Radiocarbon measurements on the carbonaceous aerosol fractions are an effective tool for aerosol source apportionment. For these measurements, a new sample preparation line (MISSMARPLE: MIlan Small-SaMple Automated Radiocarbon Preparation LinE for atmospheric aerosol) was built in Milan (Italy). MISSMARPLE can separate different carbon fractions (i.e. total carbon (TC), elemental carbon (EC)), automates the sample combustion processes and the CO2 isolation in the “combustion line”, and was designed to handle small samples, of about 50 μg carbon. The CO2 obtained in the combustion line is then reduced to graphite in the graphitization line for subsequent accelerator mass spectrometry (AMS) analysis at the INFN-LABEC in Sesto Fiorentino (Italy). MISSMARPLE was tested for reproducibility of 14C/12C ratio in primary standard samples, for background contamination by the analysis of blank samples (graphite with zero percent Modern Carbon (pMC)), and for accuracy by the analysis of IAEA-C7 for pMC(TC) and NIST RM8785 for pMC(EC) used as secondary standards. Measurements were carried out in different AMS runs. Reproducibility of 14C/12C was within 1.2%; blank values were down to 2.2 ± 0.2 pMC in the latest AMS run, and both IAEA-C7 and NIST RM8785 measurements were within 1σ with the reference value (but for one IAEA-C7 sample within 2.3σ). These results point to MISSMARPLE as a new, valuable tool for aerosol sample preparation for radiocarbon measurements to be exploited not only on traditional 24-h samples but also when small carbon quantities are available (e.g. collected at remote sites or with high temporal resolution).