title: Evaluation of boldenone formation and related steroids transformations in veal faeces by liquid chromatography/tandem mass spectrometry.
abstract: It is established that bovine urine can result positive for boldenone and androstadienedione in consequence of faecal contamination. The simple transfer of steroids to urine is one minor aspect of faecal contamination. A high de novo production of steroids in faeces after deposition and in faeces-contaminated urine is almost certainly due to microbial activity, although the precursor compounds and transformations leading to the presence of these illegal steroids are unclear. We developed a simple in vitro method - incubation of faecal matter suspended in 0.9% saline - to induce steroid transformations in faeces, and analyzed the products by liquid chromatography/tandem mass spectrometry, without the need for prior extraction. Norethandrolone was the internal standard. The linearity (R(2): 0.987-0.999), sensitivity (LODs: 0.3 to 1.0 ng/mL; LOQs: 1.0 to 3.0 ng/mL), precision (intra-day CVs: 2.6-8.2; inter-day CVs: 4.5-11.5) and accuracy (percentage recovery: 89-120%) were calculated for the studied steroids. Androstenedione, androstadienedione, alpha- and beta-boldenone, testosterone and epitestosterone transformations were investigated. Mutual interconversion of steroids was observed, although 17beta-hydroxy steroids had low stability compared with 17alpha-hydroxy and 17-keto steroids. The results suggest that this simple in vitro system may be an effective way of studying hormone transformations in faeces and, after analogue studies, in faeces-contaminated urine.
authors: Department of Veterinary Sciences and Technologies for Food Safety, University of Milan, Via Celoria 10, Milan, Italy. francesco.arioli@unimi.it
authors: Francesco
authors: F
authors: Arioli
authors: Matteo P
authors: MP
authors: Gavinelli
authors: Maria L
authors: ML
authors: Fracchiolla
authors: Alessio
authors: A
authors: Casati
authors: Marco
authors: M
authors: Fidani
authors: Emilia
authors: E
authors: Ferrer
authors: Giuseppe
authors: G
authors: Pompa
mesh_terms: D045930:Anabolic Agents / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001711:Biotransformation; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D005243:Feces / Q000737:chemistry; D066298:In Vitro Techniques; D008297:Male; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D015813:Substance Abuse Detection / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013739:Testosterone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism
chemical_list: D045930:Anabolic Agents; D013739:Testosterone; C013915:boldenone