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Rogridone hapten, artificial antigen and antibody as well as preparation method and application thereof

A technology of rosiglitazone and artificial antigen, which is applied in the field of preparation of artificial antigen and antibody, can solve the problems such as no rosiglitazone immunological detection method, and achieve the effect of high affinity, fast speed and low detection cost

Active Publication Date: 2011-06-22
华农(潮州)食品研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there is no effective immunological detection method for rosiglitazone

Method used

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  • Rogridone hapten, artificial antigen and antibody as well as preparation method and application thereof
  • Rogridone hapten, artificial antigen and antibody as well as preparation method and application thereof
  • Rogridone hapten, artificial antigen and antibody as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] (1) Preparation of rosiglitazone hapten (n=1), as shown in the following reaction formula:

[0058]

[0059] Weigh 0.43g (1mmol) of rosiglitazone (purity: 98.5%) and 0.232g (2mmol) of sodium chloroacetate (analytically pure) and place it in a round-bottomed flask, and dissolve its mixture with 5ml of absolute ethanol , while adding 2mmol potassium carbonate, reflux and stir the reaction at 60°C, add 1mmol potassium iodide as a catalyst during the reaction, and reflux and stir for 4 hours. The reaction mixture was filtered to obtain a yellow-brown filtrate, which was concentrated to obtain a brown-yellow oily substance. The brown-yellow greasy substance was passed through a silica gel column for silica gel column chromatography, and the eluate of the chloroform / methanol elution phase with a volume ratio of 8:1 was collected, and the eluate was concentrated to dryness to obtain 0.09 g of rosiglitazone hapten.

[0060] (2) The preparation of rosiglitazone artificial an...

Embodiment 2

[0068] (1) Preparation of rosiglitazone hapten (n=6), as shown in the following reaction formula:

[0069]

[0070] Take by weighing 0.43g (1mmol) of rosiglitazone (purity: 98.5%) and 0.76g (5mmol) of chloroheptanoic acid (analytically pure) sodium and place it in a round bottom flask, and its mixture is washed with 5ml of absolute ethanol Dissolve, add 5mmol potassium carbonate at the same time, reflux and stir the reaction at 50°C, add 5mmol potassium iodide as a catalyst during the reaction, reflux and stir the reaction for 5 hours, then stop the reaction. The reaction mixture was filtered to obtain a yellow-brown filtrate, which was concentrated to obtain a brown-yellow oily substance. Pass through a silica gel column for silica gel column chromatography, collect the eluate from the chloroform / methanol elution phase with a volume ratio of 7:1, concentrate the eluate to dryness, and obtain 0.08 g of rosiglitazone hapten.

[0071] (2) The preparation of rosiglitazone art...

Embodiment 3

[0076] Example 3 Preparation of Rosiglitazone Hapten (n=1), R is Br, as shown in the following reaction formula:

[0077]

[0078] Weigh 0.43g (1mmol) of Rosiglitazone (purity: 98.5%) and 0.16g (1mmol) of sodium bromoacetate (analytically pure) and place in a round bottom flask, and its mixture is dissolved with 5ml of absolute ethanol , while adding 1mmol of potassium carbonate, reflux and stirring reaction at 70°C, adding 0.75mmol of potassium iodide as a catalyst during the reaction, reflux and stirring for 6 hours, then stop the reaction. The reaction mixture was filtered to obtain a yellow-brown filtrate, which was concentrated to obtain a brown-yellow oily substance. Pass through a silica gel column for silica gel column chromatography, collect the eluate from the chloroform / methanol elution phase with a volume ratio of 6:1, and concentrate the eluate to dryness to obtain 0.10 g of rosiglitazone hapten.

[0079] (2) The preparation of rosiglitazone artificial antigen...

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Abstract

The invention discloses a rogridone hapten as well as a corresponding artificial antigen and an antibody thereof. The invention also discloses a preparation method and an application for the rogridone hapten as well as the corresponding artificial antigen and the antibody. The method comprises the following steps: adopting rogridone as a raw material which reacts with halogenated benzyl benzoate to generate the carboxyl-contained rogridone hapten; then, coupling the rogridone hapten and protein by an activated fat method or a mixed acid anhydride method to prepare the rogridone artificial antigen; and using the rogridone artificial antigen to immunize animals so as to generate the high-specificity rogridone antibody. An immunodetection method for the rogridone antibody and the rogridone artificial antigen can be applied to the on-site rapid detection of the rogridone and has an important realistic meaning to realize the rapid detection of the health food safety.

Description

technical field [0001] The present invention relates to a rosiglitazone hapten and its corresponding artificial antigen and antibody, and also relates to the preparation method and application of the rosiglitazone hapten and its corresponding artificial antigen and antibody. Background technique [0002] Rosiglitazone, also known as Avandia, its chemical name is 5-[4-[2-(N-methyl-N-(2-pyridyl)amino)ethoxy]benzyl]thiazolidine -2,4-diketone, molecular weight 429.92, almost insoluble in water, its structural formula is: [0003] [0004] Rosiglitazone is a thiazolidinedione antidiabetic drug, which is a highly selective peroxisome proliferator-activated receptor gamma (PPAR-γ) agonist. By binding to PPAR-γ, it activates fat, skeletal muscle and liver, etc. PPAR-γ in the tissue on which insulin acts increases the synthesis of various proteins, regulates the transcription of insulin-responsive genes, controls the production, transport and utilization of blood sugar, corrects ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D417/12C07K14/765C07K14/795C07K14/77C07K14/42C07K14/435C07K16/44C12N15/06C12P21/08G01N33/64G01N33/02
Inventor 孙远明雷红涛沈玉栋石敏杨金易王弘徐晓艳
Owner 华农(潮州)食品研究院有限公司
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