Analysis of multiple herbicides in soybeans using pressurized liquid extraction and capillary electrophoresis

Several herbicides commonly used on soybeans are often difficult to extract, isolate, and quantify from the complex soybean matrix at low concentrations. Typical analytical methods for herbicide residues in soybeans are single analyte procedures using HPLC or GC after chemical derivatization. In this study, method development for the analysis of six polar herbicides in soybeans was performed using pressurized liquid extraction (PLE), which is also known by the trade name, Accelerated Solvent Extraction, and capillary electrophoresis (CE). In CE, a 50 mM ammonium acetate running buffer, pH 4.75, was able to separate imazaquin (Scepter), chlorimuron-ethyl (Classic), thifensulfuron-methyl (Harmony), acifluorfen (Blazer), bentazon (Basagran), and 2,4-dichlorophenoxyacetic acid in a 75 mu m i.d., 83 cm capillary (65 cm to detector) within 30 min at 17 kV applied voltage. Chlorsulfuron (Glean) was used as an internal standard in the analysis, and detection was by UV absorbance at 240 nm in a high-sensitivity optical cell. PLE extracts required extensive cleanup prior to CE/UV analysis. Several cleanup techniques were investigated and compared, including liquid-liquid partitioning, gel-permeation chromatography, semipreparative HPLC, and solid-phase extraction with a variety of stationary and mobile phase combinations. A combination of techniques that provided the most efficient cleanup was selected in the final method. Four of the six herbicides could be determined by the method in samples fortified at tolerance levels with average recoveries of 71% and relative standard deviation (RSD) of 11%. At a higher spiking level, all of the herbicide recoveries were > 70% with %RSDs < 10%, except for acifluorfen which gave more variable recoveries.