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Chemiluminescence detection kit for sulfadimidine and preparation method of chemiluminescence detection kit

A technology for detection of sulfamethazine and chemiluminescence, which is applied in chemiluminescence/bioluminescence, biological testing, and analysis through chemical reactions of materials, etc. problem, to achieve the effect of stable kit, high sensitivity and good specificity

Inactive Publication Date: 2017-10-13
太原瑞盛生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the liquid chromatography analysis method lacks highly sensitive detectors, and the instrument operation is cumbersome, and the pretreatment is complicated and time-consuming.
The ELISA method uses horseradish peroxidase or alkaline phosphatase markers. The enzyme markers are easily inactivated, and the chromogenic substrate is easily decomposed when exposed to light. The sensitivity is low, and it has a certain cross-reactivity to compounds with similar structures. inaccurate test results
[0006] CN 200310117065.6 (2003.12) Using ELISA to detect sulfamethazine, using alkaline phosphatase labeling, its sensitivity is low, and the operation process is cumbersome
CN 201935918 U (2010.8) uses the colloidal gold method to detect the residual amount of sulfamethazine in honey. The final detection result of the kit is characterized by the color visible to the naked eye, and the error is large, and the operation is cumbersome, the process is more, and it is easier error occurred
[0007] CN 106771132 A (2016.11) discloses a chemiluminescent detection kit for sulfamethazine, the kit uses horseradish peroxidase to label sulfamethazine hapten, and the disadvantages of using horseradish peroxidase are mainly Enzymes are greatly affected by the environment: such as disinfectant, pH, and ions will affect it and affect the measurement results

Method used

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  • Chemiluminescence detection kit for sulfadimidine and preparation method of chemiluminescence detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: The formation of kit 1 and the preparation of its specific components

[0023] 1. Assembly of the kit

[0024] A chemiluminescent detection kit for sulfamethazine, which contains the following components:

[0025] Acridine ester-labeled sulfamethazine antigen;

[0026] Carboxyl magnetic beads coupled with sulfamethazine antibody;

[0027] Chemiluminescence pre-excitation solution A and chemiluminescence excitation solution B;

[0028] Sulfamethazine series standard solutions, the standard concentrations are: 0 μg / L, 0.02 μg / L, 0.1 μg / L, 0.5 μg / L, 2.5 μg / L, 12.5 μg / L, the buffer solution is Tris-HCl containing 0.5-5.0% BSA and 0.1-0.5% PC300;

[0029] The cleaning solution is specifically a Tris-HCl solution with a pH of 7.2 and a concentration of 25 mmol / L, which contains NaCl and 0.05% Tween-20 at a concentration of 0.15 mol / L.

[0030] 2. Preparation of magnetic bead-conjugated antibodies

[0031] (1) Take 1 mg of carboxyl magnetic particles in a 0...

Embodiment 2

[0056] Embodiment 2: the formation of kit 2 and the preparation of its components

[0057] 1. Assembly of the kit

[0058] A chemiluminescent detection kit for sulfamethazine, which contains the following components:

[0059] Acridinium ester-labeled sulfamethazine antibody;

[0060] Carboxyl magnetic beads coupled with sulfamethazine antigen;

[0061] Chemiluminescence pre-excitation solution A and chemiluminescence excitation solution B;

[0062] Sulfamethazine series standard solutions, the standard concentrations are: 0 μg / L, 0.02 μg / L, 0.1 μg / L, 0.5 μg / L, 2.5 μg / L, 12.5 μg / L, the buffer solution is Tris-HCl containing 0.5-5.0% BSA and 0.1-0.5% PC300;

[0063] The cleaning solution is specifically a Tris-HCl solution with a pH of 7.2 and a concentration of 25 mmol / L, which contains NaCl and 0.05% Tween-20 at a concentration of 0.15 mol / L.

[0064] 2. Preparation of magnetic bead-conjugated antigen

[0065] (1) Take 1 mg of carboxyl magnetic particles in a 0.5 mL cent...

Embodiment 3

[0077] Embodiment 3: the detection of kit

[0078] 1. Sample pretreatment

[0079] (1) Obtaining the milk test sample solution: Take 150 μL of fresh milk in a 500 μL centrifuge tube, centrifuge at 4°C for 10 min (3000 r / min), and discard the upper layer of fat. Pipette 25 μL of the centrifuged milk sample into a clean glass test tube, and add 950 μL of 0.02 mol / L phosphate buffer for dilution.

[0080] (2) Take 1 g of honey and add it to a 25 mL centrifuge tube, add 2 mL of 0.5 mol / L hydrochloric acid solution, oscillate with an oscillator until the honey is completely dissolved, warm it in hot water at 55-75°C for several minutes, add 5 mL of Sodium hydroxide with a concentration of 0.2 mol / L, adjust the pH range to 4.0-6.0, shake and mix. Add 8 mL of ethyl acetate, shake up and down for 5 min, and then centrifuge for 10 min (3000 r / min). Take 5 mL of the organic solvent in the upper layer and put it in a clean glass tube, dry it in the air, add 0.5 mL of 0.02 mol / L phosph...

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Abstract

The invention discloses a chemiluminescence detection kit for sulfadimidine and a preparation method of the chemiluminescence detection kit. The kit comprises magnetic particles coupled with sulfadimidine antigens or antibodies, an acridine ester marker, sulfadimidine series standard substances, a chemiluminescence pre-excitation solution A, a chemiluminescence excitation solution B and a cleaning solution. Compared with an existing technology for detecting sulfadimidine, the kit has the advantages of being high in sensitivity, high in signal-to-noise ratio and quick in detection. The analysis sensitivity can reach 0. 020 microgram per liter.

Description

technical field [0001] The invention relates to the technical field of food detection, in particular to a chemiluminescent detection kit for sulfamethazine in food and a preparation method thereof. Background technique [0002] Sulfamethazine, also known as 4-aminobenzenesulfonamide, its structure is: [0003] [0004] Sulfamethazine, a sulfonamide drug, is a synthetic derivative of sulfamethazine. It mainly remains in food of animal origin, such as: liver, kidney, muscle, aquatic products, milk, etc. Once sulfamethazine is ingested, it takes a long time to metabolize. Generally, the amount of food of animal origin used by people is small, and most of them will not cause acute toxicity due to residues. However, if you often eat animal-derived foods containing such drugs, it may cause chronic poisoning and damage human health. Such as affecting the body's urinary system, causing allergic reactions, inhibiting the hematopoietic system, and even leading to cancer. In ad...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N21/76
CPCG01N33/53G01N21/76
Inventor 胡雪婷常燕曹晶刘丽青杜爱铭徐兵
Owner 太原瑞盛生物科技有限公司
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