The aim of this study was to evaluate differences in milkability based on lactation peak and parity in Holstein cattle. Milk yield, actual milking time, milking time differences, occurrence of bimodal milk flow, average and partial milk flows within the first two minutes of milking were monitored for each milking during the first 120 days in milk (DIM). The Holstein cows (n = 135) were divided into groups according to the day when they achieved lactation peak (lactation peak group (LPG): 0–30 DIM; 30–60 DIM; 60–90 DIM and 90–120 DIM) and parity (primiparous and multiparous cows). The obtained results suggested that the values of milk yield were higher when cows reached the lactation peak between 30–60 DIM regardless of parity (15.62 and 19.65 kg for primiparous and multiparous cows, respectively). The shortest milking times were achieved by both primiparous (7:03 min) and multiparous (7:39 min) cows in LPG90–120. The average milk flow per min was higher in LPG30–60 and LPG90–120 for both parity groups. Primiparous cows in LPG30–60 and LPG60–90 had a lower incidence of bimodal milk flows, while multiparous cows had the highest proportion of bimodal flows in LPG90–120. High values of partial milk flows during the first 15 s of milking were achieved by primiparous cows in LPG30–60, LPG60–90 and LPG90–120, while multiparous cows had the highest values for LPG90–120. Other partial milk flows from 15 s to 2 min of milking showed a similar trend of higher values in LPG30–60 and LPG90–120 regardless of parity, except partial flow from 30 to 60 s for primiparous cow. The results showed that there is no one ideal LPG for all milkability parameters. In terms of milk yield, lactation peak is optimal between 30–60 DIM. Therefore, such animals should be considered for breeding and targeted for milking efficiency.