高效液相色谱法测定木糖母液及发酵液中单糖含量

Determination of Monosaccharides in Xylose Mother Liquor and Fermentation Solution by HPLC

建立了木糖母液及其酵母发酵液中葡萄糖和半乳糖含量的高效液相色谱测定方法。测定方法基于铅配位体交换色谱分离模式,色谱分离柱采用Sepax Carbomix Pb-NP5(300×7.8 mm ID,5μm,8%交联度)色谱柱,柱温75℃;流动相采用超纯水(100%),泵速0.55 m L·min^(-1);示差折光检测器温度30℃;木糖母液样品采用固相萃取脱色前处理,母液的发酵液样品采用乙腈沉淀蛋白前处理。在最终的方法条件下,葡萄糖和半乳糖组分在25 min内完成基线分离,在0.2~10.0 mg·m L^(-1)浓度范围内呈现良好的线性关系,方法检测限分别为0.124 mg·m L^(-1)和0.115 mg·m L^(-1)。仪器系统简单,方法重现性好,结果准确。

The determination method of glucose and galactose in xylose mother liquor and its yeast fermentation solution was established with HPLC. The method was based on seperation mode called lead ligand exchange. A Sepax Carbomix Pb-NP5（300×7.8 mm ID,5 μm,8% crosslinkage）column was used for the seperation with ultra-pure water（100%）as mobile phase. The pump rate of mobile phase was 0.55 m L·min^（-1） at a column temperature of 75 ℃. Temperature of refractive index detector（RID）was 30 ℃.Xylose mother liquor was pretreated by solid phase extraction（SPE） to remove colourings,fermentation solution was pretreated by acetonitrile to precipitate protein. Baseline separation was achieved within 25 min at final conditions. The calibration curve was linear in the mass concentration range of 0.2~10.0 mg·m L^（-1）,the LOD of glucose and galactose were 0.124 mg·m L^（-1） and 0.115 mg·m L^（-1） respectively. The instrument system was simple,and method was accurate and reproducible.