Development of a Low-Cost Spectrophotometer for Protein Determination Using the Lowry Method

Lestha Aurel Salsabilla; Dadan Hermawan; Hartiwi Diastuti; Mekar Dwi Anggraeni; Amin Fatoni

doi:10.22146/ijc.99361

Development of a Low-Cost Spectrophotometer for Protein Determination Using the Lowry Method

https://doi.org/10.22146/ijc.99361

Lestha Aurel Salsabilla⁽¹⁾, Dadan Hermawan⁽²⁾, Hartiwi Diastuti⁽³⁾, Mekar Dwi Anggraeni⁽⁴⁾, Amin Fatoni^(5*)

(1) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Jenderal Soedirman, Jl. Dr. Soeparno No. 61, Karangwangkal, Purwokerto 53123, Indonesia
(2) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Jenderal Soedirman, Jl. Dr. Soeparno No. 61, Karangwangkal, Purwokerto 53123, Indonesia
(3) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Jenderal Soedirman, Jl. Dr. Soeparno No. 61, Karangwangkal, Purwokerto 53123, Indonesia
(4) Department of Nursing, Faculty of Health Sciences, Universitas Jenderal Soedirman, Jl. Dr. Soeparno No. 61, Karangwangkal, Purwokerto 53123, Indonesia
(5) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Jenderal Soedirman, Jl. Dr. Soeparno No. 61, Karangwangkal, Purwokerto 53123, Indonesia
(*) Corresponding Author

Abstract

Affordable and accessible laboratory equipment is increasingly crucial, particularly in resource-limited settings. This research presents the development of a cost-effective spectrophotometer using an Arduino microcontroller and a TSL2591 light sensor to quantify protein concentration via the Lowry method. The device employs a 750 nm LED and a 3D-printed PLA case to house the components. Casein was used for calibration, yielding a linear detection range of 100–700 ppm with a high correlation coefficient (r = 0.9991). The limit of detection (LOD) and limit of quantification (LOQ) were 18.05 and 60.16 ppm, respectively. Precision was confirmed with a HORRAT value of 0.982. Accuracy was validated using the Wilcoxon Signed-Rank test, which showed no significant difference between the fabricated device and a commercial UV-vis spectrophotometer (p = 0.779). These findings support the reliability and reproducibility of the device for protein quantification. The project’s open-source nature encourages further development and application in educational, clinical, and field settings. This innovation supports the development of accessible, low-cost biochemical analysis tools.

Keywords

Lowry method; protein analysis; spectrophotometer alternative

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References

[1] Warner, D.L., Brown, E.C., and Shadle, S.E., 2016, Laboratory instrumentation: An exploration of the impact of instrumentation on student learning, J. Chem. Educ., 93, 1223–1231.

[2] Golwalkar, K.R., and Kumar, R., 2022, “Process Control and Instrumentation” in Practical Guidelines for the Chemical Industry: Operation, Processes, and Sustainability in Modern Facilities, Springer International Publishing, Cham, Switzerland, 167–175.

[3] Appadurai, M., Irudaya Raj, E.F., and Irudaya Rani, E.F., 2024, “Application of Self-powered Sensors and Actuators in Engineering and Medical Domains” in Self-Powered AIoT Systems, Apple Academic Press, Burlington, Canada, 27–62.

[4] Worsfold, P., Townshend, A., Poole, C.F., and Miró, M., 2019, Encyclopedia of Analytical Science, Academic Press, Cambridge, MA, US.

[5] Cardoso Rial, R., 2024, AI in analytical chemistry: Advancements, challenges, and future directions, Talanta, 274, 125949.

[6] Xiao, J., Jiang, C., Li, L., Ye, S., Zhang, S., Xiong, X., Zou, Z. and Huang, Z., 2023, UV–vis spectrophotometer and smartphone RGB dual mode detection of inorganic arsenic based on hydride generation iodine–starch system, Microchem. J., 186, 108298.

[7] Gariglio, S., Malegori, C., Menżyk, A., Zadora, G., Vincenti, M., Casale, M., and Oliveri, P., 2024, Determination of time since deposition of bloodstains through NIR and UV–vis spectroscopy – A critical comparison, Talanta, 278, 126444.

[8] Elik, A., Sarac, H., Durukan, H., Demirbas, A., and Altunay, N., 2022, Vortex assisted magnetic ionic liquid based dispersive liquid–liquid microextraction approach for determination of metribuzin in some plant samples with UV–vis spectrophotometer, Microchem. J., 2022, 181, 107809.

[9] Atsever, N., Borahan, T., Girgin, A., Selali Chormey, D., and Bakırdere, S., 2021, A simple and effective determination of methyl red in wastewater samples by UV–vis spectrophotometer with matrix matching calibration strategy after vortex assisted deep eutectic solvent based liquid phase extraction and evaluation of green profile, Microchem. J., 162, 105850.

[10] Yağmuroğlu, O., 2023, Determination of trace lead(II) in cleavers (Galium aparine) tea by UV-vis spectrophotometry after preconcentration with deep eutectic solvent/DTZ probe-based liquid-liquid microextraction, J. Food Compos. Anal., 118, 105164.

[11] Ali, H., Gupta, R., and Verma, N., 2023, Selective detection of bilirubin in blood serum using UV–vis spectroelectrochemistry technique with CeO₂-CNF/ITO transparent electrode, Electrochim. Acta, 468, 143159.

[12] Udum, Y.A., Killard, A.J., and Wagner, M., 2024, Determination of the relative sensitivity of polyaniline inks and films to deprotonation by ammonia using UV–vis spectroscopy, Eur. Polym. J., 205, 112709.

[13] Tan, W.K., Husin, Z., Yasruddin, M.L., and Ismail, M.A.H., 2023, Recent technology for food and beverage quality assessment: A review, J. Food Sci. Technol., 60 (6), 1681–1694.

[14] de Oliveira, M.R.R., Ribeiro, S.G., Mas, J.F., and Teixeira, A.S., 2021, Advances in hyperspectral sensing in agriculture: A review, Rev. Cienc. Agron., 51, e20207739.

[15] Weiman, Y., Jibin, K.P., Praveen, G.L., Thomas, S., and Kalarikkal, N., 2022, Emerging Trends in Advanced Spectroscopy, River Publishers, Gistrup, Denmark.

[16] Damit, B., and Antoine, M., 2021, “Portable biological spectroscopy: Field applications” in Portable Spectroscopy and Spectrometry, John Wiley & Sons, Hoboken, NJ, US, 545–563.

[17] Tantray, J.A., Un Nissa, N., Choh Wani, R.F., and Shafi, S.M., 2023, “Protein Estimation by Lowry’s Method” in Basic Life Science Methods: A Laboratory Manual for Students and Researchers, Academic Press, Cambridge, MA, US, 65–67.

[18] Lee, N., Shin, S.J., Chung, H.J., Kim, D.K., Lim, J., Park, H., and Oh, H.J., 2015, Improved quantification of protein in vaccines containing aluminum hydroxide by simple modification of the Lowry method, Vaccine, 33 (39), 5031–5034.

[19] Olson, B.J.S.C., 2016, Assays for determination of protein concentration, Curr. Protoc. Pharmacol., 73 (1), A-3A.1–A.3A.32.

[20] Wang, E.J., Li, W., Hawkins, D., Gernsheimer, T., Norby-Slycord, C. and Patel, S.N., 2016, HemaApp: noninvasive blood screening of hemoglobin using smartphone cameras, Proceedings of the 2016 ACM International Joint Conference on Pervasive and Ubiquitous Computing, Association for Computing Machinery, New York, NY, US, 593–604.

[21] Anggraeni, M.D., Fatoni, A., and Rahmawati, E., 2022, Non-invasive neonatal jaundice determination using smartphone camera, AIP Conf. Proc., 2553 (1), 020012.

[22] Aid, T., Kaljurand, M., and Vaher, M., 2015, Colorimetric determination of total phenolic contents in ionic liquid extracts by paper microzones and digital camera, Anal. Methods, 7 (7), 3193–3199.

[23] Fatoni, A., Supiani, S., Dwiasi, D.W., and Anggraeni, M.D., 2020, Introducing colorimetric analysis with document scanner for high school students, J. Phys.: Conf. Ser., 1494, 12026.

[24] Meng, X., Schultz, C.W., Cui, C., Li, X., and Yu, H.Z., 2015, On-site chip-based colorimetric quantitation of organophosphorus pesticides using an office scanner, Sens. Actuators, B, 215, 577–583.

[25] Fatoni, A., Aziz, A.N., and Anggraeni, M.D., 2020, Low-cost and real-time color detector developments for glucose biosensor, Sens. Bio-Sens. Res., 28, 100325.

[26] Radich, J.P., Briercheck, E., Chiu, D.T., Menon, M.P., Sala Torra, O., Yeung, C.C.S., and Warren, E.H., 2022, Precision medicine in low- and middle-income countries, Annu. Rev. Pathol.: Mech. Dis., 17, 387–402.

[27] Ganchi, F.A., and Hardcastle, T.C., 2023, Role of point-of-care diagnostics in lower- and middle-income countries and austere environments, Diagnostics, 13 (11), 1941.

[28] United Nations, 2015, Sustainable Development Goals. Goal 10: Reduce inequality within and among countries, https://www.un.org/sustainabledevelopment/inequality, accessed on June 30, 2024

[29] Harris, D.C., and Lucy, C.A., 2020, Quantitative Chemical Analysis, Macmillan Learning, New York, NY, US.

[30] Latimer, G.W., 2023, Official Methods of Analysis of AOAC International, Oxford University Press, Oxford, UK.

[31] ALKaisy, Q.H., Al-Saadi, J.S., AL-Rikabi, A.K.J., Altemimi, A.B., Hesarinejad, M.A., and Abedelmaksoud, T.G., 2023, Exploring the health benefits and functional properties of goat milk proteins, Food Sci. Nutr., 11 (10), 5641–5656.

[32] Vicente-Saez, R., and Martinez-Fuentes, C., 2018, Open Science now: A systematic literature review for an integrated definition, J. Bus. Res., 2018, 88, 428–436.

[33] Drogalis, C., Zampino, C., and Chauhan, V., 2023, Food Quality Inspection and Sorting Using Machine Vision, Machine Learning and Robotics, Proceedings of the ASME 2023 International Mechanical Engineering Congress and Exposition. Volume 3: Advanced Manufacturing, New Orleans, Louisiana, USA, October 29–November 2, 2023, V003T03A066.

[34] Laganovska, K., Zolotarjovs, A., Vázquez, M., Mc Donnell, K., Liepins, J., Ben-Yoav, H., Karitans, V., and Smits, K., 2020, Portable low-cost open-source wireless spectrophotometer for fast and reliable measurements, HardwareX, 7, e00108.

[35] Pereira, V.R., and Hosker, B.S., 2019, Low-cost (<€ 5), open-source, potential alternative to commercial spectrophotometers, PLoS Biol., 17 (6), e3000321.

[36] Balado Sanchez, C., Díaz Redondo, R.P., Fernandez Vilas, A., and Sánchez Bermúdez, A.M., 2019, Spectrophotometers for labs: A cost‐efficient solution based on smartphones, Comput. Appl. Eng. Educ., 27 (2), 371–379.

[37] Yan, J.C., Ren, J., Ren, L.L., Jian, J.M., Yang, Y., Yang, S.F., and Ren, T.L., 2019, Development of a portable setup using a miniaturized and high-precision colorimeter for the estimation of phosphate in natural water, Anal. Chim. Acta, 1058, 70–79.

[38] Safarik, I., Baldikova, E., Prochazkova, J., and Pospiskova, K., 2019, Smartphone-based image analysis for evaluation of magnetic textile solid phase extraction of colored compounds, Heliyon, 5 (12), e02995.

DOI: https://doi.org/10.22146/ijc.99361