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使用连续流接口的微径高效液相色谱/质谱联用技术分析肽混合物

Microbore HPLC/mass spectrometry for the analysis of peptide mixtures using a continuous flow interface.

作者信息

Caprioli R M, DaGue B, Fan T, Moore W T

出版信息

Biochem Biophys Res Commun. 1987 Jul 15;146(1):291-9. doi: 10.1016/0006-291x(87)90724-8.

Abstract

Microbore HPLC techniques have been combined with fast atom bombardment mass spectrometry to provide HPLC/MS capabilities for the analysis of mixtures of peptides and small proteins. The interface between the liquid chromatograph and mass spectrometer is a continuous flow direct insertion probe which contains a fused silica capillary that delivers the eluting solvent to the FAB source of the mass spectrometer at a rate of 5-10 microL/min. Data are presented for the analysis of several mixtures of peptides ranging in molecular weights from about 900 to 6000 daltons. In addition, the analysis of 100 pmol of a tryptic digest of whale myoglobin is shown where 16 of the possible 19 peptides were identified in the mass range m/z 2200-250. The advantages of this approach to HPLC/MS are a relatively high sensitivity because of the low flow rates and low background, and the ability to detect high molecular weight compounds.

摘要

微径高效液相色谱(HPLC)技术已与快原子轰击质谱联用,以实现HPLC/MS功能,用于分析肽和小蛋白质混合物。液相色谱仪和质谱仪之间的接口是一个连续流动直接插入探头,其中包含一根熔融石英毛细管,该毛细管以5-10微升/分钟的流速将洗脱溶剂输送到质谱仪的快原子轰击源。给出了对几种分子量在约900至6000道尔顿范围内的肽混合物进行分析的数据。此外,还展示了对100皮摩尔鲸肌红蛋白胰蛋白酶消化产物的分析,其中在m/z 2200-250质量范围内鉴定出了19种可能肽中的16种。这种HPLC/MS方法的优点是由于流速低和背景低而具有相对较高的灵敏度,以及能够检测高分子量化合物。

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