This is an investigation of the effect of changing the pupil diameter on the P1 amplitude and latency of the multifocal electroretinogram (mfERG). MfERGs were recorded using a custom built wide field electrophysiological system. An array of 61 empirically scaled hexagons was used to stimulate the visual field. The duration of overall recording period was 8 min, segmented into 16 intervals each lasting 30 s. A combination of mydriatics and miotics were used to pharmacologically alter the pupil size and diameters between 1 and 10 mm were measured. There was a reduction in mfERG P1 amplitude in some cases greater than 50 (mfERG P1 amplitude 53 nV at 8 mm to 25 nV at 1 mm), with a change in pupil diameter of 7 mm. The mfERG P1 latency increased in some cases by as much as 8 ms in the central 40° (mfERG P1 latency 39 ms at 8 mm to 47 ms at 1 mm). These results suggest that pupil size has significant effects on mfERG P1 amplitude and latency.