Analyzing the physiological signature of anabolic steroids in cattle urine using pyrolysis/metastable atom bombardment mass spectrometry and pattern recognition.

Abstract

Pyrolysis coupled to metastable atom bombardment (MAB) and time-of-flight mass spectrometry (TOFMS) is used for generating mass spectra from bovine urine samples obtained from cattle treated with anabolic steroids. These spectra constitute fingerprints, which can be discriminated by multivariate statistical analysis. Four main conclusions can be drawn from this work: (i) The use of different metastable gases, such as Xe*, Kr*, or N2*, as an energy-tunable ionization beamline allows control of the internal energy and the dissociation processes of the produced odd electron molecular ions, thus giving rise to complementary mass spectra fingerprints. (ii) A variable transformation depending on the biofluid matrix suitably contracts the frequency distribution of the generated data for low m/z ratios holding information related to endogenous metabolites encountered in urine. (iii) Coupling variable selection to statistical pattern recognition methods results in low error rates (< 1%) for predicting MAB mass fingerprints, especially using lineardiscriminant analysis (LDA). (iv) LDA discriminates controls from treated animals and also correlates to quantitative physiological responses induced by anabolic steroids. This work shows that Py-MAB-TOFMS could be a suitable method for complementary monitoring anabolic use in sports, medicine, and cattle breeding, as well as monitoring many other long-lasting although weak physiological disruptions.