Endogenous steroid profiling by gas chromatography-tandem mass spectrometry and multivariate statistics for the detection of natural hormone abuse in cattle

Abstract

steroid profiling by gas-chromatography tandem mass spectrometry and multivariate statistics for detection of natural hormone abuse in cattle. Food Additives and Contaminants, 2012, pp.1. <10.1080/19440049.2012 For years it is suspected that natural hormones are illegally used as growth promoters in cattle in the European Union. Unfortunately there is a lack of methods and criteria that can be used to detect the abuse of natural hormones and distinguish treated from non-treated animals. Pattern recognition of steroid profiles is a promising approach for tracing/detecting the abuse of natural hormones administered to cattle. Traditionally steroids are analyzed in urine as free steroid after deconjugation of the glucuronide (and sulphate) conjugates. Disadvantage of this deconjugation is that valuable information about the steroid profile in the sample is lost. In this study we developed a method to analyze steroids at very low concentration levels (ng.L-1) for the free steroid, glucuronide and sulphate conjugates in urine samples. This method was used to determine concentrations of natural (pro)hormones in a large population (n=620) of samples from male and female bovine animals and from bovine animals treated with testosterone-cypionate, estradiol-benzoate, dihydroepiandrosterone and pregnenolone.

Urinary Concentrations of Steroids in Bulls under Anabolic Treatment by Revalor-XS® Implant

“…Beside the residue analyses, new approaches have been proposed in recent years to prove illicit treatment with growth promoters [26]: some focused on targeted metabolite analyses [17, 18, 27] and others on untargeted proteomics [28–30] or metabolomics approaches [31–33]. Despite all being promising, none of them is definitive to establish a unique and robust set of biomarkers of treatment independently from drug administration, breed, gender, age, and physiological condition.…”

“…Consequently, several research studies have been implemented through the years to test the effect of different growth promoters administered via different routes [18, 28, 34, 35] and to evaluate residual levels and target metabolites. In this work a commercially available ear implant containing trenbolone acetate and estradiol (Revalor-XS) was tested; this formulation is used in the United States in bovine breeding and is able to release a constant amount of hormones over a time period up to 200 days eliminating the need to reimplant the device.…”

Structure elucidation and HPLC-MS/MS determination of a potential biomarker for estradiol administration in cattle

“…In recent years, new methodological approaches based on untargeted and global measurement are emerging [14]. Pattern recognition of steroid profiles has risen also as a promising approach for tracing/detecting the abuse of natural hormones and their esters administered to cattle [15][16][17]. Other approaches also include protein biomarker profiles [18] or blood chemistry [19].…”

High-resolution time-of-flight mass spectrometry fingerprinting of metabolites from cecum and distal colon contents of rats fed resistant starch

“…Tools such as principal component analysis, soft independent modeling of class analogy, partial least-squares discriminant analysis (PLS-DA), linear discriminant analysis, and artificial neural networks all allow differentiation and group prediction [26,27,34,35,39,[44][45][46]. Using these tools, difficult and complex analyses of large amounts of data can be more easily graphed and visualized.…”

“…GC-MS often utilizes electron impact (EI) ionization. The GC separation and subsequent EI fragmentation often allow metabolite identification through database searching [27,[33][34][35]. Liquid chromatography-mass spectrometry (LC-MS) is very useful for metabolite analysis because specialized columns can be used to separate either polar or non-polar compounds [25,26,36].…”