The information regarding the occurrence and abundance of lead (Pb) in PM2.5 is useful for the evaluation of air pollution status and tracing the pollution source. Herein, electrochemical mass spectrometry (EC-MS) for sequential determination of Pb species in PM2.5 samples without sample pretreatment has been developed using the combination of online sequential extraction with mass spectrometry (MS) detection. Four kinds of Pb species including water-soluble Pb compounds, fat-soluble Pb compounds, water/fat-insoluble Pb compounds, and a water/fat-insoluble Pb element were sequentially extracted from PM2.5 samples, in which water-soluble Pb compounds, fat-soluble Pb compounds, and water/fat-insoluble Pb compounds were extracted sequentially by elution using H2O, CH3OH, and EDTA-2Na as the eluent respectively, while the water/fat-insoluble Pb element was extracted by electrolysis using EDTA-2Na as the electrolyte. The extracted water-soluble Pb compounds, water/fat-insoluble Pb compounds, and water/fat-insoluble Pb element were transformed into EDTA-Pb in real time for online electrospray ionization mass spectrometry analysis, while the extracted fat-soluble Pb compounds were directly detected by electrospray ionization mass spectrometry. The advantages of the reported method include the obviation of sample pretreatment, high speed of analysis (<60 min/sample), low detection limit (0.16 pg), low sample consumption (30 μg), and high accuracy (>90%), which indicates the potential of this method for the rapid quantitative species detection of metals in environmental particulate matter samples.