Particle packed mixed‐mode chromatographic stationary phase for the separation of peptide in liquid chromatography. Issue 7 (10th February 2021)

• Record Type: Journal Article
• Authors: Ali, Faiz; Cheong, Won Jo; Rafique, Aamra; AlOthman, Zeid A.; sadia, Maria; Muhammad, Mian
• Is Part Of: Journal of separation science. Volume 44:Issue 7(2021)
• Page Start: 1430
• Page End: 1439
• Publication Date: 2021-02-10
• Subjects: mixed‐mode chromatography -- peptide separation -- polymerization reaction -- reproducibility -- silica monolith particles; Separation (Technology) -- Periodicals; Chromatographic analysis -- Periodicals; 543.089
• DOI: 10.1002/jssc.202100001
• Languages: English
• ISSNs: 1615-9306

Abstract

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.