Determination of anabolic steroids in bovine serum by liquid chromatography–tandem mass spectrometry

Abstract

In the present paper we report the LC-MS/MS determination of residues of 12 anabolic steroids in bovine serum, as an expansion of our work protocols for steroids determination in biological matrices. Steroids analyzed included α-zearalanol, β-zearalanol, α-trenbolone, β-trenbolone, methyltestosterone, α-estradiol, β-estradiol, ethynylestradiol, α-boldenone, β-boldenone, α-nortestosterone and β-nortestosterone. Following protein precipitation, serum samples were cleaned up by solid-phase extraction using Oasis HLB and Amino cartridges. Atmospheric pressure chemical ionization (APCI) in both positive and negative ionization modes was used and mass spectrometry detection was carried out in multiple reaction monitoring mode following two or (in most cases) three product ions per precursor ion. The method was validated in accordance with the Commission Decision 2002/657/EC. The decision limit (CCα) values obtained, ranged from 0.01 to 0.07 ng/ml and the detection capability (CCβ) values obtained ranged from 0.02 to 0.12 ng/ml. The recoveries ranged from 70.2% to 118.2%. The developed method is suitable for routine and confirmatory purposes such as control of illegal use in livestock production.

Medroxyprogesterone Acetate and Progesterone Measurement in Human Serum: Assessments of Contraceptive Efficacy

“…There is variety among the LC-MS/MS methods including different ionization techniques (electrospray (ESI) [2,22], atmospheric chemical ionization (APCI) [4,5,23] and detectors (ion trap, triple quadrapole) [2,4,21]. Furthermore these techniques have been optimized for a variety of sample types which include animal fat, muscle, blood, and plasma.…”

“…As such, it provides a significant risk for the population consuming the treated meats [1]. Over the years, multiple methods and techniques have been developed for measuring the levels of hormones such as MPA in animal fat, tissues, plasma, and urine [2][3][4][5]. The older techniques included enzyme immune assay [6] and gas chromatography-mass spectrometry [7] but more recently liquid chromatograph-mass spectrometry (LC-MS/MS) techniques have been developed.…”

Natural Hormones in Food-Producing Animals:Legal Measurements and Analytical Implications

“…Kaklamanos et al, 2009a;Kaklamanos et al, 2009b;Kaklamanos et al, 2011;Shao et al, 2005), all validated according to the criteria set out in Decision 2002/657/CE for banned substances. Although normally the levels of steroids that accumulate in animal tissues are lower than in other matrices, many effective methods are known currently for the determination of these compounds in muscle tissue, sometimes monitoring a wide range of anabolic compounds (Courant et al, 2008;Kaklamanos et al, 2007;Vanhaecke et al, 2011;Yang et al, 2009).…”

“…With regard to the confirmation of use of xenobiotic analogues of natural sex steroids and non-steroidal compounds, such as stilbenes and zeranol, there is an extensive range of successful methods that has been performed on different analytic matrices. These analytical procedures have made the confirmation of illicit administrations of anabolics in cattle feasible (De Brabander et al, 2007;Duffy et al, 2009;Kaklamanos et al, 2011;Noppe et al, 2008).…”

Quantification of Glucuronide Metabolites in Biological Matrices by LC-MS/MS

“…Narrow bore UHPLC capillaries and columns are especially sensitive to particulate impurities because of their finer pore diameters and dense inlet filters. Therefore, at least some sort of sample pre-treatment is necessary, like diluting the samples 1:10 with water in combination with filtering through 0.2 µm pore filters or centrifugation (McMaster 2007;Kaklamanos, Theodoridis et al 2011). Such an approach without any form of extraction greatly reduces the time needed for an analysis; however the method and system robustness parameters should be carefully monitored.…”