title: Development of a rapid method for the analysis of trenbolone, nortestosterone, and zeranol in bovine liver using liquid chromatography tandem mass spectrometry.
abstract: A rapid liquid chromatography tandem mass spectrometry method has been developed and validated for the determination of α-trenbolone, β-trenbolone, α-nortestosterone, β-nortestosterone, zeranol, and taleranol in bovine liver. The impact of liquid-liquid extraction with methyl tert-butyl ether and optimized solid phase extraction on silica cartridges significantly reduced effort and time of sample preparation. Electrospray ionization gives a significant signal increase compared with atmospheric pressure chemical ionization and atmospheric pressure photoionization. The HPLC gradient was optimized to separate isobaric analytes and matrix constituents from the hormone molecules. The optimized time and temperature of enzymatic hydrolysis of conjugated trenbolone was 4 h at 52 °C. The method validated in the range of 0.5-30 μg kg(-1) for α-trenbolone, β-trenbolone, zeranol, taleranol, and 2-30 μg kg(-1) for α-nortestosterone, β-nortestosterone. Combined uncertainty of measurements was in the range of 4%-23%. The matrix effect was negligible (1%-5%) for all analytes except of α-nortestosterone (19%). The developed method with changes concerning sample size and hydrolysis was also applied for the analysis of meat, serum, and urine samples. Graphical Abstract Determination of trenbolone, nortestosterone and zeranol in bovine liver.
authors.affiliation: Department of Food and Feed Safety, Laboratory of Food Analysis, The All-Russian State Centre for Quality and Standardization of Veterinary Drugs and Feed (VGNKI), 123022, Moscow, Russia, yuninma@gmail.com.
authors.forename: Maxim A
authors.initials: MA
authors.lastname: Yunin
authors.forename: Pavel S
authors.initials: PS
authors.lastname: Metalnikov
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Komarov
authors.forename: Alexander N
authors.initials: AN
authors.lastname: Panin
mesh_terms: D045930:Anabolic Agents / Q000032:analysis*; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004968:Estrogens, Non-Steroidal / Q000032:analysis*; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction / Q000379:methods; D008099:Liver / Q000737:chemistry*; D008738:Methyl Ethers; D009277:Nandrolone / Q000032:analysis*; D053719:Tandem Mass Spectrometry / Q000379:methods; D014204:Trenbolone Acetate / Q000032:analysis*; D015025:Zearalenone / Q000032:analysis; D015029:Zeranol / Q000032:analysis*
chemical_list: D045930:Anabolic Agents; D004968:Estrogens, Non-Steroidal; D008738:Methyl Ethers; C043243:methyl tert-butyl ether; D015025:Zearalenone; D009277:Nandrolone; D015029:Zeranol; C028226:taleranol; D014204:Trenbolone Acetate