Tandem mass spectrometry of peptides using hybrid and four-sector instruments: a comparative study.
Tandem mass spectrometry of peptides using hybrid and four-sector instruments: a comparative study.
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使用混合仪器和四扇区仪器进行肽的串联质谱分析：比较研究。
DOI：
10.1021/ac00014a024
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发表时间：
1991
影响因子：
7.4
通讯作者：
Gaskell,SJ
中科院分区：
文献类型：
--
作者：
Bean,MF;Carr,SA;Thorne,GC;Reilly,MH;Gaskell,SJ
在四区串联质谱仪和混合串联质谱仪上，比较了一系列多肽(Mr 550-2500)[+H]+离子的高能和低能Colll·斜面激活解离(CAD)产生的乘积离子光谱。被分析的最小的多肽(蛋氨酸-脑啡肽)的快原子轰击产物离子光谱非常相似，但对于较重的分析物，在碎裂方面有很大的差异。对于含有M，>；1000的多肽，从四扇区仪的高能计算机辅助设计中获得了更完整的序列信息。然而，混合质谱计上的低能量CAD能够产生部分序列信息，即使是对于所比较的最大的多肽。小肽的分析下限(约15PMOI)相似(约15PMOI)，但较大的多肽(AFR>；1000)的分析下限可用四区仪器获得。高能CAD光谱被发现具有高度的重现性，在广泛的操作条件下获得了定性相似的光谱。相反，需要仔细控制碰撞气体压力和碰撞能量，以便为低能量CAD获得良好的可重复性数据。给出了在混合仪器上获得重复性好、灵敏度好的多肽光谱的实验方法。
Product-Ion spectra produced by high-and low-energy colll· slonally activated dissociation (CAD) of [+ H]+ ions of a series of peptides (Mr 550-2500) have been compared on four-sector and hybrid tandem mass spectrometers, respectively. The fast atom bombardment product-ion spectra ob-tained for the smallest peptide analyzed (methionine-enkephalin) were remarkably similar, but substantial differences In fragmentation were observed for the heavier analytes. For peptides with M,> 1000, more complete sequence Informa-tion was obtained from high-energy CAD on the four-sector Instrument. Nevertheless, low-energy CAD on the hybrid mass spectrometer was able to produce partial sequence Information even for the largest of the peptides compared. Limits of analysis, defined as the least quantities of analyte tor which product-ion spectra of essentially uncompromised quatty could be obtained, were similar (ca. 15 pmoi) for small peptides, but lower limits were achieved for larger peptides (Afr> 1000) with the four-sector Instrument. High-energy CAD spectra were found to be highly reproducible, with qualitatively similar spectra obtained over a wide range of operating conditions. In contrast, It was necessary to care-fully control collision gas pressures and collision energies In order to obtain good reproducible data for low-energy CAD. Experimental procedures for obtaining reproducible spectra with good sensitivity for peptides on the hybrid Instrument are presented.