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Synthetic method and application of universal hapten and antigen for phthalate plasticizer

A synthesis method and technology of plasticizers, which are applied in the preparation methods of peptides, chemical instruments and methods, and cyanide reaction preparations, etc., can solve the problems such as the inability to realize multi-residue detection of phthalate plasticizers, and achieve excellent immunity. The effect of originality, high sensitivity and good universality of antibodies

Inactive Publication Date: 2014-11-05
XINJIANG ACADEMY OF AGRI & RECLAMATION SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing immunoassays are mainly aimed at specific phthalate compounds, and cannot achieve multi-residue detection of phthalate plasticizers

Method used

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  • Synthetic method and application of universal hapten and antigen for phthalate plasticizer
  • Synthetic method and application of universal hapten and antigen for phthalate plasticizer
  • Synthetic method and application of universal hapten and antigen for phthalate plasticizer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Preparation of phthalate plasticizer universal hapten

[0029] (1) Take 15g of 4-nitrophthalic acid, add it to 30mL absolute ethanol, and slowly add H while stirring 2 SO 4 , Reacted at 120°C for 7h, and distilled off the water and excess ethanol produced in the reaction. Transfer to ice water while hot, take the precipitated solid with 10% Na 2 CO 3 (pH7-8) was washed until the aqueous layer was colorless, and recrystallized with absolute ethanol to obtain light yellow crystals.

[0030] (2) Dissolve 1.5g of light yellow crystals in 200mL of benzene, add pure zinc powder, add concentrated HCL in portions, stir at room temperature for 12 hours, add cold water to the reaction solution, neutralize the reaction solution with NaOH until alkaline, and use Benzene was separated and extracted, and the extract was washed with anhydrous Na 2 SO 4 Dry, distill off benzene and recrystallize with absolute ethanol to obtain light yellow crystals, which are general ha...

Embodiment 2

[0031] Example 2 Preparation of a universal artificial antigen for phthalate plasticizers.

[0032] (1) Preparation of liquid A: Add 12 mg of phthalate plasticizer universal hapten to 2 mL of N,N-dimethylformamide (DMF), stir it under magnetic force at room temperature until fully dissolved, then drop by drop at 4°C Add 500 μL of 2.5% glutaraldehyde solution, mix well in the dark and react under magnetic stirring for 2 hours to obtain liquid A.

[0033] (2) Preparation of liquid B: Weigh 38.8 mg of bovine serum albumin, dissolve in 6 mL of carbonate buffer (pH 9.6, 0.05 M), and store at low temperature. This liquid is B liquid.

[0034] (3) Slowly add solution A to solution B in an ice-water bath, and continue to react overnight under stirring conditions to obtain a mixed solution of phthalate plasticizer universal artificial antigen.

[0035] (4) Transfer the mixed solution into a dialysis bag, dialyze with PBS buffer for 3-4 days, and change the PBS buffer twice a day. Th...

Embodiment 3

[0036] Example 3 Preparation of phthalate plasticizer antiserum.

[0037] Use the universal artificial antigen prepared in Example 2 as the immunogen, mix it with Freund's complete adjuvant in equal amounts, and select 6-8 week-old Balb / c female mice after the emulsification is complete and inject it subcutaneously at multiple points on the back of the neck, every 3 Weekly booster immunization. On the 10th day after the fourth immunization, blood was collected from the tail of the mice, and the antiserum was obtained by centrifugation.

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Abstract

The present invention provides a synthetic method and application of a universal hapten and antigen for phthalate plasticizers and relates to biological detection technology. The invention uses 4-nitrophthalic acid as a raw material, which is subjected to an esterification and reduction reaction for the formation of a hapten; the hapten is coupled with a carrier protein by a glutaraldehyde method to obtain the artificial general antigen for phthalate ester plasticizer. The successful synthesis of artificial antigen is confirmed by UV scanning, and immune animal experiments show that the artificial antigen prepared by the invention has the advantages of good immunogenicity, good selectivity of generated antibodies and high sensitivity. The antigen or antibody of the invention can be used for immune analysis of phthalate plasticizer, and satisfy the need of rapid detection of phthalate plasticizer residual in food or environmental medium, and has broad application prospects.

Description

technical field [0001] The invention relates to biological detection technology, in particular to a preparation method and application of a general artificial antigen of a phthalate plasticizer. Background technique [0002] Phthalic acid esters (Phthalic Acid Esters, PAEs) compounds are the most commonly used plasticizers in plastic production, accounting for 88% of the plasticizer market share, including about 30 compounds, the most commonly used such as phthalic acid di Methyl ester (DMP), diethyl phthalate (DEP), dibutyl phthalate (DBP), bis(2-ethylhexyl) phthalate (DEHP), etc., are mainly used in production Medical products, plastic toys, cosmetics, food packaging, agricultural mulching film, etc., due to the non-covalent combination between phthalate esters and product substrates, they are continuously released into the environment during production, processing and use, which has become a global concern. The most common type of environmental pollutants. Toxicologic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C227/04C07C229/62C07K14/765C07K14/77C07K14/795C07K1/107C07K16/44G01N33/544
Inventor 孙凤霞罗小玲彭夏雨卢春霞康立超唐宗贵方婷婷谢勇
Owner XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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