Fabrication and evaluation of a label-free piezoelectric immunosensor for sensitive and selective detection of amantadine in foods of animal origin.

A label-free piezoelectric immunosensor was fabricated and applied to the detection of the antiviral drug amantadine (AM) in foods of animal origin. Experimental parameters associated with the fabrication and measurement process were optimized and are discussed here in detail. The proposed piezoelectric sensor is based on an immunosuppression format and uses a portable quartz crystal microbalance (QCM) chip. It was found to provide a good response to AM, with a sensitivity and limit of detection (LOD) of 33.9 and 1.3 ng mL, respectively, as well as low cross-reactivity (CR, < 0.01%) with AM analogues. The immunosensor was further applied to quantify AM at three levels in spiked samples of typical foods of animal origin, and yielded recoveries of 83.2-93.4% and standard deviations (SDs, n = 3) of 2.4-4.5%, which are comparable to the results (recoveries: 82.6-94.3%; SDs: 1.7-4.2%) obtained using a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method. Furthermore, the piezoelectric immunosensing chip can be regenerated multiple (at least 20) times with low signal attenuation (about 10%). A sample analysis can be completed within 50 min (sample pretreatment: about 40 min, QCM measurement: 5 min). These results demonstrate that the developed piezoelectric immunosensor provides a sensitive, accurate, portable, and low-cost analytical strategy for the antiviral drug AM in foods of animal origin, and this label-free detection method could also be applied to analyze other targets in the field of food safety. Graphical abstract.