Interference-free and simultaneous quantification of three breast cancer-related DNAs in multiple biological fluids using excitation-emission matrix fluorescence spectroscopy combined with second-order calibration method

•Second-order calibration assisted EEMF to quantify ErbB2, TERT, and p53 in complex matrices.•ANWE can resolve highly overlapping fluorescence peaks and obtain satisfactory results.•The strategy is suitable for the anti-interference determination of multiple cancer biomarkers.•ANWE-EEMF has potential value in clinical diagnosis and cancer research.