Analysis of hormonal steroids in fish plasma and bile by coupling solid-phase extraction to GC/MS

Budzinski, H.; Devier, M. H.; Labadie, P.; Togola, A.
December 2006
Analytical & Bioanalytical Chemistry;Dec2006, Vol. 386 Issue 5, p1429
Academic Journal
An analytical procedure for the simultaneous determination of twelve endogenous steroids (testosterone, androstenedione, 17β-estradiol, estrone, pregnenolone, progesterone, dihydroandrostenedione, dihydrotestosterone, 11α-ketotestosterone, 17α-hydroxyprogesterone, 17α-hydroxypregnenolone, 17α,20β-dihydroxy-4-pregnen-3-one) in plasma and bile samples by solid-phase extraction (SPE) and gas chromatography/mass spectrometry (GC–MS) has been developed. After enzymatic hydrolysis for bile samples only, samples were concentrated and purified using two successive SPE (C18 and NH2) cartridges. Analytes were derivatized with a mixture of N-methyl- N-(trimethylsilyl)trifluoroacetamide (MSTFA) / mercaptoethanol / ammonium iodide (NH4I) and determined by GC–MS in selective ion monitoring mode. For most of the steroids monitored, recoveries were in the range 90–120% in plasma and in the range 60–70% in bile, and the reproducibility was below 10% for the complete procedure. Limits of detection obtained ranged from 0.1 to 0.4 ng/g in fish plasma and from 1.6 to 14 ng/g in fish bile. The developed method was successfully applied to the determination of plasma steroids in flounders ( Platichthys flesus) collected from two French estuaries.


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