HPLC Conditions ; Column : PolySULFOETHYL™ Aspartamide,
200 x 4.6 mm, 5 µm, 1000Å.
Mobile phase gradient 5' hold, then 0-100%B in 60'
Solvent A: 70% acetonitrile, 30% 0.1% methylphosphonic acid, 5mM NaClO4, pH 3.0
Solvent B: Solvent A plus 100mM NaClO4, pH 3.0
Flow Rate: 1.0 ml min-1 Injection volume: 80 µL
Use of a 1000Å, PolySULFOETHYL™ Aspartamide for these very hydrophobic protein molecules reduced the amount of salt necessary for elution from a strong cation exchange column. Chromatogram shows the separation achieved for Lung Surfactant Proteins from a sample containing a large excess of lipid materials (which would have complicated the use of RPC) for this assay. Use of a PolyHYDROXYETHYL™ A would also have been possible.
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Last Updated: 01/15/18