Abstract
The illegal or unlabelled addition of plant protein in milk can cause serious anaphylaxis. For sustainable food security, it is therefore important to develop a methodology to detect non-milk protein in milk products. This research aims to differentiate milk adulterated with plant protein using two-dimensional gel electrophoresis (2-DE) coupled with mass spectrometry. According to the protein spots highlighted on the gel of adulterated milk, β-conglycinin and glycinin were detected in milk adulterated with soy protein, while legumin, vicilin, and convicilin indicated the addition of pea protein, and β-amylase and serpin marked wheat protein. These results suggest that a 2-DE-based protein profile is a useful method to identify milk adulterated with soy and pea protein, with a detection limit of 4% plant protein in the total protein.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Becker D, Wieser H, Koehler P, Folck A, Muhling KH, Zorb C (2012) Protein composition and techno-functional properties of transgenic wheat with reduced alpha-gliadin content obtained by RNA interference. J Appl Bot Food Qual 85:23–33
Berggren K et al (2000) Background-free, high sensitivity staining of proteins in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gels using a luminescent ruthenium complex. Electrophoresis 21:2509–2521. https://doi.org/10.1002/1522-2683(20000701)21:12<2509::AID-ELPS2509>3.0.CO;2-9
Chalvon-Demersay T, Azzout-Marniche D, Arfsten J, Egli L, Gaudichon C, Karagounis LG, Tome D (2017) A systematic review of the effects of plant compared with animal protein sources on features of metabolic syndrome. J Nutr 147:281–292. https://doi.org/10.3945/jn.116.239574
Cordewener JHG, Luykx DMAM, Frankhuizen R, Bremer MGEG, Hooijerink H, America AHP (2009) Untargeted LC-Q-TOF mass spectrometry method for the detection of adulterations in skimmed-milk powder. J Sep Sci 32:1216–1223. https://doi.org/10.1002/jssc.200800568
Gatehouse JA, Croy RD, Boulter D (1980) Isoelectric-focusing properties and carbohydrate content of pea (Pisum sativum) legumin. Biochem J 185:497–503. https://doi.org/10.1042/bj1850497
Haasnoot W, Olieman K, Cazemier G, Verheijen R (2001) Direct biosensor immunoassays for the detection of nonmilk proteins in milk powder. J Agric Food Chem 49:5201–5206. https://doi.org/10.1021/Jf010440p
Hurkman WJ, Tanaka CK (2004) Improved methods for separation of wheat endosperm proteins and analysis by two-dimensional gel electrophoresis. J Cereal Sci 40:295–299. https://doi.org/10.1016/j.jcs.2004.09.001
Jaiswal P, Jha SN, Borah A, Gautam A, Grewal MK, Jindal G (2015) Detection and quantification of soymilk in cow-buffalo milk using Attenuated Total Reflectance Fourier Transform Infrared spectroscopy (ATR-FTIR). Food Chem 168:41–47. https://doi.org/10.1016/j.foodchem.2014.07.010
Krusa M, Torre M, Marina ML (2000) A reversed-phase high-performance liquid chromatographic method for the determination of soya bean proteins in bovine milks. Anal Chem 72:1814–1818. https://doi.org/10.1021/Ac990776m
Luykx DMAM, Cordewener JHG, Ferranti P, Frankhuizen R, Bremer MGEG, Hooijerink H, America AHP (2007) Identification of plant proteins in adulterated skimmed milk powder by high-performance liquid chromatography-mass spectrometry. J Chromatogr A 1164:189–197. https://doi.org/10.1016/j.chroma.2007.07.017
Nakamura R, Teshima R (2013) Proteomics-based allergen analysis in plants. J Proteomics 93:40–49. https://doi.org/10.1016/j.jprot.2013.03.018
Nishinari K, Fang Y, Guo S, Phillips GO (2014) Soy proteins: a review on composition, aggregation and emulsification. Food Hydrocolloids 39:301–318. https://doi.org/10.1016/j.foodhyd.2014.01.013
Parmar N et al (2014) Evaluation of physicochemical, textural, mineral and protein characteristics of kidney bean grown at Himalayan region. Food Res Int 66:45–57. https://doi.org/10.1016/j.foodres.2014.08.048
Pomastowski P, Buszewski B (2014) Two-dimensional gel electrophoresis in the light of new developments. TrAC Trends Anal Chem 53:167–177. https://doi.org/10.1016/j.trac.2013.09.010
Samoto M, Maebuchi M, Miyazaki C, Kugitani H, Kohno M, Hirotsuka M, Kito M (2007) Abundant proteins associated with lecithin in soy protein isolate. Food Chem 102:317–322. https://doi.org/10.1016/j.foodchem.2006.05.054
Sanchez L, Perez MD, Puyol P, Calvo M, Brett G (2002) Determination of vegetal proteins in milk powder by enzyme-linked immunosorbent assay: interlaboratory study. J AOAC Int 85:1390–1397
Scholl PF, Farris SM, Mossoba MM (2014) Rapid turbidimetric detection of milk powder adulteration with plant proteins. J Agric Food Chem 62:1498–1505. https://doi.org/10.1021/jf405617f
Sirtori E, Isak I, Resta D, Boschin G, Arnoldi A (2012) Mechanical and thermal processing effects on protein integrity and peptide fingerprint of pea protein isolate. Food Chem 134:113–121. https://doi.org/10.1016/j.foodchem.2012.02.073
Tzitzikas EN, Vincken JP, de Groot J, Gruppen H, Visser RG (2006) Genetic variation in pea seed globulin composition. J Agric Food Chem 54:425–433. https://doi.org/10.1021/jf0519008
Wang WQ, Moller IM, Song SQ (2012) Proteomic analysis of embryonic axis of Pisum sativum seeds during germination and identification of proteins associated with loss of desiccation tolerance. J Proteomics 77:68–86. https://doi.org/10.1016/j.jprot.2012.07.005
Yang YX, Zheng N, Yang JH, Bu DP, Wang JQ, Ma L, Sun P (2014) Animal species milk identification by comparison of two-dimensional gel map profile and mass spectrometry approach. Int Dairy J 35:15–20. https://doi.org/10.1016/j.idairyj.2013.09.008
Zarkadas CG, Gagnon C, Poysa V, Khanizadeh S, Cober ER, Chang V, Gleddie S (2007) Protein quality and identification of the storage protein subunits of tofu and null soybean genotypes, using amino acid analysis, one- and two-dimensional gel electrophoresis, and tandem mass spectrometry. Food Res Int 40:111–128. https://doi.org/10.1016/j.foodres.2006.08.005
Acknowledgements
This study was financially supported by Agro-Scientific Research in the Public Interest (201403071), the Agricultural Science and Technology Innovation Program (ASTIP-IAS12), Modern Agro-Industry Technology Research System of the People’s Republic of China (CARS-37), and the Project of Risk Assessment on Raw Milk (GJFP2016008), granted by the Chinese Ministry of Agriculture.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yang, J., Zheng, N., Yang, Y. et al. Detection of plant protein adulterated in fluid milk using two-dimensional gel electrophoresis combined with mass spectrometry. J Food Sci Technol 55, 2721–2728 (2018). https://doi.org/10.1007/s13197-018-3194-y
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13197-018-3194-y