Panagiotopoulos C, Sempéré R, Lafont R, Kerhervé P
Laboratoire de Microbiologie Marine, CNRS-INSU UMR 6117, Centre d'Océanologie de Marseille, Université de la Méditerranée, France.
J Chromatogr A. 2001 Jun 22;920(1-2):13-22. doi: 10.1016/s0021-9673(01)00697-5.
The effects of column temperature in the range 10-45 degrees C using high-performance anion-exchange chromatography (HPAEC) and pulse amperometric detection are described for the determination of monosaccharides. The influence of temperature was tested with an isocratic elution of NaOH at concentrations varying from 2.5 to 20 mM and with a post-column addition of 1 M NaOH. The results showed that small changes of temperature greatly affect retention times and resolution (Rs) of monosaccharides and particularly those of the both pairs xylose-mannose and rhamnose-arabinose which cannot be simultaneously detected at usual room temperature (approximately 25 degrees C). Our results suggest that a subambient temperature of 17 degrees C and an eluent concentration of 19 mM are the more appropriate conditions for an acceptable separation (R(s rha/ara) = 1.02, R(s man/xyl) = 0.70) in a short analytical run time (35 min). The results showed that within the range of temperatures studied, enthalpy and entropy are invariant of temperature indicating that changes in the retention processes are mainly due to temperature than other associated changes in the system. This study demonstrated the importance of controlling temperature during HPAEC of monosaccharides, both to accomplish highly reproducible retention times and to achieve optimal separation of sugars. This method gave acceptable results for detection of marine sugars.
描述了使用高效阴离子交换色谱法（HPAEC）和脉冲安培检测法在10-45摄氏度范围内的柱温对单糖测定的影响。在2.5至20 mM浓度范围内的NaOH等度洗脱以及柱后添加1 M NaOH的条件下测试了温度的影响。结果表明，温度的微小变化会极大地影响单糖的保留时间和分离度（Rs），尤其是木糖-甘露糖和鼠李糖-阿拉伯糖这两对单糖，在通常的室温（约25摄氏度）下无法同时检测到。我们的结果表明，17摄氏度的低于环境温度和19 mM的洗脱液浓度是在短分析运行时间（35分钟）内实现可接受分离（R(s rha/ara) = 1.02，R(s man/xyl) = 0.70）的更合适条件。结果表明，在所研究的温度范围内，焓和熵与温度无关，这表明保留过程中的变化主要是由温度引起的，而非系统中的其他相关变化。本研究证明了在单糖的HPAEC过程中控制温度的重要性，这对于实现高度可重复的保留时间和实现糖类的最佳分离均很重要。该方法在检测海洋糖类方面给出了可接受的结果。
