Towards an accurate method for column void volume determination using liquid chromatography-mass spectrometry

David Ribar (Author), Miha Lukšič (Author), Irena Kralj Cigić (Author)

Abstract

Prediction of analyte retention times requires prior knowledge of the column void volume, the measurement of which is still highly contested within the literature and therefore experimental based prediction is often used. In this study, we investigated deuterated acetonitrile as an isotopically labelled mobile phase component to observe its elution behaviour in a binary mixture with water at 25 different mobile phase compositions (from 5 to 95 vol. % of acetonitrile), on two stationary phases (C8 and C18), and at two temperatures (30 and 40 °C) using LC-MS. The same experimental design was additionally used for three commonly used neutral void volume markers: uracil, phloroglucinol and N,N-dimethylformamide. Temperature was observed to influence the elution of acetonitrile in an inversely proportional manner with higher temperatures coinciding with lower elution times. By utilizing a three-way ANOVA, the composition of the mobile phase has been shown to have a significant effect on deuterated acetonitrile and other investigated void volume markers, demonstrating the fact that both void volume markers and acetonitrile itself exhibit retention-like behaviour. Excess adsorption isotherms for acetonitrile were calculated using deuterated acetonitrile elution data. The comparison of void volumes, obtained with conventional neutral void volume markers, revealed the former to be 24–36% lower than the void volume obtained using deuterated acetonitrile, as an isotopically labelled mobile phase component. For a water:acetonitrile mobile phase, the minor disturbance method using deuterated acetonitrile to obtain an integral average void volume (2.08 and 2.05 mL for C18 at 30 and 40 °C, respectively and 2.16 and 2.13 mL for C8 at 30 and 40 °C, respectively) was found to be the most appropriate method for determining the elusive column void volume.

Keywords

reverzno-fazna tekočinska kromatografija;mrtvi volumen;markerji mrtvega volumna;retencijski mehanizem;modeliranje;reversed-phase liquid chromatography;void volume;void volume markers;retention mechanism;modelling;

Data

Language:	English
Year of publishing:	2023
Typology:	1.01 - Original Scientific Article
Organization:	UL FKKT - Faculty of Chemistry and Chemical Technology
UDC:	543.544.5:543.51
COBISS:	159876099
ISSN:	0021-9673
Views:	182
Downloads:	30
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	Slovenian
Secondary keywords:	reverzno-fazna tekočinska kromatografija;mrtvi volumen;markerji mrtvega volumna;retencijski mehanizem;modeliranje;
Type (COBISS):	Article
Pages:	str. 1-7
Issue:	ǂVol. ǂ1706, [article no.] 464245
Chronology:	13 Sep. 2023
DOI:	10.1016/j.chroma.2023.464245
ID:	19766071