Although the distribution of mono-, di-and trimethylated 2-methyl-2-(4,8,12-Trimethyltrideeyl) chromans (MTTCs) is well understood as an indicator of water salinity, their origin and formation mechanism are still ambiguous and under debate. In this paper, abnormally high levels of MTTCs were detected in Cenozoic saline lacustrine source rocks from the Western Qaidam Basin. Using a two-step column chromatography method, the MTTCs and naphthalenes were separated from other aromatic compounds and concentrated in one fraction, so that the stable carbon isotope compositions of these compounds could be accurately measured. Similar carbon isotope ratios for the mono-, di-and trimethylated 2-methyl-2-(4,8,12-Trimethyltrideeyl) chromans in a given sample suggest the MTTCs may share the same biological source(s). The MTTCs from the Western Qaidam Basin have similar carbon isotope compositions to primary producer-derived pristane and phytane. However, the 13 C values for the MTTCs showed significant differences with other primary producers-derived biomarkers (e.g. C 27 and C 29 steranes), probably indicating a distinct microalgae source from specific niches for the MTTC compounds. We speculate that the MTTCs distribution patterns may be controlled by the water chemistry dynamics in niches, which, in turn, is affected by hypersaline bottom water.