Centro de Diagnóstico de Enfermedades Moleculares, CEDEM, Facultad de Ciencias, Módulo 10, Universidad Autónoma de Madrid, 28049 Madrid, Spain.
Centro de Diagnóstico de Enfermedades Moleculares, CEDEM, Facultad de Ciencias, Módulo 10, Universidad Autónoma de Madrid, 28049 Madrid, Spain.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Jan 1;944:141-3. doi: 10.1016/j.jchromb.2013.10.032. Epub 2013 Nov 15.
This paper describes a full detailed high performance liquid chromatography/tandem mass spectrometry method for the identification and quantification of human urine alpha-aminoadipic semialdehyde, biomarker of pyridoxine-dependent epilepsy. The ionization mode of the electrospray interface was negative and the metabolite was detected in the multiple reaction monitoring mode. Intra-day and inter-day laboratory precision were 4.64% and 7.30%, respectively, total run time was 3.5min. The calibration curve was linear between 0.25 and 10nmol with a correlation coefficient of the calibration line (R(2)≥0.9984); the limit of quantification was 0.25nmol within the control group. This simple, fast, high reproducible and robust procedure facilitates a rapid diagnosis of patients with pyridoxine-dependent epilepsy and can also be used to confirm the elevated urinary alpha-aminoadipic semialdehyde excretion in patients with other metabolic diseases as molybdenum cofactor and isolated sulphite oxidase deficiencies.
本文描述了一种用于鉴定和定量人尿液中α-氨基己二酸半醛（吡哆醇依赖性癫痫的生物标志物）的全二维高效液相色谱/串联质谱法。电喷雾接口的电离模式为负离子，代谢物在多反应监测模式下检测。日内和日间实验室精密度分别为 4.64%和 7.30%，总运行时间为 3.5min。校准曲线在 0.25 至 10nmol 之间呈线性，校准线的相关系数（R(2)≥0.9984）；对照组的定量限为 0.25nmol。该方法简单、快速、重现性好、稳健，有利于吡哆醇依赖性癫痫患者的快速诊断，也可用于确认其他代谢性疾病（如钼辅因子和孤立的亚硫酸盐氧化酶缺乏症）患者尿液中α-氨基己二酸半醛排泄量升高。
