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Gold label test paper for rapidly detecting molybdic ions, well as preparation method and application thereof

A technology of gold standard test paper and molybdenum ions, which is applied in the fields of immunology and sanitary inspection, can solve the problems that the equipment is not easy to be equipped in general laboratories, restricts wide use, and is difficult to be widely used, so as to save testing costs and be easy to operate and control , the effect of saving detection time

Inactive Publication Date: 2015-06-10
HENAN INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have their own advantages and disadvantages. Ultraviolet spectrophotometry is simple, fast, and less disturbing, but its sensitivity is not high; electrochemical analysis is sensitive, simple, etc., and requires high technical requirements for operators; atomic absorption spectroscopy, inductively coupled plasma Bulk mass spectrometry, inductively coupled plasma atomic emission spectrometry, and hydride generation-atomic fluorescence spectrometry have the advantages of good selectivity, high measurement accuracy, simplicity, and speed, but the instruments used are relatively expensive and can only be tested in laboratories. And the technical requirements for operators are relatively strict, which limits its wide application; neutron activation analysis is an analysis method based on nuclear reactions, which has the advantages of trace, fast, accurate and non-destructive and can analyze multiple elements at the same time. However, the required equipment is not easily available in general laboratories, and it is difficult to be widely used

Method used

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  • Gold label test paper for rapidly detecting molybdic ions, well as preparation method and application thereof
  • Gold label test paper for rapidly detecting molybdic ions, well as preparation method and application thereof
  • Gold label test paper for rapidly detecting molybdic ions, well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1. Preparation of hexavalent molybdenum ion artificial immune antigen

[0048] Using the isothiocyanate method. Take 20mL molybdic acid and mix it with 10mmol / L 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES) buffer with a pH of 8.0 to form Mo 6+ Solution; Weigh 10 mg of benzyl ethylene diamine tetraacetic acid (ITCBE) isothiocyanate and dissolve in 1 mL of dimethyl sulfoxide (DMSO) to form a metal chelating agent solution; 6+ After the solution and the metal chelating agent solution are mixed, adjust the pH value of the solution to 7.4, stir for 24 h at 25°C, and rotate at 1000 r / min to form Mo 6+ -ITCBE chelated hapten mother liquor;

[0049] Weigh 20 mg of carrier protein BSA or OVA and dissolve it in 1 mL of HEPES buffer with a concentration of 10 mmol / L and a pH of 8.0 to form a carrier protein solution; take 1 mL of Mo 6+ -ITCBE chelated hapten solution was added to 1mL carrier protein solution, adjusted the pH to 9.0, stirred at 1000 r / min for 24 h at roo...

Embodiment 2

[0050] Example 2: Identification of hexavalent molybdenum ion artificial immune antigen

[0051] Determination of Mo by the quinolinic acid method 6+ -The concentration of carrier protein BSA in ITCBE-BSA. With BSA as the standard protein, the concentration detection standard curve was constructed by the quinolinic acid method. The linear equation of the BSA concentration standard curve is: y=0.0004x+0.0072, R 2 =0.9987, where y is the absorbance of the sample at a wavelength of 562nm, and x is the protein concentration of the sample.

[0052] Determination of Mo by ICP-AES 6+ -ITCBE-BSA in Mo 6+ concentration. Add 100 μg / mL Mo 6+ Standard stock solution, diluted with 2% nitric acid to a concentration gradient of 0 μg / mL, 0.25 μg / mL, 0.5 μg / mL, 1 μg / mL, 2 μg / mL, 4 μg / mL, the instrument software automatically draws the standard curve , And get the linear regression equation; the sample solution is diluted 50 times, and the measurement is performed under the optimal experimental c...

Embodiment 3

[0053] Example 3: Preparation of anti-hexavalent molybdenum ion monoclonal antibody

[0054] (1) Animal immunity. Immunize Balb / c mice with Mo 6+ -ITCBE-BSA immunized 5 female Balb / C mice of 6 weeks old, 50 μg·0.2 mL / mouse, and injected multiple points under the skin on the back. First immunization, dilute Mo with sterile PBS 6+ -ITCBE-BSA, mixed with the same amount of CFA to emulsify; boost immunity, dilute Mo with sterile PBS 6+ -ITCBE-BSA, mixed and emulsified with the same amount of IFA, the second immunization is carried out 3 weeks after the first immunization, and the immunization is carried out at 2 weeks intervals thereafter, a total of 5 immunizations, 10 days after the third immunization, the tail is removed and the blood is taken, 37 ℃ water bath Place it overnight at 4°C for 30 minutes, centrifuge at 4000 r / min 4°C for 5 minutes, take the supernatant, and store at -20°C for later use.

[0055] (2) Selection of spare mice for cell fusion. Indirect ELISA to detect Mo...

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Abstract

The invention discloses gold label test paper for rapidly detecting molybdic ions, as well as a preparation method and an application thereof. The bottom layer of the gold label test paper is a supporting layer, the middle layer of the gold label test paper is an adsorption layer, a protective film is fixedly arranged on the absorption layer, the absorption layer comprises a sample cushion, a gold label antibody combined cushion, a cellulose film layer and a handle-end absorbent cushion from a test end in sequence, and detection imprints printed by a carrier protein solution of coupling molybdic ions and contrast imprints printed by a rabbit anti-mouse IgG antibody solution are arranged on the absorbent cushion; a molybdic ion monoclonal antibody marked by colloidal gold is wrapped in the gold label antibody combined cushion. The gold label test paper can realize the rapid detection of molybdenum ion residual contamination in oil, water and food, and has the advantages of being specific, sensitive, rapid, easy and convenient, vivid and visual in result, economic, and the like; the gold label test paper can be used for screening of samples in large batch, also can be used for the rapid detection on samples in small batch, and is wide in application 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 hexavalent molybdenum ions, a preparation method and application thereof. Background technique [0002] my country is rich in molybdenum ore resources, with a total reserve of 8.4 million tons. Molybdenum is a transition metal element, and its compounds present five valence states, of which hexavalent molybdenum is the most stable and is most closely related to human life activities. Overdose will affect the quality of life. Molybdenum is one of the essential trace elements for organisms. Various tissues of the human body contain molybdenum. The total amount in the adult body is 9 mg, and the content in the liver and kidney is the highest. Molybdenum itself has no biological activity. Only when hexavalent molybdenum ions combine with purine to form a coenzyme can it have biological activity. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577
Inventor 张海棠王自良王淑云范国英姜金庆黄华国
Owner HENAN INST OF SCI & TECH
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