Simultaneous analysis of D-alanine, D-aspartic acid, and D-serine using chiral high-performance liquid chromatography-tandem mass spectrometry and its application to the rat plasma and tissues

J Pharm Biomed Anal. 2015 Nov 10:115:123-9. doi: 10.1016/j.jpba.2015.05.024. Epub 2015 Jun 16.

Abstract

A highly sensitive and selective chiral LC-MS/MS method for D-alanine, D-aspartic acid and D-serine has been developed using the precolumn derivatization reagents, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AccQ-Tag) or p-N,N,N-trimethylammonioanilyl N'-hydroxysuccinimidyl carbamate iodide (TAHS). The thus N-tagged enantiomers of the derivatized amino acids were nicely separated within 20min using the cinchona alkaloid-based zwittterionic ion-exchange type enantioselective column, Chiralpak ZWIX(+). The selected reaction monitoring was applied for detecting the target d-amino acids in biological matrices. By using the present chiral LC-MS/MS method, the three d-amino acids and their l-forms could be simultaneously determined in the range of 0.1-500nmol/mL. Finally, the technique was successfully applied to rat plasma and tissue samples.

Keywords: Chiral separation; LC-MS/MS; d-Amino acids.

MeSH terms

  • Alanine / analysis*
  • Alanine / blood
  • Animals
  • Brain / metabolism
  • Chromatography, High Pressure Liquid / methods*
  • D-Aspartic Acid / analysis*
  • D-Aspartic Acid / blood
  • Male
  • Organ Specificity
  • Rats, Sprague-Dawley
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Serine / analysis*
  • Serine / blood
  • Stereoisomerism
  • Tandem Mass Spectrometry / methods*
  • Viscera / metabolism

Substances

  • Serine
  • D-Aspartic Acid
  • Alanine