Environmental Analysis By On-Line Immunoextraction And Reversed-Phase Liquid Chromatography: Optimization Of The Immunoextraction/RPLC Interface

Authors

Mary Anne Nelson, University of Nebraska - Lincoln
Efthimia Papastavros, University of Nebraska-LincolnFollow
Maud Dodlinger, University of Nebraska - Lincoln
David S. Hage, University of Nebraska - LincolnFollow

Date of this Version

5-2007

Citation

Published in final edited form as: J Agric Food Chem. 2007 May 16; 55(10): 3788–3797. doi:10.1021/jf063286l.

Comments

Copyright Elsevier Inc. Used By Permission.

Abstract

The use of antibodies in HPLC columns for on-line immunoextraction combined with reversed-phase liquid chromatography (RPLC) is of growing interest in environmental and agricultural analysis. This technique is typically performed by using a small RPLC precolumn to capture and concentrate analytes as they elute from the immunoextraction column; however, there is little information on the conditions required for optimizing this interface. This study examined the behavior of this interface by using 2,4-dichlorophenoxyacetic acid (2,4-D) and related herbicides as model analytes. It was found that analyte dissociation from immunoextraction columns followed first-order decay and that the elution of these analytes through the immunoextraction/RPLC interface gave an exponentiallymodified Gaussian profile. Computer simulations were used to see how analyte elution through the interface changed with different dissociation and retention conditions. Several guidelines were developed from this work that could be used for developing and optimizing on-line immunoextraction/RPLC systems for other chemicals of environmental or agricultural interest.