17α-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 17α-methyltestosterone in fish feed. The present work describes a highly sensitive and robust Liquid Chromatography-Mass Spectrometry method for the quantitation of 17α-methyltestosterone in aquatic water systems using 17β-hydroxy-3β-methoxyandrost-5-en-7-one as internal standard. The method was validated in the concentration range of 0.2 to 25 ng of 17α-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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