Introduction: Rapid processing, sample analysis and accurate results are crucial for clinical data management of patients with fertility problems. The method of inductively coupled plasma mass spectrometry (ICP-MS) is suitable for measurement of a large number of metals and metalloids in biological samples upon a very low detection limit. Spectrophotometric methods are recommended for measurement of zinc levels due to the speed of analysis, accessibility and simplicity. Objective: The objective of this research is to compare the results of zinc concentrations in seminal fluid obtained using the method recommended by the WHO (spectrophotometric method with 5-Br-PAPS) with the results obtained through the ICP-MS method. Materials and methods: This research used seminal plasma samples provided by 185 respondents who were admitted to the Department for Clinical Laboratory Diagnostics of the Clinical Hospital Centre in Osijek for infertility testing over a one-year period. After adequate sample processing, zinc concentrations in seminal fluid were determined on ICP-MS analyser Agilent 7500cx (Agilent Technologies, Tokyo, Japan) and through spectrophotometric method (Sentinel Diagnostics, Milan, Italy) on biochemical analyser Beckman-Coulter AU680 (Beckman-Coulter, Brea, USA). Results: The results of the comparison between spectrophotometric and ICP-MS method for determination of zinc levels in seminal fluid showed that there is a small, yet statistically significant constant difference (around 0.08 mmol/l according to Passing-Bablok regression and around 0.16 mmol/l according to Deming regression) and a significant proportional difference (around 30.30% according to Passing-Bablok regression and around 27.98% mmol/l according to Deming regression) in the results. Conclusion: The results of the comparison between spectrophotometric and ICP-MS methods for determination of zinc levels in seminal fluid showed that the two methods are not comparable. Irrespective of the fact that ICP-MS method gives incomparably more accurate results, due to its complexity, price and unavailability, it is currently unsuitable for routine clinical use.