D-Amino acids are important biological molecules. Improved analytical methods for their resolution and quantification remain of keen interest. In this study, we investigated the use of Marfey's reagent (chiral) derivatization coupled with LC-MS/MS-based separation and detection of the resulting diastereomers for quantification of the 19 common L- and D-amino acids and glycine. Standard formic acid (pH 2)-based separations on reverse phase media were unable to separate all 19 amino acid DL pairs. In contrast, a water/acetonitrile/ammonium acetate (pH 6.5) solvent system allowed all 19 amino acid DL pairs to be chromatographically resolved on a 30 min gradient, with negative mode detection at pH 6.5 giving good sensitivity. Derivatization reaction rates between amino acids varied substantially, with overnight derivatization required for some amino acids. Chromatography at pH 6.5 combined with MS/MS quantification in negative mode demonstrated good linearity over a wide concentration range for all 20 amino acids. Matrix effects, assessed with an MRSA extract, were negligible. Marfey's derivatized analytes were stable for 24 h at room temperature. This method was demonstrated by determining the levels of these analytes in mid-log phase MRSA extracts. This approach provides for the chromatographic resolution and MS/MS-based quantification of all 20 common L- and D-amino acids in complex matrices. Graphical Abstract.

Navid J. Ayon

Amar Deep Sharma

William G. Gutheil

Journal of the American Society for Mass Spectrometry

《美国质谱学会杂志》发表的研究论文涵盖了质谱的各个方面，涵盖了质谱可以发挥作用的科学探究领域。该杂志发表的论文范围广泛，涉及质量的基础和应用光谱学。基础科目包括仪器原理、设计和演示、气相离子的结构和化学性质、热力学性质研究、离子光谱学、化学动力学、电离机制、离子碎裂理论、簇离子和势能表面。除了全文之外，该期刊还提供通讯、应用笔记、账户和观点

The Journal of the American Society for Mass Spectrometry publishes research papers covering all aspects of mass spectrometry, including its applications in various scientific inquiries. The journal features a wide range of papers that encompass both fundamental and applied spectroscopy. Fundamental topics include principles of instrumentation, design and demonstration, structure and chemical properties of gas-phase ions, studies of thermodynamic properties, ion spectroscopy, chemical kinetics, ionization mechanisms, ion fragmentation theory, cluster ions, and potential energy surfaces. In addition to full articles, the journal also offers communications, application notes, accounts, and viewpoints.

《Journal of the American Society for Mass Spectrometry》发表的研究论文涵盖质谱的各个方面，涉及质谱在科学研究中的广泛应用。该杂志的论文范围广泛，涵盖基础和应用光谱学。基础主题包括仪器原理、设计与演示、气相离子的结构与化学性质、热力学性质研究、离子光谱学、化学动力学、电离机制、离子碎裂理论、簇离子和势能表面等。此外，期刊还提供通讯、应用笔记、账户和观点等内容。

LC-MS/MS-Based Separation and Quantification of Marfey’s Reagent Derivatized Proteinogenic Amino Acid <scp>dl</scp>-Stereoisomers

Marfey’s reagent (chiral) derivatization coupled with LC-MS/MS-based separation and detection of the resulting diastereomers for quantification of the 19 common l- and d-amino acids and glycine in complex matrices is investigated.

Category	Quartile
化学	2
化学, 光谱学	2
化学, 生化研究方法	3
化学, 分析化学	3
化学, 物理化学	3

LC-MS/MS-Based Separation and Quantification of Marfey’s Reagent Derivatized Proteinogenic Amino Acid dl-Stereoisomers

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