The Fazio Laterality Inventory (FLI) is a recent measure of handedness. Although initially validated, there is still a lack of studies assessing its psychometric properties in samples outside the USA. The present study explores the validity of the Polish adaptation of the FLI. We used data gathered from a convenience sample of 727 participants. They completed the FLI and the Edinburgh Handedness Inventory to establish concurrent validity. Confirmatory factor analysis was used to investigate the factor structure of the FLI. In addition, an Item Response Theory (IRT) model for continuous item scores was also used to identify the discrimination and difficulty parameters of the FLI items. The Polish version of the FLI was characterized by good reliability indices and has high concurrent validity with the Edinburgh Handedness Inventory. We identified a bi-factorial structure for the questionnaire. The IRT analyses showed that the FLI items have good discrimination and difficulty parameters. Our study provides new insights into the properties of the Fazio Laterality Inventory.