This paper compares the prevalence of risk factors for sexual acquisition of human immunodeficiency virus infection (HIV-1) among blood donors with the risk in the general population. Sexual behaviour data collected as part of a cross-sectional survey of central London blood donors were compared, after indirect standardization for age and sex, with those collected among Greater London participants of the National Survey of Sexual Attitudes and Lifestyles (NATSSAL). Overall, male blood donors had lower risk sexual lifestyles than age-matched NATSSAL participants in that they had had fewer lifetime female sexual partners, were more likely to have had first sexual intercourse after 16 years and were less likely to have had a homosexual partner. Men who have had sex with men since 1977 were asked not to donate blood. Despite this, 3.6% of male donors had had some sexual experience with another man and 1.9% had had sexual intercourse with at least one male partner. In contrast, female donors had had more sexual partners, were more likely to have had first sexual intercourse before 16 years and had probable higher rates of sexually transmitted disease (STD) clinic attendance than NATSSAL participants, explained by the much higher proportion of single rather than married female blood donors at each age. In conclusion, male donors had lower risk sexual lifestyles than age-matched NATSSAL participants, although even low levels of homosexual contact give some cause for concern. Female donors appeared to have higher risk sexual lifestyles than the general population because a much higher proportion of the female blood donors were single. The results provide important insights for those defining blood donor referral criteria.