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<p>Enzyme-based time–temperature indicators (TTIs) have attracted significant interest because they offer high customizability, responsiveness, visual simplicity, reliability, and cost-effectiveness. This study aims to optimize three key parameters, namely enzyme activity, substrate concentration, and pH indicator level for the development of an enzymatic TTI device for the smart packaging of milk. Additionally, the study focuses on developing a smartphone app for automatic detection of milk safety. A 3D printer and wax stamping of filter paper were used for enzyme immobilization. The effects of substrate concentration (10–60%, v/v), enzyme activity (1–10 U), and pH indicator concentration (1–10%, v/v) on the performance of the TTI system were evaluated to optimize the fabrication process. A smartphone app was developed using the Thunkable workspace, with Imagga integrated as the application programming interface (API) platform. The optimal conditions for achieving high TTI performance were determined to be 4.41 U for enzyme activity, 29.34% for substrate concentration, and 4.42% for pH indicator level. Using these optimized parameters, a TTI device was developed and tested at different storage temperatures (5–45&#xa0;°C) for the smart packaging of milk. The activation energy (<i>E</i><sub>a</sub>) and coefficient of determination (<i>R</i><sup>2</sup>) of the TTI system were observed as 49.44&#xa0;kJ/mol and 0.9892, respectively. The device showed a color transition from yellow (initial) to orange (final), which indicated the milk safety status. The developed smartphone app was observed to be effective and reliable for monitoring time and temperature through image-based colorimetric analysis. It could detect and classify the freshness condition of milk as “fresh,” “borderline,” or “spoiled,” which provides useful information to end-users for decision-making regarding product safety. The results of this study suggest that the enzymatic TTI device has promising potential for application in smart milk packaging as a freshness indicator.</p>

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Development of a Time–Temperature Indicator Device for Smart Safety Monitoring of Milk Packaging with Smartphone-Assisted Detection

  • 期刊论文

  • 发表日期：

    2026年7月11日

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  • K. R. Jolvis Pou,
  • Yvan Gariepy,
  • Muthukumaran Packirisamy,
  • Vijaya Raghavan

摘要

Enzyme-based time–temperature indicators (TTIs) have attracted significant interest because they offer high customizability, responsiveness, visual simplicity, reliability, and cost-effectiveness. This study aims to optimize three key parameters, namely enzyme activity, substrate concentration, and pH indicator level for the development of an enzymatic TTI device for the smart packaging of milk. Additionally, the study focuses on developing a smartphone app for automatic detection of milk safety. A 3D printer and wax stamping of filter paper were used for enzyme immobilization. The effects of substrate concentration (10–60%, v/v), enzyme activity (1–10 U), and pH indicator concentration (1–10%, v/v) on the performance of the TTI system were evaluated to optimize the fabrication process. A smartphone app was developed using the Thunkable workspace, with Imagga integrated as the application programming interface (API) platform. The optimal conditions for achieving high TTI performance were determined to be 4.41 U for enzyme activity, 29.34% for substrate concentration, and 4.42% for pH indicator level. Using these optimized parameters, a TTI device was developed and tested at different storage temperatures (5–45 °C) for the smart packaging of milk. The activation energy (Ea) and coefficient of determination (R2) of the TTI system were observed as 49.44 kJ/mol and 0.9892, respectively. The device showed a color transition from yellow (initial) to orange (final), which indicated the milk safety status. The developed smartphone app was observed to be effective and reliable for monitoring time and temperature through image-based colorimetric analysis. It could detect and classify the freshness condition of milk as “fresh,” “borderline,” or “spoiled,” which provides useful information to end-users for decision-making regarding product safety. The results of this study suggest that the enzymatic TTI device has promising potential for application in smart milk packaging as a freshness indicator.