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Semicarbazide derivatization reagent and rapid detection card thereof

A technology of derivatization reagent and semicarbazide, which is applied in the field of semicarbazide derivatization reagent and rapid detection card, can solve the problems of derivative extraction, destruction of antibody activity, long detection time, etc., and achieve simplified detection steps, simplified separation and purification steps , The effect of reducing the detection time and operation error

Inactive Publication Date: 2016-11-09
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods mainly have the disadvantages of requiring large-scale instruments, complicated extraction and purification process of derivatives, and long detection time.
For immunological-based detection methods such as ELISA and rapid colloidal gold detection cards, there are problems such as derivative extraction, low recovery efficiency, and organic extraction reagents destroying antibody activity, which have a great impact on the stability and accuracy of detection. It is easy to produce large batch-to-batch differences and reduce product stability

Method used

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  • Semicarbazide derivatization reagent and rapid detection card thereof
  • Semicarbazide derivatization reagent and rapid detection card thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Take ractopamine-modified p-aldehyde benzoic acid as an example for illustration:

[0024] (1) Preparation of magnetic beads

[0025] Pipette 0.5mL (10mg) of carboxylated magnetic beads into a 15mL centrifuge tube and place on a magnetic separation rack until the supernatant becomes completely transparent, then carefully remove the supernatant with a pipette. Add 5mL of MES buffer, mix well and wash. Place the centrifuge tubes on the magnetic separation rack until the supernatant becomes clear, then carefully remove the supernatant with a pipette. Repeat wash three times. After the last wash, resuspend the magnetic beads in 5 mL of MES buffer. Remove EDAC from refrigerator to room temperature for 30 minutes. Weigh 16 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDAC) into a centrifuge tube containing magnetic beads, shake vigorously. Activate the reaction by placing the tube on a rotator for 30 minutes at room temperature. During the reaction...

Embodiment 2

[0035] Take digoxin-modified p-aldehyde benzoic acid as an example to illustrate:

[0036] (1) Preparation of magnetic beads

[0037] Pipette 0.5mL (10mg) of carboxylated magnetic beads into a 15mL centrifuge tube and place on a magnetic separation rack until the supernatant becomes completely transparent, then carefully remove the supernatant with a pipette. Add 5mL of MES buffer, mix well and wash. Place the centrifuge tubes on the magnetic separation rack until the supernatant becomes clear, then carefully remove the supernatant with a pipette. Repeat wash three times. After the last wash, resuspend the magnetic beads in 5 mL of MES buffer. Remove the EDAC from refrigeration to room temperature for 30 minutes. Weigh 16mg of EDAC and add it into the centrifuge tube containing magnetic beads, shake vigorously. Place the centrifuge tube on a rotary mixer to activate the reaction for 30 minutes at room temperature. During the reaction, be careful not to let the magnetic b...

Embodiment 3

[0047] Take p-aldehyde benzoic acid modified by cyanine dye 5 as an example:

[0048] (1) Preparation of magnetic beads

[0049] Pipette 0.5mL (10mg) of carboxylated magnetic beads into a 15mL centrifuge tube and place on a magnetic separation rack until the supernatant becomes completely transparent, then carefully remove the supernatant with a pipette. Add 5mL of MES buffer, mix well and wash. Place the centrifuge tubes on the magnetic separation rack until the supernatant becomes clear, then carefully remove the supernatant with a pipette. Repeat wash three times. After the last wash, resuspend the magnetic beads in 5 mL of MES buffer. Remove the EDAC from refrigeration to room temperature for 30 minutes. Weigh 16mg of EDAC and add it into the centrifuge tube containing magnetic beads, shake vigorously. Place the centrifuge tube on a rotary mixer to activate the reaction for 30 minutes at room temperature. During the reaction, be careful not to let the magnetic bead p...

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Abstract

The invention relates to a semicarbazide derivatization reagent and a rapid detection card thereof. The derivatization reagent is aldehyde benzoic acid with a specific small molecular marker, the aldehyde benzoic acid can react with the amino of a metabolite through a aldehyde group to form a metabolite-aldehyde benzoic acid derivative with a specific molecular marker, the specific molecular marker can bind with a ligand on a magnetic bead, through magnetic bead separation, a semicarbazide derivatization product marked magnetic bead can be obtained, and the magnetic bead can be added dropwise directly on a rapid chromatographic card to perform detection. The reagent provided by the invention utilizes the advantages of lateral chromatography, simplifies the separation and purification steps of furan characteristic metabolite derivatives, and reduces the detection time and operation error.

Description

technical field [0001] The invention belongs to the field of biochemical detection, and relates to a semicarbazide derivatization reagent and a rapid detection card. Background technique [0002] Semicarbazide (SEM) is a characteristic metabolite of nitrofurazone. Furacillin belongs to nitrofuran antibiotics, which has effects on both Gram-positive and negative bacteria, and is widely used as a therapeutic drug and feed additive to control diseases or epidemics in poultry, livestock, and aquaculture. Due to the certain carcinogenicity of nitrofurazone, the European Union, Australia, the United States and China have successively introduced relevant laws to restrict the use of nitrofurazone in the aquaculture industry. [0003] Furacilin is easily degraded into semicarbazide in animals, but semicarbazide is easy to combine with protein, and the combined product is stable and difficult to decompose, and can remain in animals for several weeks. Common food cooking methods such ...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/544
CPCG01N33/9446G01N33/544G01N33/558
Inventor 曾令文鲁学文方志远
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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