Analytical and Bioanalytical Chemistry

, Volume 409, Issue 27, pp 6315–6323 | Cite as

Enantioselective separation and determination of miconazole in rat plasma by chiral LC–MS/MS: application in a stereoselective pharmacokinetic study

  • Yueying Du
  • Linda Luo
  • Shuo Sun
  • Zhen Jiang
  • Xingjie Guo
Research Paper


Miconazole has one chiral center, and consists of two enantiomers. In this study, a novel chiral liquid chromatography–tandem mass spectrometry method was developed for enantioselective separation and determination of miconazole in rat plasma. For the first time, the enantioselective pharmacokinetics of miconazole was investigated by the current method. Firstly, attempts were made to separate the enantiomers in reversed-phase mode with a mobile phase that was mass spectrometry compatible. Baseline separation was achieved on a Chiralpak IC column with a mobile phase composed of acetonitrile and aqueous ammonium hydrogen carbonate (5 mM; 80:20, v/v). Data were acquired in multiple reaction monitoring mode with positive electrospray ionization by triple-quadrupole mass spectrometry. Then, overall method validation regarding the linearity, accuracy, precision, extraction recovery, matrix effect, and stability of each enantiomer was performed, and acceptable results were obtained for all of these. Finally, the method developed was applied in an enantioselective pharmacokinetic study of miconazole enantiomers in rats after oral administration of racemic miconazole at doses of 5 and 10 mg/kg. The results demonstrated that (–)-(R)-miconazole had a higher concentration than (+)-(S)-miconazole in plasma, with a ratio of 1.3–1.7 for both doses. This is the first experimental evidence of enantioselective behavior of miconazole in vivo, and provides a reference for clinical practice and encourages further research into miconazole enantioselective metabolism and drug interactions.

Graphical Abstract

A stereoselective pharmacokinetic study of the miconazole enantiomers was investigated using a novel chiral liquid chromatography–tandem mass spectrometry method. Baseline separation was achieved on Chiralpak IC column, and Chiralcel OJ column was used to collect single enantiomer. A significant difference between the two enantiomers was observed in view of the plasma concentration


Miconazole enantiomers Liquid chromatography–tandem mass spectrometry Enantioselective pharmacokinetics 


Compliance with ethical standards

All procedures involving animals were performed in accordance with the regulations of the Experimental Animal Administration (State Committee of Science and Technology of the People's Republic of China) and were approved by the Medical Ethics Committee of Shenyang Pharmaceutical University (no. SYPU-IACUC-C2016-11-23-201, no. SYPU-IACUC-C2017-6-16-201).

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Erny GL. Liquid separation techniques coupled with mass spectrometry for chiral analysis of pharmaceuticals compounds and their metabolites in biological fluids. J Pharm Biomed Anal. 2006;40:509–15.CrossRefGoogle Scholar
  2. 2.
    Brocks DR. Drug disposition in three dimensions: an update on stereoselectivity in pharmacokinetics. Biopharm Drug Dispos. 2006;27:387–406.CrossRefGoogle Scholar
  3. 3.
    Brocks DR, Mehvar R. Stereoselectivity in the pharmacodynamics and pharmacokinetics of the chiral antimalarial drugs. Clin Pharmacokinet. 2003;42:1359–82.CrossRefGoogle Scholar
  4. 4.
    Fanous J, Swed A, Joubran S. Superiority of the S,S conformation in diverse pharmacological processes Intestinal transport and entry inhibition activity of novel anti-HIV drug lead. Int J Pharm. 2015;495:660.CrossRefGoogle Scholar
  5. 5.
    FDA’s policy statement for the development of new stereoisomeric drugs. Chirality. 1992;4:338.Google Scholar
  6. 6.
    Fothergill AW. Miconazole: a historical perspective. Expert Rev Anticanc. 2006;4:171.CrossRefGoogle Scholar
  7. 7.
    Mangassánchez J, Busto E, Gotorfernández V, Malpartida F, Gotor V. Asymmetric chemoenzymatic synthesis of miconazole and econazole enantiomers. J Org Chem. 2011;76:2115–2.CrossRefGoogle Scholar
  8. 8.
    Liu YM, Yao XN, Guo XJ. Chiral separation of miconazole using Chiralpak AD-H chiral column. J Shenyang Pharm Univ. 2010;27:567–9.Google Scholar
  9. 9.
    Ali I, Aboul-Enein HY, Gaitonde VD, Singh P, Rawat MS, Singh P. Chiral separations of imidazole antifungal drugs on AmyCoat RP column in HPLC. Chromatographia. 2009;70:223–7.CrossRefGoogle Scholar
  10. 10.
    Aboul-Enein HY, Ali I. Comparative study of the enantiomeric resolution of chiral antifungal drugs econazole, miconazole and sulconazole by HPLC on various cellulose chiral columns in normal phase mode. J Pharm Biomed Anal. 2002;27:441–6.CrossRefGoogle Scholar
  11. 11.
    Hamdy DA, Brocks DR. A stereospecific high-performance liquid chromatographic assay for the determination of ketoconazole enantiomers in rat plasma. Biomed Chromatogr. 2008;22:542–7.CrossRefGoogle Scholar
  12. 12.
    Feng Z, Zou Q, Tan X, Che W, Zhang Z. Determination of fenticonazole enantiomers by LC-ESI-MS/MS and its application to pharmacokinetic studies in female rats. Arzneimittelforschung. 2011;61:587–93.Google Scholar
  13. 13.
    US Food and Drug Administration. Guidance for industry: bioanalytical method validation. Accessed 12 Sep 2013.
  14. 14.
    Ali I, Aboul-Enein HY. Enantioseparation of some clinically used drugs by HPLC using cellulose tris (3,5-dichlorophenylcarbamate) chiral stationary phase. Biomed Chromatogr. 2003;17:113.CrossRefGoogle Scholar
  15. 15.
    Ahmed M, Gwairgi M, Ghanem A. Conventional Chiralpak ID vs. capillary Chiralpak ID-3 Amylose-tris-(3-chlorophenylcarbamate)-based chiral stationary phase columns for the enantioselective HPLC separation of pharmaceutical racemates. Chirality. 2014;26:677.CrossRefGoogle Scholar
  16. 16.
    Aboul-Enein HY, Ali I. Comparison of the chiral resolution of econazole, miconazole and sulconazole by HPLC using normal-phase amylose CSPs. Anal Bional Chem. 2001;370:951–5.CrossRefGoogle Scholar
  17. 17.
    Zhang T, Nguyen D, Franco P. Reversed-phase screening strategies for liquid chromatography on polysaccharide-derived chiral stationary phases. J Chromatogr A. 2010;1217:1048–55.CrossRefGoogle Scholar
  18. 18.
    Peng LM, Farkas T. Analysis of basic compounds by reversed-phase liquid chromatography-electrospray mass spectrometry in high-pH mobile phases. J Chromatogr A. 2008;1179:131–44.CrossRefGoogle Scholar
  19. 19.
    Cirilli R, Ferretti R, Gallinella B, La TF, La RG, Silvestri R. Comparative study between the polysaccharide-based Chiralcel OJ and Chiralcel OD CSPs in chromatographic enantioseparation of imidazole analogues of fluoxetine and miconazole. J Sep Sci. 2005;28:627–34.CrossRefGoogle Scholar
  20. 20.
    Kobylifiska M, Kobylifiska K, Sobik B. High-performance liquid chromatographic analysis for the determination of miconazole in human plasma using solid-phase extraction. J Chromatogr B. 1996;685:191–5.CrossRefGoogle Scholar
  21. 21.
    Sternson LA, Pation TF, King TB. High-performance liquid chromatographic analysis of miconazole in plasma. J Chromatogr. 1982;227:223–8.CrossRefGoogle Scholar
  22. 22.
    Ahmed TA, El-Say KM, Mahmoud MF, Samy AM, Badawi AA. Miconazole nitrate oral disintegrating tablets: in vivo performance and stability study. AAPS PharmSciTech. 2012;13:760–71.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  1. 1.School of PharmacyShenyang Pharmaceutical UniversityLiaoningChina

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