Can Serum be a Match for Urine in the Regulatory Analysis of Boldenone in Cattle? A Systematic Comparison Between Detection Window, Stability, and Enzymatic Hydrolysis

Abstract

This work involved a systematic comparison between serum and urine for the monitoring of anabolic androgenic steroids in livestock. Incurred samples were collected over 120 days from crossbred steers treated with intramuscular injections containing boldenone undecylenate. Independent high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/ MS) methods were used for the assessment of the respective detection windows, which were larger for serum samples. Both matrices presented adequate performance in terms of long-term stability, assessed using an isochronous approach during 196 days at −20 °C and for five freeze−thaw cycles. The effectiveness of the enzymatic hydrolysis reaction using Helix pomatia juice was also compared. The calculated concentrations in serum samples were not statistically influenced by the deconjugation reaction. On the other hand, urine hydrolysis conditions were studied using a 3 3 Box−Behnken Design, in which a central point condition led to a satisfactory deconjugation performance. It could be observed that serum exhibited equivalent or better performance than urine for most of the evaluated criteria; thus, its inclusion in the regulatory analysis of boldenone in cattle is supported.

Multiresidue Determination of Growth Promoters in Bovine Serum using QuEChERS and UHPLC-Q-ORBITRAP: Validation and In Vivo Study

“…Thus, in both stability studies, the STN residues were stable under all of the evaluated conditions, which reinforces that the serum matrix is sufficiently stable to be used as a matrix of choice for detecting STN treatment in steers when it comes to stability. 38,42,43 The obtained concentration values for MS A , MS E , and F calc from both stability studies are presented in Table 3.…”

Determination of 18 β-Agonists in Aquatic Products Using High-Performance Liquid Chromatography Tandem Mass Spectrometry

“…The hydrolysis of β-agonist-conjugated ester residues in the muscles is necessary before extraction. 41 Their conjugates mostly exist in the form of glucuronide or sulfate. Ractopamine, tulobuterol, and salbutamol are selected to prepare the incurred samples because they can frequently form conjugate states in the organism.…”

“…The hydrolysis of β-agonist-conjugated ester residues in the muscles is necessary before extraction . Their conjugates mostly exist in the form of glucuronide or sulfate.…”

In Vivo Administration of Stanozolol in Cattle: Depletion and Stability Studies Using UHPLC-Q-Orbitrap

“…14 Stability studies concerning anabolic steroids were reported by Rocha et al, in which bovine urine and serum presented stability during 196 days at −20 °C. 19 De Wilde et al evaluated human urine for the stability study of steroid conjugates, in which a stable result was obtained after six freeze−thaw cycles and after 10 years of storage at −20 °C. 20 De Clercq et al studied bovine feces stability for six glucocorticoids; the work explored redox conditions, influence of ethanol addition, and lyophilization and also included freeze−thaw and long-term studies.…”

“…Stability studies concerning anabolic steroids were reported by Rocha et al, in which bovine urine and serum presented stability during 196 days at −20 °C . De Wilde et al evaluated human urine for the stability study of steroid conjugates, in which a stable result was obtained after six freeze–thaw cycles and after 10 years of storage at −20 °C .…”