1.2.2. HPLC of amino acids as phenylthiocarbamoyl derivatives

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Some of the general problems encountered in the analysis of amino acids derivatized with various isothiocyanates, determine both as phenylthiocarbamoyl (PTC) and as phenylthiohydantoin (PTH) derivatives, are presented. Based on our own experience and the literature associated with sample preparation, stability, optimum chromatographic and quantitation conditions are discussed. Twenty-seven PTC-AAs have been separated on seven columns, with the same gradient program, applying various temperatures and eluent flow rates, within 40 min, including equilibration time. Response values and spectral characteristics of the 27 AAs proved to be independent of the chromatographic conditions performed. Particular attention was paid to the issue of sensitive amino acids under hydrolysis (cystine/cysteine, methionine, tryptophan), and derivatization (cystine/cysteine) conditions as well. The special behavior of the PTC-cystine/cysteine has been examined by LC/API/MS measurements showing that, irrespective of the initial compound to be derivatized, the same steroisomer pair are formed at m/z=255, PTC-cysteines (≈80% of the total) and their oxidized version at m/z=287 (≈20% of the total). Whether there is an advantage of the microwave irradiation procedure is still not decided: however the hydrolysis time of lysozyme has been shortened considerably, but optimum hydrolysis conditions were not uniformly found for all essential amino acids.

Original languageEnglish
Pages (from-to)137-162
Number of pages26
JournalJournal of Chromatography Library
Volume70
Issue numberC
DOIs
Publication statusPublished - 2005

Fingerprint

Cystines
Cysteine
Cystine
Amino acids
Hydrolysis
High Pressure Liquid Chromatography
Derivatives
Amino Acids
Phenylthiohydantoin
Microwave irradiation
Isothiocyanates
Application programming interfaces (API)
Essential Amino Acids
Muramidase
Enzymes
Microwaves
Flow rate
Tryptophan
Methionine
Temperature

ASJC Scopus subject areas

  • Analytical Chemistry
  • Filtration and Separation
  • Biochemistry
  • Clinical Biochemistry

Cite this

1.2.2. HPLC of amino acids as phenylthiocarbamoyl derivatives. / Molnár-Perl, I.

In: Journal of Chromatography Library, Vol. 70, No. C, 2005, p. 137-162.

Research output: Contribution to journalArticle

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