高效液相色谱法与容量分析法测定发酵液中L-赤藓酮糖含量的比较

Comparison of high performance liquid chromatography and volumetric analysis method to determine L-erythrulose in fermentation broth

目的 建立高效液相色谱法（HPLC）测定发酵液中L-赤藓酮糖含量的方法,并与测定还原糖常用的容量分析法（VA）斐林试剂滴定法进行了比较。方法 HPLC法的色谱条件为：色谱柱：Lichrospher5-NH2（4.6mm×250mm）;柱温：32℃;流动相：甲醇—水（85∶15,V/V）;流速：1.0mL·min-1。用紫外检测器室温下检测L-赤藓酮糖,检测波长为277nm,检测器温度为35℃。结果 所得L-赤藓酮糖的线性范围为1.00~100.00g·L-1,R2为0.9999,最低检测限为0.50g·L-1,最低定量限为0.85g·L-1。F检验和t检验结果显示：在各自的线性范围内,VA法与HPLC法在95%的置信区间上存在显著性差异。结论与VA法相比,HPLC法精密度好,准确性高,不受发酵液中其它组分干扰,更适用于测定发酵液中L-赤藓酮糖的含量。

Objective A high performance liquid chromatographic （ HPLC） method has been developed for determining L-erythrulose in fermentation broth.Also,the HPLC method was compared with a volumetric analysis （ VA） method （ using Fehling reagent） .Meth-ods The chromatographic conditions were as follows：Lichrospher 5-NH2 column （4.6 mm ×250 mm） with the temperature of 32 ℃, methanol-water （85∶15,V/V） as mobile phase with the flow rate of 1.0 mL· min-1 ,L-erythrulose was detected by ultraviolet （ UV） detector at 277 nm at room temperature.Results HPLC method exhibited good linearity over the range from 1.00-100.00 g· L-1 with R2 of 0.999 9.The limit of detection and the limit of quantification for HPLC method were 0.50 g· L-1 and 0.85 g· L-1 ,respec-tively.Statistical analysis by F-test showed significant difference at 95%confidence level between VA and HPLC methods.Conclusion Compared with VA method,the HPLC methods established in this paper were simple and rapid and there is no interference from other constitutes in the fermentation broth on using the methods.The methods were much applicable for L-erythrulose determination in fer-mentation process.