Immunoaffinity stir bar for adsorbing raxofelast acid lactone compound, and preparation method and application thereof

A technology for ester compounds and rasolic acid, applied in the field of food safety testing, can solve the problems of difficult qualitative confirmation, cumbersome and time-consuming operation, short separation time, etc., achieve strong retention and concentration ability, simplify pretreatment process, and save organic solvents Effect

Inactive Publication Date: 2015-06-10
TIANJIN AGRICULTURE COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The TLC method is low in cost, but the sensitivity and selectivity are not high; the ELISA method is highly sensitive, but due to its cross-reaction characteristics, the selectivity is poor; the GC-MS method has good sensitivity and selectivity, but requires derivatization, operation It is tedious and time-consuming; HPLC method can be directly determined, but for complex matrix samples, impurities interfere greatly, and there are certain difficulties in qualitative confirmation; LC-MS / MS method does not need to achieve complete chromatographic separation of the analyte and interfering components. It can confirm the structure of the compound with higher sensitivity and accuracy, but it is expensive and often limited in practical applications
Biological sample pretreatment technologies mainly include liquid-liquid extraction and solid-phase extraction. Liquid-liquid extraction is cumbersome and requires a large amount of organic solvents. Solid-phase extraction has the advantages of less organic solvents, strong selectivity, and short separation time. At present, C18 and HLB solid-phase extraction cartridges are commonly used for the determination of zearalanol. Although common solid-phase extraction cartridges are convenient and fast to operate, they have poor specificity.
The Immunoaffinity SPME technology was first proposed in 2001, and the use of Immunoaffinity stir bar to purify rexolactone compounds has not yet been reported, and there is no commercialized Immunoaffinity stir bar for sale

Method used

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  • Immunoaffinity stir bar for adsorbing raxofelast acid lactone compound, and preparation method and application thereof
  • Immunoaffinity stir bar for adsorbing raxofelast acid lactone compound, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Preparation of zearalanol monoclonal antibody

[0043] Synthesis of ZER-BSA immunogen: Dissolve 5 mg zearalanol (α-zearalanol, ZER) in 3 mL pyridine, add 3.3 mg carboxymethoxylamine hemihydrochloride (Carboxymethoxylamine hemihydrochloride), stirred at room temperature for 24 h, dried under vacuum, added 5 mL of deionized water to dissolve the residue, and adjusted the pH to 8 with 2 mol / L NaOH. Extract with ethyl acetate to remove unreacted ZER. Then adjust the pH to 3 with 2 mol / L HCl, extract 4 times with ethyl acetate, anhydrous Na 2 SO 4 Filter, dry the extract in vacuo, and recrystallize from methanol-water solution (1:1, v / v). Dissolve 20 mg of BSA in 1.5 mL of PBS solution (0.02 mol / L, pH 7.2), dissolve about 5 mg of ZER crystals in 1 mL of 1,4-Dioxacyclohexane, and mix The two solutions were slowly added dropwise N-hydroxysuccinimide (N-Hydroxysuccinimide, NHS) and N,N-dicyclohexylcarbodiimide (N,N'-dicyclohexylcarbodiimide, DCC) solution (3 mg NHS and ...

Embodiment 2

[0059] Comparative Test

[0060]

Embodiment 3

[0062] Immunoaffinity stir bar was used for the extraction of racolactone compounds in water. The blank water sample was passed through a 0.45 μm water-based filter membrane to remove impurities, and 5 mL of the filtered water sample was accurately weighed, placed in a 10 mL centrifuge tube, an appropriate amount of risolactone compound standard solution was added, vortexed, and the Immunoaffinity glass rods were placed in the water sample after removal and mixing, and oscillated on a shaker for 30 min. Take the shaken glass rod out of the centrifuge tube, wash it with ultrapure water, put it into a 2 mL centrifuge tube filled with 700 μL of methanol, shake it on a shaker for 20 min to elute, dry it with nitrogen, and flow Phase reconstitution, 0.22 μm microporous membrane filtration, high performance liquid chromatography-mass spectrometry for separation and detection. The results showed that zearalenol, β-zearalenol, zearalenone, zearalenone, α-zearalenol and β-zearalenol c...

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PUM

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Abstract

The invention discloses an immunoaffinity stir bar for adsorbing raxofelast acid lactone compound, and a preparation method and an application thereof. The immunoaffinity stir bar for adsorbing raxofelast acid lactone compound is composed of solid phase carrier and zeranol murine monoclonal antibody coupled with the same, wherein the ratio of the coupling length of the solid phase carrier and coupled zeranol murine monoclonal antibody to the weight thereof is 1 cm: 2 mg. Through combining with a chromatography, an extracting method can effectively separate and condense the ingredients to be detected in a sample, the raxofelast acid lactone compound detection is facilitated, the disadvantages of less information amount, low quantification precision, low physico-chemical method selectivity and the like due to using the simplex immunoassay to directly detect the sample are overcome, and the analysis mechanism complementarity between the immunological technique and conventional physical and chemical technology is reflected.

Description

technical field [0001] The invention belongs to the technical field of food safety detection, and relates to an immunoaffinity stirring rod and a preparation method thereof. In particular, it relates to an immunoaffinity stirring bar for absorbing racolactone compounds, its preparation method and application. Background technique [0002] With the development of my country's economy and the improvement of people's living standards, the demand for various animal-derived foods is gradually increasing, and food safety issues have naturally become the focus of everyone's attention. In recent years, there have been many reports about the harm to human health caused by veterinary drug residues, and relevant food safety departments are also committed to the research of detection methods. Due to the complex components of biological samples, the easy combination of drugs and sample components, and the low concentration of residual drugs, it has brought great difficulties to the extr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543
CPCG01N30/06G01N30/14G01N2030/062
Inventor 李存姚凯赵思俊张伟杨琳燕金天明李建军王翠翠
Owner TIANJIN AGRICULTURE COLLEGE
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