Benavente Fernando, van der Heijden Rob, Tjaden Ubbo R, van der Greef Jan, Hankemeier Thomas
Division of Analytical Biosciences, Leiden/Amsterdam Center for Drug Research, Center for Medical Systems Biology, Leiden, The Netherlands.
Electrophoresis. 2006 Nov;27(22):4570-84. doi: 10.1002/elps.200600055.
We investigated the potential of CE coupled to electrospray MS (CE-ESI-MS) in metabolite profiling of human urine without any sample prefractionation step. A heterogeneous mixture of biologically relevant compounds covering a broad range of physicochemical properties was used to optimize separation conditions in fused-silica capillaries. A running electrolyte containing 50 mM of acetic acid and 50 mM of formic acid at pH 2.5 was used for the CE separations. A sheath-flow electrospray interface was employed for CE-ESI-MS analysis. Sheath liquids containing 80:20 v/v methanol/water with 0.1% v/v of acetic acid or 60:40 v/v isopropanol/water with 0.5% v/v of ammonia were selected for optimum detection in positive and negative ESI modes, respectively. Reproducibility and sensitivity were studied, and strategies for identification of the separated urinary compounds are suggested. We report major advantages and disadvantages of CE-ESI-MS for metabolite profiling of human body fluids. This work may be regarded as a first step in the use of CE-ESI-MS for reliable differential analysis of body fluids from healthy and diseased individuals.
我们研究了在不对尿液样本进行任何预分离步骤的情况下，毛细管电泳联用电喷雾质谱（CE-ESI-MS）用于人体尿液代谢物谱分析的潜力。使用了一种涵盖广泛物理化学性质的生物相关化合物的异质混合物来优化熔融石英毛细管中的分离条件。采用含有50 mM乙酸和50 mM甲酸、pH值为2.5的运行电解质进行毛细管电泳分离。采用鞘流电喷雾接口进行CE-ESI-MS分析。分别选择含有80:20 v/v甲醇/水和0.1% v/v乙酸的鞘液或含有60:40 v/v异丙醇/水和0.5% v/v氨的鞘液，以在正离子和负离子电喷雾模式下实现最佳检测。研究了重现性和灵敏度，并提出了鉴定分离出的尿液化合物的策略。我们报告了CE-ESI-MS用于人体体液代谢物谱分析的主要优缺点。这项工作可被视为将CE-ESI-MS用于健康个体和患病个体体液可靠差异分析的第一步。
