Integrated analytical approach in veal calves administered the anabolic androgenic steroids boldenone and boldione: urine and plasma kinetic profile and changes in plasma protein expression.
Surveillance of illegal use of steroids hormones in cattle breeding is a key issue to preserve human health. To this purpose, an integrated approach has been developed for the analysis of plasma and urine from calves treated orally with a single dose of a combination of the androgenic steroids boldenone and boldione. A quantitative estimation of steroid hormones was obtained by LC-APCI-Q-MS/MS analysis of plasma and urine samples obtained at various times up to 36 and 24 h after treatment, respectively. These experiments demonstrated that boldione was never found, while boldenone alpha- and beta-epimers were detected in plasma and urine only within 2 and 24 h after drug administration, respectively. Parallel proteomic analysis of plasma samples was obtained by combined 2-DE, MALDI-TOF-MS and muLC-ESI-IT-MS/MS procedures. A specific protein, poorly represented in normal plasma samples collected before treatment, was found upregulated even 36 h after hormone treatment. Extensive mass mapping experiments proved this component as an N-terminal truncated form of apolipoprotein A1 (ApoA1), a protein involved in cholesterol transport. The expression profile of ApoA1 analysed by Western blot analysis confirmed a significant and time dependent increase of this ApoA1 fragment. Then, provided that further experiments performed with a growth-promoting schedule will confirm these preliminary findings, truncated ApoA1 may be proposed as a candidate biomarker for steroid boldenone and possibly other anabolic androgens misuse in cattle veal calves, when no traces of hormones are detectable in plasma or urine.
Draisci
Rosa
R
Department of Food Safety and Veterinary Public Health, Istituto Superiore di Sanità, Rome, Italy.
Montesissa
Clara
C
Santamaria
Barbara
B
D'Ambrosio
Chiara
C
Ferretti
Giovanni
G
Merlanti
Roberta
R
Ferranti
Carolina
C
De Liguoro
Marco
M
Cartoni
Claudia
C
Pistarino
Erika
E
Ferrara
Lino
L
Tiso
Micaela
M
Scaloni
Andrea
A
Cosulich
M Elisabetta
ME
D000284:Administration, Oral; D045930:Anabolic Agents / Q000008:administration & dosage / Q000097:blood* / Q000652:urine*; D000730:Androstadienes / Q000008:administration & dosage / Q000097:blood* / Q000652:urine*; D000818:Animals; D001798:Blood Proteins / Q000032:analysis* / Q000378:metabolism; D002417:Cattle; D002853:Chromatography, Liquid; D003198:Computer Simulation; D015180:Electrophoresis, Gel, Two-Dimensional; D007150:Immunohistochemistry; D007700:Kinetics; D010449:Peptide Mapping; D040901:Proteomics / Q000379:methods; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D013739:Testosterone / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000097:blood / Q000652:urine; D013997:Time Factors
D045930:Anabolic Agents; D000730:Androstadienes; D001798:Blood Proteins; C006573:1,4-androstadiene-3,17-dione; D013739:Testosterone; C013915:boldenone