Liang Yu, Wu Ci, Dai Zhongpeng, Liang Zuocheng, Liang Zhen, Zhang Lihua, Zhang Yukui
Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, National Chromatographic R. & A. Center, Dalian 116023, China.
Se Pu. 2011 Jun;29(6):469-74. doi: 10.3724/sp.j.1123.2011.00469.
Due to the high throughput and high sensitivity, the hyphenation of microchip-based high performance liquid chromatography with tandem mass spectrometry has been paid much attention. In our recent work, with poly (lauryl methacrylate-co-trimethylolpropane trimethacrylate) monolithic materials prepared in microchannels as trap and separation columns, conventional micro-liquid chromatography pumps and valves for fluidic control, and a small-bore open-tube capillary attached to the outlet channel as chip-mass spectrometer (MS) interface, the microchip-based reversed-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS) platform was established, and applied for the identification of proteins. By such platform, 100 ng digest of bovine serum albumin (BSA) was successfully analyzed with the sequence coverages as 39.37%, 37.89% and 34.10% (with the relative standard deviation (RSD) of 7.3%) in three runs, separately. To evaluate the chip-to-chip reproducibility, BSA was identified by such platform with the microchips from different batches containing trap column, separation column and chip-MS interface. The obtained sequence coverage and the number of peptides identified were comparable. All these results showed high sensitivity and good reproducibility of such platform, demonstrating the great potential for rapid protein analysis.
由于具有高通量和高灵敏度，基于微芯片的高效液相色谱与串联质谱联用技术备受关注。在我们最近的工作中，以在微通道中制备的聚（甲基丙烯酸月桂酯 - 三羟甲基丙烷三甲基丙烯酸酯）整体材料作为捕集柱和分离柱，使用传统的微液相色谱泵和用于流体控制的阀门，并在出口通道连接一根小内径开口管毛细管作为芯片 - 质谱仪（MS）接口，建立了基于微芯片的反相液相色谱 - 串联质谱（RPLC - MS/MS）平台，并将其应用于蛋白质鉴定。通过该平台，在三次运行中分别成功分析了100 ng牛血清白蛋白（BSA）的酶解产物，序列覆盖率分别为39.37%、37.89%和34.10%（相对标准偏差（RSD）为7.3%）。为了评估芯片间的重现性，使用包含捕集柱、分离柱和芯片 - MS接口的不同批次微芯片，通过该平台对BSA进行鉴定。获得的序列覆盖率和鉴定出的肽段数量具有可比性。所有这些结果表明该平台具有高灵敏度和良好的重现性，展示了其在快速蛋白质分析方面的巨大潜力。
