Developed rapid and simple RP-HPLC method for simultaneous separation and quantification of bovine milk protein fractions and their genetic variants

Henrietta Buzás; Rita Székelyhidi; Gábor Szafner; Katalin Szabó; Judit Süle; Solveig Bukovics; Attila József Kovács

doi:10.1016/j.ab.2022.114939

Developed rapid and simple RP-HPLC method for simultaneous separation and quantification of bovine milk protein fractions and their genetic variants

Anal Biochem. 2022 Dec 1:658:114939. doi: 10.1016/j.ab.2022.114939. Epub 2022 Oct 4.

Authors

Henrietta Buzás¹, Rita Székelyhidi², Gábor Szafner³, Katalin Szabó³, Judit Süle³, Solveig Bukovics³, Attila József Kovács⁴

Affiliations

¹ Department of Biosystems and Food Engineering Faculty of Agricultural and Food Sciences, Széchenyi István University, 1 Vár Square, H-9200, Mosonmagyaróvár, Hungary; Hungarian Dairy Research Institute Ltd, 24 Lucsony Street, H-9200, Mosonmagyaróvár, Hungary. Electronic address: hbuzas@mtki.hu.
² Department of Food Science, Faculty of Agricultural and Food Sciences, Széchenyi István University, 15-17 Lucsony Street, H-9200, Mosonmagyaróvár, Hungary.
³ Hungarian Dairy Research Institute Ltd, 24 Lucsony Street, H-9200, Mosonmagyaróvár, Hungary.
⁴ Department of Biosystems and Food Engineering Faculty of Agricultural and Food Sciences, Széchenyi István University, 1 Vár Square, H-9200, Mosonmagyaróvár, Hungary.

PMID: 36206846
DOI: 10.1016/j.ab.2022.114939

Abstract

The aim was to develop a reliable rapid reversed-phase high-performance liquid chromatography (RP-HPLC) method to simultaneously determine the main bovine milk protein fractions, including their genetic variants. Compared to the previous studies, our method is able to separate the main protein fractions within 20 min of total run time. The method validation consisted of testing repeatability, reproducibility linearity, repeatability, and accuracy. The procedure was developed using raw individual, bulk, and commercially available heat-treated cow milk samples. The RSD of peak areas ranged from 1.43 to 3.16% within analytical day and from 3.29 to 6.70% across analytical days. The method can be applied to investigate both raw and heat-treated milk samples.

Keywords: Acetonitrile; Bis-Tris; Bronopol; DL-1,4-Dithiothreitol; Guanidine hydrochloride; PubChem CID: 2450; PubChem CID: 446094; PubChem CID: 5742; PubChem CID: 6342; PubChem CID: 6422; PubChem CID: 71474; PubChem CID: 81462; RP-HPLC; Trifluoroacetic acid (TFA); Trisodium citrate dihydrate; caseins; milk protein fractionation and quantification; whey proteins; β-casein A1- A2.

MeSH terms

Animals
Cattle
Chromatography, High Pressure Liquid / methods
Chromatography, Reverse-Phase / methods
Female
Milk Proteins* / analysis
Milk* / chemistry
Reproducibility of Results

Substances

Milk Proteins