Septic tanks treat wastewater of individual houses and small communities (up to 2000 people in Scotland) in rural and semi-urban areas and are understudied sources of surface water contamination. A multi-analyte methodology with solid phase extraction (SPE), ultra-sonic extraction, and direct injection sample preparation methods was developed to analyse a comprehensive range of emerging contaminants (ECs) including prescription and over-the-counter pharmaceuticals and related metabolites, natural and synthetic hormones, and other human wastewater marker compounds in septic tank influent and effluent, river water, suspended solids, and septic tank sludge by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). The number of quantifiable compounds in each matrix varied from 68 in septic tank wastewater to 59 in sludge illustrating its applicability across a range of matrices. Method quantification limits were 2.9 × 10-5-1.2 μg L-1 in septic tank influent, effluent and river water, with ≤0.01 μg L-1 achieved for 60% of ECs in all three water matrices, and 0.080-49 μg kg-1 in sludge. The developed method was applied to a septic tank (292 population equivalents) and the receiving river in the North-East of Scotland. Across all samples analysed, 43 of 68 ECs were detected in at least one matrix, demonstrating the method's sensitivity. The effluent concentrations suggest limited removal of ECs in septic tanks and a potential impact to river water quality for some ECs. However, further monitoring is required to better appreciate this. The developed methodology for a wide variety of ECs in a range of liquid and solid phases will allow, for the first time, a comprehensive assessment of ECs fate and removal in septic tanks, and their impact to surface water quality.