Ammonium and selected anions were determined in wastewater samples with highly complex matrices by spectrophotometry using the reagent-kit method. For this purpose, the interferents of coloured compounds and S2-, SO32-, CO32- and Cl-, which are often present in wastewater samples, were systematically investigated in the spectrophotometric determination of ammonium, nitrate, chloride, sulphate, fluoride and phosphate. After this, innovative procedures for their removal were proposed. For sample decolourization, a DEAE column was used to determine ammonium, while a Florisil column was used for the colour removal and anions' determination. S2- and CO32- were eliminated from the samples by adding HCl or HNO3, which transformed them into gases H2S and CO2. The stepwise addition of CaCl2 to the sample, adjusted to pH 8, initiated the formation of CaSO3, which was removed by filtration. Cl- was removed by the addition of Ag2O, which formed a AgCl precipitate that was removed from the solution by filtration. The accuracy of the determination was tested with spike-recovery tests, which showed recoveries for the analytes in the spiked samples ranging from 95 to 105%. The repeatability of the measurements of nitrate, chloride, sulphate and phosphate in the wastewater samples was better than ±1%, while that for the ammonium and fluoride samples was ±2 and ±5%, respectively. The data from the present investigation revealed that the developed procedures for the decolourization and stepwise removal of interferents enabled accurate spectrophotometric determination of ammonium, nitrate, chloride, sulphate, fluoride and phosphate by using cuvette tests in complex wastewater and environmental water samples.