Ethylenediaminetetraacetic acid (EDTA) is a chelating agent that is extensively used indifferent fields and industries. Multiple analytical techniques have previously been developedfor the determination of EDTA in water. This study was set up to compare different methodsas well as to optimize and implement a method for analytical determination of EDTA in processand wastewater from paper and pulp manufacturing. Three analytical methods whereinvestigated: 1) spectrophotometry, 2) gas chromatography – mass spectrometry (GC-MS) and3) reversed phase ion pair high performance liquid chromatography – UV detection (HPLCUV).Preliminary tests as well as literature studies indicated that HPLC was the best option toinvestigate further. The method was optimized by investigating complexing parameters as wellas mobile phase composition for the analysis of EDTA in waste and process water. The samplepreparation was performed by rapid complexation of EDTA with Fe(III) on a 60°C heatingblock for 30 minutes. The separation was achieved using a reversed phase column with a mobilephase containing 10 mM tetrabutylammonium bromide, 35 mM sodium formate, 6%acetonitrile at pH 3.3. The mobile phase was run isocratically with a flow rate of 1 mL/min andthe detection was carried out using a UV-detector operating at 260 nm. The method displayedgood accuracy and the limit of detection was 1.3 μg L-1. The method repeatability was testedon two effluent samples from pulp and paper manufacturing and was determined as 3.4%(n=20) and 1.7% (n=11). The optimized method was used to analyze a total of 40 process andwastewater samples from 12 different sampling locations at two paper and pulp factories, wherethe concentrations of EDTA ranged from 1.0 mg L -1 to 220 mg L -1.