Pharmacophore-guided production of high-affinity antibodies for rapid detection of antipyrine in food and environment.

The potential health risks posed by antipyrine-adulterated herbal teas and surface water underscore an urgent need for rapid detection methods. However, no rapid analytical techniques are currently available for antipyrine detection. This study proposes an effective Pharmacophore-based hapten design strategy, successfully generating a highly sensitive and selective antipyrine-specific antibody. This antibody underpins a novel immunoassay: two lateral-flow immunochromatographic methods, AuNP-LFIA and Au@PtNP-LFIA, were developed for antipyrine in herbal tea. Au@PtNP-LFIA, employing a signal-enhancement strategy, reduced the visual detection limit from 100 ng/mL to 30 ng/mL (herbal tea) and 90 ng/mL to 25 ng/mL (surface water). Validation confirmed the method's accuracy and stability, with recoveries and coefficients of variation within acceptable ranges. Additionally, the paper strips retain stability for up to 90 days at room temperature. To our knowledge, this represents the first development of such an immunoassay for antipyrine. This study introduces a novel hapten design concept and a rapid, user-friendly test for food regulatory authorities.