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Particle packed mixed‐mode chromatographic stationary phase for the separation of peptide in liquid chromatography
Author(s) -
Ali Faiz,
Cheong Won Jo,
Rafique Aamra,
AlOthman Zeid A.,
sadia Maria,
Muhammad Mian
Publication year - 2021
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.202100001
Subject(s) - chromatography , monolith , ammonium hydroxide , chemistry , packed bed , capillary electrochromatography , elution , polymerization , particle size , analytical chemistry (journal) , stationary phase , organic chemistry , catalysis , polymer
A particle‐based stationary phase has been prepared for the separation of five synthetic peptides and a mixture containing tryptic digest of cytochrome C in liquid chromatography. Particles originating from silica monolith were differentially sedimented to obtain 1–2 μm particles. A stationary phase was achieved by the coating of poly(styrene‐methacrylic acid‐N‐phenylacrylamide) copolymer onto the particles via reversible addition‐fragmentation chain transfer polymerization reaction. Stainless steel column (30 cm long and 1 mm internal diameter) was packed with stationary phase. Very high separation efficiency (ca. 351 000 plates/m) was achieved for five commercial peptides with a percent relative standard deviation of less than 1%. Protocol for the synthesis and modification of silica monolith particles has been well optimized with a good reproducibility both in particle and pore size. The column resolved about 21 peptide components from a mixture containing tryptic digest of cytochrome C, under the elution conditions of acetonitrile/15 mM ammonium format (65/35 v/v%) with a flow rate of 28 μL/min.