Sensitivity enhancement and second-dimensional information from solid phase extraction-capillary electrophoresis of entire high-performance liquid chromatography fractions.
Sensitivity enhancement and second-dimensional information from solid phase extraction-capillary electrophoresis of entire high-performance liquid chromatography fractions.
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整个高效液相色谱馏分的固相萃取毛细管电泳的灵敏度增强和二维信息。
DOI：
10.1002/elps.1150160189
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发表时间：
1995
期刊：
影响因子：
2.9
通讯作者：
Landers,JP
中科院分区：
文献类型：
--
作者：
Strausbauch,MA;Madden,BJ;Wettstein,PJ;Landers,JP
展示了一种新颖的分离技术，可在单一低 pH 毛细管区带电泳 (CZE) 分离中对整个 100 μL 高效液相色谱 (HPLC) 组分进行肽分析。该方法采用混合毛细管，该混合毛细管由包含反相填料的可拆卸入口部分和标准 CZE 裸熔石英分离毛细管组成。填充吸头允许样品以大于 20 μL/min 的流速浓缩，并以受控方式释放以进行 CZE 分离。分离效果与标准 CZE 相当，但选择性略有改变。使用模型肽的混合物，混合毛细管可将灵敏度提高 ≥ 100 倍。通过固相萃取毛细管电泳 (SPE-CE) 方法对牛血清白蛋白 (BSA) 消化获得的肽进行 HPLC 分离的五个组分进行分析，该方法被证明可以提供快速的二维信息。使用该技术计算出肽的实际检测浓度限为 1‐‐10 ng/mL。
A novel separation technique is demonstrated for peptide analysis of entire 100 μL high‐performance liquid chromatography (HPLC) fractions in a single, low pH, capillary zone electrophoresis (CZE) separation. The method employs a hybrid capillary consisting of a removable inlet section containing a reverse‐phase packing and a standard CZE bare‐fused silica separation capillary. The packed tip allows sample to be concentrated at flow rates greater than 20 μL/min and released in a controlled manner for CZE separation. Separations are comparable to standard CZE with minor modifications in selectivity. Using a mixture of model peptides, the hybrid capillary is shown to effect enhancement in sensitivity of ≥ 100‐fold. Five fractions from the HPLC separation of peptides obtained from the digestion of bovine serum albumin (BSA) were analyzed by the solid phase extraction‐capillary electrophoresis (SPE‐CE) method which was shown to provide rapid second‐dimensional information. Practical concentration limits of detection for peptides are calculated to be 1‐‐10 ng/mL with this technology.