang Weimin, Shao Hui.
The following documents are indispensable for the application of this document. For all dated references, only the dated version applies to this document. For all undated references, the latest version (including all amendments) applies to this document.
GB2761 National Food Safety Standard Limits of Mycotoxins in Food
GB5491 Sampling and Sub-sampling Methods for Grain and Oilseed Inspection
GB/T6682—2008 Specifications and Test Methods for Water Used in Analytical Laboratories
GB/T23504 Determination of Zearalenone in Food by Immunoaffinity Chromatography and Cleanup High Performance Liquid Chromatography

ICS67.060
Registration No.: 44930—2014
Grain Industry Standard of the People's Republic of China
LS/T6109-—2014
Grain and Oils Inspection
Determination of Zearalenone in Cereals
Colloidal Gold Rapid Test Card Method
Inspection of grain and oils-Detection of zearalenone in cereal-Rapidmethod of colloidal gold technique2014-04-28 Issued
State Administration of Grain
2014-06-01 Implementation
This standard was drafted in accordance with the rules given in GB/T1.1-2009. This standard is under the jurisdiction of the National Technical Committee for Standardization of Grain and Oils (SAC/TC270). LS/T6109-2014
The drafting units of this standard are: Beijing Cereals, Oils and Foodstuffs Inspection Institute, Hubei Cereals, Oils and Foodstuffs Quality Monitoring Station, Sichuan Cereals and Oils Central Monitoring Station, Liaoning Cereals and Oils Inspection and Monitoring Institute, Chongqing Cereals and Oils Quality Supervision and Inspection Station, Guangxi Cereals and Oils Quality Supervision and Inspection Station, National Grain Administration Standard Quality Center, Henan Province Shangqiu Cereals and Oils Technology Inspection Institute. The main drafters of this standard are: Guo Jian, Wang Zhengping, Liu Jian, Yang Jun, Zhu Xudong, Peng Chao, Liu Yongying, Liu Zheng, Zhang Yan, Yang Weimin, Bu Hui. 1 Scope
Cereals and Oils InspectionDetermination of Zearalenone in CerealsColloidal Gold Rapid Test Card Method
LS/T6109—2014
This standard specifies the principle, reagents and materials, instruments and equipment, sample preparation, sample determination, result determination, and positive sample confirmation of the colloidal gold rapid test card method for zearalenone. This standard applies to the rapid screening of zearalenone in cereals such as wheat and corn. 2 Normative referencesWww.bzxZ.net
The following documents are essential for the application of this document. For all references with dates, only the versions with dates apply to this document. For all references without dates, the latest versions (including all amendments) apply to this document. GB2761 National Food Safety Standard Limits of Fungal Toxins in Foods Sampling and Sample Separation Method for Inspection of Cereals and Oilseeds
GB5491
GB/T6682—2008 Specifications and Test Methods for Water Used in Analytical Laboratories GB/T23504 Determination of Zearalenone in Foods Immunoaffinity Chromatography Purification High Performance Liquid Chromatography 3 Principle
This method is based on antigen-antibody reaction, immunogold labeling technology and capillary chromatography technology. The microporous membrane is used as a solid phase carrier. The antibody that can specifically identify zearalenone is labeled with red or purple colloidal gold particles. The two color bands on the test card membrane are fixed with zearalenone antigen and the secondary antibody against mouse-derived antibody, and the test results are determined according to the color changes of the color bands. 4
Reagents and Materials
Unless otherwise specified, all reagents used are analytically pure, and the water used in the laboratory should comply with the third-grade water in GB/T6682-2008. 4.1 Zearalenone Colloidal Gold Test Card\: The performance of the test card shall meet the requirements of Appendix A. Before using test cards of different brands and batches, the performance of the test card shall be measured in accordance with Appendix A. Extraction solution: 70% methanol solution or special extraction solution for colloidal gold test cards. 4.2
Instruments and equipment
In addition to conventional laboratory instruments and equipment, the following instruments and equipment should be noted. 1) The deoxynivalenol colloidal gold test card produced by Beijing Huaan Maike Biotechnology Co., Ltd., Shanghai Kuailing Biotechnology Co., Ltd. and Nanchang Boheng Biotechnology Co., Ltd. is currently the most widely used rapid test card. This information is provided only for the convenience of users of this standard and does not represent the recognition of the product by this standard. Any test card that meets the requirements of Appendix A of this standard can be used. 1
LS/T6109-—2014
Balance: graduation value 0.01g.
Crusher.
Centrifuge: speed 4000r/min.
Oscillator.
Timer.
Centrifuge tube.
Sample preparation
Sampling and sub-sampling: According to GB5491.
6.2 Sample crushing: crush the sample (6.1) to pass through a 20-mesh sieve, and mix thoroughly to ensure uniform distribution of zearalenone.
6.3 Preparation of test solution: accurately weigh 2.00 g of the crushed and mixed sample (6.2) into a centrifuge tube, add 10 mL of 70% methanol or special extract (4.2), shake and extract for 3 minutes, centrifuge at 4000 r/min for 1 minute (or let it stand), and the supernatant is the test solution. Note: Sample preparation can also be carried out according to the requirements of the test card instruction manual. 7
Sample determination
Use a pipette to slowly add 3 drops of the test solution (6.3) to the sample well of the test card (4.1) (if no liquid moves in the test window within 30 seconds after adding the test solution, add 1 drop of the test solution). Start timing when liquid moves in the test window, and observe the test results within 5min~10min.
Note: The quality control color band must appear on the test card during each test, and the test results must be observed within 5min~10min, otherwise the test results will be biased. 8
Result judgment
If the quality control color band does not show color, it means that this test card is invalid and needs to be retested. Negative and positive samples
If the quality control color band shows color and the test color band shows color, it is judged as negative. If the quality control color band shows color and the test color band does not show color or the color is very blurred, it is judged as positive.
Confirmation of positive samples
When the positive samples screened by this method need to be confirmed, the judgment should be based on the limit standard of zearalenone specified in GB2761, and the detection method should refer to the method specified in GB/T23504 or GB2761. A.1 Technical requirements
Basic requirements
Appendix A
(Normative Appendix)
Technical requirements and verification methods of zearalenone colloidal gold rapid test card in cereals Working environment requirements
Air temperature: 20℃~30℃.
Cross-reaction
LS/T6109—2014
No cross-reaction to aflatoxins, deoxynivalenol, trochotoxins and other mycotoxins. A.1.2
Performance requirements
A.1.2.1 Appearance
The appearance shall meet the following requirements:
The surface of the test card shall be smooth and flat, with uniform color, and the fiber membrane surface shall not be exposed, and there shall be no defects such as bubbles, pitting, wrinkles, and brush marks: The spot holes shall be clean:
The text symbols and other signs shall be in a straight, clear and correct font. Accuracy
The error rate of the test results of the samples shall not exceed 15%. For samples with a zearalenone content of 50μg/kg~70g/kg, false positives are allowed in the test results, but false negatives are not allowed. A.1.2.3 Stability
The error rate of the test cards of the same batch shall not exceed 10%, and the error rate of the test cards of different batches shall not exceed 15%. A.2
Verification method
Positive sample determination
The positive samples shall be determined according to the method specified in GB2761. A.2.2
Performance determination
Accuracy
Select 10 positive samples with different concentrations within the concentration range of 50μg/kg~70Jμg/kg of zearalenone for determination and calculate the error rate.
LS/T6109—2014
A.2.2.2.1
Stability
Batch-to-batch stability
Randomly select 3 batches of products, and use the same positive sample for at least 6 determinations for each batch to calculate the error rate. A.2.2.2.2
Intra-batch stability
Randomly select 1 batch of products, and use the same positive sample for at least 6 determinations to calculate the error rate. A.2.2.3
Product failure rate
The product failure rate within the shelf life should comply with the allowable value provided by the manufacturer. A.2.2.4
Agreed error range
The agreed error range should comply with the allowable value provided by the manufacturer.
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