Hair samples continue to be used extensively for biomonitoring of mercury (Hg) exposure. Routine methods require a bundle of 100-150 hair strands and involve chemical digestion. Recently, Hg analyzers that combine combustion, gold amalgamation, and atomic absorption spectrometry (C-GA-AAS) became commercially available. This method was shown to provide quick and sensitive measurements in solid samples such as hair. The objective of this study was to validate C-GA-AAS for measuring total Hg in single hair strands as an alternative method for Hg biomonitoring. Hair samples from 12 women with a wide range of Hg exposure were obtained from two projects conducted in Brazil and Canada. A 1:1 relationship was observed between C-GA-AAS and the established cold vapor atomic absorption spectrometry (CV-AAS) for analysis of 1-cm segments from a bundle of hair. For individual hair variability, the average relative standard deviation (RSD) of Hg between hair strands was 6.5 +/- 2.8%, thus justifying the use of single hair strand for biomonitoring. With a limit of quantification of 0.10 ng of total Hg, a single hair strand can be used to assess monthly exposure. This technique will facilitate routine biomonitoring and thus help prevent Hg poisoning among the public.