TY - JOUR
T1 - Development of an online two-dimensional high-performance liquid chromatographic system in combination with tandem mass spectrometric detection for enantiomeric analysis of free amino acids in human physiological fluid
AU - Ishii, Chiharu
AU - Akita, Takeyuki
AU - Mita, Masashi
AU - Ide, Tomomi
AU - Hamase, Kenji
N1 - Funding Information:
This study was partly supported by JSPS KAKENHI Grant Number 15H05749 and 16H05080 Japan. The authors appreciate Shiseido Co., Ltd. , for their technical support and fund.
Publisher Copyright:
© 2018 Elsevier B.V.
PY - 2018/10/5
Y1 - 2018/10/5
N2 - An automated two-dimensional HPLC-MS/MS system was designed and developed for the highly selective determination of trace levels of D-amino acids. As the targets, frequently observed ones in mammalian physiological fluids, Ala, Asp, Glu, Leu, Pro and Ser, were selected because these D-amino acids are the potential biomarkers for the early and sensitive diagnoses of various diseases. The target analytes were derivatized with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), then isolated by a reversed-phase column in the first dimension. The NBD-amino acid fractions were automatically collected into the multi-loop device, and introduced into the enantioselective column in the second dimension to separate the D- and L-forms followed by detection by an MS/MS. The obtained resolution values of the enantiomers were 1.87–5.17 and the calibration lines, precision and accuracy were practically sufficient. By using the present 2D HPLC-MS/MS system, trace levels of D-amino acids in complex biological matrices were determined without disturbance by the intrinsic interfering compounds, and successfully applied to the analysis of the human clinical samples (plasma and urine).
AB - An automated two-dimensional HPLC-MS/MS system was designed and developed for the highly selective determination of trace levels of D-amino acids. As the targets, frequently observed ones in mammalian physiological fluids, Ala, Asp, Glu, Leu, Pro and Ser, were selected because these D-amino acids are the potential biomarkers for the early and sensitive diagnoses of various diseases. The target analytes were derivatized with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), then isolated by a reversed-phase column in the first dimension. The NBD-amino acid fractions were automatically collected into the multi-loop device, and introduced into the enantioselective column in the second dimension to separate the D- and L-forms followed by detection by an MS/MS. The obtained resolution values of the enantiomers were 1.87–5.17 and the calibration lines, precision and accuracy were practically sufficient. By using the present 2D HPLC-MS/MS system, trace levels of D-amino acids in complex biological matrices were determined without disturbance by the intrinsic interfering compounds, and successfully applied to the analysis of the human clinical samples (plasma and urine).
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U2 - 10.1016/j.chroma.2018.07.076
DO - 10.1016/j.chroma.2018.07.076
M3 - Article
C2 - 30104059
AN - SCOPUS:85051264327
SN - 0021-9673
VL - 1570
SP - 91
EP - 98
JO - Journal of Chromatography A
JF - Journal of Chromatography A
ER -
