Puribond Sesame Aflatoxin Total Detection Solution
[Imported sesame detected aflatoxin exceeded the standard] At the beginning of the new year, the Huangdao Inspection and Quarantine Bureau released the recent import and export inspection and quarantine situation, and the detection of aflatoxin in the imported sesame was in concern. Sesame is one of the main oil crops in China, and sesame is rich in nutrients and has become the main material for many foods. China has become a major producer and consumer of sesame. At present, China's annual consumption and processing of sesame needs about 1.1 million tons, but the annual production of sesame is only 650,000-700,000 tons. The shortage of production will lead to a significant increase in China's sesame imports in the first nine months of 2013. Customs, entry-exit inspection and quarantine departments and other departments have gradually increased the amount of sesame detection.
However, sesame is also one of the crops susceptible to A. flavus, and aflatoxin has a high affinity for sesame products. Aflatoxin has strong carcinogenicity and toxicity. The infection and production of Aspergillus flavus not only occurs in the cultivation, maturity and harvest of sesame products, but also in the drying, storage and subsequent processing, storage and transportation of the harvest. It will also occur during the process. China's sesame cultivation area is large and the area is extensive. At present, the sesame cultivation areas in rural areas of China have not yet implemented the record management or certification qualifications, and the relevant state departments cannot conduct complete control. In addition, the sesame imported from China also has quality problems. Recently, the Inspection and Quarantine Bureau continuously detected that the imported sesame was unqualified, and the aflatoxin exceeded the standard.
As one of the leading suppliers of mycotoxin testing technology and product services, Pribolab integrates the original Beijing Tellabs testing application laboratory, focusing on the services of mycotoxin testing products and technical solutions, with a view to rapid Propose solutions. Aiming at the detection of the total amount of aflatoxin in sesame, a complete solution was established with stability, accuracy, speed and simplicity.
Reference standard : According to the national standard "GB/T18979-2003" food aflatoxin determination standards.
(1) Immunoaffinity column-high performance liquid chromatography : GB/T18979-2003 national standard method
1. Â Required equipment and supplies
High performance liquid chromatography
Column: Mycotoxin-specific chromatography column (PRC-18, Pribolab).
Immunoaffinity column: PriboFast® aflatoxin total immunoaffinity column (3ml, 25pcs /box, IAC-011-3, Pribolab, fast column type ) .
High-speed homogenizer: up to 22000rpm (EQ-WR-1L, Pribolab).
Glass fiber filter paper: PriboFast® glass fiber filter paper (GMF-110 , Pribolab )
PriboFast® eight-stage pump flow operator ( Pribolab for controlling the flow rate of the immunoaffinity column )
Standard: aflatoxin mixed standard (2μg/ml aflatoxin B1, G1 and 0.5μg/ml aflatoxin B2, G2 (4/1/4/1), STD#1081, Pribolab)
2. Sample pretreatment: Prebun offers different treatment options for grain, oil, oil, etc. (For detailed solutions, please contact Puribang, Puri State has developed an optimized treatment plan for different products)
3. Immunoaffinity column purification :
Enrichment: the extract passes through the immunoaffinity column at a flow rate of 2-3 mL/min until 2-3 mL of air passes through the column;
Washing: 10 mL of 0.1% Tween/PBS wash, wash the column twice with 10 mL of water, and pass 2-3 mL of air through the column.
Elution: Elution was carried out by adding 1.0 mL of chromatographic grade methanol, incubating for more than 30 seconds, and passing the column at a flow rate of 1-2 mL/min, and collecting all the eluents for detection.
4. High performance liquid chromatography analysis
(two) enzyme-linked immunosorbent assay
Method principle
Direct competition ELISA, pre-coated on the microplate the antibodies are specific for aflatoxin aflatoxin antigen insolens, added to the sample (or standard solution aflatoxin) and horseradish peroxidase-labeled. The antigen in the sample or standard solution competes with the antigen pre-coated on the plate well to bind to the enzyme-labeled antibody. Unbound enzyme-labeled antibodies are removed upon washing. Add TMB coloring solution and read the absorbance. Sample absorbance value and its total contained amount of aflatoxin negatively correlated with the standard curve to obtain total amount of the toxin aflatoxin sample.
2. Reagents and materials: ELISA kit (PriboFast R aflatoxin total kit EKT-011 , Pribolab )
3. Sample preparation: Sesame and its products: crush, dissolve, centrifuge and take the supernatant for testing.
4. Detection steps
4.1 The immune reaction was carried out for 30 min; the plate was washed 5 times; the color reaction was carried out for 30 min; the reaction was terminated, and the OD value was read by the microplate reader.
5. Expression of analysis results
The results were interpreted using a microplate reader.
(3) Rapid detection method of immune gold standard
Method principle
The aflatoxin total rapid detection strip uses the principle of competitive inhibition immunochromatography. The total amount of aflatoxin in the sample binds to the colloidal gold-labeled specific anti-aflatoxin monoclonal antibody in the enzyme-labeled well, inhibiting the antibody. Binding to the AF-BSA conjugate on the NC membrane detection line. If the total content of aflatoxin in the sample is greater than 0.5 ug/L, the test line does not show color and the result is positive; otherwise, the test line is red and the result is negative.
2. Reagents and materials
Colloidal gold test strip: PriboStrip TM aflatoxin total immunogold standard rapid test strip ( PR-011 , Pribolab )
3. Analytical procedure: After the sesame and its products are dissolved, the supernatant is taken for testing. Pipette the sample into the well and read the results for 8-15 minutes. In the case of C-line color development: T-line color development is negative or deeper than C-line. The T line is lighter than the C line, or no coloration is a positive result.
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