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

The study presents a novel procedure for robust detection and quantification of nine anabolic steroids in horse urine, a pivotal advancement in doping control. Utilizing ultra-high-performance liquid chromatography with tandem mass spectrometry, the method shows a high recovery rate and a low coefficient of variation, indicating its efficiency and reliability.

Development of New Procedure

• The researchers set out to develop a new, quick and simple analytical protocol for the simultaneous detection and quantification of nine anabolic steroids: deslorelin, dexamethasone sodium phosphate, prednisolone, methylprednisolone, stanozolol, boldenone, nandrolone, dexamethasone isonicotinate and altrenogest. These substances are often misused in horse doping to enhance performance.
• The detection method deployed the use of ultra-high-performance liquid chromatography coupled with tandem mass spectrometry technique (UHPLC-MS/MS), a well-recognized and highly efficient tool in chemical analysis.

Implementation of the Procedure

• A total amount of 400 μl of sample was evaporated, restored and then introduced to the UHPLC-MS/MS for analysis.
• No specific details were given by the abstract regarding specific experimental conditions or the exact procedure to evaporate and restore the samples. However, this groundwork preparation is key in ensuring the integrity and representativeness of the sample being analyzed.

Results and Validation

• The proposed method was thoroughly validated, showing impressive and consistent outcomes. Specifically, it exhibited a recovery rate (a measure of how much of the initially injected sample was retained in the process) which was higher than 89.12%, indicating a high efficiency of the analytical process.
• The procedure also demonstrated a coefficient of variation lower than 6.02%. The coefficient of variation is a measure of the relative variability of the data, thus a lower coefficient signifies a reliable and consistent method.
• The range of correlation coefficients of the examined compound’s calibration curves was 0.9955-0.9997, indicating an exceptionally high degree of reliability in the analytical method.
• The detection and quantification limits of the method were within the range of 0.1 and 0.25 μg/l respectively, demonstrating ability to trace even trace amounts of the substances.

Conclusions

• This new procedure offers a powerful tool for doping control in horses, with the potential to significantly improve accuracy and reliability in anabolic steroid detection and quantification.