分散固相萃取结合毛细管电泳-激光诱导荧光法测定血清中甲基化精氨酸

doi:10.7523/j.issn.2095-6134.2020.04.005

中国科学院大学学报 ›› 2020, Vol. 37 ›› Issue (4): 465-472.DOI: 10.7523/j.issn.2095-6134.2020.04.005

分散固相萃取结合毛细管电泳-激光诱导荧光法测定血清中甲基化精氨酸

谢士杰¹, 杨泽诚¹, 周洲², 蔺亚辉², 苏保满², 丁永胜¹

1 中国科学院大学生命科学学院, 北京 100049;
2 中国医学科学院/北京协和医学院国家心血管病研究中心, 北京阜外医院检验中心, 北京 100044

收稿日期:2018-12-17 修回日期:2019-05-08 发布日期:2020-07-15
通讯作者: 丁永胜
基金资助:
Supported by the National Nature Science Foundation of China (31571481,31771585)

Determination of methylated arginines in serum by a dispersive solid-phase extraction coupled with capillary electrophoresis-laser induced fluorescence detection

XIE Shijie¹, YANG Zecheng¹, ZHOU Zhou², LIN Yahui², SU Baoman², DING Yongsheng¹

1. College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China;
2. National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College&Testing Center, Fuwai Hospital, Beijing 100044, China

Received:2018-12-17 Revised:2019-05-08 Published:2020-07-15
Supported by:
Supported by the National Nature Science Foundation of China (31571481,31771585)

摘要/Abstract

摘要： 介绍一种分散固相离子交换萃取方法，结合毛细管电泳-激光诱导荧光检测法，测定血清样品中不对称二甲基精氨酸、对称二甲基精氨酸和单甲基精氨酸。将提取甲基化精氨酸的树脂粒加入到4-氯-7-硝基苯并呋喃乙腈溶液和碱性硼酸钠溶液（pH=10.5）的混合溶液中，于60℃水浴中衍生化。在80 mmol/L磷酸盐溶液（pH=2.0）分离电解质和20 kV分离电压条件下，这3种衍生物在熔融石英毛细管中获得基线分离。根据FDA生物分析方法验证指南，进行方法验证，结果满意。利用该方法测定了当地医院血清样品中的3种甲基化精氨酸含量。

关键词: 甲基化精氨酸, 分散固相萃取, 4-氯-7-硝基苯并呋喃, 毛细管电泳-激光诱导荧光

Abstract: We describe a dispersive solid-phase ion exchange extraction method combined with capillary electrophoresis-laser induced fluorescence detection for the determination of asymmetric dimethylarginine, symmetric dimethylarginine, and monomethylarginine in serum samples. The methylated arginine-extracted resin beads were added into a mixed solution of 4-chloro-7-nitrobenzofuran acetonitrile solution and alkaline sodium borate solution (pH=10. 5) for the derivatization in a 60℃ water bath. The three derivatives were subjected to baseline separation in a fused silica capillary under the conditions of a separation electrolyte of 80 mmol/L phosphate (pH=2. 0) and a separation voltage of 20 kV. On the basis of the FDA bioanalytical method validation guidance, the method validation was conducted and the results were satisfactory. This method was used to determine the three methylated arginines in the serum samples from local hospital.

Key words: methylated arginines, dispersive solid-phase extraction, 4-chloro-7-nitrobenzofurazan, capillary electrophoresis-laser induced fluorescence

中图分类号:

O657.1

谢士杰, 杨泽诚, 周洲, 蔺亚辉, 苏保满, 丁永胜. 分散固相萃取结合毛细管电泳-激光诱导荧光法测定血清中甲基化精氨酸[J]. 中国科学院大学学报, 2020, 37(4): 465-472.

XIE Shijie, YANG Zecheng, ZHOU Zhou, LIN Yahui, SU Baoman, DING Yongsheng. Determination of methylated arginines in serum by a dispersive solid-phase extraction coupled with capillary electrophoresis-laser induced fluorescence detection[J]. , 2020, 37(4): 465-472.

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分散固相萃取结合毛细管电泳-激光诱导荧光法测定血清中甲基化精氨酸

Determination of methylated arginines in serum by a dispersive solid-phase extraction coupled with capillary electrophoresis-laser induced fluorescence detection

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