Method of assisting to identify irradiated lipid-containing foods

A technology for assisting identification and food, applied in material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of a large number of organic solvents, expensive instruments, long extraction and purification time, etc., and achieve the effect of satisfying the detection range

Inactive Publication Date: 2013-06-26
INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for assisting the identification of irradiated fat-containing food, so as to overco

Method used

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  • Method of assisting to identify irradiated lipid-containing foods
  • Method of assisting to identify irradiated lipid-containing foods
  • Method of assisting to identify irradiated lipid-containing foods

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preparation example Construction

[0063] Preparation of epsilon subunit polyclonal antibody:

[0064] Using the purified ε subunit as the immune antigen, 2 rabbits of 2 kg were immunized. Quantitatively take out according to the immunization dose (0.5mg antigen per rabbit), add an equal volume of Freund’s (in)complete adjuvant, pump back and forth with a syringe, and make a water-in-oil (W / O) emulsion, back , Multi-point injection under the skin of the neck. The established immunization procedure was as follows: for the first immunization, the antigen was emulsified with complete Freund's adjuvant, 0.5ml / mouse, and the leg muscle was injected; for the second immunization two weeks later, the antigen was emulsified with incomplete Freund's adjuvant. The third immunization was carried out at an interval of 10 days. 10 days after the third immunization, the ear vein blood was collected to separate the serum, and the serum antibody titer was detected by indirect ELISA method.

[0065] Preparation of 2-oxo-cyclo...

Embodiment 1

[0076] Embodiment 1, the selection of extraction solvent

[0077] Most of the reported sample pretreatment methods use the traditional Soxhlet extraction method. Using n-hexane as the extractant to extract the fat in fat-containing irradiated food, and after purification by homemade Florisil column, GC-MS analysis and determination were carried out. This method takes a long time to extract and purify and consumes a large amount of organic solvent; The critical fluid extraction (SFE) method is fast and convenient for extracting 2-ACBs, and the extraction time is 30 to 60 minutes, but the SFE equipment is expensive and not widely used; the accelerated extraction method (ASE) method uses an accelerated solvent extractor, and ethyl acetate is used as The extractant extracts fat under high temperature and high pressure. The extract is added to acetonitrile and placed at -20°C to filter the fat, and then purified with a 1g silica gel column. The content of 2-ACBs is analyzed by GC-M...

Embodiment 2

[0090] Embodiment 2, construction of molecular motor biosensor

[0091] 15 μL of photosynthetic chromophores (chromatophores with F 0 f 1 -ATP synthase) (50mg / mL) mixed with 8μL (0.5mg / ml) biotinylated ε subunit polyclonal antibody, diluted to 1mL with PBS buffer, then incubated at 37°C for 60min, the excess The epsilon subunit polyclonal antibody was removed by centrifugation at 40,000 rpm for 20 min at 4°C; the precipitate was redissolved with 500 μL of PBS buffer, then 7.5 μL (0.1 mg / mL) of streptavidin was added, and diluted to 1 mL with PBS buffer, Incubate at 37°C for 10 minutes, remove excess streptavidin by centrifuging at 40,000 rpm for 20 minutes at 4°C; reconstitute the pellet with 500 μL of PBS buffer, and then add 28 μl (0.036 mg / ml) of biotinylated 2- DCB polyclonal antibody, and diluted to 1mL with PBS buffer, incubated at 37°C for 10min, excess 2-DCB polyclonal antibody was removed by centrifugation at 40,000 rpm at 4°C for 20min; then the precipitate was rec...

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Abstract

The invention discloses a method of assisting to identify irradiated lipid-containing foods. The method comprises the steps of: (1) sequentially connecting a biotinylated epsilon subunit clonal antibody, streptavidin and a biotinylated 2-dodecyl cyclobutanone polyclonal antibody on epsilon subunit of F0F1-ATP (Adenosine Triphosphate) synthase to obtain a molecular motor biosensor; (2) extracting lipid-containing foods which are not irradiated by using a solvent to obtain a solvent extracting solution a, carrying out reaction on the solvent extracting solution a and the molecular motor biosensor under the condition with the existence of a start buffer solution, adding a PBS (Phosphate Buffer Solution) into a reaction system to stop the reaction, uniformly mixing a reaction solution of the reaction system and a fluorescein/luciferase solution, and detecting a mixture by using an ultra-weak luminescence instrument to obtain the fluorescence intensity of the lipid-containing foods which are not irradiated; and (3) extracting the lipid-containing foods to be identified by using a solvent to obtain a solvent extracting solution b, and carrying out the same operations in the step (2) on the solvent extracting solution b to obtain the fluorescence intensity of the lipid-containing foods to be identified, thus realizing identification of the lipid-containing foods. With the adoption of the method of identifying the irradiated lipid-containing foods, the detection time can be shortened to be about 10mins, and in addition, the detection sensitivity of 2-DCB (Dodecyl Cyclobutanone) can reach 10-4 micrograms/mL, thus satisfying the existing detection range of the irradiated lipid-containing foods.

Description

technical field [0001] The invention relates to a method for assisting identification of irradiated fat-containing food. Background technique [0002] Irradiation technology has been widely used to improve food hygiene and safety and extend shelf life. Since the commercialization of irradiated food in 1990, the irradiated food industry has developed into an important international trade (Stewart, E.M. In Food Irradiation Principals and Applications. Molins, R.A., Ed., New York: Wiley Interscience, 2001: 347-386 ). In order to enforce irradiated food labeling regulations and increase consumer confidence, a reliable and rapid method for the detection of irradiated food is needed. [0003] 2-Alkylcyclobutanones (2-ACBs) are specific radiolysis products of fat-containing foods and can be used as markers to detect fat-containing irradiated foods (Obana H, Furuta M, Tanaka Y. Evaluation of 2-dodecylcyclobutanone as an irradiation dose indicator in fresh irradiated ground beef.J...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 哈益明赵月亮李安李伟明王锋李庆鹏靳婧
Owner INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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