Characterization and determination of bovine immunoglobulin G subtypes in milk and dairy products by UPLC-MS

Jingshun Zhang; Qin Chen; Ying Zhou; Yunzhen Zhang; Yiping Ren; Lianliang Liu

doi:10.1016/j.foodchem.2022.133170

Characterization and determination of bovine immunoglobulin G subtypes in milk and dairy products by UPLC-MS

Food Chem. 2022 Oct 1:390:133170. doi: 10.1016/j.foodchem.2022.133170. Epub 2022 May 7.

Authors

Jingshun Zhang¹, Qin Chen², Ying Zhou³, Yunzhen Zhang², Yiping Ren⁴, Lianliang Liu⁵

Affiliations

¹ Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, China. Electronic address: jshunzhang@163.com.
² Ningbo University, College of food and pharmaceutical sciences, Deep Processing Technology Key Laboratory of Zhejiang Province Animal Protein Food, Ningbo University, Ningbo 315832, Zhejiang Province, PR China.
³ Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, China; Zhejiang University of Technology, School of food science and technology, Hangzhou 310014, China.
⁴ Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, China. Electronic address: yipingren@yeah.net.
⁵ Ningbo University, College of food and pharmaceutical sciences, Deep Processing Technology Key Laboratory of Zhejiang Province Animal Protein Food, Ningbo University, Ningbo 315832, Zhejiang Province, PR China. Electronic address: hahaliang408@126.com.

PMID: 35597093
DOI: 10.1016/j.foodchem.2022.133170

Abstract

In this study, ultra-high performance liquid chromatography mass spectrometry (UPLC-MS) method was established for the characterization and quantitative determination of immunoglobulin G (IgG) subtypes (IgG1, IgG2, IgG3) in bovine dairy products. High-resolution mass spectrometry (HRMS) was applied to qualitatively confirm the theoretical peptides with specificity, enzymatic hydrolysis curve and stability among in heavy chain constant (CH1, CH2 and CH3) regions. The characteristic peptides VHNEGLPAPIVR, EPSVFIFPPKPK, GLPAPIVR, VVSALR were screened to quantitative analysis bovine IgG1, IgG2, IgG3 and the total amount of bovine IgG1 and IgG3, respectively. Isotope-labeled peptides were obtained by isotope dimethylation reaction, which aimed to correct the matrix effects. The results showed that the recovery was between 98.7% and 103.5%, and the precision of inter-day and intra-day was less than 6.8%. Moreover, this method had good linearity (R² ≥ 0.999). Therefore, this research provided an effective method for quantitatively detecting bovine IgG subtypes in milk and dairy products.

Keywords: Characteristic peptides; Enzymatic hydrolysis; High-resolution mass spectrometry (HRMS); Immunoglobulin G; Isotope dimethylation; Ultra-high performance liquid chromatography (UPLC).

MeSH terms

Animals
Cattle
Chromatography, High Pressure Liquid / methods
Chromatography, Liquid
Immunoglobulin G / analysis
Isotopes / analysis
Milk* / chemistry
Peptides / analysis
Tandem Mass Spectrometry* / methods

Substances

Immunoglobulin G
Isotopes
Peptides