Fondazione Policlinico Universitario A. Gemelli IRCCS, Rome, Italy.
Fondazione Policlinico Universitario A. Gemelli IRCCS, Rome, Italy; Università Cattolica del Sacro Cuore, Rome, Italy.
Clin Chim Acta. 2023 Mar 15;543:117306. doi: 10.1016/j.cca.2023.117306. Epub 2023 Mar 27.
Arginine metabolism is involved in the regulation of several biological processes. Many liquid chromatography tandem-mass spectrometry methods for the determination of arginine and its metabolites have been developed but they are time consuming and imply long pre-analytical procedures. The purpose of this study was to develop a rapid method for the simultaneous determination of arginine, citrulline, ornithine, symmetric and asymmetric dimethylarginine and monomethylarginine in human plasma.
The pre-analytical procedure consisted in a simple deproteinization. The chromatographic separation was performed using hydrophilic interaction liquid chromatography. Analytes detection was performed with a triple quadrupole equipped with electrospray ion source operating in positive ion mode. Mass spectrometry experiments were conducted in multiple reaction monitoring mode.
Recovery ranged from 92.2 to 108.0%. The within-run imprecision and between-run imprecision ranged from 1.5 to 6.8 % and 3.8 to 11.9%, respectively. Carry over and matrix effect did not affect quantitative analysis. Extraction recovery was between 95 and 105 %. Stability after pre-analytical procedure was tested and all the metabolites were stable after 48 h at 4 °C. In conclusion, our novel method allow a rapid and easy determination of arginine and its metabolites both for research and clinical routine use.
精氨酸代谢参与了多种生物过程的调节。已经开发出许多用于测定精氨酸及其代谢物的液相色谱串联质谱法，但这些方法耗时且需要进行长时间的前处理。本研究旨在开发一种快速测定人血浆中精氨酸、瓜氨酸、鸟氨酸、对称和非对称二甲基精氨酸和单甲基精氨酸的方法。
前处理步骤包括简单的蛋白沉淀。采用亲水相互作用液相色谱进行色谱分离。采用电喷雾源正离子模式的三重四极杆进行分析物检测。质谱实验采用多重反应监测模式进行。
回收率范围为 92.2%至 108.0%。批内精密度和批间精密度范围分别为 1.5%至 6.8%和 3.8%至 11.9%。无交叉污染和基质效应。提取回收率在 95%至 105%之间。检测了前处理后的稳定性，所有代谢物在 4°C 48 小时内均稳定。总之，我们的新方法可以快速、简便地测定精氨酸及其代谢物，适用于研究和临床常规用途。
