GH/T 1315-2020 in English

VALID

Determination of casein in honey—Enzyme-linked immunosorbent assay method

Issued on:2020-12-07
Implemented on:2021-03-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $110.00
$107.00

Standard No:	GH/T 1315-2020
Document status:	VALID
Title in English:	Determination of casein in honey—Enzyme-linked immunosorbent assay method
Title in Chinese:	蜂蜜中酪蛋白的测定酶联免疫法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2020-12-07
Implemented on:	2021-03-01
ICS Classification:	67.180.10-Sugar and sugar products
Chinese Classification:	X31-Sugar refining
Related Keywords:	enzyme-linked immunosorbent assay method introduction technical background enzyme-linked immunosorbent assay vitex honey samples ridascreen® fast casein casein content
Related Topics:	enzyme protein immunity Immunization Honey Immunization mass spectrometry immunoassay casein honey ELISA ELISA immunity? Test immunity Rapid Immunization Protein+ protease method protease law mass spectrometry immunoassay microgram+protein+ casein sample protease on hand 666 ELISA Sodium content determination method in honey v8 protease protein assay How to detect protease Protease detection method pronase How to detect protease ELISA Enzyme-linked samples Enzyme cheese casein+ microwell strip immunomass spectrometry Caseinase Enzyme Immunization Mass spectrometry and immunoassays mass spectrometry immunoassay Chemiluminescent ELISA porin Immunogenic ELISA bacterial protease ELISA plate porin chemical enzyme linked Immuno-EM moisture in honey Casein Luminosity GH 2 enzyme-linked method medium density protein Collagenase III gh/t Enzyme Immunization microgram protein various proteases enzyme-linked method indirect ELISA ELISA and Chemiluminescence human ELISA centrosomal protein ELISA double antibody Enzyme-linked system honey sample protein standard Annexin protein in sludge enzyme immunoassay ELISA testosterone antigen ELISA Proteases in Food enzymatic tyrosol protein in seeds Enzyme-linked immunosorbent immunoassay and immune response test Chemiluminescent ELISA ELISA protein assay homogeneous enzyme immunoassay ELISA IgG ELISA protease Immunoglobulinase radioimmunoassay Methods for determining the amino acid sequence of proteases Protein cross-linking ELISA capture method Action site of neutral protease protein kinase stn7 protease cleavage process Application of protease in flour The difference between ribozymes and proteases Caseinase activity Characteristics of enzyme-linked immunoassay methodology Elastase action Disadvantages of enzyme-linked immunoassay NF1 immunotherapy Precautions for enzyme-linked immunoassay Protein quantification after immunoprecipitation enzyme-linked immunoassay chemiluminescence assay A method for measuring enzyme activity of protease Protease enzyme activity assay method Where enzyme-linked immunoassay detects proteases Enzyme-linked immunoassay results determination method Common methods for measuring protease activity Method for determination of protease activity in dairy products Methods for protease isolation Protease processing Preparation method of casein solution for measuring protease activity Protease determination method Determination method of aluminum content in honey Methods for Determining Collagen Cross-Links Determination method of chymotrypsin content Vaccine impurity protein detection method Is enzymatic method an immunological method? Main production methods of protein enzymes Methods for detecting immunoglobulins in food Protease activity detection method azocasein Quantitative detection method of protein in drugs Protein detection methods to avoid non-protein Enzyme-linked immunoassay method validation Serine protease assay method Principles of several enzyme-linked immunoassay methods Enzyme protein content determination method Quantitative Immunology Industry What are the types of enzyme-linked immunoassay methods? Caseinase assay method Enzyme detection methods Immunological methods radioactive enzyme-linked immunoassay Chymotrypsin assay method What methods does immunization include? Method for detecting protease using microplate reader Protease assay method Specific protein industry Characterization methods of keratinase gh/t 1161 Protease Country Methods for determining the isoelectric point of proteins

本文件规定了蜂蜜中酪蛋白的酶联免疫测定方法。
本文件适用于洋槐蜜、油菜蜜、椴树蜜、荆条蜜中酪蛋白的测定。

Introduction

Technical background and evolution of standards

my country's bee product safety standard system is constantly improving. GH/T 1315-2020 established a special detection method for casein in honey for the first time. Compared with traditional electrophoresis, enzyme-linked immunosorbent assay has the characteristics of high sensitivity (1.5 mg/kg) and strong specificity, which can effectively identify honey adulterated with milk protein.

Core Detection Principle

Adopting the "sandwich" ELISA principle:

Steps	Reactants	Temperature conditions	Time control
Antigen-antibody binding	Casein + coated antibody	20-25℃	10min
Enzyme-labeled antibody binding	HRP-labeled secondary antibody	20-25℃	10min
Color reaction	TMB substrate	Avoid light 20-25℃	10min

Key Instruments and Reagents Specifications

A refrigerated centrifuge (≥3000g) and microplate reader (450nm) must be used. The kit recommends RIDASCREEN® FAST Casein, and the standard gradient is 0-13.5µg/mL.

Actual Application Case

A testing agency conducted three parallel determinations on vitex honey samples and found that the casein content was 2.37mg/kg (RSD=8.2%), which met the precision requirements (≤20%).

Analysis of key points of operation

Sample extraction: The 60℃ water bath needs to be precisely controlled at ±1℃. After centrifugation, take the supernatant and dilute it 3 times
Plate washing control: Add 250µL of washing buffer each time, and tap the plate evenly
Color termination: The reading must be completed within 10 minutes after adding the stop solution

Implementation suggestions

Laboratories should focus on:

Six concentration points (0-13.5µg/mL) must be included when establishing the standard curve
Negative quality control of the kit must be carried out simultaneously for each batch of tests
For high-concentration samples (＞40.5mg/kg), they should be appropriately diluted and retested

More samples: DL/T 5154-2012 GB 18306-2015 JC/T 2453-2018

Sample only — not a preview of GH/T 1315-2020

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