Integrated analytical approach in veal calvesadministered the anabolic androgenic steroidsboldenone and boldione: urine and plasma kineticprofile 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-QMS/
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 a- and b-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 mLC-ESI-IT-MS/MS procedures. A specific protein, poorly represented
in normal plasmasamples collected before treatment,was found upregulated even 36 h after
hormone treatment.Extensivemassmapping 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 byWestern blot analysis confirmed a significant and time dependent
increase of thisApoA1 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
