Objectives: The aim of this study has been to investigate serum activities of liver enzymes in workers exposed to sub-TLV levels of dimethylformamide (DMF). Material and Methods: Seventy-two workers and 72 healthy controls participated in the study. All subjects underwent complete physical examinations and abdominal ultrasound examination. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and c-glutamyl transpeptidase (c-GT) were determined by an auto-chemistry analyzer. The data of airborne concentrations of DMF was obtained from the local Center of Disease Control and Prevention. The level of urine N-acetyl-S-(N-methylcarbamoyl)cysteine (AMCC) was measured by means of high-performance liquid chromatography. Results: Time weighted average (TWA) concentration of the DMF in workplace was 18.6 (range: 9.8–36.2) mg/m3. The concentration of the AMCC in workers’ urine was 28.32 (range: 1.8–58.6) mg/l and 9 workers’ AMCC exceeded the biological exposure index (40 mg/l). Thirty-one workers reported gastrointestinal symptoms (abdominal pain, nausea, anorexia) and 10 workers reported headache, dizziness and/or palpitation in the exposed group. Serum analysis revealed that both the mean of serum activities of liver enzymes (ALT, AST and c-GT) and the percentage of workers with abnormal liver function were significantly higher in the exposed group as compared to the controls. Conclusions: Dimethylformamide can cause liver damage even if air concentration is in the sub-threshold limit value (sub-TLV) level. The protection of skin contact against the exposure to the DMF might be a critical issue as far as the occupational health is concerned.