高等学校化学学报

高等学校化学学报

• 研究论文 • 上一篇    下一篇

在线固相萃取净化-液相色谱-高分辨质谱法测定海洋沉积物中的L型和D型氨基酸

何秀平1,2, 李芳1, 陈军辉1,2, 辛明1,2, 王保栋1,2   

  1. 1.自然资源部第一海洋研究所, 青岛市现代分析与近海生态环境安全保障重点实验室 2.青岛海洋科技中心, 海洋生态与环境科学功能实验室
  • 收稿日期:2025-07-12 修回日期:2025-09-29 网络首发:2025-10-14 发布日期:2025-10-14
  • 通讯作者: 陈军辉 E-mail:jhchen@fio.org.cn
  • 基金资助:
    全球变化与海气相互作用（二期）专项（GASI-01-ATP-STwin）资助

Determination of L- and D-Amino Acids in Marine Sediments Using Online Solid-Phase Extraction Cleanup Coupled with Liquid Chromatography-High-Resolution Mass Spectrometry

HE Xiuping1,2, LI Fang1, CHEN Junhui1,2, XIN Ming1,2, WANG Baodong1,2   

  1. 1. Qingdao Key Laboratory of Analytical Technology Development and Offshore Eco-Environment Conservation, The First Institute of Oceanography, Ministry of Natural Resources 2. Laboratory for Marine Ecology and Environmental Science, Qingdao Marine Science and Technology Center
  • Received:2025-07-12 Revised:2025-09-29 Online First:2025-10-14 Published:2025-10-14
  • Contact: CHEN Junhui E-mail:jhchen@fio.org.cn
  • Supported by:
    Supported by the Global Change and Air-Sea Interaction II (GASI-01-ATP-STwin)

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摘要： 氨基酸（L-氨基酸和D-氨基酸）是指示海洋沉积环境有机质来源及成岩过程的重要生物标志物, 因此, 开发精准检测海洋沉积物中主要手性氨基酸的方法尤为重要. 本文采用在线固相萃取（SPE）结合液相色谱-飞行时间质谱联用技术（LC-TOF/MS）, 建立了一种同步测定海洋沉积物中17种L-氨基酸和7种D-氨基酸的新方法. 采用C18保护柱为在线SPE柱, 填料粒径为1.8 μm的超高效反相C18柱为分析柱, 基于酸性和碱性双流动相分离体系, 实现海洋沉积物中各种L/D-氨基酸的高效分离, 采用高分辨电喷雾TOF/MS 全扫描模式下在线检测. 结果表明, 海洋沉积物样品经盐酸水解处理后稀释5倍, 进行衍生化处理, 采用在线SPE-LC-TOF/MS直接进样分析可实现各种L/D-氨基酸的准确测定. 各种氨基酸的加标回收率在66.1% ~ 106.8%之间, 相对标准偏差（RSD）为2.40%~12.3%, 并且具有较宽的线性范围, 相关系数（R2）均大于0.990. 与传统的液相色谱法相比, 该方法仅需少量（0.01~0.10 g）沉积物样品, 是深海及大洋热液区沉积物中手性氨基酸生物标志物检测的有效方法. 采用本方法对西印度洋表层沉积物中的L/D-氨基酸进行了检测, 所有样品中均检出了17种L-氨基酸和7种D-氨基酸, 总含量在1.69~19.30 μmol/g（干重）范围内, 并且发现海底热液活动对深海沉积环境中有机碳、氮的循环有重要影响。

关键词: 在线固相萃取, 高分辨质谱, 海洋沉积物, D-氨基酸, 深海热液喷口

Abstract: Amino acids (both L- and D-enantiomers) serve as crucial biomarkers for indicating the sources of organic matter and diagenetic processes within marine sedimentary environments. Consequently, developing methods for their precise detection in marine sediments is of significant importance. This study established a novel method for the simultaneous determination of 17 L-amino acids and 7 D-amino acids in marine sediments using online solid-phase extraction (SPE) coupled with liquid chromatography-time-of-flight mass spectrometry (LC-TOF/MS). A C18 guard column served as the online SPE column, while an ultra-high-performance reversed-phase C18 analytical column (1.8 μm particle size) was employed for separation. Utilizing dual acidic and alkaline mobile phase systems, efficient separation of various L- and D-amino acids was achieved. Detection was performed online using high-resolution electrospray ionization TOF/MS in full scan mode. The results demonstrate that accurate quantification of various L- and D-amino acids can be achieved by direct injection analysis using online SPE-LC-TOF/MS following hydrochloric acid hydrolysis of marine sediment samples, 5-fold dilution, and derivatization. The spiked recoveries for the various amino acids ranged from 66.1% to 106.8%, with relative standard deviations (RSD) between 2.40% and 12.3%. The method exhibited wide linear ranges, and all correlation coefficients (R2) were greater than 0.990. Compared to conventional liquid chromatography methods, this approach requires only a small sediment sample amount (0.01~0.10 g), making it an effective method for detecting chiral amino acid biomarkers in deep-sea and oceanic hydrothermal vent sediments. Applying this method to analyze L- and D-amino acids in surface sediments from the Western Indian Ocean revealed the presence of all 17 L-amino acids and 7 D-amino acids in all samples, with total concentrations ranging from 1.69 to 19.30 μmol/g (dry weight). Furthermore, the analysis indicated that submarine hydrothermal activity significantly influences the cycling of organic carbon and nitrogen in the deep-sea sedimentary environment.

Key words: Online solid phase extraction, High resolution mass spectrometry, Marine sediments; D-amino acids, Deep-sea hydrothermal vents

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