Magnetic particle chemiluminescence kit for detecting sudan I, and applications thereof

A technology of chemiluminescence reagents and kits, applied in chemiluminescence/bioluminescence, analysis by making materials react chemically, and measuring devices, etc., can solve problems such as poor stability of reagents, reaction time and temperature effects, and achieve detection The effect of low time, fast detection and high sensitivity

Inactive Publication Date: 2013-02-13
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reaction time and temperature have a great influence on the enzyme activity, and the stability of the reagent is poor

Method used

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  • Magnetic particle chemiluminescence kit for detecting sudan I, and applications thereof
  • Magnetic particle chemiluminescence kit for detecting sudan I, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The preparation of the concrete component of embodiment one kit

[0035] 1) Synthesis of Sudan Red Hapten

[0036] The Sudan red hapten was synthesized by the succinic anhydride method.

[0037] 2) Preparation of Sudan Red Artificial Antigen

[0038] The Sudan red hapten and human serum albumin (HSA) were coupled to the immunogen by the mixed anhydride method.

[0039] 3) Preparation of monoclonal antibodies

[0040] Animal immunization: Immunize Balb / c mice with the immunogen at a dose of 100 μg / mouse to produce antiserum.

[0041] Cell fusion and cloning: Splenocytes from immunized Balb / c mice were fused with SP2 / 0 myeloma cells at a ratio of 9:1 to obtain hybridoma cell lines of monoclonal antibodies.

[0042] Cell cryopreservation and recovery: the hybridoma cells were made into 1×10 cryopreservation medium 9 cells / ml for long-term storage in liquid nitrogen. When recovering, take out the cryopreservation tube, put it into a 37°C water bath to thaw quickly, re...

Embodiment 2

[0053] The formation of embodiment two kits

[0054] Construct the magnetic particle direct competition chemiluminescence detection kit for detecting Sudan red, so that it contains the following components:

[0055] Fluorescent marker for FITC-labeled Sudan Red monoclonal antibody

[0056] Luminescent marker of ABEI-labeled Sudan Red hapten

[0057] Separation reagent of paramagnetic nanobeads coated with goat anti-FITC monoclonal antibody

[0058] Sudan red standard solution (0ng / ml, 0.02ng / ml, 0.05ng / ml, 1.0ng / ml, 2.0ng / ml, 5.0ng / ml), the standard diluent is pH7.2, 0.03%NaN 3 , 0.1mol / L PBS buffer solution. The percentage content is a mass percentage content.

[0059] The concentrations of the Sudan Red quality control solution are 0.03ng / ml and 3.0ng / ml respectively, and the dilution solution of the quality control substance is pH7.2, 0.03%NaN 3 , 0.1mol / L PBS buffer solution. The percentage content is a mass percentage content.

[0060] Concentrated lotion pH7.2-7.4...

Embodiment 3

[0061] The detection of Sudan Red in the actual sample of embodiment three

[0062] 1. Sample pretreatment

[0063] Processing method of food samples (water base, oil base and powder) with Sudan red as food additive

[0064] Weigh 2 g of sample into a centrifuge tube, add 10 ml of acetonitrile, shake vigorously for 10 min, and centrifuge at room temperature above 3000 rpm for 10 min. Pipette 5ml of the supernatant and dry it under nitrogen at 50°C. Add 4ml of n-hexane, vortex for 30s to dissolve the dry residue, add 1ml of 1M NaOH, vortex for 15s, centrifuge at room temperature 3000rpm for 10min, pipette 1ml of the supernatant and dry it with nitrogen at 50°C, add 0.5ml of DMF to fully dissolve the dry residue , take 50 μl and add 950 μl diluted reconstitution solution and mix well to obtain the sample solution to be tested, sample dilution factor: 20.

[0065] 2. Detection with kit and analysis of results

[0066] Pipette 20μl-100μl standard or sample respectively, then a...

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Abstract

The present invention relates to a magnetic particle chemiluminescence kit for detecting sudan I. The kit comprises a luminous marker, a fluorescein marker, a standard substance, a quality control substance and a separation reagent, wherein the luminous marker is isoluminol luminous marker labeled sudan I hapten, the fluorescein marker is fluorescein labeled sudan I monoclonal antibody or fluorescein derivative labeled sudan I monoclonal antibody, and the separation reagent is sheep anti-FITC monoclonal antibody coated paramagnetic nanometer microbeads. The present invention further relates to a method for detecting sudan I in foods of animal origin by using the kit, wherein the method provides characteristics of high sensitivity, high specificity and rapid detection for sudan I detection.

Description

technical field [0001] The invention relates to a direct chemiluminescence detection kit and a detection method thereof. In particular, the magnetic particle competition direct chemiluminescence detection kit for the detection of abamectin drug residues in samples such as chili oil, chili sauce, and chili powder. technical background [0002] Sudan red (Sudan I) is a kind of artificially synthesized red dye, which is often used as an industrial dye, and is widely used in the color enhancement of solvents, oils, waxes, gasoline, as well as in the enhancement of shoes and floors. After Sudan, the dye contains "azobenzene". When "azobenzene" is degraded, "aniline" will be produced, which is a moderately toxic carcinogen. Excessive "aniline" inhaled into the human body may cause tissue hypoxia, poor breathing, damage to the central nervous system, cardiovascular system and other organs, and even lead to infertility. Because of its serious carcinogenicity, allergenicity and gen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N33/577
Inventor 何方洋冯才伟马孝斌李勇朱亮亮韩锐
Owner BEIJING KWINBON BIOTECH
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