Burkitt William, O'Connor Gavin
LGC, Queens Road, Teddington TW11 0LY, UK.
Rapid Commun Mass Spectrom. 2008 Dec;22(23):3893-901. doi: 10.1002/rcm.3794.
A system to perform automated hydrogen/deuterium exchange mass spectrometry measurements was constructed using an XYZ robotic autosampler that was capable of performing solvent manipulations and a 4.7 T Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer. The system included features such as the first demonstration of a 'dual column' high-performance liquid chromatography (HPLC) setup, and a novel digestion strategy. The performance of the system, in terms of the repeatability and reproducibility of the measurement of protein hydrogen/deuterium exchange, was assessed over a 2-month period. The sensitivity of the measurement of hydrogen exchange towards several parameters was assessed, which allowed their impact on the reproducibility to be discussed. The parameters assessed were the temperature of the HPLC columns and switching valves, the temperature of the quench solutions, the pH of the mobile phase, the pH of the quenched solution, the acid used in the mobile phase and the analytical column used.
利用一台能够进行溶剂操作的XYZ机器人自动进样器和一台4.7 T傅里叶变换离子回旋共振（FT-ICR）质谱仪构建了一个用于进行自动化氢/氘交换质谱测量的系统。该系统具有多项特性，例如首次展示了“双柱”高效液相色谱（HPLC）装置以及一种新颖的消化策略。在为期2个月的时间里评估了该系统在蛋白质氢/氘交换测量的重复性和再现性方面的性能。评估了氢交换测量对几个参数的灵敏度，从而能够讨论这些参数对再现性的影响。评估的参数包括HPLC柱和切换阀的温度、淬灭溶液的温度、流动相的pH值、淬灭后溶液的pH值、流动相中使用的酸以及所使用的分析柱。
