Enantiomer-Specific Stable Carbon and Nitrogen Isotopic Analyses of Underivatized Individual l- and d-Amino Acids by HPLC + HPLC Separation and Nano-EA/IRMS.

IF 6.7 1区化学 Q1 CHEMISTRY, ANALYTICAL

Analytical Chemistry Pub Date : 2024-11-15 DOI:10.1021/acs.analchem.4c02851

Yuchen Sun, Thomas M Blattmann, Yoshinori Takano, Nanako O Ogawa, Yuta Isaji, Naoto F Ishikawa, Naohiko Ohkouchi

{"title":"Enantiomer-Specific Stable Carbon and Nitrogen Isotopic Analyses of Underivatized Individual l- and d-Amino Acids by HPLC + HPLC Separation and Nano-EA/IRMS.","authors":"Yuchen Sun, Thomas M Blattmann, Yoshinori Takano, Nanako O Ogawa, Yuta Isaji, Naoto F Ishikawa, Naohiko Ohkouchi","doi":"10.1021/acs.analchem.4c02851","DOIUrl":null,"url":null,"abstract":"We developed a new method for stable carbon and nitrogen isotopic (δ13C and δ15N) analysis of underivatized amino acid (AA) enantiomers simultaneously, based on high-performance liquid chromatography (HPLC) separation and off-line isotopic measurement. l- and d-Enantiomers of each AA were isolated using a ReproSil Chiral-AA column, purified by wet chemical procedure, and analyzed for δ13C and δ15N values with a nanomol-scale elemental analyzer/isotope-ratio mass spectrometry (nano-EA/IRMS) system. We successfully achieved the separation of l- and d-enantiomers of 15 proteinogenous AAs, with all l-enantiomers eluting before respective d-enantiomers. The δ13C and δ15N values of AA enantiomers were consistent before and after HPLC separation, demonstrating that this analytical method conserves isotopic information. By coupling this column with a multidimensional HPLC system for isolating individual AAs, we analyzed l- and d-AAs in a natural sample, peptidoglycan isolated from Gram-positive bacterium Bacillus subtilis. Results show a surprisingly large 15N-depletion, up to 20‰, in d-glutamic acid relative to its l-counterpart. The first example, to our knowledge, of δ13C and δ15N analyses of underivatized AA enantiomers is expected to contribute to various research areas in the future.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":null,"pages":null},"PeriodicalIF":6.7000,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Analytical Chemistry","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1021/acs.analchem.4c02851","RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}

引用次数: 0

Abstract

We developed a new method for stable carbon and nitrogen isotopic (δ¹³C and δ¹⁵N) analysis of underivatized amino acid (AA) enantiomers simultaneously, based on high-performance liquid chromatography (HPLC) separation and off-line isotopic measurement. l- and d-Enantiomers of each AA were isolated using a ReproSil Chiral-AA column, purified by wet chemical procedure, and analyzed for δ¹³C and δ¹⁵N values with a nanomol-scale elemental analyzer/isotope-ratio mass spectrometry (nano-EA/IRMS) system. We successfully achieved the separation of l- and d-enantiomers of 15 proteinogenous AAs, with all l-enantiomers eluting before respective d-enantiomers. The δ¹³C and δ¹⁵N values of AA enantiomers were consistent before and after HPLC separation, demonstrating that this analytical method conserves isotopic information. By coupling this column with a multidimensional HPLC system for isolating individual AAs, we analyzed l- and d-AAs in a natural sample, peptidoglycan isolated from Gram-positive bacterium Bacillus subtilis. Results show a surprisingly large ¹⁵N-depletion, up to 20‰, in d-glutamic acid relative to its l-counterpart. The first example, to our knowledge, of δ¹³C and δ¹⁵N analyses of underivatized AA enantiomers is expected to contribute to various research areas in the future.

Abstract Image

查看原文本刊更多论文

通过高效液相色谱+高效液相色谱分离和纳米EA/IRMS对未充分活化的单个l-和d-氨基酸进行对映体特异性稳定碳氮同位素分析。

我们开发了一种基于高效液相色谱（HPLC）分离和离线同位素测量的新方法，可同时对未充分活化的氨基酸（AA）对映体进行稳定的碳氮同位素（δ13C 和 δ15N）分析。使用 ReproSil Chiral-AA 色谱柱分离出每种 AA 的 l-和 d-对映体，采用湿化学方法纯化，并使用纳摩尔级元素分析仪/同位素比质谱（nano-EA/IRMS）系统分析其 δ13C 和 δ15N 值。我们成功地分离了 15 种蛋白源 AA 的 l-和 d-对映体，所有 l-对映体均先于各自的 d-对映体洗脱。AA对映体的δ13C和δ15N值在HPLC分离前后是一致的，表明这种分析方法保留了同位素信息。通过将这种色谱柱与用于分离单个 AA 的多维 HPLC 系统联用，我们分析了从革兰氏阳性细菌枯草杆菌（Bacillus subtilis）中分离出来的天然样品肽聚糖（peptidoglycan）中的 l-AAs 和 d-AAs。结果表明，相对于 l-谷氨酸，d-谷氨酸中的 15N 丢失量惊人，高达 20‰。据我们所知，这是对未充分活化的 AA 对映体进行δ13C 和 δ15N 分析的第一个实例，有望在未来的各个研究领域做出贡献。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Analytical Chemistry 化学-分析化学

CiteScore

12.10

自引率

12.20%

发文量

1949

审稿时长

1.4 months

期刊介绍： Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.