Method for detecting fish parvalbumin through liquid chromatography tandem mass spectrometry

Abstract

The invention provides a method for detecting fish parvalbumin through liquid chromatography tandem mass spectrometry. The method comprises the following steps: step 1: choosing a characteristic peptide fragment; step 2: making a standard curve: taking the concentration value of a quantitative peptide fragment standard product as the x-coordinate and a quantitative daughter ion peak area as the y-coordinate to draw the standard curve, and obtaining an equation; step 3: processing and implementing enzymolysis on a to-be-detected fish meat sample to obtain a liquid supernatant, implementing HPLC-MRM-MS/MS detection on the liquid supernatant, obtaining the daughter ion peak area of the quantitative peptide fragment in the fish meat sample, resolving the concentration of the quantitative peptide fragment in the sample, and further obtaining the concentration of the parvalbumin in the sample. The invention aims at establishing a liquid chromatography tandem mass spectrometry method, and themethod has the advantages of accuracy, precision, sensitivity and the like.

Description

technical field [0001] The invention belongs to a method for detecting food allergens, and in particular relates to a method for detecting parvalbumin, the main allergen of fish, by liquid chromatography tandem mass spectrometry. Background technique [0002] Although there are many reports on the detection methods of parvalbumin in fish at home and abroad, there are mainly enzyme-linked immunoassay (ELISA), polymerase chain reaction (PCR) and liquid chromatography tandem mass spectrometry (HPLC-MS / MS). . Among them, the ELISA method is the most commonly used method. It is based on the principle of antigen-antibody specific binding, and quantitatively detects allergens through double-antibody sandwich method, indirect method and competition method. However, it has the disadvantages of poor repeatability and specificity: the same species However, the results of ELISA kits from different manufacturers are quite different, mainly due to the different sources and titers of anti...

AI Technical Summary

Problems solved by technology

Among them, the ELISA method is the most commonly used method. It is based on the principle of antigen-antibody specific binding, and quantitatively detects allergens through double-antibody sandwich method, indirect method and competition method. However, it has the disadvantages of poor repeatability and specificity: the same species However, the results of ELISA kits from different manufacturers are quite different, mainly due to the different sources and titers of antibodies; in addition, matrix interference in food will also affect the accuracy of the ELISA method. False positives will occur when the antibody is bound to a foreign protein, and false negatives will appear when the endogenous interfering substances in the matrix affect the binding of the allergen to the antibody, especially after the food is processed (cooking, frying, high pressure), the allergen is Modification, the structure changes so that it cannot specifically bind to the antibody

So far, domestic and foreign research on the detection of parvalbumin in fish meat by liquid chromatography tandem mass spectrometry has mainly focused on the qualitative aspect of parvalbumin, and no quantitative research has been reported.

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