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Generation of Enantiomeric Amino Acids during Acid Hydrolysis of Peptides Detected by the Liquid Chromatography/Tandem Mass Spectroscopy
Author(s) -
Miyamoto Tetsuya,
Sekine Masae,
Ogawa Tetsuhiro,
Hidaka Makoto,
Homma Hiroshi,
Masaki Haruhiko
Publication year - 2010
Publication title -
chemistry and biodiversity
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.427
H-Index - 70
eISSN - 1612-1880
pISSN - 1612-1872
DOI - 10.1002/cbdv.200900309
Subject(s) - chemistry , amino acid , racemization , hydrolysis , enantiomer , chromatography , peptide , hydrolysate , acid hydrolysis , tandem mass spectrometry , mass spectrometry , organic chemistry , biochemistry
The number of reports indicating the occurrence of D ‐amino acids in various proteins and natural peptides is increasing. For a usual detection of peptidyl D ‐amino acids, proteins or peptides are subjected to acid hydrolysis, and the products obtained are analyzed after cancellation of the effect of amino acid racemization during the hydrolysis. However, this method does not seem reliable enough to determine the absence or presence of a small amount of innate D ‐amino acids. We introduce a modification of an alternative way to distinguish true innate D ‐amino acids from those artificially generated during hydrolysis incubation. When model peptides ( L ‐Ala) 3 , D ‐Ala‐( L ‐Ala) 2 are hydrolyzed in deuterated hydrochloric acid (DCl), only newly generated D ‐amino acids are deuterated at the α ‐H‐atom. Both innate D ‐amino acids and artificially generated ones are identified by the combination of high‐performance liquid chromatography and liquid chromatography/tandem mass spectrometry equipped with a chiral column. When a peptide containing D ‐Phe residues was analyzed by this method, the hydrolysis‐induced conversion to L ‐Phe was similarly identified.