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Hence, these methods can assist in investigating the potential application of hair analysis and serum analysis to provide complementary information when the urinary excretion of stanozolol and 3 ? -hydroxystanoozlol is impaired. To our knowledge, the detection of stanozolol and 3 ? -hydroxystanozolol in rat hair, urine and serum at such low concentration levels using LC-MS/MS, has been reported here for the first time. Using the newly developed methods presented here, future research can carry out in vivo studies to further investigate stanozolol metabolism, thus making an important step towards understanding of the array of factors that may confound urinalysis results. Also, these methods can be extended by analysing human hair, urine and serum samples in tandem to provide a pattern of drug use and this can be useful for testing doping with stanozolol and other commonly abused AAS. 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