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使用加热液滴接口将液相色谱与基质辅助激光解吸电离质谱联用。

Combining liquid chromatography with MALDI mass spectrometry using a heated droplet interface.

作者信息

Zhang Boyan, McDonald Chris, Li Liang

机构信息

Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada T6G 2G2.

出版信息

Anal Chem. 2004 Feb 15;76(4):992-1001. doi: 10.1021/ac034934s.

Abstract

A novel interfacing technology is described to combine solution-based separation techniques such as liquid chromatography (LC) with matrix-assisted laser desorption ionization (MALDI) mass spectrometry. The interface includes a transfer tube having an inlet and an outlet, the inlet being adapted to accept the LC effluents and the outlet being adapted to form continuously replaced, hanging droplets of the liquid stream, and a MALDI sample plate mounted below the outlet of the transfer tube for collecting the droplets. The liquid stream in the transfer tube is heated to a temperature sufficient to cause partial evaporation of the carrier solvent from the hanging droplets. The droplets are dislodged to the MALDI plate, which is heated to above the boiling point of the carrier solvent to cause further evaporation of the carrier solvent from the collected droplets. It is found that analytes can be fractionated and deposited to a sample spot of 0.8 mm in diameter when a liquid flow rate of up to 50 microL/min and a fractionation interval of 1 min/spot are used. Flow rate of up to 200 microL/min can be used with a deposition sample spot of 2.4 mm in diameter on a commercial MALDI target. This heated droplet interface does not introduce sample loss, and the detection sensitivity of LC/MALDI is similar to that of standard MALDI, i.e., low femtomoles for peptide analysis with a microliter sample deposition. It is compatible with microbore and narrow-bore column separation, thus allowing the injection of a larger amount of sample for separation and analysis, compared to a capillary column LC/MALDI system. The detection dynamic range is shown to be in the order of 10(6) for peptide mixture analysis, which is 4 orders of magnitude greater than standard MALDI. The application of this interface for combining LC with MALDI MS/MS is demonstrated in the proteome analysis of water-soluable protein components of E. coli K12 extracts.

摘要

描述了一种新型接口技术,用于将基于溶液的分离技术(如液相色谱(LC))与基质辅助激光解吸电离(MALDI)质谱相结合。该接口包括一个具有入口和出口的传输管,入口适于接收LC流出物,出口适于形成连续替换的液流悬滴,以及一个安装在传输管出口下方用于收集液滴的MALDI样品板。传输管中的液流被加热到足以使载体溶剂从悬滴中部分蒸发的温度。液滴被转移到MALDI板上,MALDI板被加热到高于载体溶剂的沸点以促使载体溶剂从收集的液滴中进一步蒸发。结果发现,当使用高达50微升/分钟的液体流速和1分钟/点的分馏间隔时,分析物可以被分馏并沉积到直径为0.8毫米的样品点上。在商业MALDI靶上,高达200微升/分钟的流速可用于直径为2.4毫米的沉积样品点。这种加热液滴接口不会导致样品损失,LC/MALDI的检测灵敏度与标准MALDI相似,即对于微升样品沉积的肽分析为低飞摩尔级。它与微径和窄径柱分离兼容,因此与毛细管柱LC/MALDI系统相比,允许注入更多的样品进行分离和分析。对于肽混合物分析,检测动态范围显示为10(6)量级,比标准MALDI大4个数量级。在大肠杆菌K12提取物的水溶性蛋白质组分的蛋白质组分析中证明了该接口在将LC与MALDI MS/MS相结合方面的应用。

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