title: Liquid chromatographic determination of oestradiol in serum by pre-column derivatization with dansyl chloride or laryl chloride and peroxyoxalate chemiluminescence detection.
abstract: Column liquid chromatography with chemiluminescence detection is used to determine oestradiol in serum. Oestradiol is labelled by means of a two-phase derivatization procedure with either dansyl chloride or laryl chloride. After derivatization the excess reagent is removed on a short amino-bonded column: the sulphonyl chloride functional group reacts with the amino groups causing removal of the label; the oestradiol derivatives are not retained. Chromatography of oestradiol derivatives is performed with methanol-water mixtures as eluent. Chemical excitation is carried out by adding bis(2-nitrophenyl)oxalate and hydrogen peroxide dissolved in acetonitrile to the column eluate. Linearity is observed (r = 0.9998; n = 7) over at least three decades (10(-6)-10(-9) M) for both dansyl chloride and laryl chloride. A detection limit of 5 x 10(-10) M (50 fmol injected) has been achieved and the dansyl derivatization method has been applied to the determination of oestradiol in spiked serum.
authors.affiliation: Department of Analytical Chemistry, Free University, Amsterdam, The Netherlands.
authors.forename: P J
authors.initials: PJ
authors.lastname: Kwakman
authors.forename: D A
authors.initials: DA
authors.lastname: Kamminga
authors.forename: U A
authors.initials: UA
authors.lastname: Brinkman
authors.forename: G J
authors.initials: GJ
authors.lastname: de Jong
mesh_terms: D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid / Q000379:methods; D003619:Dansyl Compounds; D004958:Estradiol / Q000097:blood*; D010070:Oxalates; D012235:Rhodamines
chemical_list: D003619:Dansyl Compounds; D010070:Oxalates; D012235:Rhodamines; C022027:lissamine rhodamine B; D004958:Estradiol; C044622:peroxyoxalate; C029038:dansyl chloride