Populations of the endangered white-clawed crayfish (Austropotamobius pallipes) have rapidly declined in distribution and density in recent decades as a result of invasive crayfish, disease and habitat degradation. The species is thought to be particularly sensitive to water chemistry, and has been proposed as a bio-indicator of water quality. Here we detail the results of a systematic review of the literature regarding the chemistry of waterbodies inhabited by white-clawed crayfish, along with a wide-scale field study of the chemistry of crayfish-inhabited waterbodies in the UK. We use these data to examine potentially significant variables influencing crayfish distribution. Several variables appear to have thresholds that affect crayfish distribution; crayfish presence was associated with high dissolved oxygen, low conductivity, ammonium, sodium, and phosphate, and to a lesser extent low sulphate, nitrate, and total suspended solids. Some variables (magnesium, potassium, sodium, sulphate, nitrate, and total suspended solids) may be tolerated at moderate to high concentrations in isolation (indicated by the presence of some populations in high levels of these variables), but suites of chemical conditions may act synergistically in situ and must be considered together. Recent efforts to conserve white-clawed crayfish have included relocations to Ark Sites; novel protected habitats with reduced risk of the introduction of disease, invasive crayfish and habitat degradation. We use our findings to propose the first detailed guidelines for common water chemistry variables of potential Ark Sites for the conservation of the species throughout its European range.