Analysis of hormonal steroids in fish plasma and bile by coupling solid-phase extraction to GC/MS.
An analytical procedure for the simultaneous determination of twelve endogenous steroids (testosterone, androstenedione, 17beta-estradiol, estrone, pregnenolone, progesterone, dihydroandrostenedione, dihydrotestosterone, 11alpha-ketotestosterone, 17alpha-hydroxyprogesterone, 17alpha-hydroxypregnenolone, 17alpha,20beta-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 (C(18) and NH(2)) cartridges. Analytes were derivatized with a mixture of N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) / mercaptoethanol / ammonium iodide (NH(4)I) 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.
Budzinski
H
H
Laboratoire de Physico- et Toxico-Chimie (LPTC), UMR 5472 CNRS, Université Bordeaux I, 351 cours de la libération, 33405 Talence Cedex, France. h.budzinski@lptc.u-bordeaux1.fr
Devier
M H
MH
Labadie
P
P
Togola
A
A
D000818:Animals; D001646:Bile / Q000737:chemistry*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D008968:Molecular Conformation; D017686:Oncorhynchus mykiss; D010949:Plasma / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052617:Solid Phase Microextraction / Q000379:methods*; D012997:Solvents / Q000737:chemistry; D013256:Steroids / Q000032:analysis*; D013696:Temperature
D012997:Solvents; D013256:Steroids