CONTINUOUS BEDS FOR MICROCHROMATOGRAPHY - REVERSED-PHASE CHROMATOGRAPHY

Simple and cost-effective methods for the preparation of microcolumns (i.d. 0.025-0.32 mm) for reverse-phase chromatography are described. T he procedure includes (1) synthesis in the column tube of a continuous bed matrix from a monomer solution (piperazine diacrylamide, methacry lamide) containing allyl glycidyl ether and 2-hydroxyethyl methacrylat e and (2) linking of C18 ligands by reacting 1,2-epoxyoctadecane with the epoxy and hydroxy groups in the matrix. The derivatization can be accomplished within 20 min. The columns prepared in this way showed hi gh performance in the separation of proteins and peptides and permitte d short analysis times (100 s). (C) 1996 Academic Press, Inc.