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- Title
Particle packed mixed‐mode chromatographic stationary phase for the separation of peptide in liquid chromatography.
- Authors
Ali, Faiz; Cheong, Won Jo; Rafique, Aamra; AlOthman, Zeid A.; sadia, Maria; Muhammad, Mian
- Abstract
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.
- Subjects
LIQUID chromatography; ADDITION polymerization; PHASE separation; PACKINGS (Chromatography); CYTOCHROME c; PEPTIDOMIMETICS; TANDEM mass spectrometry
- Publication
Journal of Separation Science, 2021, Vol 44, Issue 7, p1430
- ISSN
1615-9306
- Publication type
Article
- DOI
10.1002/jssc.202100001