High red blood cell (RBC) thiopurine methyltransferase (TPMT) activity is associated with a higher relapse rate in children with acute lymphoblastic leukaemia on 6-mercaptopurine therapy. RBC TPMT activity is subject to genetic polymorphism and inter-ethnic variation. Higher TPMT activity in male subjects has previously been reported in RBC and liver tissue, but only in non-healthy subjects. In this healthy, drug-free study group the gender difference in the RBC TPMT high activity subgroup was confirmed with 8.3% higher TPMT activity in male subjects (n = 105).