Simultaneous quantitation of 9 anabolic and natural steroidal hormones in equine urine by UHPLC-MS/MS triple quadrupole

A new fast and easy analytical procedure for the simultaneous detection and quantification of 9 anabolic steroids (deslorelin, dexamethasone sodium phosphate, prednisolone, methylprednisolone, stanozolol, boldenone, nandrolone, dexamethasone isonicotinate and altrenogest) in horse urine for doping control have been developed by using the ultra-high-performance liquid chromatography coupled with tandem mass spectrometry technique (UHPLC-MS/MS). A total amount of 400 μl of sample was evaporated, restored and injected in the UHPLC-MS/MS. The proposed method was fully validated showing a recovery higher than 89.12% and a coefficient of variation lower than 6.02%. The correlation coefficients range of the analyzed compound's calibration curves was 0.9955–0.9997, and the limits of detection and quantification were in the range of 0.1 and 0.25 μg/l, respectively. •A new fast and sensitive method for 9 anabolic steroids in urine has been set-up.•High sensitivity and accuracy have been obtained.•The method could be applied for anti-doping controls of horse racing.