Optimization of in-line fritless solid-phase extraction for capillary electrophoresis–mass spectrometry

In this study, in-line frit-free solid-phase extraction (SPE) has been studied for the preconcentration of analytes prior to analysis by capillary electrophoresis–mass spectrometry (CE–MS). The mixed-mode sorbent Oasis HLB was selected for the trapping of compounds of different polarity. Using 2-ethylidene-1,5-dimethyl-3,3-diphenylpirrolidine (EDDP), dihydrocodeine and codeine as test compounds, SPE parameters such as the pH of the sample and composition of the washing and elution solvent were optimized. Trapping of the analytes was optimal at pH 8.0 or higher. For efficient elution of the SPE micro column, 85% of methanol in water with 2% (v/v) acetic acid was used, which also prevented current break down in subsequent CE analysis. CE resolution of the test compounds was highest for background electrolytes (BGEs) with a pH above 8. For optimal analysis, samples were 1:1 diluted with carbonate buffer (1 M, pH 8.0) prior to analysis, BGE was 60 mM ammonium acetate buffer (pH 10.0), and the injected sample volume was 60 μl (i.e., 30 capillary volumes). Good recoveries were found: 101% for EDDP, 88% for codeine and 90% for dihydrocodeine. Intraday RSDs for migration time and peak areas were below 0.56% and 15%, respectively. Peak widths at half height obtained with SPE–CE–MS were 12 s for EDDP, 3.7 s for dihydrocodeine and 7.4 s for codeine, and were comparable to those for CE–MS. LODs were 0.22 pg/ml for EDDP, 2.1 pg/ml for dihydrocodeine and 24 pg/ml for codeine. It is concluded that the applied fritless in-line preconcentration construct proved to be highly useful for improving the sensitivity of CE while maintaining separation.

► An integrated SPE–CE–MS system is presented.
► In-line frit-free SPE is used to facilitate efficient trapping.
► In-line SPE and separation conditions are studied and optimized.
► Achieved LODs are in the low pg/ml range.