A simplified method allowing the determination of trace concentrations (ng/ml) of hexachlorocyclohexanes in human sera suitable for the analysis of large numbers of samples has been developed. Comparison of acid (conc. H2SO4,) and basic (5 M KOH) digestion has shown that good recoveries are obtained with the former when an internal standard, 1,2,4,5-tetrabromobenzene (TBB), is used to correct for volatilization losses. Good separation between alpha-, beta- and gamma-HCH isomers, TBB, hexachlorobenzene and some interfering compounds is obtained with DB-5 columns. The use of electron-capture detection (ECD) or negative ion chemical ionization mass spectrometry (NICI-MS) in the selected ion monitoring mode (m/z 71) provides sufficient sensitivity for quantitative determination. ECD is the most sensitive method for beta-HCH and NICI-MS for the other isomers. GC-NICI-MS is needed for the unambiguous determination of delta-HCH due to the coelution with one interfering compound. The method has been successfully applied to the analysis of a series of 625 samples collected in a population situated near the effluents of an organochlorinated solvent factory. The results have shown that beta-HCH is selectively accumulated in human sera and prompt to the preferential investigation of the toxic effects of this isomer in humans and mammals.