Journal of Analytical Chemistry

, Volume 68, Issue 11, pp 986–991 | Cite as

Validation of an analytical LC-MS/MS method in human plasma for the pharmacokinetic study of atomoxetine

  • C. I. Choi
  • C. G. Jang
  • J. W. Bae
  • S. Y. Lee


This study aimed to validate a sensitive and reliable analytical method for the pharmacokinetic study of atomoxetine in human plasma by liquid chromatography-electrospray ionization-tandem mass spectrometry. Metoprolol was used as an internal standard. After liquid-liquid extraction with methyl t-butyl ether, the supernatant was evaporated. The residue was then reconstituted and an aliquot was injected into the high performance liquid chromatographic system. Separation was performed on a Phenomenex Luna C18 column (2.0 mm × 100 mm, 3 μm particles) with a mobile phase of 10 mM ammonium formate buffer: methanol = 10: 90 (v/v). Tandem mass spectrometry was performed in the electrospray ionization positive ion mode using the multiple reaction monitoring mode for quantification. The mass transition pairs of m/z 256 → 44 for atomoxetine and m/z 268 → 116 for the internal standard were used. The flow rate of the mobile phase was 0.25 mL/min and the retention times of atomoxetine and the internal standard were found to be 1.0 and 0.9 min, respectively. The calibration curve for atomoxetine was linear in the concentration range of 1–750 ng/mL (r 2 = 0.9992) with a lower limit of quantification of 1 ng/mL. The mean accuracy for atomoxetine was 93–102%. The coefficients of variation (precision) in the intra- and inter-day validation for atomoxetine were 4.0–6.8 and 1.1–9.6%, respectively. The pharmacokinetic parameters of atomoxetine were evaluated after administration of a 40-mg single oral dose to twelve healthy male volunteers. The mean AUC0–24 h, C max, T max and T 1/2 for atomoxetine were 1.9 ± 0.8 μg h/mL, 0.34 ± 0.11 μg/mL, 1.0 ± 0.5 h and 3.9 ± 1.3 h, respectively.


atomoxetine LC-ESI-MS/MS human plasma pharmacokinetics 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Wong, D.T., Threlkeld, P.G., Best, K.L., and Bymaster, F.P., J. Pharmacol. Exp. Ther., 1982, vol. 222, no. 1, p. 61.Google Scholar
  2. 2.
    Gehlert, D.R., Gackenheimer, S.L., and Robertson, D.W., Neurosci. Lett., 1993, vol. 157, no. 2, p. 203.CrossRefGoogle Scholar
  3. 3.
    Bymaster, F.P., Katner, J.S., Nelson, D.L., Hemrick-Luecke, S.K., Threlkeld, P.G., Heiligenstein, J.H., Morin, S.M., Gehlert, D.R., and Perry, K.W., Neuropsychopharmacology, 2002, vol. 27, no. 5, p. 699.CrossRefGoogle Scholar
  4. 4.
    Sauer, J.M., Ring, B.J., and Witcher, J.W., Clin. Pharmacokinet., 2005, vol. 44, no. 6, p. 571.CrossRefGoogle Scholar
  5. 5.
    Sauer, J.M., Ponsler, G.D., Mattiuz, E.L., Long, A.J., Witcher, J.W., Thomasson, H.R., and Desante, K.A., Drug Metab. Dispos., 2003, vol. 31, no. 1, p. 98.CrossRefGoogle Scholar
  6. 6.
    Belle, D.J., Ernest, C.S., Sauer, J.M., Smith, B.P., Thomasson, H.R., and Witcher, J.W., J. Clin. Pharmacol., 2002, vol. 42, no. 11, p. 1219.CrossRefGoogle Scholar
  7. 7.
    Chalon, S.A., Desager, J.P., Desante, K.A., Frye, R.F., Witcher, J., Long, A.J., Sauer, J.M., Golnez, J.L., Smith, B.P., Thomasson, H.R., and Horsmans, Y., Clin. Pharmacol. Ther., 2003, vol. 73, no. 3, p. 178.CrossRefGoogle Scholar
  8. 8.
    Sauer, J.M., Long, A.J., Ring, B., Gillespie, J.S., Sanburn, N.P., DeSante, K.A., Petullo, D., VandenBranden, M.R., Jensen, C.B., Wrighton, S.A., Smith, B.P., Read, H.A., and Witcher, J.W., J. Pharmacol. Exp. Ther., 2004, vol. 308, no. 2, p. 410.CrossRefGoogle Scholar
  9. 9.
    Witcher, J.W., Long, A., Smith, B., Sauer, J.M., Heilgenstein, J., Wilens, T., Spencer, T., and Biederman, J., J. Child. Adolesc. Psychopharmacol., 2003, vol. 13, no. 1, p. 53.CrossRefGoogle Scholar
  10. 10.
    Cui, Y.M., Teng, C.H., Pan, A.X., Yuen, E., Yeo, K.P., Zhou, Y., Zhao, X., Long, A.J., Bangs, M.E., and Wise, S.D., Br. J. Clin. Pharmacol., 2007, vol. 64, no. 4, p. 445.CrossRefGoogle Scholar
  11. 11.
    Zhu, H.J., Wang, J.S., Donovan, J.L., DeVane, C.L., Gibson, B.B., and Markowitz, J.S., J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 2007, vol. 846, nos. 1–2, p. 351.CrossRefGoogle Scholar
  12. 12.
    Patel, C., Patel, M., Rani, S., Nivsarkar, M., and Padh, H., J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 2007, vol. 850, no. 1–2, p. 356.CrossRefGoogle Scholar
  13. 13.
    Guo, W., Li, W., Guo, G., Zhang, J., Zhou, B., Zhai, Y., and Wang, C., J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 2007, vol. 854, nos. 1–2, p. 128.CrossRefGoogle Scholar
  14. 14.
    Mattiuz, E.L., Ponsler, G.D., Barbuch, R.J., Wood, P.G., Mullen, J.H., Shugert, R.L., Li, Q., Wheeler, W.J., Kuo, F., Conrad, P.C., and Sauer, J.M., Drug Metab. Dispos., 2003, vol. 31, no. 1, p. 88.CrossRefGoogle Scholar
  15. 15.
    Mullen, J.H., Shugert, R.L., Ponsler, G.D., Li Q., Sundaram, B., Coales, H.L., Yakupkovic, J.E., Lelacheur, R.M., Wheeler, W.J., Belas, F.J., and Sauer, J.M., J. Pharm. Biomed. Anal., 2005, vol. 38, no. 4, p. 720.CrossRefGoogle Scholar
  16. 16.
    Rybak, M.E., Parker, D.L., and Pfeiffer, C.M., J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 2008, vol. 861, no. 1, p. 145.CrossRefGoogle Scholar
  17. 17.
    Ring, B.J., Gillespie, J.S., Eckstein, J.A., and Wrighton, S.A., Drug Metab. Dispos., 2002, vol. 30, no. 3, p. 319.CrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2013

Authors and Affiliations

  • C. I. Choi
    • 1
  • C. G. Jang
    • 1
  • J. W. Bae
    • 1
    • 2
  • S. Y. Lee
    • 1
  1. 1.School of PharmacySungkyunkwan University SuwonSuwonRepublic of Korea
  2. 2.College of PharmacyKeimyung University DaeguDaeguRepublic of Korea

Personalised recommendations