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Coupling compound of Norfloxacin, preparation process and application thereof

A technology of norfloxacin and conjugates, which is applied in the field of antibiotic drug immunodetection, can solve the problems of no antibody preparation kit, short shelf life and high price, and achieves the effect of making up for a long time and saving testing time.

Inactive Publication Date: 2006-11-15
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After searching, there have been research reports on the synthesis of norfloxacin, bovine serum albumin and ovalbumin conjugates in China, but no research on antibody preparation and development of corresponding kits has been carried out.
Moreover, most of the ELISA kits for domestic detection of norfloxacin drug residues are purchased abroad, with short shelf life and high price, which are far from meeting the detection needs. Therefore, it is very necessary to study the synthesis of immunogens and antibody preparation of norfloxacin

Method used

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  • Coupling compound of Norfloxacin, preparation process and application thereof
  • Coupling compound of Norfloxacin, preparation process and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] (1) Preparation of liquid A: Weigh 10.00 mg of norfloxacin, 10.40 mg of hydroxysuccinimide, and 21.66 mg of EDC, dissolve them in 3ml of dimethylformamide, and react to form the active intermediate of norfloxacin and EDC. body, spare;

[0040](2) Preparation of cBSA: Dissolve 18.03mg of ethylenediamine in 20ml of phosphate buffer solution with a pH of 7.40 and a concentration of 0.01M at 0-4°C, adjust the pH to 7.40 with concentrated hydrochloric acid; weigh 1000.00 mg BSA (molecular weight: 68,000) and 57.51 mg EDC, then added to ethylenediamine solution, stirred and reacted at 20°C for 2 hours; The dialyzed solution was stirred and dialyzed for 70 hours, and then dialyzed with distilled water for 24 hours, and the dialysate was replaced every 6 hours; the dialyzed solution was centrifuged at 13,000 rpm at 0-4°C for 15 minutes, and the supernatant was taken; Clear liquid, obtain white powder solid cBSA, standby;

[0041] (3) Preparation of solution B: dissolve cBSA i...

Embodiment 2

[0046] (1) Preparation of A: Weigh 10.00 mg of norfloxacin, 13.00 mg of hydroxysuccinimide, and 43.32 mg of EDC, dissolve them in 4.6 ml of dimethylformamide, and react to form the active intermediate of norfloxacin and EDC. body, spare;

[0047] (2) Preparation of cBSA: Dissolve 18.03mg of ethylenediamine in 20ml of phosphate buffer solution with a pH of 7.40 and a concentration of 0.01M at 0-4°C, adjust the pH to 7.40 with concentrated hydrochloric acid; weigh 1000.00 mg BSA (molecular weight: 68,000) and 57.51 mg EDC, then added to ethylenediamine solution, stirred and reacted at 25°C for 4 hours; The dialyzed solution was stirred and dialyzed for 72 hours, and then dialyzed with distilled water for 30 hours, and the dialysate was replaced every 6 hours; the dialyzed solution was centrifuged at 13,000 rpm at 0-4°C for 15 minutes, and the supernatant was taken; freeze-dried Clear liquid, obtain white powder solid cBSA, standby;

[0048] (3) Preparation of solution B: disso...

Embodiment 3

[0053] (1) Preparation of liquid A: Weigh 10.00 mg of norfloxacin, 20.81 mg of hydroxysuccinimide, and 64.99 mg of EDC, dissolve them in 10 ml of dimethylformamide, and react to form the active intermediate of norfloxacin and EDC. body, spare;

[0054] (2) Preparation of cBSA: Dissolve 18.52mg of ethylenediamine in 20ml of phosphate buffer solution with a pH of 7.56 and a concentration of 0.02M at 0-4°C, adjust the pH to 7.56 with concentrated hydrochloric acid; weigh 1000.00 mg BSA (molecular weight: 67,000) and 57.51 mg EDC, then added to ethylenediamine solution, stirred and reacted at 22°C for 3 hours; The dialyzed solution was stirred and dialyzed for 80 hours, and then dialyzed with distilled water for 20 hours, and the dialysate was replaced every 6 hours; the dialyzed solution was centrifuged at 13,000 rpm at 0-4°C for 15 minutes, and the supernatant was taken; freeze-dried Clear liquid, obtain white powder solid cBSA, standby;

[0055] (3) Preparation of solution B:...

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Abstract

A conjugate of norfloxacin is prepared from norfloxacin semi-antigen and the bovine serum albumin (or egg albumin) as the carrier able to generate immunogenicity through coupling. It can be used for preparing the reagent kit for the enzyme-linked immunoassay of norfloxacin.

Description

technical field [0001] The invention relates to a conjugate of quinolone antibiotics and a preparation method and application thereof, in particular to a norfloxacin conjugate and a preparation method and application thereof. The invention belongs to the field of antibiotic immunoassay. Background technique [0002] The following titles involved in the present invention apply to the entire specification and claims: [0003] BSA: bovine serum albumin (Bovine Serum Albumin), Sigma company product [0004] PBS: Phosphate Buffered Saline (0.01M, pH=7.40) [0005] Sephadex-G75: dextran gel, product of Sigma [0006] cBSA: bovine serum albumin modified with ethylenediamine [0007] Dialysis membrane: product of United Carbide [0008] Norfloxacin: China Veterinary Drug Administration [0009] EDC: Ethyl[3-(dimethylamino)propyl]carbodiimide, referred to as EDC, a product of Sigma [0010] Norfloxacin (norfloxacin) belongs to the quinolone antibiotics. Because of its broad a...

Claims

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Application Information

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IPC IPC(8): C07K14/435C07D401/04C07K1/00C07K16/18G01N33/53
Inventor 郗日沫卢圣欣张玉兰
Owner SHANDONG UNIV
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