RATIONALE: Collisional cross sections (CCS) are an important characteristic of gas-phase ions that are measured using ion mobility-mass spectrometry (IMS). Typically, CCS measurements are performed with drift-tube IMS or travelling-wave IMS. However. in a high-field asymmetric waveform ion mobility (FAIMS) device, ion heating effects make CCS determination more challenging. This research explores whether CCS can be predicted with microscale FAIMS by using known CCS standards. METHODS: An Owlstone ultraFAIMS microscale FAIMS spectrometer was coupled to an Orbitrap Exactive mass spectrometer. Two different CCS standard mixtures (tetraalkylammonium halides [TAAHs] and poly-DL-alanine oligomers) were used to evaluate the system's potential to determine CCS. Test peptide bradykinin acetate and substance P were used to evaluate CCS determination accuracy for singly and doubly charged peptide species using external calibration with a series of poly-DL-alanine peptides for +1, +2 charge states. RESULTS: Calibrations with excellent correlation coefficients (R CONCLUSIONS: Distinct relationships were observed between CCS and transmission CF with both calibrants. Optimum accuracy was obtained at DF 240-260 Td. At lower DF, accuracy is reduced by insufficient resolution of analyte ions from solvent cluster adducts, while at higher DF values, poor transmission becomes a factor. Nevertheless, these data suggest microscale FAIMS can conduct CCS measurements with reasonable accuracy when the compound being measured has similar structural features to the CCS standards used.