Isocratic separation of PTH-amino acids at picomole level by reverse-phase HPLC in the presence of sodium dodecylsulfate.
The phenylthiohydantoin (PTH) derivatives of protein amino acids have been separated by reverse-phase high performance liquid chromatography (HPLC) on a fully end-capped C18 column using an isocratic solvent system. The developing solvent was 0.01 M sodium acetate buffer (pH 4.5) containing 39.5% acetonitrile and 0.02% sodium dodecylsulfate (SDS). With an automated liquid chromatography equipped with a dual-channel detector, operating at 254 and 313 nm, the present isocratic separation system was quite useful for routine microanalysis of PTH-amino acids released with a "gas-phase" sequencer. The time for one run was approximately 23 min and the limit of analysis approximately 2.5 pmol of a PTH-amino acid.
D000596:Amino Acids / Q000302:isolation & purification*; D001322:Autoanalysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006827:Hydantoins / Q000302:isolation & purification*; D008832:Microchemistry; D010669:Phenylthiohydantoin / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate; D013997:Time Factors
D000596:Amino Acids; D006827:Hydantoins; D010669:Phenylthiohydantoin; D012967:Sodium Dodecyl Sulfate