JZUS - Journal of Zhejiang University SCIENCE

Journal of Zhejiang University SCIENCE B 2011 Vol.12 No.6 P.460-467

Analysis of 19-nortestosterone residue in animal tissues by ion-trap gas chromatography-tandem mass spectrometry

Author(s): Jin-qing Jiang, Lei Zhang, Guang-ling Li, Hai-tang Zhang, Xue-feng Yang, Jun-wei Liu, Ren-feng Li, Zi-liang Wang, Jian-hua Wang
Affiliation(s): College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China, College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, China, College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, China
Corresponding email(s): wangzl_2008@yahoo.com.cn, jhwang1948@sina.com
Key Words: 19-Nortestosterone (19-NT), Gas chromatography-tandem mass spectrometry (GC-MS), Animal tissues

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Abstract: A rapid sample treatment procedure for the gas chromatography-tandem mass spectrometry (GC-MS) determination of 19-Nortestosterone (19-NT) in animal tissues has been developed. In our optimized procedures, enzymatic hydrolysis with β-glucuronidase from Escherichia coli was performed in an acetate buffer (pH 5.2, 0.2 mol/L). Next, the homogenate was mixed with methanol and heated at 60 °C for 15 min, then placed in an ice-bath at −18 °C for 2 h. After liquid-liquid extraction with n-hexane, the analytes were subjected to a normal-phase solid phase extraction (SPE) C₁₈ cartridge for clean-up. The dried organic extracts were derivatized with heptafluorobutyric anhydride (HFBA), and then the products were injected into GC-MS. Using electron impact mass spectrometry (EI-MS) with positive chemical ionization (PCI), four diagnostic ions (m/z 666, 453, 318, and 306) were determined. A standard calibration curve over the concentration range of 1–20 ng/g was reached, with Y=467084X−68354 (R²=0.9997) for 19-NT, and the detection limit was 0.3 ng. When applied to spiked samples collected from bovine and ovine, the recoveries ranged from 63% to 101% with relative standard deviation (RSD) between 2.7% and 8.9%. The procedure is a highly efficient, sensitive, and more economical method which offers considerable potential to resolve cases of suspected nandrolone doping in husbandry animals.

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