High-performance liquid chromatography of amino acids, peptides and proteins. LXIX. Evaluation of retention and bandwidth relationships of myosin-related peptides separated by gradient elution reversed-phase high-performance liquid chromatography.
The gradient elution behaviour of ten peptide analogues encompassing the primary amino acid sequences adjacent to the serine-19 phosphorylation site of myosin light chain has been investigated using a 30-nm pore diameter n-butylsilica stationary phase and 0.1% trifluoroacetic acid-water-acetonitrile mobile phases. Quantitative expressions derived from linear solvent strength theory and general plate height theory have been used to assess the influence of gradient time and flow-rate on the relative retentions and bandwidths of these peptides. The retention behaviour of these peptide analogues was found to closely mirror the predictions of linear solvent strength gradient elution theory. Although the experimentally observed band-width changes over a conventional range of gradient conditions (i.e. tG = 30-60 min) also correlated with bandwidth changes predicted on the basis of plate theory derived for low molecular weight organic molecules, for very steep and very shallow gradients divergencies from theory were observed. The significance of these discrepancies is discussed in relation to the influence of amino acid sequence changes rather than composition of these peptide analogues. These data allow the importance of sequential effects on the chromatographic behaviour and in particular, the resolution optimisation of these closely related peptides to be evaluated.
D000596:Amino Acids / Q000302:isolation & purification; D000818:Animals; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D005895:Gizzard, Avian / Q000032:analysis; D007202:Indicators and Reagents; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism; D009218:Myosins / Q000302:isolation & purification*; D010455:Peptides / Q000302:isolation & purification*; D011487:Protein Conformation; D011506:Proteins / Q000302:isolation & purification; D012997:Solvents
D000596:Amino Acids; D007202:Indicators and Reagents; D010455:Peptides; D011506:Proteins; D012997:Solvents; D009219:Myosin-Light-Chain Kinase; D009218:Myosins