Human error remains a critical concern in aviation safety, contributing to 70-80% of accidents despite technological advancements. While physiological measures show promise for error detection in laboratory settings, their effectiveness in dynamic flight environments remains underexplored. Through live flight trials with nine commercial pilots, we investigated whether established error-detection approaches maintain accuracy during actual flight operations. Participants completed standardized multi-tasking scenarios across conditions ranging from laboratory settings to straight-and-level flight and 2G manoeuvres while we collected synchronized physiological data. Our findings demonstrate that EEG-based classification maintains high accuracy (87.83%) during complex flight manoeuvres, comparable to laboratory performance (89.23%). Eye-tracking showed moderate performance (82.50\%), while ECG performed near chance level (51.50%). Classification accuracy remained stable across flight conditions, with minimal degradation during 2G manoeuvres. These results provide the first evidence that physiological error detection can translate effectively to operational aviation environments.