Determination of 17α-Methyltestosterone in Freshwater Samples of Tilapia Farming by High Performance Liquid Chromatography


17α-methyltestosterone is used to induce the sex reversal of Tilapia sp. to obtain cultures mono-sex to an economically viable. This practice may lead to environmental contamination and problems in human health. Therefore methods need to be developed to detect residues of 17α-methyltestosterone in aqueous matrices. A simple high-performance liquid chromatographic method using ultraviolet detection (245 nm) and testosterone as internal standard has been developed for the monitoring 17α-methyltestosterone in freshwater samples of tilapia aquaculture. The method described involves limited sample preparation as it includes a filtration followed by a single solid-phase extraction step using C18 cartridge. Validation data indicated that the HPLC-UV method for 17α-methyltestosterone determination in the concentration range of 50 - 2000 μg/L provided good linearity, sensitivity, accuracy and precision. Method performance was efficiently applied to monitoring the freshwater samples of fish ponds and the surrounding aquatic channels.

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I. Barbosa, S. Lopes, R. Oliveira, I. Domingues, A. Soares and A. Nogueira, "Determination of 17α-Methyltestosterone in Freshwater Samples of Tilapia Farming by High Performance Liquid Chromatography," American Journal of Analytical Chemistry, Vol. 4 No. 4, 2013, pp. 207-211. doi: 10.4236/ajac.2013.44026.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] W. L. Gale, M. S. Fitzpatrick, M. Lucero, W. M. Contreras-Sanchez and C. B. Schreck, “Masculinization of Nile Tilapia (Oreochromis niloticus) by Immersion in Androgens Original,” Aquaculture, Vol. 178, No. 3-4, 1999, pp. 349-357. doi:10.1016/S0044-8486(99)00136-2
[2] J. A. Beardmore, G. C. Mair and R. I. Lewis, “Monosex Male Production in Finfish as Exemplified by Tilapia: Applications, Problems, and Prospects,” Aquaculture, Vol. 197, No. 1-4, 2001, pp. 283-301. doi:10.1016/S0044-8486(01)00590-7
[3] T. J. Pandian and S. G. Sheela, “Hormonal Induction of Sex Reversal in Fish,” Aquaculture, Vol. 138, No. 1-4, 1995, pp. 1-22. doi:10.1016/0044-8486(95)01075-0
[4] J. S. Abucay and G. C. Mair, “Hormonal Sex Reversal of Tilapias: Implications of Hormone Treatment Application in Closed Water Systems,” Aquaculture Research, Vol. 28,No. 11,1997, pp. 841-845. doi:10.1111/j.1365-2109.1997.tb01008.x
[5] L. R. Curtis, F. T. Diren, M. D. Hurley, W. K. Seim and R. A. Tubb, “Disposition and Elimination of 17α-Methyltestosterone in Nile Tilapia (Oreochromis niloticus),” Aquaculture, Vol. 99, No. 1-2, 1991, pp. 193-201. doi:10.1016/0044-8486(91)90298-L
[6] L. Andersen, R. Goto-Kazeto, J. M. Trant, J. P. Nash, B. Korsgaard and P. Bjerregaard, “Short-Term Exposure to Low Concentrations of the Synthetic Androgen Methyltestosterone Affects Vitellogenin and Steroid Levels in Adult Male Zebrafish (Danio rerio),” Aquatic Toxicology, Vol. 76, No. 3-4, 2006, pp. 343-352. doi:10.1016/j.aquatox.2005.10.008
[7] S. Wason, G. Pohlmeyer-Esch, C. Pallen, X. Palazzi, G. Espuña and R. Bars, “17α-Methyltestosterone: 28-Day Oral Toxicity Study in the Rat Based in the ‘Enhanced OECD Test Guideline 407’ to Detect Endocrine Effects,” Toxicology, Vol. 192, No. 2-3, 2003, pp. 119-137. doi:10.1016/S0300-483X(03)00265-8
[8] D. Teichert-Coddington, B. Manning, J. C. Eya and D. Brock, “Concentration of 17μ-Mehtyltestosterone in Hormone-Treated Feed: Effects of Analytical Technique, Fabrication, and Storage Temperature,” Journal of the World Aquaculture Society, Vol. 31, No. 1, 2000, pp. 42-50. doi:10.1111/j.1749-7345.2000.tb00696.x
[9] W. Van Thuyne and F. T. Delbeke, “Validation of a GCMS Screening Method for Anabolizing Agents in Aqueous Nutritional Supplements,” Journal of Chromatography Science, Vol. 43, No. 1, 2005, pp. 2-6.
[10] C. A. Goudie, “Extraction of a Synthetic Androgen from Fish Muscle and Quantitation by High Performance Liquid Chromatography,” Steroid, Vol. 44, No. 3, 1984, pp. 241-252. doi:10.1016/0039-128X(84)90005-9
[11] E. A. I. Daeseleire, A. De Guesquière and C. H. Van Peteghem, “Multiresidue Analysis of Anabolic Agents in Muscle Tissues and Urines of Cattle by GC-MS,” Journal of Chromatography Science, Vol. 30, No. 10, 1992, pp. 409- 414.
[12] B. L. Lampert and J. T. Stewart, “Determination of Anabolic Steroids and Zeranol in Human Serum by Isocratic Reverse Phase HPLC on Silica,” Journal of Liquid Chromatography, Vol. 12,No.16,1989, pp. 3231-3249.
[13] M. H. Blokland, H. J. van Rossum, H. A. Herbold, S. S. Sterk, R. W. Stephany and L. A. Van Ginkel, “Metabolism of Methyltestosterone, Norethandrolone and Methylboldenone in a Heifer,” Analytica Chimica Acta, Vol. 529, No. 1-2, 2005, pp. 317-323. doi:10.1016/j.aca.2004.10.064
[14] P. Regal, C. Nebot, B. I. Vásquez, A. Cepeda and C. A. Fente, “Determination of the Hormonal Growth Promoter 17α-Methyltestosterone in Food-Producing Animals: Bovine Hair Analysis by HPLC-MS/MS,” Meat Science, Vol. 84, No. 1, 2010, pp. 196-201. doi:10.1016/j.meatsci.2009.08.047
[15] R. Chiba and Y. Ishii, “Simultaneous Determination of Yohimbine Hydrochloride, Strychnine Nitrate and Methyltestosterone by Ion-Pair High-Performance Liquid Chromatography,” Journal of Chromatography A, Vol. 588, No. 1-2, 1991, pp. 344-347. doi:10.1016/0021-9673(91)85044-G
[16] A. Cappiello, G. Famiglini, F. Mangani, P. Palma and A. Siviero, “Nano-High-Performance Liquid Chromatography-Electron Ionization Mass Spectrometry Approach for Environmental Analysis,” Analytica Chimica Acta, Vol. 493, No. 2, 2003, pp. 125-136. doi:10.1016/S0003-2670(03)00868-7
[17] A. Marwah, P. Marwah and H. Lardy, “Development and Validation of a High Performance Liquid Chromatography Assay for 17α-Methyltestosterone in Fish Feed,” Journal of Chromatography B, Vol. 824, No. 1-2, 2005, pp. 107-115. doi:10.1016/j.jchromb.2005.07.005

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