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High pressure liquid chromatography in the analysis of underivatised peptides using a sensitive and rapid procedure
Author(s) -
Hancock W.S.,
Bishop C.A.
Publication year - 1976
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(76)80830-7
Subject(s) - library science , chemistry , medicine , computer science
High pressure liquid chromatography (HPLC) has been used in the analysis of PTH amino acids [ 1,2] PTC-peptides [3] some growth promoting peptides [4] and peptide antibiotics [5-71. A major improvement would be achieved if no derivatisation of the peptides was necessary and a rapid, sensitive, low cost method could be employed. To achieve this HPLC was tested on an analytical scale with the possibility that methods developed could be scaled up to a preparative system. Because of the relative complexity of even small peptide molecules in terms of partition coefficient, polarity, hydrophobicity, and bulkiness [8] a series of column supports were investigated with a view to finding a system which both retained samples and allowed their ready elution. The reverse phase mode (liquid-liquid chromatography) was found to best satisfy these criteria. The most common and economical polar solvents used for reversed phase liquid-liquid chromatography, water, methanol and acetonitrile have the additional advantage of allowing spectrophotometry in the range 205-225 mm. In this range the peptide bond has sufficient absorption to allow quantitative detection in the nmol range.