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A new sensitive determination method of estradiol in plasma using peroxyoxalate ester chemiluminescence combined with an HPLC system
Author(s) -
Yamada Hiroyuki,
Kuwahara Yasuhiro,
Takamatsu Yasuo,
Hayase Tetsuo
Publication year - 2000
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/1099-0801(200008)14:5<333::aid-bmc992>3.0.co;2-o
Subject(s) - peroxyoxalate , chemiluminescence , chemistry , oxalate , chromatography , high performance liquid chromatography , detection limit , extraction (chemistry) , dansyl chloride , ethyl acetate , derivatization , organic chemistry
A new sensitive determination method of estradiol in a plasma sample using peroxyoxalate ester chemiluminescence was developed. Estradiol, which was extracted by liquid–liquid extraction using ethyl acetate from plasma, was derivatized with dansyl‐chloride (DNS‐Cl) and separated by reverse‐phase HPLC. The performance of four oxalates, bis(trichlorophenyl)oxalate (TCPO), bis(2,4‐dinitrophenyl)oxalate (DNPO), bis(pentafluorophenyl)oxalate (PFPO), and bis[4‐nitro‐2‐(3,6,9‐trioxadecyloxycarbonyl)phenyl] oxalate (TDPO), were evaluated using the static system, and DNPO was found to have the most sensitive and stable chemiluminescence at a H 2 O 2 concentration of 30 m M . HPLC‐chemiluminescence system using DNPO for the determination of estradiol was established. The detection limit of dansylated‐estradiol (DNS‐E2) was 15 fmol (4 pg) in the standard solution and 44 fmol (12 pg) in the rat plasma sample at S / N = 3. Copyright © 2000 John Wiley & Sons, Ltd.