The change of metabolic pathways is recognized as the key to disease discovery prompting the development of ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS)-based quantitative platforms to explore the dynamic metabolite profiles of organisms. In this study, a liquid chromatography method based on cyanopropyl (CN) was developed. By adjusting the pH environment of the column, we achieved the elution of 51 metabolites spanning the most comprehensive set of biological pathways currently known. Offering rapid chromatography, efficient separation, and green chemistry benefits, the method encompasses nucleosides and nucleotides, the oxidative-redox metabolome, the glycolysis pathway, the pentose phosphate pathway, the purine de novo pathway, amino acids, and neurological disorder-related metabolites. The mass spectrometry was equipped with electrospray ionization in both positive and negative modes with scheduled multiple reactions monitoring. The validation of the method involved a comprehensive assessment of linearity, accuracy, precision, and matrix effect. The linear range was from 1.0 to 2000 ng mL^–1 with a high correlation coefficient (r > 0.99). The LOD ranged from 0.1 to 10 ng mL^–1, and the LOQ ranged from 0.1 to 25 ng mL^–1. The overall recovery ranged from 81.3% to 117.8%, with RSD < 15.1%. Subsequently, an analysis of metabolites was conducted in dSH-SY5Y neuroblastoma cells with 6-hydroxydopamine, a commonly used neurotoxin in neurodegenerative diseases. The results demonstrate that neurotoxin-induced mitochondrial damage significantly altered related analytes, corroborating previous estimates and validating the feasibility and reliability of the bioanalytical platform.

中文翻译：

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使用基于氰丙基的液相色谱串联质谱推进靶向代谢组学


代谢途径的变化被认为是疾病发现的关键，促使开发基于超高效液相色谱-串联质谱 （UHPLC-MS/MS） 的定量平台，以探索生物体的动态代谢物谱。本研究开发了一种基于氰丙基 （CN） 的液相色谱方法。通过调节色谱柱的 pH 环境，我们实现了 51 种代谢物的洗脱，涵盖了目前已知的最全面的生物途径。该方法具有快速色谱、高效分离和绿色化学优势，包括核苷和核苷酸、氧化还原代谢组、糖酵解途径、磷酸戊糖途径、嘌呤从头途径、氨基酸和神经系统疾病相关代谢物。质谱仪配备了正离子和负离子模式的电喷雾电离，并预定了多次反应监测。该方法的验证涉及对线性、准确度、精密度和基质效应的全面评估。线性范围为 1.0 至 2000 ng mL^–1，具有高相关系数 （r > 0.99）。LOD 范围为 0.1 至 10 ng mL^–1，LOQ 范围为 0.1 至 25 ng mL^–1。总回收率范围为 81.3% 至 117.8%，RSD < 为 15.1%。随后，用 6-羟基多巴胺（神经退行性疾病中常用的神经毒素）对 dSH-SY5Y 神经母细胞瘤细胞中的代谢物进行了分析。结果表明，神经毒素诱导的线粒体损伤显著改变了相关分析物，证实了之前的估计并验证了生物分析平台的可行性和可靠性。