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Enzyme-linked immunoassay method for aflatoxin in tobacco and tobacco products

A technology for aflatoxin and tobacco products, which is applied in measuring devices, instruments, scientific instruments, etc., to achieve the effects of simple operation steps, accelerated sample detection speed, and high sensitivity

Active Publication Date: 2013-04-24
CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no relevant literature and patent report on the determination method of mycotoxins in tobacco and tobacco products in my country

Method used

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  • Enzyme-linked immunoassay method for aflatoxin in tobacco and tobacco products
  • Enzyme-linked immunoassay method for aflatoxin in tobacco and tobacco products
  • Enzyme-linked immunoassay method for aflatoxin in tobacco and tobacco products

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Experimental program
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specific Embodiment approach

[0036] Below in conjunction with accompanying drawing, the present invention will further detail the specific technological process as follows:

[0037] 1. Sample pretreatment: Weigh about 20 grams of tobacco leaf samples or tobacco products. Samples not to be analyzed immediately should be stored in the refrigerator. The collected tobacco leaf samples or shredded tobacco samples were dried at 40°C, then crushed and passed through a 40-mesh sieve, and mixed thoroughly before sampling.

[0038] 2. Extraction of aflatoxin in the sample: Weigh 5g (accurate to 0.1g) of smoke powder sample and 0.5g of sodium chloride into a 100mL conical flask with a stopper. Add 20 mL of 80% methanol in water. Close the lid and vortex on a shaker at high speed for 5 minutes. Allow the sample to settle by standing for 3 minutes. Filter 10 mL of the extract with glass fiber filter paper. Collect the filtrate into a clean container. Take 5mL of extract, add 20mL of water to dilute, mix well and ...

example 1

[0054] 1. Reagents and instruments:

[0055] Microplate reader (MD, USA); 96-well microplate plate (Corning, USA), aflatoxin, lecithin (PC), cephalin (PE), horseradish peroxidase (HRP), bovine serum albumin ( BSA), carbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), and tetramethylbenzidine (TMB) were purchased from SIGMA. Rabbit anti-aflatoxin antibody was purchased from Glory Science (Glory Science, Texas, USA). The rest of the reagents were of analytical grade, and the water used was double distilled water.

[0056] 2. Sample pretreatment and extraction of aflatoxin: Weigh about 5 pieces of tobacco leaf samples, cut the tobacco leaf samples into pieces no larger than 3cm×3cm with scissors, and place the cut samples on a balance to weigh to an accuracy of 0.1g. After recording the weighing value, the tobacco leaf samples were placed in a weighing bottle and dried at 40°C, then crushed and passed through a 40-mesh sieve, and mixed thoroughly before sampling. We...

example 2

[0060] According to the method described in Example 1, the aflatoxin contents of tobacco leaf sample B and cut tobacco sample C were measured to be 0 μg / mL and 0 μg / mL, respectively.

[0061] Add aflatoxin standard solution to the tobacco leaf sample without aflatoxin, then carry out sample pretreatment, extraction of aflatoxin, determination of enzyme-linked immunoassay, and calculate the recovery rate according to the added amount and measured value. The results are shown in Table 1. Judging from the recovery rate of standard solution of high, medium and low different concentration levels, the recovery rate is between 99.6%-102.0%, and the relative deviation is less than 6%, which shows that the recovery rate of this method is high and the repeatability is high. better.

[0062]

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Abstract

The invention relates to an enzyme-linked immunoassay method for aflatoxin in tobacco and tobacco products, which is applicable to precise measurement of aflatoxin in tobacco leaf samples and cut tobacco samples. The method disclosed by the invention comprises the steps of pretreating samples, extracting aflatoxin in the samples, preparing standard working solution, optimizing a detection method, establishing an enzyme-linked immunoassay method and the like. By adopting the method, the content of aflatoxin in tobacco and tobacco products can be detected rapidly and accurately. The method has the advantages of simplicity in experimental operation, less nonspecific adsorption, few instrument and equipment investment, low measurement cost and accuracy and reliability in measurement result.

Description

Technical field: [0001] The invention relates to a technique for measuring aflatoxin (AFT) in tobacco and tobacco products, in particular to an enzyme-linked immunoassay method for aflatoxin in tobacco and tobacco products. Background technique: [0002] Tobacco is a major commercial crop in many parts of the world. However, in the links of primary processing, storage and transportation of tobacco leaves, tobacco factory production and sales of tobacco products, once the environmental conditions such as temperature, humidity and moisture are suitable, it is easy to breed various microorganisms and cause mildew and deterioration of tobacco leaves, resulting in a decrease in the yield of tobacco leaves and their products. Reduced, quality decreased, resulting in significant economic losses. In the tobacco industry, controlling the mildew of tobacco leaves is not only to reduce the economic loss caused by mildew of tobacco leaves, but also to ensure the safety of cigarette p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
Inventor 陈欢刘彤侯宏卫韩书磊庞永强姜兴益李中皓张洪非李雪刘楠胡清源
Owner CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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