Dutch North Sea helicopter operations are characterized by multiple sector flights to offshore platforms under difficult environmental conditions. In the context of a Ministry of Transport program to improve safety levels of helicopter operations, we assessed effects of pre-duty sleep, pre-duty travel time, and workload factors on the alertness and vigilance of pilots. Data of 24 pilots comprising 224 duty days were analyzed. Pilots performed 10-min test sessions after wake up, pre-duty, halfway-duty, end-duty, and at bedtime during normal duty rosters. Test sessions included completion of a vigilance task, vigor and sleepiness ratings, and questions on sleep and operational characteristics. Pilots wore an actometer to objectify sleep data. Vigor scores were high and sleepiness levels were low during the entire flight duty periods (FDPs), while vigilance was impaired only 6.8% in the course of the FDPs. Pre-duty sleep before morning duties was 1.5 h shorter than sleep before duties starting after midday. Longer pre-duty travel time was correlated with shorter pre-duty sleep and lower vigilance levels during duty. During the FDPs, pilots maintained alertness and vigilance levels that may be considered safe in terms of alertness-related flight safety. This favorable outcome may be attributed to reasonable length of FDPs, favorable circadian start and end times of duties, sufficient opportunities for restorative pre-duty sleep, and relatively good weather conditions. Appropriate FDP scheduling is an important measure to optimize alertness of helicopter pilots who have to cope with adverse environmental conditions and limited landing and air traffic control facilities.