Rapid test by liquid chromatography/tandem mass spectrometry to evaluate equine urine reactivity towards 17beta-OH steroids.

The research article presents a rapid testing method to assess bacterial contamination in horse urine samples, which can affect the integrity of 17beta-OH steroid assessments. It also provides recommendations for optimal storage of these samples.

Research Methodology

• The research was undertaken to develop a liquid chromatography/tandem mass spectrometry (LC/MS/MS) method to evaluate bacterial contamination in equine urine, which is a marker of poor storage conditions.
• The researchers utilized Norethandrolone as the internal standard for their experiment, and evaluated the linearity, sensitivity, precision, and accuracy of the developed method.
• They explored the oxidation of 17beta-OH for testosterone, nandrolone, trenbolone, and boldenone, in the presence of bacterial growth.
• The study also examined possible reactions with alpha-epimers like alpha-boldenone and epitestosterone to substantiate the stereoselectivity of the reaction.

Test Process

• A speedy test was executed by spiking one of the four 17beta-OH steroids into diluted equine urine samples. In the context of this study, spiking refers to the deliberate addition of known quantities of a steroid into a urine sample for testing purposes.
• In the presence of bacterial activity, these spiked steroids were transformed into their respective ketone forms.
• The method allowed for direct injection of these spiked and diluted samples into the LC/MS system, thereby eliminating the need for prior extraction.

Findings and Recommendations

• The researchers discovered that freezing the samples at -18 degrees Celsius provided the optimal storage condition.
• They further recommended that urine samples be analyzed as quickly as possible after being thawed.
• This swift analysis would momentarily halt any possible bacterial degradation of the equine urine sample, making it suitable for qualitative or quantitative analysis of 17beta-OH steroids.