Background: Prevalence of non-alcoholic fatty liver disease (NAFLD) in the German population is 20–30%. Liver biopsy to detect non-alcoholic steatohepatitis (NASH) let alone for NAFLD monitoring is not feasible. Current practice regards elevated serum concentrations of liver enzymes as indicator for NAFLD or NASH. In this study we analyzed if an adjustment of the upper limit of normal (ULN) for serum liver enzymes can improve their diagnostic accuracy. Methods: Data from 363 morbidly obese patients (42.5±10.3 years old; mean BMI: 52±8.5 kg/m²), who underwent bariatric surgery were retrospectively analyzed. All patients had histologically confirmed NAFL or NASH (NAS and SAF). Results: In 121 women (45%) and 45 men (46%) elevated values for at least one serum parameter (ALT, AST, γGT) were present. The serum concentrations of ALT (p <0.0001), AST (p <0.0001) and γGT (p = 0.0023) differed significantly between NAFL and NASH, independent of classification method (NAS, SAF). Concentrations of all three serum parameters correlated significantly positively with the NAS and the SAF score, with correlation coefficients between 0.33 (ALT/NAS) and 0.40 (GGT/SAF). The AUROCs to separate NAFL and NASH by liver enzymes achieved a maximum of 0.70 (ALT applied to NAS-based classification). For 95% specificity the ULN for ALT would be 47.5 U/l; for 95% sensitivity, the ULN for ALT would be 17.5 U/l, resulting in 62% uncategorized patients. Conclusion: ALT, AST and GGT are unsuitable for non-invasive screening or diagnosis of NAFL or NASH. Utilizing liver enzymes as an indicator for NAFLD/NASH should generally be questioned.