In order to determine volatile fatty acids (VFA) in human blood, gas chromatographic analyses were performed after blood samples had been pre-treated by a vacuum distillation procedure with subsequent evaporation. Results of the reproducibility study, investigated by pre-treating five aliquots of one human serum sample showed C.V. values ranging from 7.4 to 18.2. Normal serum VFA values were determined in individual serum samples collected from healthy subjects and compared to those present in serum from patients undergoing gall-bladder surgery. The serum VFA values were comparable in the two groups. In the surgical patients, blood was also collected from the portal circulation. All portal serum VFA values, except that of iso-butyric acid, were higher than those found in the samples collected from the peripheral arm vein. VFA values were also determined in serum specimens obtained from blood collected from the arm vein of patients with extensive cirrhosis of the liver. The VFA values showed marked individual variations and were higher than those found in peripheral samples from healthy subjects and patients undergoing gall-bladder surgery, but were lower than those found in samples from the portal circulation from the surgical patients.