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Terbutaline conjugate and its preparation method and uses

A technology of terbutaline and conjugates, applied in the field of terbutaline conjugates and its preparation, to achieve the effect of saving inspection time and making up for the long time-consuming

Inactive Publication Date: 2007-11-28
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Clenbuterol is the most commonly used drug as a growth-promoting agent, but with the increase of detection of clenbuterol hydrochloride, illegal users began to use other substitutes, because terbutaline and clenbuterol hydrochloride Terbutaline has a similar growth-promoting effect, and terbutaline has become one of the main substitutes after clenbuterol hydrochloride. There are relatively few studies on terbutaline at home and abroad, so the immunogen of terbutaline is studied. Synthesis and antibody preparation are very necessary

Method used

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  • Terbutaline conjugate and its preparation method and uses
  • Terbutaline conjugate and its preparation method and uses
  • Terbutaline conjugate and its preparation method and uses

Examples

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Embodiment 1

[0040] (1) Preparation of solution A: Weigh 50 mg cBSA (0.735 μmol) and dissolve it in 3 mL of 50 mM carbonate (pH=10.7) buffer solution, then add 13.8 μL (72.9 μmol) of 1,4-butanediol to the solution Ether (BDE), reacted at room temperature for 20 hours to obtain a colorless and clear solution A for subsequent use;

[0041](2) Preparation of solution B: Weigh 30 mg (109.4 μmol) of terbutaline sulfate and dissolve it in 1.5 mL of 50 mM carbonate (pH=10.7) buffer solution, blow solution A into nitrogen, and then dissolve terbutaline sulfate The Lin solution was added dropwise to the A solution, and reacted at room temperature for 20 hours to obtain a colorless and clear solution B.

[0042] (3) Stir the dialysis solution B for 70 to 80 hours at 0-4°C with a phosphate buffer solution with a pH of 7.40 and a concentration of 0.01M, then dialyze with distilled water for 20 to 30 hours, and change the dialysate every 6 hours;

[0043] (4) Freeze-dry the dialyzed solution to obtain...

Embodiment 2

[0046] (1) Preparation of solution A: Weigh 50 mg cBSA (0.735 μmol) and dissolve it in 3 mL of 50 mM carbonate (pH=11.1) buffer solution, then add 16.7 μL (88.2 μmol) of BDE to the solution, and react at room temperature for 22 hours , to obtain a colorless and clear solution A, for subsequent use;

[0047] (2) Preparation of solution B: Weigh 25 mg (91.2 μmol) of terbutaline sulfate and dissolve it in 1.0 mL of 50 mM carbonate (pH=11.1) buffer solution, blow solution A into nitrogen, and then dissolve terbutaline sulfate The Lin solution was added dropwise to the A solution, and reacted at room temperature for 24 hours to obtain a colorless and clear solution B.

[0048] (3) Stir the dialysis solution B for 70-80 hours at 0-4°C with a phosphate buffer solution with a pH of 7.50 and a concentration of 0.01M, and then dialyze with distilled water for 20-30 hours, changing the dialysate every 6 hours;

[0049] (4) Freeze-dry the dialyzed solution to obtain a white terbutaline c...

Embodiment 3

[0051] (1) Preparation of solution A: Weigh 60 mg cBSA (0.882 μmol) and dissolve it in 4 mL of 50 mM carbonate (pH=10.9) buffer solution, then add 25.0 μL (132.3 μmol) of BDE to the solution, and react at room temperature for 24 hours , to obtain a colorless and clear solution A, for subsequent use;

[0052] (2) Preparation of solution B: Weigh 40 mg (146 μmol) of terbutaline sulfate and dissolve it in 3 mL of 50 mM carbonate (pH=10.9) buffer solution, blow solution A into nitrogen, and then dissolve the terbutaline sulfate solution Add dropwise to solution A, and react at room temperature for 24 hours to obtain colorless and clear solution B.

[0053] (3) Stir the dialysis solution B for 70 to 80 hours at 0-4°C with a phosphate buffer solution with a pH of 7.30 and a concentration of 0.02M, and then dialyze with distilled water for 20 to 30 hours, changing the dialysate every 6 hours;

[0054] (4) Freeze-dry the dialyzed solution to obtain a white terbutaline conjugate, and ...

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Abstract

The invention discloses a coupling matter of terbutaline in general formula (I), constituted in coupling by terbutaline hapten and carrier matter generating immunogenicity, which is bovine serum albumin or ovalbumin. Whereinto, n is the number of terbutaline molecules combined with a bovine serum albumin molecule. Said n is integer from 1 to 15. cBSA is activated bovine serum albumin and the range of molecular weight is 66KDa-69KDa. The invention also discloses the preparation method of said coupling matter. Terbutaline is connected with carrier matter generating immunogenicity to combine the coupling matter which can induce animal immune system to generate antibody. By immunizing New Zealand white rabbit the coupling matter of terbutaline in invention prepares antiserum with value of more than 1:50,000. The lowest measuring limitation is 0.3ppb and IC50 is 9.25ppb. The invention is convenient, quick, special and accurate. It provides a foundation for preparing ELISA kit for detecting of terbutaline.

Description

technical field [0001] The present invention relates to a β-stimulant conjugate and its preparation method and application, in particular to a terbutaline conjugate and its preparation method and application. It belongs to the field of β-stimulant immunoassay. Background technique [0002] The following titles involved in the present invention apply to the entire specification and claims: [0003] Bovine Serum Albumin (BSA): Product of Sigma [0004] Ovalbumin (OVA for short): a product of Sigma [0005] Phosphate Buffered Saline (PBS for short) (0.01M, pH=7.40) [0006] Dialysis membrane: Bioshorp products [0007] Terbutaline Sulfate: China National Institute for the Control of Pharmaceutical and Biological Products [0008] 1,4-Butanediol diglycidyl ether (1,4-Butanediol diglycidyl ether, referred to as BDE): Sigma company product [0009] Hydroxysuccinimide (NHS for short): a product of Sigma [0010] Ethyl[3-(dimethylamino)propyl]carbodiimide (EDC for short): pro...

Claims

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Application Information

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IPC IPC(8): G01N33/531G01N33/53
Inventor 郗日沫张玉兰卢圣欣刘围赵承彪
Owner SHANDONG UNIV
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