A kind of synthetic method of complete antigen of eugenol

A technology of complete antigen and synthesis method, applied in the field of synthesis of eugenol complete antigen, can solve the problem of no eugenol enzyme-linked immunosorbent assay method and the like, and achieve the effect of simple preparation method and good immune titer

Active Publication Date: 2021-09-14
CHINESE ACAD OF FISHERY SCI
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  • Abstract
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Problems solved by technology

However, there is no relevant report on eugenol enzyme-linked immunosorbent assay (ELISA) at home and abroad.

Method used

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  • A kind of synthetic method of complete antigen of eugenol
  • A kind of synthetic method of complete antigen of eugenol
  • A kind of synthetic method of complete antigen of eugenol

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Embodiment Construction

[0030] The present invention will be further described below in conjunction with the accompanying drawings.

[0031] see as Figure 1-Figure 5 As shown, the technical solution adopted in this embodiment is:

[0032] 1. Dissolve 9 mg of eugenol standard substance in 30 mL of sodium hydroxide solution (17.6%, m / V), add 5 mg of chloroacetic acid, stir and react at 90° C. for 6 hours, and cool to room temperature; Adjust the pH of the solution to 1 with hydrochloric acid; filter the solution and discard the filtrate to obtain the crude product; transfer the crude product to a boiling water bath for dissolution, and recrystallize and purify 3 times to obtain the carboxylated hapten. The synthetic route and product refer to figure 1 ;

[0033] 2. The obtained eugenol hapten was identified by liquid chromatography tandem mass spectrometry, and the primary mass spectrum and secondary mass spectrum of the eugenol hapten were as follows: figure 2 and image 3 Shown, the molecular w...

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Abstract

A method for synthesizing eugenol complete antigen, the invention relates to the technical field of chemical synthesis; preparation of eugenol carboxylated hapten; weighing eugenol carboxylated hapten, dissolving in 1mL PBS buffer; weighing 12mg carrier protein Dissolve in 1mL PBS buffer to obtain a carrier protein solution, then add the hapten solution dropwise to the carrier protein solution to obtain a mixed solution of hapten and carrier protein; dissolve 5mg EDC in 0.5mL PBS buffer to obtain an EDC solution , and then add the EDC solution dropwise to the mixed solution of hapten and carrier protein obtained in the previous step while stirring, continue to ice-bath for 15 minutes, shake and react at room temperature for 1 hour, and dialyze with PBS after the reaction; identification of the complete antigen of eugenol . The prepared eugenol complete antigen makes up the blank in the technical field of eugenol immunological detection method, and lays a foundation for the further development of eugenol ELISA detection method.

Description

technical field [0001] The invention relates to the technical field of chemical synthesis, in particular to a method for synthesizing complete eugenol antigens. Background technique [0002] Eugenol (eugenol, cas number 97-53-0) is a phenylpropanoid compound, which can be extracted from various plants such as clove tree and cassia laurel. For the use of eugenol in the aquatic field, the practices around the world are not uniform. The United States has not approved the use of eugenol as an aquatic anesthetic, but it is allowed as a food additive and a temporary analgesic in dental operations; Japan and Taiwan allow cloves Phenol is used in aquatic products, the dipping dose is 50-200 μg / mL, the drug withdrawal period is 7 days, and the maximum residue level (MRL) is set at 50 μg / kg. my country's "National Food Safety Standards for the Use of Food Additives" (GB 2760-2014) stipulates that eugenol can be used as an additive in food, but there are no relevant policies, regulati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/765C07K14/77C07K1/107
CPCC07K14/765C07K14/77
Inventor 李晋成韩刚刘欢
Owner CHINESE ACAD OF FISHERY SCI
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