肌醇酮与相应代谢产物表现出类似于色氨酸、一种必需氨基酸和神经递质血清素的生物活性。肌醇酮途径的失调参与了神经退行性/神经精神疾病和2型糖尿病以及癌症。因此，肌醇酮相关代谢产物的测量将有助于更全面了解肌醇酮途径在疾病发病机制中的重要性。通过超高效液相色谱串联质谱法（UPLC-MS/MS）对色氨酸、血清素、吡啶酸、喹啉酸、3-羟基肌醇酮、肌醇酮、3-羟基间苯二甲酸、肌醛酸以及烟酸和氧化还原辅酶NAD+等物质在异质矩阵中进行了分析。经过验证，所描述的方法被应用于测量小鼠组织和细胞系统中的天然代谢物浓度，并监测了经色氨酸-2,3-二氧化酶抑制剂680C91处理后的途径变化。此外，该方法被评估其能够通过单样本代谢物提取程序并入多组学方法中的能力。 本研究开发了一种简单而敏感的UPLC-MS/MS方法，可同时定量测定四种生物样品中高达10种肌醇酮途径相关代谢产物。在运行时间为6.5分钟内，成功进行了肌醇酮相关代谢物的色谱分离，包括同分异构体烟酸和吡啶酸，而无需衍生化。验证参数包括日内精密度（＜14.8%），平均准确度（102.4% ± 12.9%）和线性检测范围超过三个数量级，表明该方法具有可靠性。根据被研究样品基质的不同，大多数代谢物在原始小鼠和细胞培养样品中都能成功检测并定量。此外，该方法还能监测色氨酸-2,3-二氧化酶抑制剂对细胞系统中肌醇酮途径的影响，并可用于使用同一细胞提取物的多重分析。 所描述的UPLC-MS/MS方法为肌醇酮途径代谢产物的同时定量提供了简单工具。由于其适用于许多生理基质，该方法在与疾病相关的实验环境中具有广泛应用。版权所有 © 2023 Elsevier B.V. 保留所有权利。

Kynurenine and respective metabolites exhibit bioactivity as well as tryptophan, an essential amino acid, and the neurotransmitter serotonin. Dysregulations in the kynurenine pathway are involved in neurodegenerative/neuropsychiatric disorders and diabetes mellitus type 2 but also in cancer. Therefore, measurements of kynurenine-related metabolites will improve the general understanding for kynurenine pathway relevance in disease pathogenesis.Tryptophan, serotonin, picolinic acid, quinolinic acid, 3-OH-kynurenine, kynurenine, 3-OH-anthranilic acid, kynurenic acid, anthranilic acid as well as nicotinic acid and the redox cofactor NAD+ were analyzed in heterogeneous matrices by ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). After validation, the described method was applied for measurements of native metabolite concentrations in murine tissues and cellular systems including pathway-shift monitoring after treatment with the tryptophan-2,3-dioxygenase-inhibitor 680C91. In addition, the method was evaluated for its ability for integration into multi-omics approaches using a single sample metabolite extraction procedure.A simple and sensitive UPLC-MS/MS method for simultaneous quantification of up to 10 kynurenine-related metabolites in four biological matrices was developed. Within a run time of 6.5 min, chromatographic separation of kynurenine-related metabolites, including the isomers nicotinic acid and picolinic acid, was achieved without derivatization. Validation parameters, including interday precision (<14.8%), mean accuracy (102.4% ± 12.9%) and linear detection ranges of more than three orders of magnitude, indicate method reliability. Depending the investigated sample matrix, the majority of metabolites were successfully detected and quantified in native murine and cell culture derived sample materials. Furthermore, the method allowed to monitor the impact of a tryptophan-2,3-dioxygenase-inhibitor on kynurenine pathway in a cellular system and is suitable for multi-assay analyses using aliquots from the same cell extract.The described UPLC-MS/MS method provides a simple tool for the simultaneous quantification of kynurenine pathway metabolites. Due to its suitability for many physiological matrices, the method provides wide application for disease-related experimental settings.Copyright © 2023 Elsevier B.V. All rights reserved.