Steroid determination in fish plasma using capillary electrophoresis

Bykova, L.; Archer-Hartmann, S. A.; Holland, L. A.; Iwanowicz, L. R.; Blazer, V. S.

Abstract:

A capillary separation method that incorporates pH-mediated stacking is employed for the simultaneous determination of circulating steroid hormones in plasma from Perca flavescens (yellow perch) collected from natural aquatic environments. The method can be applied to separate eight steroid standards: progesterone, 17??,20??-dihydroxypregn-4-en-3-one, 17??-hydroxyprogesterone, testosterone, estrone, 11-ketotestosterone, ethynyl estradiol, and 17??-estradiol. Based on screening of plasma, the performance of the analytical method was determined for 17??,20??-dihydroxypregn-4-en-3-one, testosterone, 11-ketotestosterone, and 17??-estradiol. The within-day reproducibility in migration time for these four steroids in aqueous samples was ???2%. Steroid quantification was accomplished using a calibration curve obtained with external standards. Plasma samples from fish collected from the Choptank and Severn Rivers, Maryland, USA, stored for up to one year were extracted with ethyl acetate and then further processed with anion exchange and hydrophobic solid phase extraction cartridges. The recovery of testosterone and 17??-estradiol from yellow perch plasma was 84 and 85%, respectively. Endogenous levels of testosterone ranged from 0.9 to 44ng/ml, and when detected 17??,20??-dihydroxypregn-4-en-3-one ranged from 5 to 34ng/ml. The reported values for testosterone correlated well with the immunoassay technique. Endogenous concentrations of 17??-estradiol were ???1.7ng/ml. 11-Ketotestosterone was not quantified because of a suspected interferant. Higher levels of 17??,20??-dihydroxypregn-4-en-3-one were found in male and female fish in which 17??-estradiol was not detected. Monitoring multiple steroids can provide insight into hormonal fluctuations in fish. ?? 2010 SETAC.

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Article

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70037552

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Bykova, L.; Archer-Hartmann, S. A.; Holland, L. A.; Iwanowicz, L. R.; Blazer, V. S.

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1956

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English

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Environmental Toxicology and Chemistry

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Steroid determination in fish plasma using capillary electrophoresis; 2010; Article; Journal; Environmental Toxicology and Chemistry; Bykova, L.; Archer-Hartmann, S. A.; Holland, L. A.; Iwanowicz, L. R.; Blazer, V. S.