We studied adult Peruvian Boobies (Sula variegata) on two islands in northern Peru to classify males and females using DNA-based techniques. We used this information to (1) assess the extent of size dimorphism in this species, (2) identify males and females using discriminant functions of external characters, and (3) validate use of voice as a reliable method for identifying male and female Peruvian Boobies in the field. Female Peruvian Boobies were 19 heavier and their culmens and wings were 3 and 4% larger than males, respectively. A discriminant function that included body weight and wing chord correctly classified 90% of the birds. Alternatively, 88% of correct identification of males and females was obtained with a function that incorporated only wing chord. Whistles were performed exclusively by males (25/25 of cases), whereas grunts or goose-like honk vocalizations were performed only by females (24/24 of cases). The female-larger size dimorphism of Peruvian Boobies is intermediate in comparison to other Sula boobies. Calls and biometry provide a fast, reliable, and inexpensive method for classifying most adult Peruvian Boobies as males or females in the field. We recommend a hierarchical system for classification of male and female Peruvian Boobies: (1) use of vocalizations, (2) use of body weight and wing chord when the bird did not vocalize and was weighed immediately after daybreak or before the first feeding trip of the day, and (3) use of wing chord only when there is uncertainty in temporal variations of body weight. © 2009 by the Wilson Ornithological Society.