Collect. Czech. Chem. Commun. 1989, 54, 953-966
https://doi.org/10.1135/cccc19890953

Determination of the dead volume of columns in reversed phase liquid chromatography

Ivan Víta, Jan Fähnrichb and Milan Poplb

a Laboratorní přístroje, 162 03 Prague 6
b Department of Analytical Chemistry, Prague Institute of Chemical Technology, 166 28 Prague 6

Abstract

The method for the determination of the dead volume of RPLC columns by measuring the retention volume of phloroglucinol is compared with the method based on the retention data of homologous series of organic solutes and that based on the use of isotopically labelled components of the mobile phase. The use of homologous series is tedious and moreover, satisfactory results are not always obtained. The phloroglucinol method is found well suited to replacing the method based on the use of isotopically labelled components of the mobile phase; poorer results with phloroglucinol are only obtained when the mobile phase contains more than 40% (v/v) water, for the Separon SE polymeric sorbent, and in alkaline medium.