Abstract We investigated relationships between sedimentary solvent-extractable long-chain alkenone (LCA) concentration and composition and environmental factors in a suite of endorheic lakes from inland Spain. LCAs were found in 14 of the 54 lakes examined, with concentrations comparable with those from previously published lacustrine settings. The composition of LCAs in our sites, however, contrast from the majority of those previously reported from lake environments; in our study the tri-unsaturated component is the most abundant component at most sites where LCAs are detected, and C 38:3 is the most abundant LCA in the majority of sites. LCA occurrence appears to be restricted to brackish-hypersaline sites and C 37 LCAs are absent above a salinity of ∼40 g L −1 suggesting a salinity control on LCA-producing organisms in these sites. Low concentrations of C 37 LCA components means U 37 k and U 37 k ′ temperature indices are generally not applicable. Instead we find good relationships between C 38 components and (in particular mean autumn) temperature and the strongest LCA–temperature relationships are found when using a combination of all C 37 and C 38 compounds. We propose a new alkenone temperature index for lakes with elevated salinity and where the C 38 components dominate the LCA distributions. This is expressed as U 3738 k = 0.0464 × MAutAT - 0.867 ( r 2 = 0.80, n = 13). In this paper, we provide the first account of sedimentary LCA distributions from lakes in inland Spain, extending the range of environments within which these compounds have been found and highlighting their significance as indicators of both salinity and temperature in saline, endorheic lake environments. This has important implications for extending the potential role of LCAs as palaeoclimatic indicators in lacustrine environments.