Immune colloidal gold method for detecting diclofenac illegally added in Chinese patent medicament

A technology of diclofenac and colloidal gold, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., and can solve the problems of undiscovered detection of diclofenac

Active Publication Date: 2013-08-28
SHAOGUAN COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] After searching the literature, no related reports were found on the detection of diclofenac by colloidal gold immunochromatography

Method used

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  • Immune colloidal gold method for detecting diclofenac illegally added in Chinese patent medicament
  • Immune colloidal gold method for detecting diclofenac illegally added in Chinese patent medicament
  • Immune colloidal gold method for detecting diclofenac illegally added in Chinese patent medicament

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preparation example Construction

[0051] Preparation of immunochromatographic test strips: On a 60mm×300mm single-sided adhesive PVC board, paste glass cellulose membrane, glass cellulose membrane immobilized with gold-labeled anti-diclofenac antibody, and diclofenac-coated egg in parallel. Nitrocellulose membrane, absorbent glass cellulose membrane for albumin conjugate and goat anti-rabbit IgG antibody. Use a strip cutter to cut the pasted PVC board longitudinally into 5mm×60mm test strips.

[0052] Sample test and result judgment method, insert the test strip into the sample to be tested, stay for 5 seconds, take it out and lay it flat, or use a dropper to drop 3-4 drops of liquid sample on the sample pad, and observe the result after 10 minutes. Goat anti-rabbit IgG and diclofenac-coupled OVA conjugates were sprayed on the quality control line (C) and test line (T) of the NC membrane respectively, and the test sample containing diclofenac moved to the other end through the S end according to the principle ...

Embodiment 1

[0055] The immune antigen and the coating antigen were synthesized by the improved active ester method. Dissolve 0.05g of the hapten in 1.0mL of anhydrous DMF, then add 0.016g of DCC and 0.009g of NHS, react overnight at room temperature, centrifuge to take the supernatant overnight, and separate Add 15 mg / mL BSA and OVA carbonate buffer solution (pH8.5) dropwise, and react with magnetic stirring for 4 hours; dialyze with normal saline for 72 hours to remove unreacted diclofenac, change the dialysate every 6 hours, and freeze in -20°C freezer.

[0056] Synthetic diclofenac-bovine serum albumin conjugates were used to immunize New Zealand white rabbits. Each white rabbit was first immunized with 0.2mg / 0.2mL immune antigen plus an equal amount of Freund's complete adjuvant emulsified and then subcutaneously injected. 28 days after the initial immunization, 0.1mg / 0.1mL immunization antigen was emulsified with Freund's incomplete adjuvant and injected subcutaneously to carry o...

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Abstract

The invention relates to an immune colloidal gold method for detecting diclofenac illegally added in a Chinese patent medicament. The method comprises the following steps of: coupling the diclofenac with bovine serum albumin (BSA) by using an active ester method to synthesize an artificial immune antigen, and immunizing an animal by using the artificial immune antigen to prepare a specific diclofenac antibody; coupling the diclofenac with OVA (ovalbumin) to synthesize a detection antigen for constructing an immune detection method; and marking the separated and purified diclofenac antibody to nano colloidal gold, then immobilizing the diclofenac antibody, the diclofenac detection antigen and a goat-anti-rabbit IgG (immunoglobulin G) on a vector respectively, and performing semi-quantitative assay determination of the diclofenac according to a competitive inhibitory immuno-chromatography principle, wherein through the established detection system, the minimum detection limit of the diclofenac illegally added in a health-care product or the Chinese patent medicament is 2.0mug / kg, and quick detection can be completed within 5 minutes. By adopting the method, the nano technology and the immune technology are applied in the field of safety detection of food and medicaments, and illegal addition of the diclofenac is quickly detected in one step.

Description

technical field [0001] The invention belongs to the technical field of food and drug safety detection, and relates to a diclofenac immunocolloidal gold chromatography detection method for detecting illegally added chemicals in health care products and Chinese patent medicines. Background technique [0002] The chemical name of diclofenac is 2-[(2,6-dichloro-3-methylphenyl)amino]benzoic acid, which belongs to non-steroidal anti-inflammatory drugs. It has anti-inflammatory, analgesic and antipyretic effects. It is used for Rheumatoid arthritis, adhesive spondylitis, non-inflammatory joint pain, arthritis, non-articular rheumatism, pain caused by non-articular inflammation, various neuralgia, cancer pain, post-traumatic pain and various inflammations fever etc. But there are certain side effects, which are clinically prescribed drugs and must be taken under the guidance of a doctor. [0003] Rheumatic diseases are mainly characterized by pain in joints, muscles, soft tissues,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/531
Inventor 郭杰标
Owner SHAOGUAN COLLEGE
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