Colorimetric protein assays are useful tools in determining concentration of proteins used in the

study of proteomics and other fields of biochemistry such as biopharmaceuticals [1]. It uses
spectrophotometric device to rapidly determine the concentration of protein from absorbance readings,
relative to a standard, or using an assigned extinction coefficient [1].

Biuret protein assay is an example of copper-ion based colorimetric assay which reaction involves
the chelation of cupric ion with the nitrogen on the protein’s peptide bond in an alkaline solution [2]. The
reaction then reduces cupric ion (Cu2+) to cuprous ion (Cu+) and the copper-protein interaction produces
a purple color-complex. [2]

According to Beer-Lambert’s law, the amount of color produced and absorbed is proportional to
the amount of proteins present in the sample. The absorption of the protein complex can be read
theoretically at 540nm in a spectrophotometer [3].

[1] Noble, J., & Bailey, M. (2009). Quantitation of Protein. Methods in Enzymology, 73-95.
doi:https://doi.org/10.1016/S0076-6879(09)63008-1 Retrieved at
https://www.ncbi.nlm.nih.gov/pubmed/19892168

[2] Moore, J. C., DeVries, J. W., Lipp, M., Griffiths, J. C. and Abernethy, D. R. (2010), Total Protein
Methods and Their Potential Utility to Reduce the Risk of Food Protein Adulteration. Comprehensive
Reviews in Food Science and Food Safety, 9: 330–357. doi:10.1111/j.1541-4337.2010.00114.x Retrieved
at http://onlinelibrary.wiley.com/enhanced/exportCitation/doi/10.1111/j.1541-4337.2010.00114.x

[3] Chang, S. K., & Zhang, Y. (2017). Protein analysis. In Food analysis (pp. 315-331). Springer, Cham.
Retrieved at https://link.springer.com/chapter/10.1007/978-3-319-45776-5_18