Clinical Biochemistry Department, Institut de Recerca Sant Joan de Déu, Barcelona, Spain.
CIBERER-Instituto de Salud Carlos III, Barcelona, Spain.
Metabolomics. 2018 May 25;14(6):76. doi: 10.1007/s11306-018-1374-4.
Amino acid analysis in biological fluids is essential for the study of inborn errors of metabolism (IEM) and other diseases.
Our aim was to develop a UPLC-MS/MS procedure for the analysis of 25 amino acids and identification of 17 related compounds.
Sample treatment conditions were optimized for plasma, urine, cerebrospinal fluid (CSF) and dried blood spots. Amino acids and related compounds were analyzed on a Waters ACQUITY UPLC H-class instrument with a reversed-phase C-18 column using water and acetonitrile with 0.1% formic acid as the mobile phases (run time = 9 min). The detection was performed with a Waters Xevo TQD triple-quadrupole mass spectrometer using positive electrospray ionization in the multiple reaction monitoring mode.
The method linearity, intra-assay and inter-assay precision, detection limit, quantification limit and trueness analysis displayed adequate results in both physiological and pathological conditions. Method comparison was performed between UPLC-MS/MS and ion exchange chromatography (IEC) with ninhydrin derivatization, and the methods showed good agreement, except for 4-hydroxyproline, aspartate and citrulline. Paediatrics age-related reference values in plasma, urine and CSF were established and patients with different IEM were easily identified.
We report a modified UPLC-MS/MS procedure for the analysis of 42 amino acids and related compounds in different specimens. The method is fast, sensitive and robust, and it has been validated to be an alternative to the traditional IEC procedure as the routine method used in metabolic laboratories. The method greatly decreases the run time of the analysis while displaying good metrological results.
在生物体液中进行氨基酸分析对于研究先天性代谢错误（IEM）和其他疾病至关重要。
我们旨在开发一种 UPLC-MS/MS 程序，用于分析 25 种氨基酸并鉴定 17 种相关化合物。
优化了血浆、尿液、脑脊液（CSF）和干血斑的样品处理条件。使用 Waters ACQUITY UPLC H 类仪器，在反相 C-18 柱上，以水和含 0.1%甲酸的乙腈作为流动相（运行时间=9 分钟）分析氨基酸和相关化合物。采用 Waters Xevo TQD 三重四极杆质谱仪，在正电喷雾电离模式下的多反应监测模式进行检测。
该方法在生理和病理条件下均显示出良好的线性、日内和日间精密度、检测限、定量限和准确度分析结果。UPLC-MS/MS 与离子交换色谱（IEC）和茚三酮衍生化法进行方法比较，两种方法除 4-羟脯氨酸、天冬氨酸和瓜氨酸外，均具有良好的一致性。建立了不同标本中血浆、尿液和 CSF 中儿童年龄相关的参考值，并且能够容易地识别出患有不同 IEM 的患者。
我们报告了一种改良的 UPLC-MS/MS 程序，用于分析不同标本中的 42 种氨基酸和相关化合物。该方法快速、灵敏且稳健，已被验证可替代传统的 IEC 程序作为代谢实验室的常规方法。该方法大大缩短了分析的运行时间，同时显示出良好的计量学结果。
