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High performance liquid chromatographic analysis of six conjugated and unconjugated estrogens in serum
Author(s) -
Su Syang Y.,
Shiu Gerald K.,
Simmons John,
Viswanathan C. T.,
Skelly Jerome P.
Publication year - 1992
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/bmc.1130060604
Subject(s) - chemistry , estrone , chromatography , sodium hydroxide , acetonitrile , reagent , tetrabutylammonium hydroxide , high performance liquid chromatography , sodium , conjugated system , hormone , organic chemistry , biochemistry , polymer
Six estrogens—estrone, equilin, estradiol, sodium estrone sulphate, sodium equilin sulphate and sodium 17‐α‐dihydroequilin sulphate in dog serum were successfully separated and quantified by a high performance liquid chromatographic method developed in our laboratories. The mobile phase was optimized by studying the effects of an organic modifier (acetonitrile) and an ion pairing reagent (tetrabutylammonium hydroxide). The serum sample work‐up procedure was designed to recover both conjugated and unconjugated estrogens. The optimum method involved acetonitrile to precipitate serum proteins/peptides and extract estrogens. Residues were reconstituted in 50% acetonitrile in water for injection. The detection limits for these six estrogens via UV detection ranged from 0.5 to 5 ng on‐column with a signal‐to‐noise ratio ( S / N ) of 10 for a 20 μL injection and via fluorescence 0.1 ng on‐column for 17‐β‐estradiol. Validation data are included for all six estrogens.