Antonio Carla, Pinheiro Carla, Chaves Maria Manuela, Ricardo Cândido Pinto, Ortuño Maria Fernanda, Thomas-Oates Jane
Department of Chemistry, University of York, Heslington, York YO10 5DD, UK.
J Chromatogr A. 2008 Apr 11;1187(1-2):111-8. doi: 10.1016/j.chroma.2008.02.010. Epub 2008 Feb 8.
This work reports the development and application of a negative ion mode online LC-ESI-MS method for studying the effect of water deficit on the carbohydrate content of Lupinus albus stems, using a porous graphitic carbon (PGC) stationary phase and an ion trap mass spectrometer. Using this method, separation and detection of several water soluble carbohydrates, ranging from mono-, di-, and oligosaccharides (raffinose, stachyose, and verbascose) to sugar alcohols was achieved in approximately 10 min. This on-line PGC-LC-ESI-MS method shows good linearity with correlation coefficients R(2)>0.99, selectivity, short analysis time, and limits of detection (LOD) ranging from 0.4 to 9 pmol for sugars and 4-20 pmol for sugar alcohols. This PGC-LC-ESI-MS method is sensitive and allowed us to detect even small alterations in carbohydrate levels in L. albus stems that resulted from a mild/early water deficit (nmol g(-1)DW). This paper describes details of our method and its application to the quantitative analysis of water soluble underivatised carbohydrates extracted from L. albus stem tissues that have been subjected to early and severe water deficit conditions, followed by a rewatering period.
本研究报道了一种负离子模式在线液相色谱 - 电喷雾电离质谱法的开发与应用，该方法采用多孔石墨化碳（PGC）固定相和离子阱质谱仪，用于研究水分亏缺对白羽扇豆茎中碳水化合物含量的影响。使用该方法，在约10分钟内实现了从单糖、二糖和寡糖（棉子糖、水苏糖和毛蕊花糖）到糖醇等多种水溶性碳水化合物的分离和检测。这种在线PGC - LC - ESI - MS方法具有良好的线性，相关系数R(2)>0.99，选择性高，分析时间短，糖类的检测限（LOD）为0.4至9皮摩尔，糖醇的检测限为4至20皮摩尔。这种PGC - LC - ESI - MS方法灵敏度高，使我们能够检测到白羽扇豆茎中因轻度/早期水分亏缺而导致的碳水化合物水平的微小变化（纳摩尔克(-1)干重）。本文描述了我们的方法细节及其在定量分析从经历早期和严重水分亏缺条件并随后复水的白羽扇豆茎组织中提取的水溶性未衍生化碳水化合物方面的应用。
