Highly sensitive time-resolved fluorometric determination of estrogens by high-performance liquid chromatography using a beta-diketonate europium chelate.
A new fluorescent europium chelate labeling reagent, 5-(4"-chlorosulfo-1',1"-diphenyl-4'-yl)-1,1,1,2,2-pentafluoro-3,5-pentanedione (CDPP), was synthesized for the time-resolved fluorometric detection of HPLC. The label can be directly bound to amino or phenolic hydroxyl groups of analytes with its chlorosulfonyl group, and the labeled analytes are separated on a HPLC column. After separation, EuCl(3), TOPO (tri-n-octylphosphine oxide), and Triton X-100 were added by post-column introduction to the eluent, and the fluorescence of the europium chelate was measured with the time-resolved fluorometric detector. Estrone (E1), 17beta-estradiol (E2), ethynylestradiol (EE2) and estriol (E3) were measured with the detection limits of 0.65, 0.65, 0.65 and 0.60 ng/ml, respectively. The recovery for river water samples was in the range of 86.0-105.1% with the RSD of 1.9-5.8%. The method was applied to the analysis of a river water sample and estrone (E1) was determined to be 2.1 ng/l. The results and processing have been compared with those of a GC-MS method and a high degree of correlation (r> or =0.98) was observed.
Matsumoto
Kazuko
K
Department of Chemistry, Advanced Research Center for Science and Engineering, Waseda University, Shinjuku-ku, Tokyo 169-8555, Japan. kmatsu@mn.waseda.ac.jp
Tsukahara
Yoshie
Y
Uemura
Tomonari
T
Tsunoda
Kinichi
K
Kume
Hidehiro
H
Kawasaki
Seiji
S
Tadano
Jutaro
J
Matsuya
Takeshi
T
D002138:Calibration; D002614:Chelating Agents / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004967:Estrogens / Q000032:analysis*; D005063:Europium / Q000737:chemistry*; D005466:Fluorocarbons; D009682:Magnetic Resonance Spectroscopy; D010420:Pentanes / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
C461491:5-(4''-chlorosulfo-1',1''-diphenyl-4'-yl)-1,1,1,2,2-pentafluoro-3,5-pentanedione; D002614:Chelating Agents; D004967:Estrogens; D005466:Fluorocarbons; D010420:Pentanes; D005063:Europium