ESI/ion trap/ion mobility/time-of-flight mass spectrometry for rapid and sensitive analysis of biomolecular mixtures
ESI/ion trap/ion mobility/time-of-flight mass spectrometry for rapid and sensitive analysis of biomolecular mixtures
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DOI：
10.1021/ac9809175
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发表时间：
1999-01-15
影响因子：
7.4
通讯作者：
Clemmer, DE
中科院分区：
文献类型：
--
作者：
Henderson, SC;Valentine, SJ;Clemmer, DE
离子阱/离子迁移率/飞行时间质谱技术是一种快速、灵敏的分析多肽/蛋白质混合物的方法。在这种方法中，离子阱被用来将从混合物中电喷出来的离子聚集到浓缩的包中。离子包被注入到一个漂移管中，其中混合物的组分根据通过缓冲气体的迁移率的差异被分离。离开漂移管的离子在飞行时间质谱仪中分散，以测定质量电荷比（m/z）。气相分离策略减少了质谱中的拥塞，实验迁移率补充了分配峰的m/z测量。该方法的应用实例，以鉴定肽（从胰蛋白酶消化）和分离电荷状态分布的电喷雾混合物含有泛素和肌红蛋白被提出。从细胞色素c和肌红蛋白等蛋白质的胰蛋白酶消化中观察到的大多数肽可以在1分钟内从获得的数据中识别出来；对已知成分混合物的研究表明，单个成分的检出限类似于0.5-3 pmol。讨论了可能影响所观察到的分布的因素，如在阱中的储存时间、用于迁移率实验的注入电压以及离子截面随电荷状态的变化。
An ion trap/ion mobility/time-of-flight mass spectrometry technique is shown to be a rapid and sensitive means of analyzing peptide/protein mixtures. In this approach, an ion trap is used to accumulate ions that have been electrosprayed from a mixture into concentrated packets. The ion packets are injected into a drift tube where components of the mixture are separated based on differences in mobility through a buffer gas. Ions that exit the drift tube are dispersed in a time-of-flight mass spectrometer for mass-to-charge (m/z) determination. The gas-phase separation strategy reduces congestion in the mass spectrum, and experimental mobilities complement m/z measurements in assigning peaks. Examples of the application of the approach to identification of peptides (from tryptic digests) and to separation of charge-state distributions from electrospray of a mixture containing ubiquitin and myoglobin are presented. Most peptides that are observed from tryptic digests of proteins such as cytochrome c and myoglobin can be identified from data that are acquired in under 1 min; studies of mixtures with known compositions indicate that detection limits are similar to 0.5-3 pmol for individual components. Factors that may influence the distributions that are observed, such as storage time in the trap, injection voltages used for the mobility experiment, and variations in ion cross section with charge state, are discussed.