The most appropriate systems for treatment of metal-contaminated waters are bioreactors with microbial biofilms. A horizontal rotating tubular bioreactor (HRTB) was studied for its applicability for removing copper, iron, nickel, and zinc (Cu, Fe, Ni, and Zn) from wastewater. Monitoring of the concentrations of Cu, Fe, Ni, and Zn by a fast, simple, onsite method was needed to make decisions for further optimization. The UV-VIS spectrophotometric quantification of Cu, Fe, Ni, and Zn using sodium diethyldithiocarbamate, 1,10-phenathroline, dimethylglyoxime, and 2-{[alpha-(2-Hydroxy-5-sulfophenylazo)-benzylidene]-hydrazino}-benzoic acid monosodium salt (=zincon monosodium salt) as reagents, respectively, was optimized and validated. The limits of quantification were 0.14, 0.12, 0.21, and 0.03 mg/L for Cu, Fe, Ni and Zn, respectively. The recovery for all elements was between 98 and 104%, the uncertainty of measurement was less than 6%. Depending on the reactor parameters applied, metal removals from 40 to more than 90% could be obtained.