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Nowadays, the component content of assisted reproductive culture medium and their test methods are unclear in the quality standards. We need to establish the test methods to fill this vacancy, so as to control the quality of assisted reproductive culture medium effectively.To establish an evaluation method for determination of fructose and glucose in assisted reproductive culture medium, thereby based on which to establish the quality standards.High performance liquid chromatography-differential refractive index detector was adopted and Rezex RCM-Monosaccharide Ca2+ (8%) (300.0 mm×7.8 mm) column was used. The mobile phase was ultrapure water at the flow rate of 0.6 mL/min and the temperature of 80 °C.Resolution of the peaks of glucose and fructose was 3.2. The linear ranges of glucose and fructose were 30.1-502 mg/L (r=0.999 8) and 102-408 mg/L (r=0.999 8), respectively. The relative standard deviation of reproducibility test was 0.17% and 0.40%, respectively. The relative standard deviation of stability test was 0.22% and 0.73%, respectively. The glucose group and fructose group average recovery rates were 1.22% and 1.38%, respectively. The methodology of High performance liquid chromatography-differential refractive index detector accorded with the requirements. The glucose contents of samples 1 and 2 were 96.7 mg/L and 99.6 mg/L, respectively. The fructose contents of samples 1 and 2 were 208.5 mg/L and 197.4 mg/L, respectively. A reliable, simple, and accurate method was provided for the quality control of assisted reproductive culture medium, which fills the domestic vacancy in the quality standards for assisted reproductive culture medium.chromatography; quality control; fructose; glucose; research methodthe National Key Technology R&D Program of China, No. 2012BAI22B01Sun X, Huang YL, Ke LN. Determination of fructose and glucose in assisted reproductive culture medium by high performance liquid chromatography-differential refractive index detector. Zhongguo Zuzhi Gongcheng Yanjiu. 2014;18(30):4873-4877.