葡萄糖直接发酵产2-酮基-L-古龙酸途径中代谢产物的HPLC分析

建立了利用高效液相色谱对葡萄糖发酵产2-酮基-L-古龙酸体系中的4种代谢产物D-葡萄糖酸、2-酮基-D-葡萄糖酸、2,5-二酮基-D-葡萄糖酸和2-酮基-L-古龙酸的定量测定方法，分析条件为：300 mm′7.8 mm Aminex HPX-87H柱，柱温30℃，流动相为50 mmol/L硫酸，流速0.2 mL/min，RID-10A示差检测器，以丙二酸为内标物. 结果表明，4种物质分离较好. 该方法对2-酮基-D-葡萄糖酸、2,5-二酮基-D-葡萄糖酸和2-酮基-L-古龙酸3种代谢物的检测平均误差分别为1.53%, 0.89%和1.84%.

Analysis of Metabolites from Direct Fermentation of D-Glucose to 2-Keto-L-Gulonic Acid by HPLC

An HPLC-based method for quantitative determination of the 4 metabolites of gluconic acid (GlcA), 2-keto-D-gluconic acid (2-KDG), 2,5-diketo-D-gluconic acid (2,5-DKG) and 2-keto-L-gulonic acid (2-KLG) from direct fermentation of D-glucose to 2-KLG by using HPLC was established. The optimum separation of the 4 substances could be achieved under the conditions of using Aminex HPX-87H column (size 300 mm ′7.8 mm) with the temperature of 30℃, sulfuric acid (50 mmol/L) as mobile phase at the flow rate of 0.2 mL/min, RID-10A detector, and malonic acid as internal standard. The analytical results show that the method has average analytical deviation of 1.53%, 0.89% and 1.84% on 2-KDG, 2,5-DKG and 2-KLG, respectively.