Objectives: Glutathione S-transferases (GSTs) are detoxifying enzymes for a number of substrates, including some food compounds. Selected GST polymorphisms have been proven to significantly affect enzymatic activity; however, it is unclear whether this altered metabolism influences dietary composition. The objective of this study was to locate the correlation between GST polymorphisms and selected nutritional parameters, namely, fiber and vitamin C intake. Methods: This study was conducted on a cohort of 472 individuals (mean age 45.26 years; mean body mass index [BMI] 32.36) from the South Moravian region of the Czech Republic. Basic anthropometrical parameters were measured and no association was found for the selected polymorphisms. Polymorphisms in GSTA1, GSTM1, and GSTT1 were genotyped using a polymerase chain reaction (PCR)-based methodology. Food intake was monitored using a self-administered 7-day questionnaire that was subsequently analyzed with a special focus on vitamin C intake, fiber intake, and total energy intake. Results: For GSTA1 and GSTM1 polymorphisms, an association was observed with fiber intake. Though no association was found with vitamin C intake, mean vitamin C intake was found to be higher than recommended daily values. No association was found with either daily energy intake or anthropometric parameters. Conclusion: Based on our results, GST polymorphisms seem to affect dietary composition; however, they have no effect on total energy intake or any association with obesity.