An ultra-high-performance tandem mass spectrometry method to quantify tryptophan metabolites in aqueous humor of primary angle-closure glaucoma patients

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A Pub Date : 2025-04-03 DOI:10.1016/j.chroma.2025.465939

Xingang Fu , Weiwei Guo , Yuheng Cheng , Lin Li

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Abstract

This study presented the development and validation of a robust ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous quantification of tryptophan (TRP) and its nine metabolites in aqueous humor (AH) to explore the regulation of the TRP metabolic pathway in primary angle-closure glaucoma (PACG). The optimized UPLC-MS/MS method demonstrated good linearity (R² > 0.99), sensitivity (LLMI: 0.11 – 1.31 ng/mL), precision (CVs: 2.18 % and 12.88 %), recovery rates (85.06 % – 105.74 %), bench-top, long-term and on-instrument stabilities (CVs: 2.35 % – 6.88 %). The validated UPLC-MS/MS method was applied to AH samples from PACG patients with cataract and cataract-alone patients. The results showed that kynurenine and 3-hydroxykynurenine concentrations were significantly increased in the AH of the PACG group, indicating up-regulated indoleamine 2,3-dioxygenase activity and a metabolic shift towards the production of the neurotoxic metabolites within the kynurenine pathway. These findings underscore the potential involvement of TRP metabolism in PACG pathogenesis.

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超高效能串联质谱法定量原发性闭角型青光眼患者房水中色氨酸代谢物

本研究建立并验证了超高效液相色谱-串联质谱（UPLC-MS/MS）同时定量水相中色氨酸（TRP）及其9种代谢物的方法，以探讨原发性闭角型青光眼（PACG）中色氨酸代谢途径的调控。优化后的UPLC-MS/MS方法线性良好(R²>；0.99)、灵敏度（LLMI: 0.11 ~ 1.31 ng/mL）、精密度（cv: 2.18% ~ 12.88%）、回收率（85.06% ~ 105.74%）、台台稳定性、长期稳定性和仪器稳定性（cv: 2.35% ~ 6.88%）。将经过验证的UPLC-MS/MS方法应用于PACG合并白内障和单纯性白内障患者的AH样品。结果显示，PACG组AH中犬尿氨酸和3-羟基犬尿氨酸浓度显著升高，表明吲哚胺2,3-双加氧酶活性上调，并在犬尿氨酸途径中向产生神经毒性代谢物的代谢转变。这些发现强调了TRP代谢在PACG发病机制中的潜在参与。

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来源期刊

Journal of Chromatography A 化学-分析化学

CiteScore

7.90

自引率

14.60%

发文量

742

审稿时长

45 days

期刊介绍： The Journal of Chromatography A provides a forum for the publication of original research and critical reviews on all aspects of fundamental and applied separation science. The scope of the journal includes chromatography and related techniques, electromigration techniques (e.g. electrophoresis, electrochromatography), hyphenated and other multi-dimensional techniques, sample preparation, and detection methods such as mass spectrometry. Contributions consist mainly of research papers dealing with the theory of separation methods, instrumental developments and analytical and preparative applications of general interest.