Exposure to nitrous oxide in surgical theaters was evaluated for duration, numbers, and types of surgical procedures. The concentration of the gas in the air was 92-444 ppm. Before and after the surgical sessions, samples of urine and expired air were collected from surgical theater personnel for gas determination. Nitrous oxide concentrations in urine and in expired air showed a good correlation with gas concentration in the air (r = 0.760 and r = 0.921, respectively). Moreover, a good correlation (r = 0.823) between gas concentration in urine and that in expired air was also found. A biological threshold limit value (TLV) of 20.6 micrograms/liter for urine and of 29.6 ppm for expired air was calculated, based on the limit of 50 ppm in the air proposed by the American Conference of Governmental Industrial Hygienists (ACGIH). Other biological TLVs corresponding to higher proposed limits (200 and 500 ppm) were also calculated.