Improving the performance of MALDI-TOF in oligonucleotide analysis using a new SDIFA technology.
Improving the performance of MALDI-TOF in oligonucleotide analysis using a new SDIFA technology.
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使用新的 SDIFA 技术提高 MALDI-TOF 在寡核苷酸分析中的性能。
DOI：
10.1021/ac0007231
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发表时间：
2000
影响因子：
7.4
通讯作者：
Fan,Y
中科院分区：
文献类型：
--
作者：
Guo,B;Wang,S;Fan,Y
本文提出了一种新技术SDIFA，用于提高线性基质辅助激光解吸电离飞行时间质谱在寡核苷酸分析中的质量分辨率。与目前使用的延迟提取方法不同，SDIFA允许样品保持器与离子提取/加速电隔离，并选择性地对部分解吸离子进行采样，从而减小初始速度分布并提高分辨率。此外，还提出了一种改善TOF空间聚焦的方法。对于高达62 mer的寡核苷酸，获得了同位素限制的质量分辨率，并且在19.2 kDa处，真实的仪器分辨率达到1800。还证明了使用单个采集参数设置在大质量范围内获得了优异的分辨率。SDIFA的性能比DE更稳定、重现性更好，对激光功率、样品点、样品基质、延迟时间和提取场强等实验条件的依赖性更小。这种增强的数据采集简易性是自动化光谱采集的关键。
A new technology termed SDIFA is developed to improve the mass resolution of linear matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in oligonucleotide analysis. Unlike the currently used delayed extraction method, SDIFA allows electrical isolation of the sample holder from ion extraction/acceleration and selectively samples part of the desorbed ions, thereby reducing the initial velocity distribution and improving resolution. In addition, a method was introduced to improve the space focusing of TOF. Isotope-limited mass resolution was obtained for oligonucleotides of up to 62 mer, and the true instrumental resolution reaches to 1800 at 19.2 kDa. It was also demonstrated that excellent resolution was obtained across a large mass range using a single setting of acquisition parameters. This feature allows unambiguous identification of multiple A/T heterozygote samples in a mass range of 5200−7800 Da. Moreover, compared with DE, the performance of SDIFA was more stable, reproducible, and less dependent on the experimental conditions including the laser power, sample spots, sample substrates, delayed time, and extraction field strength. This enhanced ease of data acquisition is the key to automated spectrum acquisition.