A robust analytical method has been developed and validated by use of high-performance liquid chromatography inductively coupled plasma mass spectrometry with Dynamic Reaction Cell (DRC) technology that separates seven arsenic (As) species in human urine: arsenobetaine (AB), arsenocholine, trimethylarsine oxide (TMAO), arsenate (As(V)), arsenite (As(III)), monomethylarsonate, and dimethylarsinate. A polymeric anion-exchange (Hamilton PRP X-100) column was used for separation of the species that were detected at m/z 75 by ICP-DRC-MS (PerkinElmer SCIEX ELAN DRCII) using 10% hydrogen-90% argon as the DRC gas. The internal standard (As) is added postcolumn via an external injector with a sample loop. All analyte peaks were baseline-separated except AB and TMAO. Analytical method limits of detection for the various species ranged from 0.4 to 1.7 microg L(-1) as elemental As. As(III) conversion to As(V) was avoided by adjusting the urine sample to <pH 6. Analyses of the National Institute of Standards and Technology standard reference material (SRM) 2670 and 2670a elevated and National Institute for Environmental Studies certified reference material (CRM) no. 18 for arsenic species yielded results within the certified SRM-CRM limits for As species; likewise, the sum of all species compared favorably to SRM 2670 and 2670a target values for total As. This As speciation method is now being used in a production mode for the analysis of a US population survey, the National Health and Nutrition Examination Survey, as well as for other biomonitoring studies of As exposure. This method meets our requirement for sample throughput of 2,000-3,000 sample analyses per year.