Choosing an unsuitable bicycle saddle increases the saddle pressure and discomfort during cycling. Women contract sports injuries more easily than men during cycling owing to their anatomy. To investigate the effect of saddle widths on the saddle pressure in female cyclists. Ten healthy women with an average age of 20.7 ± 1.3 years, height of 162 ± and 5.9 cm, weight of 56.1 ± 7.5 kg, and a sciatic bone width of 15.5 ± 1.4 cm were recruited for this study. The distributions of saddle pressure for four different saddle widths (i.e., narrow, moderate, wide, and self-chosen) were recorded using a saddle pressure mat. Participants were instructed to pedal steadily with a frequency of 90 RPM and a load of 150 watts. Thirty seconds of riding data was randomly retrieved for analysis. The trials were conducted with a counter-balanced design to minimize random errors. One-way repeated measures ANOVA was used to compare the saddle pressure of different saddle widths, and the significance level was set at α = 0.05. When wide saddles were used, the maximum and average pressure on the right surface of the posterior ischium were lower than those with narrow (p = 0.001, p = 0.012) and moderate (p = 0.016, p = 0.019) saddles. The area of pressure on the pubic bone was smaller when using a wide saddle than when using narrow (p = 0.005) and moderate (p = 0.018) saddles, and the area of pressure on the right posterior sciatic bone was larger under the wide saddle than under the narrow (p = 0.017) and moderate (p = 0.036) saddles. The average force was greater with the moderate saddle than with the wide (p = 0.008) and self-chosen (p = 0.025) saddles. Using a saddle with a width that is longer than the width of the cyclist’s ischium by 1 cm can effectively improve the distribution of saddle pressure during riding, while providing better comfort.