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This method uses calibration samples containing all 13 analytes with polarity switching between positive and negative modes to maximise sensitivity for each substrate, this has resulted in an improvement of up to 100 fold assay sensitivity over previously published methods. Linearity, selectivity, accuracy and precision have been assessed and shown to perform to acceptable standards [4]. This single analytical technique will provide a significant saving of time and resource in the conduct of future in vitro DDI studies.