Two-dimensional separation method for analysis of Bacillus subtilis metabolites via hyphenation of micro-liquid chromatography and capillary electrophoresis
Two-dimensional separation method for analysis of Bacillus subtilis metabolites via hyphenation of micro-liquid chromatography and capillary electrophoresis
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DOI：
10.1021/ac035039b
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发表时间：
2004-03-01
影响因子：
7.4
通讯作者：
Nishioka, T
中科院分区：
文献类型：
--
作者：
Jia, L;Liu, BF;Nishioka, T
建立了一种用于分析枯草芽孢杆菌代谢产物的新型双向分离方法。第一维为微液相色谱(LC)，第二维为毛细管电泳法(CE)。采用动态pH连接和扫描等浓缩策略，实现了两个维度的选择性对接，有利于代谢物的检测。为了进行系统评价，采用梯度模式的微型LC对含有54种标准代谢物的人工样品进行了疏水性分离。用毛细管区带电泳法结合动态pH连接分离早洗脱组分，用扫描胶束电动毛细管色谱分离晚洗脱组分。用两种毛细管电泳法对中间组分进行分析。在最佳条件下，人工样品中的所有组分均能得到很好的分离。将该方法应用于枯草杆菌代谢产物的分析。鉴定出了一些关键的代谢物。该方法为分离含有不同性质化合物的复杂生物样品提供了极大的潜力。
A novel two-dimensional separation method, which hyphenated chromatography and electrophoresis, was developed for analysis of Bacillus subtilis metabolites. Micro-liquid chromatography (LC) with a monolithic silica-ODS column was used as the first dimension, from which the effluent fractions were further analyzed by capillary electrophoresis (CE) acting as the second dimension. Concentration strategies, namely, dynamic pH junction and sweeping, were selectively employed to interface the two dimensions, which proved to be beneficial for the detection of metabolites. For system evaluation, an artificial sample containing 54 standard metabolites was separated according to their hydrophobicity by micro-LC with gradient mode. The early-eluting fractions were separated by capillary zone electrophoresis in combination with dynamic pH junction, while the late-eluting fractions were separated by sweeping micellar electrokinetic chromatography. The middle fractions were analyzed by both modes of CE. Under the optimum conditions, all the components in the artificial sample could be well resolved. The method was applied to profile B. subtilis metabolites. Some crucial metabolites were identified. This method provided great potential for resolving complex biological samples containing compounds having different characteristics.