高效阴离子交换色谱-积分脉冲安培检测法分析糖类化合物影响因素的研究

Study on Some Factors Affecting Separation and Detection of Carbohydrates by High-performance Anion-exchange Chromatography with Integrated Pulsed Amperometric Detection

研究了高效阴离子交换色谱-积分脉冲安培检测法分离测定糖类化合物的一些影响因素。这些因素包括NaOH淋洗液浓度、色谱柱温度和积分脉冲安培检测的采样电位。实验用AminoPac PA10阴离子交换柱分离糖类化合物。当NaOH淋洗液浓度增大时,糖类化合物的保留值减小,并有洗脱顺序的变化。糖的保留只表现为放热行为,但是每种糖的保留受温度的影响程度不同,即当温度改变时每种糖保留时间的变化程度不同。一般规律是随糖分子增大(依单糖、双糖、三糖、四糖顺序),其保留时间的变化程度增大。通过改变温度可以改善两种相近洗脱糖之间的分离状况。糖的范特霍夫(Van’t Hoff)曲线具有良好的线性关系。积分脉冲安培检测的采样电位增大,糖检测的峰面积增大。通过提高采样电位,可以改善糖的检测。

Some factors influencing the separation and detection of carbohydrates by high-performance anion-exchange chromatography with integrated pulsed amperometric detection were investigated. These factors include sodium hydroxide eluent concentration, column temperature, and sampling step potential. Separation was carried out using an AminoPac PA 10 column. When sodium hydroxide eluent concentration was increased, the retention values of carbohydrates were decreased and the elution orders of some sugars were changed. Sugars retention exhibited only exothermic behavior. However, individual sugars showed different relative changes in retention time with increased temperature. There is the general regularity, which is the variation of retention time increases as the sequence of monosaccharide, disaccharide, trisaccharide and tetrasaccharide when the column temperature is increased. Selectivity changes of sugars, including elution order reversal, were also achieved by varying temperature. Thelinearityof the Van＇t Hoff plot for carbohydrates is satisfactory. Detection sensitivity of carbohydrates can be improved by increasing the sampling step potential.