Sewage contamination is a principal concern in water quality management as pathogens in sewage can cause diseases and lead to detrimental health effects in humans. This study examines the distribution of seven sterol compounds, namely coprostanol, epi-coprostanol, cholesterol, cholestanol, stigmasterol, campesterol, and β-sitosterol in filtered and particulate phases of sewage treatment plants (STPs), groundwater, and river water. For filtered samples, solid-phase extraction (SPE) was employed while for particulate samples were sonicated. Quantification was done by using gas chromatography-mass spectrometer (GC-MS). Faecal stanols (coprostanol and epi-coprostanol) and β-sitosterol were dominant in most STP samples. Groundwater samples were influenced by natural/biogenic sterol, while river water samples were characterized by a mixture of sources. Factor loadings from principal component analysis (PCA) defined fresh input of biogenic sterol and vascular plants (positive varimax factor (VF)1), aged/treated sewage sources (negative VF1), fresh- and less-treated sewage and domestic sources (positive VF2), biological sewage effluents (negative VF2), and fresh-treated sewage sources (VF3) in the samples. Association of VF loadings and factor score values illustrated the correlation of STP effluents and the input of biogenic and plant sterol sources in river and groundwater samples of Linggi. This study focuses on sterol distribution and its potential sources; these findings will aid in sewage assessment in the aquatic environment.