Determination of Fumaric Acid in Fermentation Broth of Rhizopus oryzae by Ion-exclusion Liquid Chromatography

[Objective] The aim was to correctly determine the content of fumaric acid in fermentation broth of Rhizopus oryzae(R.oryzae).[Method] Under the chromatogram conditions of column temperature of 65 ℃,mobile phase 5 mmol/L of sulphuric acid solution and velocity of 0.8 ml/min,the ion-exclusion liquid chromatography was used to determine the content of fumaric acid in fermentation broth of R.oryzae.[Result] The retention time of fumaric acid was 10.450 min,the other impurities did not interfere determination,the ion-exclusion liquid chromatography had good linear relation when the mass concentration of fumaric acid was 0.5-5.0 g/L,the relative standard deviation was 0.667%,and the recovery was over 98%.When the fermenting time was 12,48 and 72 h resp.,the concentration of fumaric acid in fermentation broth of R.oryzae was 5.712,36.420,56.449 g/L resp.,and RSD was 1.040%,0.667% and 0.430% resp.[Conclusion] The ionexclusion liquid chromatography could determine the content of fumaric acid in fermentation broth of R.oryzae rapidly,with good reproducibility,high accuracy and reliable result.