Metabolic and Newborn Screening Laboratory, Department of Paediatrics, University of Szeged.
Department of Neurology, University of Szeged, Hungary.
J Chromatogr A. 2021 Jan 4;1635:461775. doi: 10.1016/j.chroma.2020.461775. Epub 2020 Dec 2.
Concurrent measurement of tyrosine, tryptophan and their metabolites, and other co-factors could help to diagnose and better understand a wide range of metabolic and neurological disorders. The two metabolic pathways are closely related to each other through co-factors, regulator molecules and enzymes. By using high performance liquid chromatography coupled to electrospray ionization triple quadrupole mass spectrometry, we present a robust, selective and comprehensive method to determine 30 molecules within 20 min using a Waters Atlantis dC18. The method was validated according to the guideline of European Medicines Agency on bioanalytical method validation. Analytical performance met all the EMA requirements and the assay covered the relevant clinical concentrations. Linear correlation coefficients were all >0.998. Intra-day and inter-day accuracy were between 80-119% and 81-117%, precision 1-19% respectively. The method was applied to measure TYR, TRP and their metabolites, and other neurologically important molecules in human serum and CSF samples. The assay can facilitate the diagnosis and is suitable for determination of reference values in clinical laboratories.
同时测量酪氨酸、色氨酸及其代谢物和其他辅助因子有助于诊断和更好地理解广泛的代谢和神经紊乱。这两种代谢途径通过辅助因子、调节分子和酶密切相关。本研究采用高效液相色谱-电喷雾三重四极杆质谱联用技术，建立了一种在 20 min 内分析测定 30 种物质的可靠、选择性和全面的方法，使用 Waters Atlantis dC18 色谱柱。该方法按照欧洲药品管理局（EMA）关于生物分析方法验证的指南进行了验证。分析性能符合 EMA 的所有要求，检测范围涵盖了相关的临床浓度。线性相关系数均>0.998。日内和日间精密度分别为 1-19%和 81-117%，准确度为 80-119%。该方法应用于测量人血清和脑脊液样本中的 TYR、TRP 及其代谢物和其他神经重要分子。该检测方法有助于诊断，适用于临床实验室确定参考值。
