Development and validation of an analytical method for the determination of 17β-estradiol residues in muscle of tambaqui (Colossoma macropomum Cuvier, 1818) by LC-MS/MS and its application in samples from a fish sexual reversion study.

Tambaqui (Colossoma macropomum Cuvier, 1818) is the main native fish species farmed in Brazil, and 17β-estradiol (E) is a natural steroid hormone commonly used for the production of female fish monosex population, which, in tambaqui, shows a higher growth rate than the male. Thus, to assess whether the fish meat of treated tambaqui contains hormonal residue levels, a high-performance liquid chromatography coupled with electrospray tandem mass spectrometry (LC-MS/MS) method for the determination of E residues in fish muscle was developed and validated. A QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) dispersive solid phase extraction method was used for the sample preparation. The chromatographic separation was performed in a Poroshel EC-18 reverse phase column. The mobile phase was a mixture of acetonitrile with 0.01% ammonium hydroxide (A) and water with 0.01% ammonium hydroxide (B). The ratio of A:B phases was 60:40 (v/v) used in an isocratic mode. The method validation was performed according to Commission Decision 2002/657/EC and Veterinary International Conference Harmonization (VICH GL49). Since matrix effects were observed, matrix-matched analytical curves are recommended for quantitation. The linearity, selectivity, intraday and interday precision, accuracy, decision limit, detection capability, and detection and quantitation limits of the method are reported. The limits of detection and quantitation were 0.3 ng/g and 1.0 ng/g, respectively. At these limits and slaughtering fish 7 months after the end of the treatment, the muscle of tambaqui did not show detectable hormone residue level. Thus, consumption of tambaqui edible tissue from fish treated with E for the purpose of sexual reversion is unlikely to represent a risk associated with the exposure of human subjects as residue levels of this hormone are not detected in the fish muscle.