Acetonitrile (CH<sub>3</sub>CN) is one of the volatile organic compounds (VOC) and a potential tracer of biomass burning. We evaluated the capability of using observations derived from the Superconducting Submillimeter-Wave Limb-Emission Sounder (SMILES) on the International Space Station (ISS) to measure CH<sub>3</sub>CN profiles. The error in a CH<sub>3</sub>CN vertical profile from the Level-2 research (L2r) product version 3.0.0 was estimated by both theoretical error analysis and compared with other instrumental measurements. We estimated the systematic and random errors to be ~5.8 ppt (7.8 %) and 25 ppt (60 %) for a single observation at 15.7 hPa, respectively, in the Tropics, where the CH<sub>3</sub>CN measurements are enhanced. The major source of systematic error was a pressure broadening, and its contribution to the total systematic error was approximately 60 % in the middle stratosphere (15.7–4.8 hPa). The random error decreased to less than 40 % after averaging 10 profiles in the pressure range of 28.8–1.6 hPa. The total error due to uncertainties in other molecular spectroscopic parameters was comparable (2.8 ppt) to those of CH<sub>3</sub>CN spectroscopic parameters. We compared the SMILES CH<sub>3</sub>CN profiles with those of the Microwave Limb Sounder (MLS) on the Aura satellite (version 4.2). The SMILES CH<sub>3</sub>CN values were consistent with those from MLS within the standard deviation (1 σ) of the MLS observations. The difference between the SMILES and MLS CH<sub>3</sub>CN profiles increased with altitude and was within 20–35 ppt (20–260 %) at 15.7–1.6 hPa. We observed discrepancies of 5–10 ppt (10–30 %) between the SMILES CH<sub>3</sub>CN profiles observed by different spectrometers, so we do not recommend merging SMILES CH<sub>3</sub>CN profiles derived from the different spectrometers. We found that SMILES CH<sub>3</sub>CN VMR in the upper stratosphere has a seasonal maximum in February, which is consistent with the fact that biomass burning events are highest from December–March.