Selenium species were determined using HPLC/ICPMS and HPLC/vapor generation/ICPMS in the urine from seven human volunteers investigated at background selenium concentrations and at slightly elevated concentrations after ingestion of 200 microg Se as a selenite supplement. Trimethylselenonium ion (TMSe) was present, together with selenosugars, in the urine samples, a result that dispels recent doubts about its possible previous misidentification with a cationic selenosugar. Although TMSe was present as only a trace metabolite in urine from five of the seven volunteers (0.02-0.28 microg Se/L, equivalent to 1-5% of the sum of selenosugars and TMSe), it was a significant metabolite (up to 4.6 microg Se/L, 22%) in one volunteer, and it was the major identified metabolite (up to 15 microg Se/L, 53%) in another volunteer. This marked individual variability in the formation of TMSe was maintained in a duplicate investigation of urine from the same seven volunteers.