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.
Department of Food Safety and Veterinary Public Health, Istituto Superiore di Sanità, Rome, Italy.
Rosa
R
Draisci
Clara
C
Montesissa
Barbara
B
Santamaria
Chiara
C
D'Ambrosio
Giovanni
G
Ferretti
Roberta
R
Merlanti
Carolina
C
Ferranti
Marco
M
De Liguoro
Claudia
C
Cartoni
Erika
E
Pistarino
Lino
L
Ferrara
Micaela
M
Tiso
Andrea
A
Scaloni
M Elisabetta
ME
Cosulich
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