Isocratic Separation of PTH-Amino Acids at Picomole Level by Reverse-Phase HPLC in the Presence of Sodium Dodecylsulfate1
- 1 February 1985
- journal article
- research article
- Published by Oxford University Press (OUP) in The Journal of Biochemistry
- Vol. 97 (2) , 701-704
- https://doi.org/10.1093/oxfordjournals.jbchem.a135108
Abstract
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 (pH4.5) 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 -23 min and the limit of analysis ∼2.5 pmol of a PTH-amino acid.Keywords
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