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Preparation and evaluation of a hydrophilic poly(2-hydroxyethyl methacrylate-co-N,N'-methylene bisacrylamide) monolithic column for pressurized capillary electrochromatography.

Electrophoresis (2013-02-14)
Jintian Cheng, Xiadi Chen, Yimin Cai, Yu He, Zongbao Chen, Zian Lin, Lan Zhang
ABSTRACT

A polar polymethacrylate-based monolithic column was introduced and evaluated as a hydrophilic interaction CEC stationary phase. The monolithic stationary phase was prepared by in situ copolymerization of a neutral monomer 2-hydroxyethyl methacrylate and a polar cross-linker N,N'-methylene bisacrylamide in a binary porogenic solvent consisting of dodecyl alcohol and toluene. The hydroxyl and amino groups at the surface of the monolithic stationary phase provided polar sites which were responsible for hydrophilic interactions. The composition and proportion of the polymerization mixture was investigated in detail. The mechanical stability and reproducibility of the obtained monolithic column preformed was satisfied. The effects of pH and organic solvent content on the EOF and the separation of amines, nucleosides, and narcotics on the optimized monolithic column were investigated. A typical hydrophilic interaction CEC was observed on the neutral polar stationary phase. The optimized monolithic column can obtain high-column efficiencies with 62,000-126,000 theoretical plates/m and the RSDs of column-to-column (n = 9), run-to-run (n = 5), and day-to-day (n = 3) reproducibility were less than 6.3%. The calibration curves of these five narcotics exhibited good linearity with R in the range of 0.9959-0.9970 and linear ranges of 1.0-200.0 μg/mL. The detection limits at S/N = 3 were between 0.2 and 1.2 μg/mL. The recoveries of the separation of narcotics on the column were in the range of 84.0-108.6%. The good mechanical stability, reproducibility, and quantitation capacity was suitable for pressure-assisted CEC applications.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
N,N′-Methylenebisacrylamide solution, suitable for electrophoresis, 2% in H2O
Sigma-Aldrich
N,N′-Methylenebis(acrylamide), 99%
Sigma-Aldrich
2-Hydroxyethyl methacrylate, ≥99%, contains ≤50 ppm monomethyl ether hydroquinone as inhibitor
Sigma-Aldrich
2-Hydroxyethyl methacrylate, contains ≤250 ppm monomethyl ether hydroquinone as inhibitor, 97%
Sigma-Aldrich
2-Hydroxyethyl methacrylate, embedding medium (for microscopy)
Sigma-Aldrich
N,N′-Methylenebisacrylamide, powder, for molecular biology, suitable for electrophoresis, ≥99.5%
Sigma-Aldrich
N,N′-Methylenebisacrylamide, suitable for electrophoresis (after filtration or allowing insolubles to settle)