Separation of peptides by reverse-phase high-performance liquid chromatography using propyl- and cyanopropylsilyl supports.
The separation of peptides and proteins by reverse-phase high-performance liquid chromatography with cyanopropylsilyl and large-pore propylsilyl supports, together with aqueous trifluoroacetic acid/acetonitrile gradients, was studied. Operating parameters (trifluoroacetic acid concentration, flow rate, and gradient slope) were evaluated using different enzymatic digests of horse cytochrome c and bovine serum albumin. Peptides ranging in size from five amino acids to 68 kDa could be separated on the propylsilyl column in a single chromatographic run. The cyanopropylsilyl column is suitable as a supplement to the use of the large-pore column for medium size (5-20 amino acids) peptides. The chromatographic supports and conditions presented here offer a simple, sensitive, and rapid separation system for a wide size range of peptides and proteins. They extend the versatility of separation methodology for these molecules.
D002851:Chromatography, High Pressure Liquid; D003574:Cytochrome c Group / Q000032:analysis; D008970:Molecular Weight; D009570:Nitriles; D010455:Peptides / Q000302:isolation & purification*; D012212:Rheology; D012710:Serum Albumin, Bovine / Q000032:analysis; D012821:Silanes; D012825:Silicon; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet; D014269:Trifluoroacetic Acid
D003574:Cytochrome c Group; D009570:Nitriles; D010455:Peptides; D012821:Silanes; D012997:Solvents; D012710:Serum Albumin, Bovine; D014269:Trifluoroacetic Acid; D012825:Silicon