TY - JOUR
T1 - Chiral Derivatization Enables High-Resolution Ion Mobility Spectrometry of 30 Amino Acid Enantiomers
AU - Yan, Chi
AU - Chen, Xingyu
AU - Gao, Wenqing
AU - Tang, Chunlan
AU - Du, Liwen
AU - Xie, Chengyi
AU - Xu, Feng
AU - Tang, Keqi
AU - Yu, Jiancheng
N1 - Publisher Copyright:
© XXXX American Chemical Society.
Funding Information:
This work was supported by National key research and development program (2023YFC3304203), Yangtze River Delta Science and Technology Innovation Community Joint Research Project (2023CSJGG1800), the Key Research and Development Program of Zhejiang Province (2025C01200(SD2), 2024C03266, 2025C02235, and 2024C03101), Hangzhou Science and Technology Development Project (20231203A18), and the Ningbo Science and Technology Project (2022Z241, 2023Z132, 2023Z168, 2024Z230, 2024S047, and 2023S151). The authors gratefully thank all the participants and professionals contributing to this study.
PY - 2025/12/26
Y1 - 2025/12/26
N2 - Chiral analysis of amino acid enantiomers is essential due to their frequently divergent biological activities. However, ion mobility spectrometry–mass spectrometry (IMS–MS) cannot directly resolve underivatized amino acid enantiomers due to their identical physicochemical properties. This study developed a derivatization-based analytical method utilizing (S)-N-(4-nitrophenoxycarbonyl) phenylalanine methoxyethyl ester ((S)-NIFE) as a chiral derivatization agent to achieve the stereoselective modification of amino acids. The resulting diastereomers form metal ion adducts, which enhance collision cross section differences and enable separation via trapped ion mobility spectrometry and time-of-flight mass spectrometry (TIMS–MS). The differential impact of alkali metal ion adduction (Na+, K+, Rb+, and Cs+) on chiral separation was investigated, revealing that Na+ adducts provide optimal enantioselective recognition (average resolution, Rpp = 2.02). The established method achieved separation for 30 chiral amino acids, demonstrating faster speed, broader coverage, and superior resolution compared with existing approaches. When applied to rat blood samples from chlorfenapyr-induced toxicity models, the method successfully detected d-Arg, d-Ile, d-Leu, d-Phe, and d-Met, confirming its practical utility in toxicological diagnostics and biomarker discovery.
AB - Chiral analysis of amino acid enantiomers is essential due to their frequently divergent biological activities. However, ion mobility spectrometry–mass spectrometry (IMS–MS) cannot directly resolve underivatized amino acid enantiomers due to their identical physicochemical properties. This study developed a derivatization-based analytical method utilizing (S)-N-(4-nitrophenoxycarbonyl) phenylalanine methoxyethyl ester ((S)-NIFE) as a chiral derivatization agent to achieve the stereoselective modification of amino acids. The resulting diastereomers form metal ion adducts, which enhance collision cross section differences and enable separation via trapped ion mobility spectrometry and time-of-flight mass spectrometry (TIMS–MS). The differential impact of alkali metal ion adduction (Na+, K+, Rb+, and Cs+) on chiral separation was investigated, revealing that Na+ adducts provide optimal enantioselective recognition (average resolution, Rpp = 2.02). The established method achieved separation for 30 chiral amino acids, demonstrating faster speed, broader coverage, and superior resolution compared with existing approaches. When applied to rat blood samples from chlorfenapyr-induced toxicity models, the method successfully detected d-Arg, d-Ile, d-Leu, d-Phe, and d-Met, confirming its practical utility in toxicological diagnostics and biomarker discovery.
U2 - 10.1021/acs.analchem.5c06382
DO - 10.1021/acs.analchem.5c06382
M3 - Journal article
C2 - 41451537
SN - 0003-2700
SP - 1
EP - 8
JO - Analytical Chemistry
JF - Analytical Chemistry
ER -
