Improving LC-MS sensitivity through increases in chromatographic performance: Comparisons of UPLC-ES/MS/MS to HPLC-ES/MS/MS
Improving LC-MS sensitivity through increases in chromatographic performance: Comparisons of UPLC-ES/MS/MS to HPLC-ES/MS/MS
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DOI：
10.1016/j.jchromb.2005.05.037
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发表时间：
2005-10-25
影响因子：
3
通讯作者：
Doerge, DR
中科院分区：
文献类型：
--
作者：
Churchwell, MI;Twaddle, NC;Doerge, DR
最近的技术进步使得反相色谱介质的粒径为 1.7 μm，并且液体处理系统可以在更高的压力下操作此类色谱柱。这项技术被称为超高效液相色谱 (UPLC)，在分析测定的分辨率、速度和灵敏度方面具有显着的理论优势，特别是与能够高速采集的质谱仪结合使用时。本文通过对 UPLC 的几种方法进行并列比较，探讨了 LC-MS 性能的差异，这些方法之前针对基于 HPLC 的多种分析物的分离和定量进行了优化，具有最大通量。总体而言，UPLC 在方法灵敏度、速度和分辨率方面产生了显着改进。 UPLC 的灵敏度提高了 10 倍（被发现与分析物相关），并且在峰分离相当的条件下，方法速度提高了 5 倍。 UPLC 的色谱分离度明显提高，峰宽普遍较窄，而且 HPLC 无法分离非对映异构体。总体而言，UPLC 在 LC-MS 方法灵敏度、速度和分辨率方面的改进表明，分析方法可以取得进一步的进步，从而为假设驱动的研究增加显着的价值。 (c) 2005 Elsevier B.V. 保留所有权利。
Recent technological advances have made available reverse phase chromatographic media with a 1.7 mu m particle size along with a liquid handling system that can operate such columns at much higher pressures. This technology, termed ultra performance liquid chromatography (UPLC), offers significant theoretical advantages in resolution, speed, and sensitivity for analytical determinations, particularly when coupled with mass spectrometers capable of high-speed acquisitions. This paper explores the differences in LC-MS performance by conducting a side-by-side comparison of UPLC for several methods previously optimized for HPLC-based separation and quantification of multiple analytes with maximum throughput. In general, UPLC produced significant improvements in method sensitivity, speed, and resolution. Sensitivity increases with UPLC, which were found to be analyte-dependent, were as large as 10-fold and improvements in method speed were as large as 5-fold under conditions of comparable peak separations. Improvements in chromatographic resolution with UPLC were apparent from generally narrower peak widths and from a separation of diastereomers not possible using HPLC. Overall, the improvements in LC-MS method sensitivity, speed, and resolution provided by UPLC show that further advances can be made in analytical methodology to add significant value to hypothesis-driven research. (c) 2005 Elsevier B.V. All rights reserved.