Department of Hygiene, Third Faculty of Medicine, Charles University, Ruská 87, 100 00 Prague 10, Czech Republic.
Institute of Organic Chemistry and Biochemistry, The Czech Academy of Sciences, Flemingovo n. 2, 166 10 Prague 6, Czech Republic.
Int J Mol Sci. 2021 Mar 15;22(6):2970. doi: 10.3390/ijms22062970.
Asymmetric and symmetric dimethylarginines are toxic non-coded amino acids. They are formed by post-translational modifications and play multifunctional roles in some human diseases. Their determination in human blood plasma is performed using capillary electrophoresis with contactless conductivity detection. The separations are performed in a capillary covered with covalently bonded PAMAPTAC polymer, which generates anionic electroosmotic flow and the separation takes place in the counter-current regime. The background electrolyte is a 750 mM aqueous solution of acetic acid with pH 2.45. The plasma samples for analysis are treated by the addition of acetonitrile and injected into the capillary in a large volume, reaching 94.5% of the total volume of the capillary, and subsequently subjected to electrophoretic stacking. The attained LODs are 16 nm for ADMA and 22 nM for SDMA. The electrophoretic resolution of both isomers has a value of 5.3. The developed method is sufficiently sensitive for the determination of plasmatic levels of ADMA and SDMA. The determination does not require derivatization and the individual steps in the electrophoretic stacking are fully automated. The determined plasmatic levels for healthy individuals vary in the range 0.36-0.62 µM for ADMA and 0.32-0.70 µM for SDMA.
不对称和对称二甲基精氨酸是有毒的非编码氨基酸。它们是通过翻译后修饰形成的,在一些人类疾病中发挥着多功能作用。它们在人血浆中的测定采用毛细管电泳-非接触式电导检测法进行。分离是在涂有共价键合的 PAMAPTAC 聚合物的毛细管中进行的,该聚合物会产生阴离子电渗流,分离则在逆流模式下进行。背景电解质为 pH 值为 2.45 的 750 mM 乙酸水溶液。用于分析的血浆样品通过添加乙腈进行处理,并以大体积注入毛细管,达到毛细管总体积的 94.5%,随后进行电泳浓缩。测定的检出限分别为 ADMA 的 16nm 和 SDMA 的 22nm。两种异构体的电泳分辨率值为 5.3。所开发的方法对 ADMA 和 SDMA 的血浆水平测定具有足够的灵敏度。该测定不需要衍生化,电泳浓缩的各个步骤均实现了自动化。健康个体的血浆水平测定结果显示,ADMA 的范围为 0.36-0.62µM,SDMA 的范围为 0.32-0.70µM。