The composition of toxaphene residuesand the enantiomeric ratios of the major congeners were studied in the top 10 slices (1935–1992) of a sediment core collected from Hanson Lake in the Yukon Territories (Canada). This lake was treated with toxaphene as a picsicide in July 1963. Electron‐capture negative ion mass spectrometry and electron‐capture detection were applied in combination with enantioselective gas chromatography using modified cyclodextrin columns. Of the 10 toxaphene peaks identified in each of the sediment slices, two congeners, 2‐exo,3‐endo,6‐exo,8,9,10‐hexachlorobornane (B6‐923 or Hx‐Sed) and 2‐endo,3‐exo, 5‐endo,6‐exo, 8,9,10‐heptachlorobornane (B7‐1001 or Hp‐Sed), were the most abundant. Each of the three chiral stationary phases used in this study was successful at resolving the enantiomers in at least of one of the two congeners. Among them was permethylated β‐cyclodextrin (Chirasil‐Dex), which, up until now, has never been used for enantiomer separation of compounds of technical toxaphene. B6‐923 and another less abundant, as of yet unknown, hexachlorobornane were racemic in all sediment core slices. For B7‐1001, the latter eluting enantiomer was always more predominant. Furthermore, the enantiomeric ratio was found to consistently decrease with time from approximately 0.8 in the early slices (∼1950) to 0.7 in the slice representing 1992, implying that the first eluted B7‐1001 enantiomer is less persistent than the second. B6‐923 and B7‐1001 are most likely metabolites of the more highly chlorinated congeners. A structural evaluation provided evidence that reductive dechlorination at carbons with geminal chlorine atoms is the primary microbial degradation pathway in sediments from Hanson Lake.