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

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.