Ashok Marwah Xu Nianzu Padma Marwah Terence P. Barry


17a-Methyltestosterone is used as feed additive to manipulate the gender of fish for aquaculture. Earlier a simple, yet specific and robust validated high performance liquid chromatographic method has been developed for the determination of 17a-methyltestosterone in fish feed. The present work describes a highly sensitive and robust Liquid Chromatography-Mass Spectrometry method for the quantitation of 17a-methyltestosterone in aquatic water systems using 17â-hydroxy-3â-methoxyandrost-5-en-7-one as internal standard. The method was validated in the concentration range of 0.2 to 25 ng of 17a-methyltestosterone on column leading to a limit of quantitation of 0.08 ppb or 0.08 mg/L in water, and has potential to increase the limit of detection and quantitation by an order of magnitude, if required.


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High-performance liquid chromatography-Mass spectrometry, Electrospray ionization, 17a- Methyltestosterone, Solid phase extraction, Method validation

Beardmore, J.A., Mair, G.C. and Lewis, R.I. (2001). Monosex male production in finfish as exemplified by tilapia: applications, problems, and prospects. Aquaculture, 197: 283–301.
Cappiello, A., Famiglini, G., Mangani, F., Palma, P.and Siviero, A. (2003). Nano high-performance liquid chromatography– electron ionization mass spectrometry approach for environmental analysis. Anal. Chim. Acta., 493: 125-136.
Center for Veterinary Medicine (CVM), Food and Drug Administration (FDA), Bioanalytical method validation, guidance for industry (http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm070107.pdf)
Chang, H., Wu, S., Hu, J., Mari, A. and Kunikane, S. (2008). Trace analysis of androgens and progestogens in environmental waters by ultra-performance liquid chromatography-electrospray tandem mass spectrometry. J . Chromatography A, 1195: 329-354.
Coddington, D.T., Manning, B., Eya, J. and Brock, D. (2000). Concentration of 17a-methyltestosterone in hormone treated feed: effects of analytical technique, fabrication, and storage temperature. J. World Aquaculture Soc., 31: 42-45.
Cravedi, J.P. and Delous G. J (1991). Use of an immobilized enzyme reactor for the analysis of residues of 17a- methyltestosterone in trout by high-performance liquid chromatography. Chromatography, 564: 461-67.
Gale, W.L., Fitzpatrick, M.S., Lucero, M., Contreras-Sanchez, W.M. and Schreck, C.B., (1999). Masculinization of Nile tilapia (Oreochromis niloticus) by immersion in androgens. Aquaculture, 178: 349–357.
Hauser, B., Deschner T., and Boesch, C. (2008). Development of a liquid chromatography-tandem mass spectrometry method for the determination of 23 endogenous steroids in small quantities of primate urine. J Chrom. B, 862, 100-112
Ermer, Joachim and Miller, John H. McB. (2005). Method validation in pharmaceutical analysis: a guide to best practice.
Lagana, A., Fago, G., Marino, A. and Santarelli, D. (2001). Liquid chromatography tandem mass spectrometry applied to the analysis of natural and synthetic steroids in environmental waters. Analytical Letters, 34: 913-926.
Marwah, A., Marwah, P. and Lardy H. A. (2005). Development and validation of a high performance liquid chromatography assay for 17á-methyltestosterone in fish feed. J. Chrom. B, 824: 107–115.
Marwah, P., Marwah A., Kneer, N. and Lardy H. A. (2001). Ergosteroids IV: Synthesis and biological activity of steroid glucuronosides, ethers, and alkylcarbonates. Steroids, 66: 581–595.
Swartz , Michael E. and Krull, Ira S. (1997), Marcel and Dekker, New York analytical method development and validation.
Regal, P., Nebot, C., Vázquez, B.I., Cepeda, A., and Fente, C.A. (2010). Determination of the hormonal growth promoter 17á-methyltestosterone in food-producing animals: Bovine hair analysis by HPLC–MS/MS. Meat Science, 84: 196-201.
Rosing, H., Man, W. Y., Doyle, E., Bult, A., and Beijnen, J. H. (2000). Bioanalytical liquid chromatographic method validation. a review of current practices and procedures. J Liquid Chromatography & Related Technologies, 23: 329-354.
Sun, L., Liu, Y., Chu, X. and Jin-Ming, L. (2010). Trace analysis of fifteen androgens in environmental waters by LC-ESIMS- MS combined with solid-phase disk extraction cleanup. Chromatographia, 71: 867-873.
Tölgyesi, Á., Verebey, Z., Sharma, V. K., Kovacsics, L. and Fekete, J. (2010). Simultaneous determination of corticosteroids, androgens, and progesterone in river water by liquid chromatography-tandem mass spectrometry. Chemosphere, 78: 972-9.
Shah, V. P., Midha, K. K., Findlay, J. W. A., Hill, H. M., Hulse, J. D., McGilveray, I. J., McKay, G., Miller, K. J., Patnaik, R. N., Powell, M. L., Tonelli, A., Viswanathan, C. T., and Yacobi, A. (2000). Bioanalytical method validation-a revisit with a decade of progress. Pharmaceutical Research, 17, 1551-57.
Walker, C.J., Cowan, D.A., James, V.H.T., Lau, J.C.Y., and T. Kicman, A.T. (2009). Doping in sport-2. Quantification of the impurity 19-norandrostenedione in pharmaceutical preparations of norethisterone. Steroids, 74: 335-340.
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Marwah, A., Nianzu, X., Marwah, P., & Barry, T. P. (2010). Development and validation of a High performance liquid chromatography- Mass spectrometry method for 17a-methyltestosterone in Aquatic water systems. Journal of Applied and Natural Science, 2(2), 182–189. https://doi.org/10.31018/jans.v2i2.117
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