Use of a triclosan monoclonal antibody and/or triclocarban monoclonal antibody in the detection of triclosan and/or triclocarban

A technology for cloning antibodies and triclosan alone, applied in the field of immunoassays, can solve problems such as unsatisfactory sample purification and shortened analysis time

Active Publication Date: 2020-08-28
BEIJING CENT FOR DISEASE PREVENTION & CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection method provided by the invention has high sensitivity, precision, recovery rate, LOD and LOQ can meet the requirements of detection and analysis, effectively solve the problem of unsatisfactory sample purification in pretreatment, reduce sample matrix interference, shorten analysis time, and improve recovery Rate

Method used

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  • Use of a triclosan monoclonal antibody and/or triclocarban monoclonal antibody in the detection of triclosan and/or triclocarban
  • Use of a triclosan monoclonal antibody and/or triclocarban monoclonal antibody in the detection of triclosan and/or triclocarban
  • Use of a triclosan monoclonal antibody and/or triclocarban monoclonal antibody in the detection of triclosan and/or triclocarban

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 3

[0074] Example 1 Preparation of Triclosan Monoclonal Antibody (TCS mAb)

[0075] 1. Preparation of triclosan hapten formula I compound

[0076] Dissolve 424mg of triclosan in 7mL of N,N-dimethylformamide in a 50mL round-bottomed flask, add 48mg of sodium hydride in batches under magnetic stirring in an ice bath, and remove the ice bath when there are no more bubbles. React at room temperature for 40 minutes. Slowly add 229 mg of methyl 4-bromomethylbenzoate, and monitor the progress of the reaction with a thin-layer chromatography plate. Put it into a 40°C oil bath to react, keep spotting the plate, and stop heating when the reaction is complete. The reaction solution was extracted twice with ethyl acetate, and the organic phase was collected. The organic phase was evaporated to dryness, and 12 mL of ethyl acetate was added to the bottle to dissolve the residue. Silica gel weighing 2 times the weight of the residue was added to it, and evaporated to dryness to prepare the...

Embodiment 2 3

[0133] Example 2 Preparation of triclocarban monoclonal antibody (TCC mAb)

[0134] 1, the preparation of triclocarban hapten (formula II)

[0135] Weigh equal amounts of 3,4-dichlorophenyl isocyanate and methyl 4-aminophenylbutyrate into a 50 mL round bottom flask. Add 10 mL of dichloromethane and react under nitrogen protection. Thin-layer chromatography monitors the reaction. After confirming that the reaction is complete, put the reaction solution in a water bath at 40°C and evaporate to dryness. After weighing the product, dissolve it in 7 mL of methanol, add 1 mol / L NaOH solution at 2 times the amount of the substance, and put it in 40°C. Heat and monitor hydrolysis with TLC plates. After the hydrolysis is complete, adjust the pH to 2 with 6mol / L HCl. The reaction solution was extracted twice with 50 mL of ethyl acetate. The aqueous phase of extraction and the organic phase TLC spot plate respectively, after confirming that there is no target product in the aqueous...

Embodiment 3

[0144] Example 3 Test the specificity of TCC mAb and TCS mAb to analyte (TCC and / or TCS)

[0145] The IC50 of structurally similar compounds was determined by enzyme-linked immunosorbent assay, and the cross-reactivity rate (CR) of the prepared TCC mAb and TCS mAb was evaluated.

[0146] 1) Experimental steps

[0147] Coating buffer (13mM Na 2 CO 3 and 35mM NaHCO 3 , pH 9.6) diluted coating agent (100 μL per well), added to a 96-well plate, and incubated overnight at 4°C. After washing 3 times with washing buffer (0.01M PBS, pH 7.4, 0.05% Tween 20), 150 μL of blocking buffer (0.01 M phosphate buffer, pH 7.4, containing 0.5% BSA, 0.05% Tween 20) blocked the free The active ingredient 1h, 37 ℃. Wash with washing buffer 3 times, and then dry at 37°C for 1 h. Add 50 μL of TCC, TCS or structurally similar standards and 50 μL of antibodies diluted with PBS to the wells in sequence, and then incubate at 37°C for 30 min. After washing with washing buffer for 5 times, 100 μL o...

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Abstract

The invention provides the application of a triclosan monoclonal antibody and / or trichlorocaraban monoclonal antibody to detecting triclosan and / or triclocarban, in particular, a method for detecting trace triclosan and / or triclocarban in a sample. A detection method applying immunoaffinity column-ultra-high performance liquid chromatography tandem mass spectrometry is adopted. A matrix in an immunoaffinity column is coupled with the monoclonal antibodies of the triclosan and triclocarban. The detection method provided by the invention has high sensitivity; the precision, recovery rate, LOD and LOQ of the detection method can meet the requirements of detection analysis. With the detection method adopted, the problem that the purification of a sample is not ideal in pretreatment is solved; the interference of the matrix of the sample is reduced; analysis time is shortened; a recovery rate is improved; trace triclosan and / or triclocarban can be specifically detected in complex matrixes, especially various foods; and false positive detection results are avoided.

Description

technical field [0001] The invention relates to the technical field of immunoassay, in particular to the use of a triclosan monoclonal antibody and / or triclocarban monoclonal antibody in the detection of triclosan and / or triclocarban, more specifically to a food Immunoaffinity Column-Ultra Performance Liquid Chromatography Tandem Mass Spectrometry Method for Detection of Medium and Trace Triclosan and / or Triclocarban. Background technique [0002] Endocrine disrupting chemicals (Environmental endocrine disrupting chemicals, EDCs) are exogenous chemical substances that exist in the environment and interfere with the human or animal endocrine system. Circulation and accumulation of fat in the body lead to abnormal physiological systems such as the normal reproduction, development, immunity, and behavioral activities of the organism. Both triclosan (TCS) and triclocarban (triclocarban, TCC) are endocrine disruptors, which are widely added to personal care products and daily ch...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/94G01N33/531G01N30/02C07C59/68C07C27/02C07C275/42C07C273/18C07K14/765C07K16/44
CPCC07C59/68C07C275/42C07K14/765C07K16/44C07K19/00G01N30/02G01N33/531G01N33/9446
Inventor 邵兵姚凯张晶温凯杨蕴嘉沈建忠
Owner BEIJING CENT FOR DISEASE PREVENTION & CONTROL
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