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Enzyme-linked immunosorbent assay for measuring content of lincomycin hydrochloride in food

A technology of lincomycin hydrochloride and enzyme-linked immunosorbent adsorption, which is applied in the direction of measuring devices, analytical materials, color/spectral characteristics measurement, etc., can solve the problem that ultraviolet detectors cannot achieve low content detection, increase cumbersome operation, and weak ultraviolet absorption and other problems, to achieve the effect of low test cost, large test volume, and simple sample processing

Inactive Publication Date: 2010-06-09
SICHUAN UNIV
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Problems solved by technology

[0003] The method for detecting lincomycin hydrochloride is mainly high-performance liquid chromatography (HPLC), but because it has only weak ultraviolet absorption at 200-220nm, it cannot meet the requirements of low-level detection with ultraviolet detectors
Gas chromatography (GC) often requires derivatization, which increases the complexity of the operation and the sensitivity is not high
In recent years, liquid chromatography-tandem mass spectrometry (LC-MS / MS) has been used for the detection of lincomycin hydrochloride. Although the sensitivity is high, the instrument is expensive, the pretreatment is complicated, and the operation of professionals is also limited. Application in on-site inspection and sample screening

Method used

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  • Enzyme-linked immunosorbent assay for measuring content of lincomycin hydrochloride in food
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  • Enzyme-linked immunosorbent assay for measuring content of lincomycin hydrochloride in food

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Embodiment

[0034] 1. Preparation of modified lincomycin hydrochloride

[0035] Lincomycin is a small molecule hapten, which cannot directly immunize animals to produce antibodies, but it can be chemically modified, so that the modified product has a connecting bridge whose end group is an active group, so as to be connected with a carrier protein to prepare an immunogen. In order to maintain the characteristic structure of the lincomycin molecule to the greatest extent, the present invention adopts a novel hapten modification method. On the basis of protecting the hydroxyl groups at the 3, 4, and 7 positions, the hydroxyl groups at the 2 positions are protected. Participate in the modification reaction, and finally obtain a hapten derivative with only a terminal carboxyl arm connected to the 2-position of the lincomycin molecule.

[0036] Preparation of modified lincomycin hydrochloride

[0037] Step 1: Dissolve 3-5g lincomycin hydrochloride (LIN) and 5-8mL p-methoxybenzaldehyde in 15-2...

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Abstract

The invention discloses an enzyme-linked immunosorbent assay (EISA) for measuring the content of lincomycin hydrochloride in food. The method is characterized in that a novel hapten modification method is used to synthesize a modifier of lincomycin hydrochloride, and the modifier is linked to the protein to prepare an immunogen and an envelope antigen. A polyclonal antibody of rabbit anti-lincomycin hydrochloride is acquired by an immune animal. The antibody is used for the immunoassay of lincomycin hydrochloride; the concentration range of a standard curve is 1-1000ng / mL, IC50 is 23.7-29.3ng / mL, the cross reactivity of the antibody and the lincomycin hydrochloride is 18.9%, and the antibody and other seven popular antibiotics or hormones hardly have cross reaction. Six kinds of market-sold foods are extracted by using 0.01mol / L HCL after being labeled; an extract is properly diluted and measured by ELISA, the labeling recovery rate is 76.6-117.6%, and the relative standard deviation is 1.7-34.6%. The seven labeled samples are analyzed by using the HPLC and the ELISA methods which have better pertinence (R2=0.9756 and n=7). The EISA established in the invention has reliability, high sensitivity and strong specificity and can be used for detecting the content of lincomycin hydrochloride in food.

Description

technical field [0001] The invention relates to an enzyme-linked immunosorbent assay method (ELISA) for determining the content of lincomycin hydrochloride (LIN) in food, which belongs to the research field of food safety supervision or food analysis. Background technique [0002] Lincomycin hydrochloride (LIN), commonly known as lincomycin or lincomycin, belongs to lincosamine antibiotics, and is a kind of high-efficiency broad-spectrum antibiotic firstly obtained from streptomyces lin variable culture fluid by American Mason et al. It has a good effect on Gram-positive cocci. In addition to being mainly used clinically for various infections caused by sensitive bacteria, it is also widely used in animal feeding and treatment of animal diseases, and can be used as a growth promoter for broiler chickens. Although lincomycin hydrochloride is not very toxic, it is still potentially harmful to humans and animals, such as causing pseudomembranous colitis and bacterial drug resi...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N21/31
Inventor 邓安平王悦秋杨红
Owner SICHUAN UNIV
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