Previous research has shown that food deprivation enhances the acoustic startle reflex when it is elicited during presentation of visual food cues. Frustrative nonreward may explain this effect, since visual food cues are also rated to be more appetitive and arousing during food deprivation. However, the impact of menstrual cycle and sex on this effect remains unclear, and it is also not known whether this effect is influenced by hunger and motivation to eat. According to a within-study design, 20 healthy women in different menstrual cycle phases and 14 healthy men participated twice, in normal and food-deprived conditions. After 18 h of food deprivation, acoustic startle was attenuated by appetitive nonfood foreground pictures, but enhanced by presentation of food pictures. No differences between menstrual cycle phases and sexes appeared. The effect correlated with hunger changes, suggesting that motivational factors play a role.