开发一种快速、稳健的液相色谱串联质谱法,用于检测小鼠血清和组织中的 13 种内源性氨基酸以及氧化三甲胺。

IF 1.8 4区 医学 Q4 BIOCHEMICAL RESEARCH METHODS Biomedical Chromatography Pub Date : 2024-12-01 Epub Date: 2024-10-10 DOI:10.1002/bmc.6010
Didi Hu, Xudong Liu, Ying Yao, Shijie Wei, Hongyan Ji, Yang Yang, Jing Chen, Linwei Chen
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引用次数: 0

摘要

本研究旨在建立一种 HILIC-MS/MS 方法,以同时测定小鼠生物样本中 13 种内源性氨基酸和氧化三甲胺的含量。质谱分析采用电喷雾离子源。采用 Dikma Inspire Hilic 色谱柱(2.1 × 100.0 mm,3 μM),分离时间为 20 分钟。在 MRM 模式下,正离子模式的响应值令人满意。定量限的准确度为-12.59%至7.89%,精密度为1.77%至14.00%,日间和日内精密度、基质效应、回收率和稳定性均可接受。随后,该测定法被成功地用于测量生物样本中决定因子的浓度。可以观察到这些代谢物的个体差异和组织分布差异。氨基酸在脾脏中的含量始终最高,而在肝脏中的含量最低。丙氨酸是血清中含量最高的氨基酸,牛磺酸在所有组织中的含量都最高。三甲胺氧化物的含量仍然很低,尤其是在肝脏样本中。
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Development of a rapid and robust hydrop interaction liquid chromatography tandem mass spectrometry method for the detection of 13 endogenous amino acids as well as trimethylamine oxide in serum and tissues of the mice.

This work aimed to establish an HILIC-MS/MS method to simultaneously determine the levels of 13 endogenous amino acids and trimethylamine oxide in the biological samples from the mice. Electrospray ion source was used for the analysis of mass spectrometry. The 20 min separation was applied in a Dikma Inspire Hilic column (2.1 × 100.0 mm, 3 μM). Positive ion mode under an MRM model gave a satisfying response value. The limits of quantitation were evaluated by accuracy from -12.59% to 7.89% and precision from 1.77% to 14.00% as well as acceptable interday and intraday precision, matrix effect, recovery, and stability. Later, the assay was successfully used to measure the concentrations of the determinands in the biological samples. Individual and tissue distribution differences for these metabolites were observable. The amino acids had a consistent highest content in the spleens, while the lowest levels were found in the livers. Alanine was the most abundant amino acid in the serum, and taurine kept the highest content in all of the tissues. Trimethylamine oxide remained low level, especially in the liver samples.

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来源期刊
Biomedical Chromatography
Biomedical Chromatography 生物-分析化学
CiteScore
3.60
自引率
5.60%
发文量
268
审稿时长
2.3 months
期刊介绍: Biomedical Chromatography is devoted to the publication of original papers on the applications of chromatography and allied techniques in the biological and medical sciences. Research papers and review articles cover the methods and techniques relevant to the separation, identification and determination of substances in biochemistry, biotechnology, molecular biology, cell biology, clinical chemistry, pharmacology and related disciplines. These include the analysis of body fluids, cells and tissues, purification of biologically important compounds, pharmaco-kinetics and sequencing methods using HPLC, GC, HPLC-MS, TLC, paper chromatography, affinity chromatography, gel filtration, electrophoresis and related techniques.
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