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Separation of peptides with an aqueous mobile phase by temperature‐responsive chromatographic column
Author(s) -
Liu Zongjian,
Liang Yanli,
Geng Fangfang,
Ge Chen,
Ullah Kaleem,
Lv Fang,
Dai Rongji,
Zhang Yukui,
Deng Yulin
Publication year - 2012
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.201200247
Subject(s) - chromatography , chemistry , peptide , aqueous solution , differential scanning calorimetry , selectivity , phase (matter) , copolymer , reversed phase chromatography , high performance liquid chromatography , analytical chemistry (journal) , polymer , organic chemistry , biochemistry , physics , thermodynamics , catalysis
Peptide separation technology is significant and is still an analytical challenge in proteomic studies. We report a simple preparation method for poly( N ‐isopropylacrylamide) grafted silica through the copolymerization of N ‐isopropylacrylamide with acetyl moieties immobilized on the silica surfaces. Differential scanning calorimetry results indicated that the prepared silica exhibited a sharp phase transition at 35.03°C. Silica grafted with poly( N ‐isopropylacrylamide) was evaluated as a temperature‐responsive chromatography medium for the separation of peptides using 0.2 M NaCl solution as a mobile phase. Results indicated that at 10°C, the peptides were not resolved, but baseline separation with prolonged retention time at 50°C was attained. Particularly, a mixture of four peptides was efficiently separated within 8 min. The theoretical plate number of every peptide was more than 2500, and the resolutions were more than 3.40. The increased selectivity of the temperature‐responsive column resulted from the temperature‐modulated hydrophobic interaction with peptides. The retention times of these peptides were related to their hydrophobicities. This protocol provided a reliable set of chromatographic tool usable across all research and development applications that required isolation and analysis of peptides. It may represent a step forward in the complex analysis of hydrophobic and other proteins.