Implications of Extra-column Effects for Targeted or Untargeted Microflow LC-MS

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00179906%3A_____%2F25%3A10499792" target="_blank" >RIV/00179906:_____/25:10499792 - isvavai.cz</a>

Alternative codes found

RIV/00216208:11160/25:10499792

Result on the web

<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=R6prtYdox5" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=R6prtYdox5</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/acsmeasuresciau.5c00015" target="_blank" >10.1021/acsmeasuresciau.5c00015</a>

Result language

angličtina

Original language name

Implications of Extra-column Effects for Targeted or Untargeted Microflow LC-MS

Original language description

Mass spectrometry (MS) has changed our understanding of health, disease, and the environment through untargeted analyses where entire molecular classes are investigated. These techniques generate huge amounts of data which when processed by statistical tools can identify important molecular features or biomarkers. The complexities of these samples are not compatible with direct introduction to the MS system and require a high-resolution separation step, typically low flow liquid chromatography (LC), prior to MS. LC columns that can produce adequate linear velocities at these low flow rates are small in volume making their results susceptible to resolution loss in extra-column volumes. Here, we investigate the implications of the extra-column effects in five LC-MS systems with triple quadrupole and orbitrap mass analyzers. The extra-column volume of these systems in their standard configuration ranged from 26.4 to 78.1 mu L which we reduced to 9.57 to 18.7 mu L by optimizing the fluidics. The effects of this volume reduction were assessed by studying a hydrolyzed protein sample in a proteomics environment where the intensity of the largest MS peak was improved by 1.8-3.8x. Additionally, the number of molecular features detected in the protein sample improved by up to 7.5x. The relationship between extra-column volumetric variance and flow rate shows that broadening will become much larger for MS detectors at higher flow rates, unlike a traditional small volume UV detector. The methods, applications, and theoretical insights in this work can be used to improve the mass spectrometric results of any LC-MS system.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10406 - Analytical chemistry

Project

—

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2025

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

ACS Measurement Science au

ISSN

—

e-ISSN

2694-250X

Volume of the periodical

5

Issue of the periodical within the volume

3

Country of publishing house

US - UNITED STATES

Number of pages

13

Pages from-to

332-344

UT code for WoS article

001461930300001

EID of the result in the Scopus database

2-s2.0-105002762947