GB/T 38163-2019 in English

VALID

Determination of allergenic protein—LC-MS/MS method

Issued on:2019-10-18
Implemented on:2019-10-18
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $250.00
$243.00

Standard No:	GB/T 38163-2019
Document status:	VALID
Title in English:	Determination of allergenic protein—LC-MS/MS method
Title in Chinese:	常见过敏蛋白的测定　液相色谱-串联质谱法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2019-10-18
Implemented on:	2019-10-18
ICS Classification:	07.080-Biology. Botany. Zoology
Chinese Classification:	A40-Basic Subject in general
Professional Classification:	GB-National Standard
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Commonly used protein concentration determination methods What are the commonly used protein separation methods? 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《GB/T 38163-2019常见过敏蛋白的测定液相色谱-串联质谱法》由TC387（全国生化检测标准化技术委员会）归口，主管部门为国家标准化管理委员会。

Introduction

Analysis of standard technical framework

Technical elements	GB/T 38163-2019	International general method	Innovation
Detection objects	7 types of allergen proteins	Single-type allergen detection	Multi-target simultaneous screening
Feature markers	14 characteristic peptides	Intact protein detection	TrypsinEnzymatic digestion strategy
Quantitative linear range	5-1600mg/kg	50-1000mg/kg	Sensitivity increased by 10 times

Core technology innovation

Characteristic peptide screening system

The 14 characteristic peptide databases established in the standard (see Table A.1) have the characteristics of strong specificity:

Milk α-S1 casein: FFVAPFPEVFGK (m/z 692.9→920.5)
Peanut Ara h 3/4: RPFYSNAPQEIFIQQGR (m/z 684.4→601.3)

Mass spectrometry parameter optimization

Take triple quadrupole mass spectrometer as an example, the key parameter settings are:

Ionization mode	ESI+
Collision energy	20-48eV (adjusted according to the peptide segment)
Ion source temperature	500℃

Method validation data

Performance index comparison

Allergens	Detection limit (mg/kg)	Quantification limit (mg/kg)	Recovery range (%)
Milk	1.0	5.0	75.6-108.5
Peanut	2.5	10.0	70.9-216.0

Matrix Interference Solution

Using ultrafiltration centrifuge tubes (10kD molecular weight cutoff) and Tris-HCl buffer system, we can effectively eliminate:

Sugar interference in biscuit matrix
Polyphenols in chocolate

Implementation Recommendations

Critical Control Points

Enzyme hydrolysis time is strictly controlled at 4±0.5h (37℃)
Use freshly prepared DTT solution (≤1 month)
Mass spectrometer calibration must meet the resolution ≥12000

Standardized operation

It is recommended to establish a LC-MS/MS system suitability test:

Sensitivity	S/N≥10 (1pg reserpine)
Mass accuracy	≤3ppm

More samples: DL/T 5154-2012 JJG 21-2008 JGJ 94-2008

Sample only — not a preview of GB/T 38163-2019

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