10.57647/j.ijic.2025.1604.17

Evaluation of Enzyme Activity Measurement Methods: Application of a Glucometer as a Rapid and Low-Cost Alternative to HPLC

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  • Saloomeh Shoaei Naeeni1,
  • Bijan Bambai*2,
  • Bagher Yakhchali3,
  • Parvaneh Saffarian1,
  • Zarrin Minuchehr2,
  1. Department of Biology, SR.C., Islamic Azad University, Tehran, Iran
  2. Systems Biotechnology Department, National Institute of Genetic Engineering and Biotechnology (NIGEB), Shahrak-e Pajoohesh, Km 15, Tehran - Karaj Highway, Tehran, Iran
  3. Energy and Environmental Biotechnology Department, National Institute of Genetic Engineering and Biotechnology (NIGEB), Shahrak-e Pajoohesh, Km 15, Tehran - Karaj Highway, Tehran, Iran

Received: 2025-10-11

Revised: 2025-11-16

Accepted: 2025-12-22

Published in Issue 2025-12-30

Copyright (c) 2025 Saloomeh Shoaei Naeeni, Bijan Bambai, Bagher Yakhchali, Parvaneh Saffarian, Zarrin Minuchehr (Author)

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

How to Cite

Shoaei Naeeni, S., Bambai, B., Yakhchali, B., Saffarian, P., & Minuchehr, Z. (2025). Evaluation of Enzyme Activity Measurement Methods: Application of a Glucometer as a Rapid and Low-Cost Alternative to HPLC. International Journal of Industrial Chemistry, 16(4). https://doi.org/10.57647/j.ijic.2025.1604.17
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Abstract

Glucose isomerase (GI) catalyzes the reversible conversion of glucose to fructose and plays a major role in producing high-fructose corn syrup (HFCS). Accurate and rapid measurement of GI activity is essential for process optimization and quality control. This study compared two analytical methods—high-performance liquid chromatography (HPLC) and a portable glucometer—for determining GI activity in recombinant Escherichia coli BL21 cultures grown under various conditions. Enzyme activity was quantified by monitoring glucose consumption and fructose production. Statistical analysis, including regression and independent t-tests, revealed a strong correlation between glucometer and HPLC results (R² = 0.96, p = 0.0001) with no significant difference in mean activities. The findings demonstrate that the glucometer, despite its simplicity and low cost, provides reliable data comparable to HPLC. This approach provides a rapid and cost-effective alternative for routine glucose isomerase activity screening in laboratory and industrial settings.

Keywords

  • Enzymatic assay,
  • Fructose,
  • Glucose isomerase,
  • Glucometer,
  • HPLC,
  • Quantification
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References

  1. Bhosale S H,. Rao M B, Deshpande V V. Molecular and Industrial Aspects of Glucose Isomerase. Microbiol Rev 1996; 60:280–300. http//:doi.org/10.1128/mr.60.2.280-300.1996
  2. Nam K H, Glucose Isomerase: Functions, Structures, and Applications. Appl Sci 2022; 12:428. http//:doi.org/10.3390/app12010428
  3. Patel H, Hajoori M, Glucose isomerase: an Enzyme of Industrial Significance. Int J Pharm Res Appl . 2023; 8:2312–2322. http//:doi.org/10.35629/7781-080323122322
  4. Singh R S, Chauhan K, Singh R P, Enzymatic Approaches for the Synthesis of High Fructose Syrup BT - Plant Biotechnology: Recent Advancements and Developments. In: Gahlawat S K, Salar R K, Siwach P, Duhan J S, Kumar S, Kaur P (eds). Springer Singapore, Singapore. 2017. pp 189–211. http//:doi.org/10.1007/978-981-10-4732-9_10
  5. Galant A L, Kaufman R C, Wilson J D, Glucose: Detection and analysis. Food Chem. 2015; 188:149–160. http//:doi.org/10.1016/j.foodchem.2015.04.071
  6. Carballo S, Zingarello F A, Maestre S E, Todolí J L, Prats M S, Optimisation of Analytical Methods for the Characterisation of Oranges, Clementines and Citrus Hybrids Cultivated in Spain on the Basis of Their Composition in Ascorbic Acid, Citric Acid and Major Sugars. Int J Food Sci Technol. 2014; 49:146–152. http//:doi.org/10.1111/ijfs.12289
  7. Gangola M P, Jaiswal S, Khedikar Y P, Chibbar R N .A Reliable and Rapid Method for Soluble Sugars and RFO Analysis in Chickpea Using HPAEC–PAD and its Comparison with HPLC–RI. Food Chem. 2014; 154:127–133. http//:doi.org/10.1016/j.foodchem.2013.12.085
  8. Ma J, Hou X, Zhang B, Wang Y, He L , The Analysis of Carbohydrates in Milk Powder by a New “Heart-Cutting” Two-Dimensional Liquid Chromatography Method. J Pharm Biomed Anal. 2014; 91:24–31. http//:doi.org/10.1016/j.jpba.2013.11.006
  9. Yuen V G, McNeill J H. Comparison of the Glucose Oxidase Method for Glucose Determination by Manual Assay and Automated Analyzer. J Pharmacol Toxicol Methods. 2000; 44:543–546. http//:doi.org/10.1016/S1056-8719(01)00117-4
  10. Hall M B, Keuler N. Factors Affecting Accuracy and Time Requirements of a Glucose Oxidase-Peroxidase Assay for Determination of Glucose. J AOAC Int. 2008; 92:50–60. http//:doi.org/10.1093/jaoac/92.1.50
  11. Wei J, Qiang L, Ren J, Tang F, Meng X. Fluorescence Turn-Off Detection of Hydrogen Peroxide and Glucose Directly Using Carbon Nanodots as Probes. Anal Methods. 2014; 6:1922–1927. http//:doi.org/10.1039/C3AY41837E
  12. Pfützner A, Mitri M, Musholt P B, Sachsenheimer D, Borchert M, Yap A, Forst T. Clinical Assessment of the Accuracy of Blood Glucose Measurement Devices. Curr Med Res Opin. 2012; 28:525–531. http//:doi.org/10.1185/03007995.2012.673479
  13. Diabetic Outlet, Ten Common Problems You May Face When Using a Glucose Meter at Home. https://diabeticoutlet.com/common-errors-with-the-use-of-glucose-meters/. Accessed 4 Aug 2025
  14. Weinstock R S, Glucose Monitoring in the Management of Nonpregnant Adults with Diabetes Mellitus. https://www.uptodate.com/contents/search. Accessed 4 Aug 2025
  15. Agamatrix, The Truth Behind the Numbers: What Affects Blood Glucose Meter Accuracy? https://agamatrix.com/blog/blood-glucose-meter-accuracy/.Accessed 4 Aug 2025
  16. Bhasin S, Modi H. Optimization of Fermentation Medium for the Production of Glucose Isomerase Using Streptomyces sp. SB-P1. Biotechnol Res Int. 2012; 2012:874152. http//:doi.org/10.1155/2012/874152
  17. Darwin A E. The Determination of Glucose Concentration in Anaerobic Acidification Cultures by Portable Glucose Monitoring System. Asian J Chem. 2019; 31(4): 763-776. http//:doi.org/10.14233/ajchem.2019.21593
  18. Singh R S, Singh T, Pandey A. Microbial Enzymes—An Overview. In: R.S. Singh, R.R. Singhania, A. Pandey, C.B.T.A. Larroche (eds) Biomass, Biofuels, Biochemicals. Elsevier. 2019; pp 1–40
  19. Otokunefor O, Ogu R, Comparing the Glucose Results by Glucometer and Laboratory Methods: A Prospective Hospital Based Study. J Adv Med Med Res. 2018; 26:1–7. http//:doi.org/10.9734/JAMMR/2018/40923