Caprioli R M, Moore W T, DaGue B, Martin M
Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston 77030.
J Chromatogr. 1988 Jun 29;443:355-62. doi: 10.1016/s0021-9673(00)94807-6.
Microbore high-performance liquid chromatographic (HPLC) techniques have been combined with fast atom bombardment mass spectrometry (FAB-MS) for the mass-specific detection of mixtures of peptides produced by proteolytic hydrolysis of proteins. The continuous-flow FAB interface has been utilized for direct coupling of the microbore HPLC system and the mass spectrometer. Conditions are reported for the effective separation of 100 pmol of peptides at flow-rates of 5 microliters/min with acetonitrile gradients and 1-mm I.D. C8 columns. A comparison is also made between columns of 5 and 25 cm lengths for the separation of peptide mixtures. Data are presented for the separation of peptides on a slurry-packed C18 fused-silica capillary column with the continuous-flow HPLC-FAB-MS interface at flow-rates of 3 microliters/min.
微径高效液相色谱（HPLC）技术已与快原子轰击质谱（FAB-MS）相结合，用于对蛋白质经蛋白水解产生的肽混合物进行质量特异性检测。连续流FAB接口已用于微径HPLC系统与质谱仪的直接联用。本文报道了在乙腈梯度和内径为1mm的C8柱上，以5微升/分钟的流速有效分离100皮摩尔肽的条件。还比较了5厘米和25厘米长的柱子对肽混合物的分离效果。给出了在流速为3微升/分钟的情况下，使用连续流HPLC-FAB-MS接口在装填有浆液的C18熔融石英毛细管柱上分离肽的数据。
