A new sensitive determination method of estradiol in plasma using peroxyoxalate ester chemiluminescence combined with an HPLC system.
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 mM. 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.
Yamada
H
H
Kashima Pharmaceuticals Research and Development Center, Mitsubishi-Tokyo Pharmaceuticals Inc., 14 Sunayama, Hasaki-machi, Kashima-gun, Ibaraki 314-0255, Japan. 3000795@mitsubishi-pharm.co.jp
Kuwahara
Y
Y
Takamatsu
Y
Y
Hayase
T
T
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004952:Esters; D004958:Estradiol / Q000097:blood*; D008163:Luminescent Measurements; D010070:Oxalates / Q000737:chemistry*; D051381:Rats; D012680:Sensitivity and Specificity
D004952:Esters; D010070:Oxalates; D004958:Estradiol; C044622:peroxyoxalate