Journal
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
Volume 81, Issue 9, Pages 1681-1686Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1080/09168451.2017.1340090
Keywords
d-amino acid; d-proline; o-phthalaldehyde (OPA); N-acetyl- l-cysteine (NAC); 1-(9-fluorenyl)ethyl chloroformate (FLEC)
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Funding
- JSPS KAKENHI [16K07852, 15H04539]
- Kitasato University Research Grant for Young Researchers
- Grants-in-Aid for Scientific Research [16K07852, 15H04539] Funding Source: KAKEN
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This work describes a method for the simultaneous determination of primary d- and l-amino acids and secondary amino acids such as d- and l-proline. In order to remove interferences in the simultaneous determination of primary and secondary amines, the primary amines were derivatized with o-phthalaldehyde/N-acetyl-l-cysteine (OPA/NAC) and subsequently with 1-(9-fluorenyl)ethyl chloroformate (FLEC) for secondary amines, in a pre-column separation derivatization technique. These fluorescent diastereomers of the amino acids were obtained within 3min at room temperature and determined simultaneously by changing wavelengths during analysis in a single eluting run in the high-performance liquid chromatography column. This method, referred to as the two-step labelling method, is effective for the simultaneous determination of d- and l-amino acids.
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