Gold-labeled test strip for rapid detection of chromium ions as well as preparation method and application thereof

A technology of gold standard test paper and chromium ions, which is applied in the fields of immunology and hygienic inspection, can solve the problems of inability to operate on site, small amount of sample screening, cumbersome and time-consuming detection, etc., and achieves easy operation and control, low detection cost, and high detection efficiency. The effect of simple method

Inactive Publication Date: 2013-11-27
HENAN INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In August 2011, the Qujing chromium slag pollution incident, the hexavalent chromium content detected in the groundwater outlet exceeded the standard by more than 200 times; in April 2012, the chromium-excessive medicinal capsule incident caused economic losses of more than 16 billion yuan
[0004] At present, the established food chromium detection techniques mainly include graphite furnace atomic absorption spectrometry (GFAAS), flame atomic absorption spectrometry (FAAS), inductively coupled

Method used

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  • Gold-labeled test strip for rapid detection of chromium ions as well as preparation method and application thereof
  • Gold-labeled test strip for rapid detection of chromium ions as well as preparation method and application thereof
  • Gold-labeled test strip for rapid detection of chromium ions as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment one, Cr 3+ Preparation and Identification of Ionic Artificial Immunization Antigen

[0052] (1) Antigen preparation

[0053] (1) Weigh 10 mg chelating agent ITCBE and dissolve it in 1 mL DMSO to form a metal chelating agent solution; weigh 18.1 mgCrCL 3 ·6H 2 O was dissolved in 1 mL of HEPES buffer with a concentration of 10 mmol / L and a pH value of 8.0 to form Cr 3+ solution; the metal chelating agent solution and Cr 3+ The solution was mixed and the pH value was adjusted to 7.0 with NaOH, and then placed on a shaker at room temperature for 12 h to form Cr 3+ - ITCBE chelate hapten solution;

[0054] (2) Weigh 20 mg of carrier protein BSA and dissolve in 1 mL of HEPES buffer with a concentration of 10 mmol / L and a pH value of 8.0 to form a carrier protein solution;

[0055] (3) Take 1 mLCr 3+ -ITCBE chelate hapten solution was added to 1 mL carrier protein solution, and the pH value was adjusted to 9.0 with NaOH, reacted on a shaker at room temperatur...

Embodiment 2

[0060] Embodiment two, anti-Cr 3+ Preparation and identification of ionic monoclonal antibodies

[0061] (1) Anti-Cr 3+ Ionic monoclonal antibody preparation

[0062] (1) Immunization of animals. with Cr 3+ -ITCBE-BSA was used to immunize 5 Balb / c mice at a dose of 50 μg·0.2 mL / mouse, and injected subcutaneously at multiple points on the back. For the first immunization, Cr was diluted with sterile phosphate buffered saline (PBS) 3+ -ITCBE-BSA, emulsified with an equal volume of complete Freund's adjuvant (CFA); for booster immunization, dilute Cr with sterilized PBS 3+ -ITCBE-BSA, emulsified with an equal amount of Freund's incomplete adjuvant (IFA), the second immunization was performed 3 weeks after the first immunization, and then immunized once every 3 weeks, a total of 5 immunizations, 10 days after the third immunization Blood was collected by docking the tail, placed in a water bath at 37°C for 30 min, placed overnight at 4°C, centrifuged at 800 r / min at 4°C for ...

Embodiment 3

[0073] Embodiment three, the preparation of chromium ion immunogold standard test paper

[0074] (1) Colloidal gold solution preparation. Prepared by the citrate reduction method, take a 1000 mL Erlenmeyer flask, add 975 mL of double distilled water to boil, add 15 mL of trisodium citrate and continue heating for 5 min, add 10 mL of chloroauric acid with a mass concentration of 1%, and continue heating to the solution It is orange-red, and after cooling to room temperature, store it at 4°C for later use;

[0075] (2) Preparation of gold-labeled antibody. Centrifuge the monoclonal antibody solution to be labeled at 10,000 r / min for 30 min, and use 0.1 mol / L K 2 CO 3 Adjust the pH value of the colloidal gold solution to 8.2, add the monoclonal antibody solution with a concentration of 0.2 mg / mL dropwise under magnetic stirring, incubate at room temperature for 20 min, centrifuge at 10,000 r / min for 30 min, discard the supernatant, and use the precipitate for 0.02 mol / L Na 2...

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Abstract

The invention discloses a gold-labeled test strip for rapid detection of chromium ions as well as a preparation method and an application thereof. The gold-labeled test strip is characterized in that a bottom layer is a support layer, a middle layer is an absorption layer, a protection film is fixed on the absorption layer, the absorption layer is sequentially provided with a sample pad, a gold-labeled antibody binding pad, and a cellulose membrane layer from a test end as well as a water absorption pad at a handle end, detection prints printed by a carrier protein solution coupled with the chromium ions are arranged on the cellulose membrane layer, and contrast prints printed by rabbit-anti-mouse or goat-anti-mouse IgG (Intravenous Gamma Globulin) antibody solution are arranged on the cellulose membrane layer; colloidal gold-labeled chromium ion monoclonal antibodies are coated in the gold-labeled antibody binding pad. The gold-labeled test strip can be used for rapidly detecting pollution residues of the chromium ions in soil, water and food, has the advantages of specificity, sensitivity, rapidness, simplicity, convenience, visual and intuitional result and the like, not only can be used for screening large-batch samples, but also can be used for rapidly detecting small-batch samples and is wide in applicable range.

Description

technical field [0001] The invention belongs to the technical field of immunology and hygienic inspection, and in particular relates to a gold standard test paper for rapid detection of chromium ions, a preparation method and application thereof. Background technique [0002] Chromium is an element that widely exists in nature, distributed in soil, air, water and organisms, mainly in the form of trivalent chromium (Cr 3+ ) and hexavalent chromium (Cr 6+ ) exists in the form of Chromium is one of the essential trace elements for humans and animals, Cr 3+ Has the activity of glucose tolerance factor (GTF), participates in the regulation of cholesterol and glucose metabolism in organisms, Cr 3+ Lack of it will cause fat deposition on the inner wall of blood vessels in the body and lead to arteriosclerosis. But Cr 3+ and Cr 6+ Another toxic substance, Cr 6+ The toxicity is Cr 3+ 100 times more toxic, many studies have confirmed that Cr 3+ Carcinogenic and mutagenic; Cr ...

Claims

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Application Information

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IPC IPC(8): G01N33/577
Inventor 张海棠王自良王淑云姜金庆范国英丁函
Owner HENAN INST OF SCI & TECH
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