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

http://doi.org/10.1631/jzus.B1000301


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


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. Analysis of 19-nortestosterone residue in animal tissues by ion-trap gas chromatography-tandem mass spectrometry[J]. Journal of Zhejiang University Science B, 2011, 12(6): 460-467.

@article{title="Analysis of 19-nortestosterone residue in animal tissues by ion-trap gas chromatography-tandem mass spectrometry",
author="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",
journal="Journal of Zhejiang University Science B",
volume="12",
number="6",
pages="460-467",
year="2011",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1000301"
}

%0 Journal Article
%T Analysis of 19-nortestosterone residue in animal tissues by ion-trap gas chromatography-tandem mass spectrometry
%A Jin-qing Jiang
%A Lei Zhang
%A Guang-ling Li
%A Hai-tang Zhang
%A Xue-feng Yang
%A Jun-wei Liu
%A Ren-feng Li
%A Zi-liang Wang
%A Jian-hua Wang
%J Journal of Zhejiang University SCIENCE B
%V 12
%N 6
%P 460-467
%@ 1673-1581
%D 2011
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1000301

TY - JOUR
T1 - Analysis of 19-nortestosterone residue in animal tissues by ion-trap gas chromatography-tandem mass spectrometry
A1 - Jin-qing Jiang
A1 - Lei Zhang
A1 - Guang-ling Li
A1 - Hai-tang Zhang
A1 - Xue-feng Yang
A1 - Jun-wei Liu
A1 - Ren-feng Li
A1 - Zi-liang Wang
A1 - Jian-hua Wang
J0 - Journal of Zhejiang University Science B
VL - 12
IS - 6
SP - 460
EP - 467
%@ 1673-1581
Y1 - 2011
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1000301


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.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1]Bagatell, C.J., Bremner, W.J., 1996. Androgens in men-uses and abuses. N. Engl. J. Med., 334(11):707-714.

[2]Bagnati, R., Fanelli, R., 1991. Determination of 19-nortestosterone, testosterone and trenbolone by gas chromatography-negative-ion mass spectrometry after formation of the pentafluorobenzyl carboxymethoxime-trimethylsilyl derivatives. J. Chromatogr. A, 547(1-2):325-334.

[3]Bahrke, M.S., Yesalis, C.E., 2004. Abuse of anabolic androgenic steroids and related substances in sport and exercise. Curr. Opin. Pharmacol., 4(6):614-620.

[4]Barrón, D., Barbosa, J., Pascual, J.A., Segura, J., 1996. Direct determination of anabolic steroids in human urine by on-line solid-phase extraction/liquid chromatography/ mass spectrometry. J. Mass Spectrom., 31(3):309-319.

[5]Brännvall, K., Bogdanovic, N., Korhonen, L., Lindholm, D., 2005. 19-Nortestosterone influences neural stem cell proliferation and neurogenesis in the rat brain. Eur. J. Neurosci., 21(4):871-878.

[6]Bricout, V., Wright, F., 2004. Update on nandrolone and norsteroids: how endogenous or xenobiotic are these substances? Eur. J. Appl. Physiol., 92(1-2):1-12.

[7]Chen, Y., Liu, N., Liu, X.X., Li, K., 2004. Effects of nandrolone phenylpropionate on fibroblasts after injury in rats. J. Sichuan Univ. (Med. Sci. Ed.), 35(4):508-511 (in Chinese).

[8]Choi, M.H., Chung, B.C., Lee, W., Lee, U.C., Kim, Y., 1999. Determination of anabolic steroids by gas chromatography/ negative-ion chemical ionization mass spectrometry and gas chromatography/negative-ion chemical ionization tandem mass spectrometry with heptafluorobutyric anhydride derivatization. Rapid Commun. Mass Spectrom., 13(5):376-380.

[9]Conneely, G., Aherne, M., Lu, H., Guilbault, G.G., 2007. Electrochemical immunosensors for the detection of 19-nortestosterone and methyltestosterone in bovine urine. Sens. Actuators B, 121(1):103-112.

[10]Daeseleire, E., Vandeputte, R., Peteghem, C.V., 1998. Validation of multi-residue methods for the detection of anabolic steroids by GC-MS in muscle tissues and urine samples from cattle. Analyst, 123(12):2595-2598.

[11]de Boer, D., Bensink, S.N., Borggreve, A.R., van Ooijen, R.D., Maes, R.A.A., 1995. Profiling 19-norsteroids. I-tandem mass spectrometric characterization of the heptafluorobutyryl ester and pentadnorobenzyloxime heptafluorobutyryl ester derivatives of 19-nortestosterone using collisionally activated dissociation. J. Mass Spectrom., 30(3):497-504.

[12]Delbeke, F.T., Desmet, N., Debackere, M., 1995. The abuse of doping agents in competing body builders in Flanders (1988–1993). Int. J. Sports Med., 16(1):66-70.

[13]Draisci, R., Palleschi, L., Ferretti, E., Lucentini, L., Delli Quadri, F., 2003. Confirmatory analysis of 17β-boldenone, 17α-boldenone and androsta-1,4-diene-3,17-dione in bovine urine by liquid chromatography-tandem mass spectrometry. J. Chromatogr. B, 789(2):219-226.

[14]Dubois, M., Taillieu, X., Colemonts, Y., Lansival, B., de Graeve, J., Delahaut, P., 1998. GC-MS determination of anabolic steroids after multi-immunoaffinity purification. Analyst, 123(12):2611-2616.

[15]Gasparini, M., Assini, W., Bozzoni, E., Tognoli, N., Dusi, G., 2007. Development and validation of a liquid chromatography-tandem mass spectrometry method for the separation of conjugated and unconjugated 17α- and 17β-boldenone in urine sample. Anal. Chim. Acta, 586(1-2):154-162.

[16]Hartmann, S., Steinhart, H., 1997. Simultaneous determination of anabolic and catabolic steroid hormones in meat by gas chromatography-mass spectrometry. J. Chromatogr. B, 704(1-2):105-117.

[17]Heitzman, R.J., 1976. The effectiveness of anabolic agents in increasing rate of growth in farm animals: report on experiments in cattle. Environ. Qual. Saf. Suppl., 5:89-98.

[18]Houghton, E., Oxley, G.A., Moss, M.S., Evans, S., 1978. Studies related to the metabolism of anabolic steroids in the horse: a gas chromatographic mass spectrometric method to confirm the administration of 19-nortestosterone or its esters to horses. Biol. Mass Spectrom., 5(2):170-173.

[19]Kohler, R.M., Lambert, M.I., 2002. Urine nandrolone metabolites: false positive doping test? Br. J. Sports Med., 36(5):325-329.

[20]Le Bizec, B., Bryand, F., Gaudin, I., Monteau, F., Poulain, F., Andre, F., 2002. Endogenous nandrolone metabolites in human urine: preliminary results to discriminate between endogenous and exogenous origin. Steroids, 67(2):105-110.

[21]Petersen, H.W., Christensen, F., Vad, M., 1994. Method validation in relation to accuracy and precision for the gas chromatography-mass spectrometric analysis of 19-nortestosterone in urine with heptafluorobutyric anhydride and N-methyl-N-(trimethylsilyl) trifluoroacetamide. Analyst, 119(12):2627-2630.

[22]Roda, A., Manetta, A.C., Portanti, O., Mirasoli, M., Guardigli, M., Pasini, P., Lelli, R., 2003. A rapid and sensitive 384-well microtitre format chemiluminescent enzyme immunoassay for 19-nortestosterone. Luminescence, 18(2):72-78.

[23]Roig, M., Segura, J., Ventura, R., 2007. Quantitation of 17β-nandrolone metabolites in boar and horse urine by gas chromatography-mass spectrometry. Anal. Chim. Acta, 586(1-2):184-195.

[24]Saito, K., Yagi, K., Ishizaki, A., Kataoka, H., 2010. Determination of anabolic steroids in human urine by automated in-tube solid-phase microextraction coupled with liquid chromatography-mass spectrometry. J. Pharm. Biomed. Anal., 52(5):727-733.

[25]Sauer, M.J., Samuels, T.P., Howells, L.G., Seymour, M.A., Nedderman, A., Houghton, E., Bellworthy, S.J., Anderson, S., Coldham, N.G., 1998. Residues and metabolism of 19-nortestosterone laurate in steers. Analyst, 123(12):2653-2660.

[26]van Poucke, C., van Peteghem, C., 2002. 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, 772(2):211-217.

[27]Ventura, R., Roig, M., Pérez, B., López, S., Medina, M., Bosch, J., Segura, J., 2008. Detection of the administration of 17β-nortestosterone in boars by gas chromatography/ mass spectrometry. Rapid Commun. Mass Spectrom., 22(12):1863-1870.

[28]Yamada, M., Aramaki, S., Kurosawa, M., Kijima-Suda, I., Saito, K., Nakazawa, H., 2008. Simultaneous doping analysis of main urinary metabolites of anabolic steroids in horse by ion-trap gas chromatography–tandem mass spectrometry. Anal. Sci., 24(9):1199-1204.

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