An automated analyzer for methylated arginines in rat plasma by high-performance liquid chromatography with post-column fluorescence reaction

被引:29
作者
Dobashi, Y
Santa, T
Nakagomi, K
Imai, K
机构
[1] Univ Tokyo, Grad Sch Pharmaceut Sci, Bunkyo Ku, Tokyo 1130033, Japan
[2] Toyama Med & Pharmaceut Univ, Fac Pharmaceut Sci, Toyama 9300194, Japan
关键词
D O I
10.1039/b106828h
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A fully automated analyzer for methylated L-arginine metabolites [N,N-dimethyl-L-arginine (ADMA), N-methylarginine (NMMA) and N,N'-dimethyl-L-arginine (SDMA)] by high-performance liquid chromatography with post-column fluorescence derivatization was developed. This system consists of an on-line extraction. a separation on a reversed phase ion-pair chromatograph. a post-column derivatization by o-phthaladehyde (OPA) and thiol reaction. and fluorescence detection. NMMA. ADMA and SDMA were separated in 40 min with isocratic elution by a combination of octanoate and cyclohexane carboxylate as ion-pair reagents. The eluate was monitored at 450 nm with excitation at 337 nm. The calibration curves for NMMA. ADMA and SDMA showed linearity over the range from 0.05 mumol l(-1) (0.5 pmol on column) to 5.0 mumol l(-1) (50 pmol on column). This method does not require any time-consuming pre-treatment and requires only 10 mul of plasma sample for assay.
引用
收藏
页码:54 / 59
页数:6
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