Determination of Cortisol, Cortisone, Prednisolone and Prednisone in Bovine Urine by Liquid Chromatography–electrospray Ionisation Single Quadrupole Mass Spectrometry

Abstract

A quantitative LC-ESI single quadrupole MS method for the determination of cortisol (F), cortisone (E), prednisolone (PL), and prednisone (PN) in bovine urine has been developed and validated. After addingflumethasone as internal standard, the samples were subjected tofiltration, deconjugation, and solid-phase extraction, while the chromatographic separation was achieved using a Restek Ultra II Allure Biphenyl column with isocratic mobile phase. The analytes were detected after negative electrospray ionization using SIM mode. In order to obtain spectra with maximum intensities of at least one of the three characteristic ions, (Mþformate) , (M-H), and [(M-H) -CH 2 0]~, an individual optimization of MS parameters for each corticosteroid was set up. MS data was acquired in the three-ion selected monitoring mode and the ion ratios between chosen diagnostic ions were used in order to increase the specificity. Calibration graphs were linear and the intra-day and intermediate precision was estimated as RSD values which were less than 17%. For F and E, obtained values indicated negligible absolute matrix effects (10 3 % and 98%, respectively). The method was applied to real samples, and basal levels ofFandE were preliminarily evaluated, while PL and PN were not detected.

Tetrahydro-metabolites of cortisol and cortisone in bovine urine evaluated by HPLC–ESI-mass spectrometry

“…Literature data on natural corticosteroid levels are available almost exclusively for human urine [1] or plasma [19], while there is little information on bovine urine. We have recently published a method for measuring F and E in untreated cow and bovine urine [20], but no studies regarding the amount of F and E tetrahydro-metabolites or their metabolism in cattle are available. This work, although it is only a preliminary study, may provide a useful methodological contribution to the future definition of cortisol, cortisone and their tetrahydro-metabolite urinary baseline values in order to obtain indirect evaluations of the HSDs activity.…”

Detection of selected corticosteroids and anabolic steroids in calf milk replacers by liquid chromatography–electrospray ionisation – Tandem mass spectrometry

“…The use of steroids in food-producing animals for therapeutic purposes is regulated (corticosteroids) or banned (anabolic ste-roids) in the European Union; however, their use as growth pro-moters cannot be excluded (Pavlovic et al, 2012). Cortisol, cortisone, testosterone and epitestosterone are endogenous, pred-nisolone (Bertocchi et al, 2013), boldenone (Chiesa et al, 2014) and nandrolone (Glenn Kennedy et al, 2009) are considered pseu-doendogenous steroids while dexamethasone and trenbolone are well-known synthetic steroids.…”

“…There have been just a few studies in the literature on powdered milk e infant formulae for human use (Romero-González, Aguilera-Luiz, Plaza-Bolan ~os, Frenich, &Vidal, 2011 andZhan et al, 2013) and only one that described a multi-residue method for detecting 17 selected veterinary hormones in six different powdered in-gredients derived from bovine milk used modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) sample preparation (Ehling & Reddy, 2013). Other researchers devised a method for the detection of eight corticosteroids in milk replacers, through C18 SPE, but with relatively high detection limits (Fiori, Pierdominici, Longo, & Brambilla, 1998).…”

Study on cortisol, cortisone and prednisolone presence in urine of Chianina cattle breed

“…The inhibitory effects of PL and dexamethasone could eventually be evaluated by analysis of the ratio of CL to CN. Therefore, the evaluation of natural corticosteroids may be considered a tool for defining a condition regarding the abuse of therapeutic agents (Pavlovic et al., ).…”