Journal article Open Access

Improving LC–MS sensitivity through increases in chromatographic performance: Comparisons of UPLC–ES/MS/MS to HPLC–ES/MS/MS

Churchwell, M.; Twaddle, N.; Meeker, L.; Doerge, D.


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    <subfield code="a">Recent technological advances have made available reverse phase chromatographic
media with a 1.7 micron 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.</subfield>
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    <subfield code="a">Twaddle, N.</subfield>
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    <subfield code="a">10.1016/j.jchromb.2005.05.037</subfield>
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