Screening and confirmatory GC-MS methods for the detection of trenbolone in bovine urine

• Autorzy: Wozniak B. , Matraszek-Zuchowska I. , Semeniuk S. , Klopot A. , Zmudzki J.
• Języki publikacji: EN

Abstrakty

EN

Sensitive and selective methods for the screening (GC-MS) and confirmatory analysis (GC-MS/MS) of 17α- and 17ß- trenbolone in bovine urine were developed. In the first stage of the analysis, the enzymatic hydrolysis of trenbolone metabolites with glucuronidase AS-HP in acetate buffer (pH 5.2) solution was carried out. Free compounds were extracted from urine with diethyl ether. For the purification of the extract solid phase, extraction with C18 and NH2 columns was applied. The evaporated extract was subjected to two derivatisation steps; the first with MSTFA/12 solution and second with MSTFA. The separation of the analytes on HP-5 ms capillary column was conducted, llie methods were validated according to the Commission Decision 2002/657/EC. For GC-MS method, CCα and CCß were 0.21-0.36 µg L⁻¹ for 17α- trenbolone and 0.20-0.34 µg L⁻¹ for 17β- trenbolone, while for GC-MS/MS method the values were lower and amounted to 0.15-0.25 µg L⁻¹ for 17α- trenbolone and 0.20-0.34 µg L⁻¹ for 17ß-trenbolone. Method recoveries in spiked samples ranged from 86%-111% with standard deviation lower than 25% for both detection techniques.

• Wydawca: -
• Czasopismo: Bulletin of the Veterinary Institute in Puławy
• Rocznik: 2013
• Tom: 57
• Numer: 4
• Opis fizyczny: p.559-566,fig.,ref.

Twórcy

autor

Wozniak B.

• Department of Pharmacology and Toxicology National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Matraszek-Zuchowska I.

• Department of Pharmacology and Toxicology National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Semeniuk S.

• Department of Pharmacology and Toxicology National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Klopot A.

• Department of Pharmacology and Toxicology National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Zmudzki J.

• Department of Pharmacology and Toxicology National Veterinary Research Institute, 24-100 Pulawy, Poland

Bibliografia

• 1. ARO SOP 475B. Method validation using ResVal®. Validation according to EC/2002/657. National Institute of Public Health and Environment, the Netherlands 2004. 1-17.
• 2. Blackwell B.R., Cai Q., Smith P.N., Cobb G.P.: Liquid chromatography-tandem mass spectrometry analysis of 17α-trenbolone. 17ß-trenbolone and trendione in airborne particulate matter. Talanta 2011, 85, 1317-1323.
• 3. Blokland M.H., Zoontjes P.W., Sterk S.S., Stephany R.W., Zweigenbaum J., Van Ginkel L.A.: Confirmatory analysis of trenbolone using accurate mass measurement with LC/TOF-MS. Anal Chim Acta 2008, 618, 86-93.
• 4. Council Directive 96/22/EC, Off J Eur Commun 1996, L 125, 3.
• 5. CRL Guidance Paper (7 December 2007). http://www.rivm.nl/ bibliotheek/digitaaldepot/crlguidance2007.pdf.
• 6. European Community Commission Decision 2002/657/EC, Off J Eur Commun 2002, L221, 8.
• 7. European Community. Council Directive 96/23/EC. Off J Eur Commun 1996, L125, 10.
• 8. Fedeniuk R.W., Boison J.O., MacNeil J.D.: Validation of a gas chromatography-mass spectrometry method for the determination of pg/ml levels of 17ß-estradiol and 17ß-trenbolone in bovine serum. J Chromatogr B 2004, 802, 307-315.
• 9. Gasparini M., Curatolo M., Assini W., Bozzoni E., Tognoli N., Dusi G.: Confirmatory method for the determination of nandrolone and trenbolone in urine samples using immunoaffinity cleanup and liquid chromatography-tandem mass spectrometry. J Chromatogr A 2009. 1216, 8059-8066.
• 10. Hansen M., Jacobsen N.W., Nielsen F.K., Björklund E., Styrishave B., Halling-Sørensen B.: Determination of steroid hormones in blood by GC-MS/MS. Anal Bioanal Chem 2011, 400, 3409-3417.
• 11. Impens S., De Wasch K., Cornelis M., De Brabander H.F.: Analysis on residues of estrogens, gestagens and androgens in kidney fat and meat with gas chromatography - tandem mass spectrometry. J Chromatogr A 2002, 955, 281-293.
• 12. Impens S., Van Loco J., Degroodt J.M., De Brabander H.: A downscaled multi residue strategy for detection of anabolic steroids in bovine urine using gas chromatography tandem mass spectrometry (GC- MS3). Anal Chim Acta 2007, 586, 43-48.
• 13. Jouquey A., Mouren M., Salmon J.: Analytical methods for trenbolone. In: Anabolics in Animal Production, edited by E. Meissonnier, Office International des Epizooties, Paris, 1983. pp. 423-440.
• 14. Kootstra P.R., Zoontjes P.W., Van Tricht E.F., Sterk S.S.: Multiresidue screening of a minimum package of anabolic steroids in package urine with GC-MS. Anal Chim Acta 2007, 586, 82-92.
• 15. Marchand P., Le Bizec B., Gade C., Monteau F., André F.: Ultra trace detection of a wide range of anabolic steroids in meal by gas chromatography coupled to mass spectrometry. J Chromatogr A, 2000, 867, 219-233.
• 16. Matuszewski B.K., Constanzer M.L., Chavez-Eng C.M.: Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS. Anal Chem 2003, 75, 3019-3030.
• 17. Maume D., Le Bizec B., Marchand P., Montrade M.P., André F.: N-Methyl-N-alkylsilyltrifluoroacetamide-I-2 as a new derivati- zation reagent for anabolic steroid control. Analyst 1998, 123, 2645-2648.
• 18. Noppe H., Le Bizec B., Verheyden K., De Brabander H.F.: Novel analytical methods for the determination of steroid hormones in edible matrices. Anal Chim Acta 2008, 611, 1-16.
• 19. Opinion of the Scientific Committee on Veterinary Measures Relating to Public Health: Assessment of potential risk to human health from hormone residues in bovine meat and meat products. 30 April 1999. http://europa.eu.int/genino/query_en.htm.
• 20. Taylor P.J.: Matrix effects: the Achilles heel of quantitative high -performance liquid chromatography-electrospray-tandem mass spectrometry. Clin Biochem 2005, 38, 328-334.
• 21. Van Poucke C., Van Peteghem C.: Development and validation of a multi-analyte method for the detection of anabolic steroids in bovine urine with liquid chromatography-tandem mass spectrometry. J Chromatogr B 2002, 772, 211-217.
• 22. Woźniak B., Matraszek-Żuchowska I., Żmudzki J.: Determination of 17ß-oestradiol and testosterone in bovine serum with gas chromatography-mass spectrometry. Bull Vet Inst Pulawy 2011, 55, 755-759.

Identyfikatory

DOI

10.2478/bvip-2013-0096