To study the effect of pupil dilation on a biometric iris recognition (BIR) system for personal authentication and identification. A prospective, non-randomized, single-center cohort study was conducted on patients who reported for a routine eye check-up from November 2017 to November 2019 (2 years). An iris scanning device "IRITECH-MK2120U" was used to initially enroll the undilated eyes. Baseline scans were taken after matching with the enrolled database. All eyes were topically dilated and matched again with the enrolled database. The Hamming distance (a measure of disagreement between two iris codes) and recognition status were recorded from the device output, and eyes were evaluated by slit-lamp ophthalmoscopy with special emphasis on pupil shape, size, and texture. All 321 enrolled eyes matched after topical dilation. The pupil size had a significant effect on Hamming distance with a P value <0.05. There were no false matches. A correct recognition rate of 100% was obtained after dilation. No loss of iris texture or pupil shape was observed after dilation. A BIR system is a reliable method for identification and personal authentication after pupil dilation. Topically dilated pupils are not a cause for non-recognition of iris scans.