Purpose: To detect fatty acid metabolism of healthy volunteers and determine the influence of age and gender on the 13C-Hiolein breath test. Methods: Twelve healthy volunteers were given 113g standard diet containint 3 mL Hiolein of 15 mg 13C. Breath samples were taken at baseline and at 30 or 60 min intervals after the test meal. Results: The results of cumulative precent dose of 13C recovery (cPDR) at 3,6,24 h and maximal PDR (PDRpeak) were 4.02%, 11.26%, 22.18% and 3.02%, the time of PDRpeak was (5.96+-1.21)h. Both age and gender had no different from all normal values (P>0.05). Conclusions: The age and gender did not influence the cPDR of 13C-Hiolein breath test by isotope ratio mass spectrometry.