The endogenous content of serotonin in human platelets has been used in many clinical studies to indicate platelet activation. A decrease in platelet serotonin compared with controls has been regarded as an indicator of platelet activation. However, the results published are difficult to compare, because of huge variations in endogenous serotonin between control groups in different investigations. This is likely to be because of lack of standardization. Several factors that influence the endogenous platelet serotonin content were studied in more than 200 blood donors. The most important factor was the total g force of the centrifugation used to isolate platelets. Also the age and sex of platelet donors, number of platelets in platelet-rich plasma, and mean platelet volume influenced normal serotonin values. Using a standardized centrifugation procedure (2700 g min) the mean endogenous serotonin was 2.80 nmol/10(9) platelets in young women and 2.58 in elderly women, and 2.67 in young men and 2.30 in elderly men. The differences both for age and sex were statistically significant. Endogenous platelet serotonin shows intrapersonal stability over time, since endogenous platelet serotonin did not change on repeated venepuncture for 9 weeks. Factors such as age, sex and isolation procedure must therefore be considered when endogenous platelet serotonin are studied in relation to disease and treatment.