The stable o-phthalaldehyde-ferrocene/6-ferrocenyl-1-hexanethiol pre-column derivatization high-performance liquid chromatography of dimethylarginine by novel dual fluorescence and electrochemical detector

被引:4
作者
Suzuki, Katsuhiro [1 ]
Katayama, Masatoki [2 ]
Takamatsu, Kiyoshi [3 ]
Kaneko, Satoru [3 ]
Ishikawa, Hiromichi [4 ]
Tsukimura, Takahiro [2 ]
Togawa, Tadayasu [2 ]
机构
[1] Astellas Pharma Inc, Drug Discovery Res, Drug Metab Res Labs, Yodogawa Ku, Osaka 5328514, Japan
[2] Meiji Pharmaceut Univ, Dept Funct Bioanal, Tokyo 2048588, Japan
[3] Ichikawa Gen Hosp, Tokyo Dent Coll, Dept Obstet & Gynecol, Ichikawa, Chiba 2720824, Japan
[4] Ichikawa Gen Hosp, Tokyo Dent Coll, Sch Med, Dept Urol, Ichikawa, Chiba 2720824, Japan
关键词
dual detection; fluorescence detection; pre-column derivatization HPLC; electrochemical detection; asymmetric dimethylarginine; AMINO-ACIDS; L-ARGININE; PLASMA; SERUM; ADMA;
D O I
10.1002/bmc.2823
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Monomethylarginine, asymmetric dimethylarginine and symmetric dimethylarginine were separated on a Wakopak Combi ODS with an acetonitrile100mm potassium phosphate buffer (pH 7.0; 1:1, v/v). Dimethylarginines were derived from o-phthalaldehyde for the fluorescence detector and from 6-ferrocenyl-1-hexanethiol for the electrochemical detector. The detection limits of the dimethylarginines in spiked plasma were 0.30.5 pmol by electrochemical detection and 12 pmol by fluorescence detection. The detection limits were improved over 30 times by electrochemical detection and 10 times by fluorescence detection compared with previous reports. In previous derivatization liquid chromatography, the reaction solutions, o-phthalaldehyde, 2-mercaptethanol and dimethylarginines were unstable and required quick derivatization at 4 degrees C. By our proposed pre-column methods, the dimethylarginines were derivatized at room temperature and the fluorescent products were stable for 6h. The manipulation performance was greatly advanced compared with previous LC reports. This is the first report on stable and sensitive dimethylarginines by dual detection. The selectivity was also improved by dual detection. The proposed method was applied to preliminary monitoring of dimethylargines in plasma and urine. Copyright (c) 2012 John Wiley & Sons, Ltd.
引用
收藏
页码:535 / 538
页数:4
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