Nothing Special   »   [go: up one dir, main page]
Skip to main content
Springer Nature Link
Log in

Performance of Commercial Colorimetric Assays for Quantitation of Total Soluble Protein in Thermally Treated Milk Samples

  • Published:
Food Analytical Methods Aims and scope Submit manuscript

Abstract

Commercial colorimetric protein assays have been widely used, but their performance for protein quantitation in thermally processed milk samples has not been investigated. This study evaluated four colorimetric assays for quantitation of soluble protein in nonfat milk samples that were subjected to different degrees of heat treatment. Copper ion-based Pierce™ bicinchoninic acid protein assay (BCA) and Pierce™ modified Lowry protein assay (modified Lowry) consistently registered higher protein levels than those determined by dye binding-based Pierce™ Coomassie Plus (Bradford) assay (Coomassie Plus) or Pierce™ 660-nm protein assay (660-nm). When milk samples were autoclaved or dry-heated at ≥ 121 °C for 10 min, reduction in soluble protein content was reflected in measurements of Coomassie Plus and 660-nm assays but was not indicated by BCA or modified Lowry assays. In samples dry-heated at 176 °C, while no protein was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Coomassie Plus or 660-nm, BCA, and modified Lowry indicated protein levels that were 215.7 and 79.4%, respectively, of those of unheated samples. Analysis of lactose samples showed that lactose may contribute to a 13.4 or 1.0% overestimation in BCA or modified Lowry measurement of protein content in unheated dry milk samples. This study demonstrated that quantitation of protein in thermally treated milk samples using copper ion-based BCA and modified Lowry assays could be significantly affected by lactose and, to a greater degree, other reducing substances formed during heating likely via the caramelization and Maillard reaction pathways. In contrast, dye binding-based Coomassie Plus and 660-nm assays were not affected and may be more accurate for measurement of protein content in thermally treated milk samples.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Abernethy G, Higgs K (2013) Rapid detection of economic adulterants in fresh milk by liquid chromatography–tandem mass spectrometry. J Chromatogr A 1288:10–20

    Article  CAS  Google Scholar 

  • Andrews G (1989) Lactulose in heated milk. Bull Int Dairy Fed 238:45–52

    CAS  Google Scholar 

  • Bergqvist Y, Karlsson L, Fohlin L (1989) Total protein determined in human breast milk by use of coomassie brilliant blue and centrifugal analysis. Clin Chem 35:2127–2129

    Article  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Chan F, Reineccius GA (1998) The reaction kinetics for the formation of isovaleraldehyde, 2-acetyl-1-pyrroline, di(H)di(OH)-6-methylpyranone, phenylacetaldehyde, 5-methyl-2-phenyl-2-hexenal, and 2-acetylfuran in model systems. In: Labuza TP, Reineccius GA, Monnier VM, O'Brien J, Baynes JW (eds) Maillard reactions in chemistry, food and health. Woodhead Publishing, Cambridge, pp 131–139

    Google Scholar 

  • Chang KC (1998) Protein analysis. In: Nielsen SS (ed) Food analysis, 2nd edn. Aspen Publishers, Gaithersburg, pp 237–263

    Google Scholar 

  • Finete VLM, Gouvêa MM, Marques FF, Netto ADP (2013) Is it possible to screen for milk or whey protein adulteration with melamine, urea and ammonium sulphate, combining Kjeldahl and classical spectrophotometric methods? Food Chem 141:3649–3655

    Article  CAS  Google Scholar 

  • Friedman M (1996) Food browning and its prevention: an overview. J Agric Food Chem 44:631–653

    Article  CAS  Google Scholar 

  • Fu TJ, Maks N (2013) Impact of thermal processing on ELISA detection of peanut allergens. J Agric Food Chem 61:5649–5658

    Article  CAS  Google Scholar 

  • Fu TJ, Maks N, Banaszewski K (2010) Effect of heat treatment on the quantitative detection of egg protein residues by commercial enzyme-linked immunosorbent assay test kits. J Agric Food Chem 58:4831–4838

    Article  CAS  Google Scholar 

  • Gendel SM, Zhu J (2013) Analysis of U.S. Food and Drug Administration food allergen recalls after implementation of the food allergen labeling and consumer protection act. J. Food Prot 76:1933–1938

    Article  Google Scholar 

  • Gerosa S, Skoet J (2012) Milk availability: trends in production and demand and medium-term outlook. FAO Corporate Document Repository 12:1–40

    Google Scholar 

  • Holsinger VH (1997) Physical and chemical properties of lactose. In: Fox PF (ed) Advanced dairy chemistry Vol. 3. lactose, water, salts and vitamins, Chapman & Hall, London, UK, pp 1-38

  • Jackson LS, Al-Taher FM, Moorman M, Devries JW, Tippett R, Swanson KM, Fu TJ, Salter R, Dunaif G, Estes S, Albillos S, Gendel SM (2008) Cleaning and other control and validation strategies to prevent allergen cross-contact in food-processing operations. J Food Prot 71:445–458

    Article  Google Scholar 

  • Kaanane A, Labuza TP (1989) The Maillard reaction in foods. Prog Clin Biol Res 304:301–327

    CAS  PubMed  Google Scholar 

  • Kamizake NK, Gonçalves MM, Zaia CT, Zaia DA (2003) Determination of total proteins in cow milk powder samples: a comparative study between the Kjeldahl method and spectrophotometric methods. J Food Compos Anal 16:507–516

    Article  CAS  Google Scholar 

  • Keller RP, Neville MC (1986) Determination of total protein in human milk: comparison of methods. Clin Chem 32:120–123

    Article  CAS  Google Scholar 

  • Kjeldahl J (1883) Neue methode zur bestimmung des stickstoffs in organischen körpern. Z Anal Chem 22:366–382

    Article  Google Scholar 

  • Kroh LW (1994) Caramelisation in food and beverages. Food Chem 51:373–379

    Article  CAS  Google Scholar 

  • Kusunoki H, Okuma K, Hamaguchi L (2012) Estimation of lactose interference in vaccines and a proposal of methodological adjustment of total protein determination by the Lowry method. Jpn J Infect Dis 65:489–494

    Article  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    CAS  Google Scholar 

  • Moore JC, DeVries JW, Lipp M, Griffiths JC, Abernethy DR (2010) Total protein methods and their potential utility to reduce the risk of food protein adulteration. Compr Rev Food Sci Food Saf 9:330–357

    Article  CAS  Google Scholar 

  • Morales FJ, Romero C, Jiménez-Pérez S (1997) Chromatographic determination of bound hydroxymethylfurfural as an index of milk protein glycosylation. J Agric Food Chem 45:1570–1573

    Article  CAS  Google Scholar 

  • Nascimento CF, Santos PM, Pereira-Filho ER, Rocha FR (2017) Recent advances on determination of milk adulterants. Food Chem 221:1232–1244

    Article  CAS  Google Scholar 

  • Nursten H (2011) Lactose and oligosaccharides| maillard reaction. In: Fuquay JW, Fox PF, McSweeney PLH (eds) Encyclopedia of dairy sciences, 2nd edn. Elsevier, New York, pp 217–235

    Chapter  Google Scholar 

  • O’Brien J (2009) Non-enzymatic degradation pathways of lactose and their significance in dairy products. In: McSweeney PLH, Fox PF (eds) Advanced dairy chemistry: lactose, water, salts and minor constituents, 3rd edn. Springer, New York, pp 231–294

    Chapter  Google Scholar 

  • O'Brien JM, Morrissey PA (1989) The Maillard reaction in milk products. In: Fox PF (ed) Monograph on heat-induced changes in milk, vol 238. International Dairy Federation, Brussels, pp 53–61

    Google Scholar 

  • Pan GG, Melton LD (2007) Nonenzymatic browning of lactose and caseinate during dry heating at different relative humidities. J Agric Food Chem 55:10036–10042

    Article  Google Scholar 

  • Paschke A, Besler M (2002) Stability of bovine allergens during food processing. Ann Allergy Asthma Immunol 89:16–20

    Article  CAS  Google Scholar 

  • Ribadeau-Dumas B, Grappin R (1989) Milk protein analysis. Lait 69:357–416

    Article  CAS  Google Scholar 

  • Sharma K, Paradakar M (2010) The melamine adulteration scandal. Food Security 2:97–107

    Article  Google Scholar 

  • Smith PK, Krohn RI, Hermanson G, Mallia A, Gartner F, Provenzano M, Fujimoto E, Goeke N, Olson B, Klenk D (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150:76–85

    Article  CAS  Google Scholar 

  • Taylor SL, Hefle SL, Farnum K, Rizk SW, Yeung J, Barnett ME, Busta F, Shank FR, Newsome R, Davis S, Bryant CM (2006) Analysis and evaluation of food manufacturing practices used to address allergen concerns. Compr Rev Food Sci Food Saf 5:138–157

    Article  CAS  Google Scholar 

  • Thermo Fisher Scientific (2010) Thermo scientific pierce protein assay technical handbook. Version 2. https://www.thermofisher.com/content/dam/LifeTech/Images/integration/1602063_PAssayHB_122910.pdf. Accessed 5 August 2019

  • U.S. Food and Drug Administration (2006) Approaches to establish thresholds for major food allergens and for gluten in food. https://www.fda.gov/food/food-labeling-nutrition/approaches-establish-thresholds-major-food-allergens-and-gluten-food. Accessed 5 August 2019

  • Verheul FEAM, Bosch MJA, Cornelissen PJHC (1986) Simplified and rapid methods for the determination of protein, fat and lactose in human milk and the energy intake by the breast-fed infant. J Clin Chem Clin Biochem 24:341–346

    CAS  PubMed  Google Scholar 

Download references

Funding

This work was supported by Cooperative Agreement U19FD005322 between the U.S. Food and Drug Administration and the Institute for Food Safety and Health.

Author information

Authors and Affiliations

  1. Institute for Food Safety and Health, Illinois Institute of Technology, 6502 South Archer Road, Bedford Park, IL, 60501, USA

    Yingshuang Lu

  2. Division of Food Processing Science and Technology, U.S. Food and Drug Administration, 6502 South Archer Road, Bedford Park, IL, 60501, USA

    Tong-Jen Fu

Authors
  1. Yingshuang Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tong-Jen Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tong-Jen Fu.

Ethics declarations

Conflict of Interest

Yingshuang Lu declares that she has no conflict of interest. Tong-Jen Fu declares that she has no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed Consent

Not applicable.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lu, Y., Fu, TJ. Performance of Commercial Colorimetric Assays for Quantitation of Total Soluble Protein in Thermally Treated Milk Samples. Food Anal. Methods 13, 1337–1345 (2020). https://doi.org/10.1007/s12161-020-01748-w

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12161-020-01748-w

Keywords

Search

Navigation