Cooling sensation biosensor coupled with the Chou-Talalay method for quantitative assessment of interaction patterns in binary cooling agent mixtures.

Modulating cooling sensation is crucial for enhancing consumer appeal in food and daily chemical products. Conventional methods struggle to quantify cooling sensation and its synergistic effects, especially in mimicking human perception. Here, we developed a highly sensitive cell-based biosensor for in vitro cooling sensation assessment. This biosensor successfully quantified menthol, WS-3, and WS-23. Notably, the Chou-Talalay method was integrated with the biosensor to quantitatively investigate synergistic and antagonistic effects among these cooling agents. Our strategy revealed that interactions were highly dependent on both combinations and ratios; for instance, menthol/WS-3 (1:3) showed pronounced synergism, while certain WS-3/WS-23 ratios (1:3) exhibited antagonism. Sensory evaluation strongly correlated with biosensor measurements, confirming the high predictive validity of our method. This biomimetic strategy establishes a new paradigm for physiologically relevant cooling sensation analysis. Moreover, this integrated biosensor and the Chou-Talalay method also holds significant potential for exploring other taste analyses.