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

作　　者：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 

机构地区：[1]College of Veterinary Medicine, Northwest A &F University, Yangling 712100, China [2]College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, China [3]College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, China

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2011年第6期460-467,共8页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：Project supported by the Eleventh Five-Year Plan for National Science and Technology of China (No.2006BAK02A21/1);the Henan Innovation Project for University Prominent Research Talents (No.2010HASTIT026),China

摘　　要：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 coliwas performed in an acetate buffer （pH 5.2, 0.2 mol/L） Next, the homogenate was mixed with methanol and heated at 60℃ for 15 min, then placed in an ice-bath at -18℃ for 2 h. After liquid-liquid extraction with n-hexane, the analytes were subjected to a normal-phase solid phase ex- traction （SPE） C18 cartridge for clean-up. The dded organic extracts were derivatized with heptafluorobutydc anhydride （HFBA）, and then the products were injected into GC-MS. Using electron impact mass spectrometry （El-MS） with positive chemical ionization （PCI）, four diagnostic ions （mlz 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 （R^2=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.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 ℃ for 15 min,then placed in an ice-bath at 18 ℃ 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.

关 键 词：19-Nortestosterone （19-NT） Gas chromatography-tandem mass spectrometry （GC-MS） Animal tissues 

分 类 号：S859[农业科学—临床兽医学]