Void volume markers in reversed-phase and biomimetic liquid chromatography

A comparative investigation of 15 void volume marker candidates was carried out on two reversed-phase (RP), two immobilized artificial membrane (IAM) and two immobilized protein columns, namely Human Serum Albumin (HSA) and alpha(1)-acid-glycoprotein (AGP), using different mobile phases in respect to pH and buffer composition. Pycnometric analysis of the 6 chromatographic columns was also employed for reasons of comparison. The results revealed the possible overestimation of stationary phase void volume if a general void volume marker is used under different chromatographic conditions. In particular, overestimation of the investigated columns' void volumes was a common phenomenon when using acidic eluents or High Purity Water (HPW) as mobile phase. For this purpose, a classification of the recommended chemicals according to the column type or the pH of the mobile phase is suggested for the accurate determination of retention factors. (C) 2010 Elsevier B.V. All rights reserved.