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Simultaneous determination of five anti-epilepsy drugs in human plasma using liquid chromatography-mass spectrometry

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Abstract

A new liquid chromatography-mass spectrometry method for the determination of carbamazepine, clonazepam, alprazolam, estazolam and phenytoin in human plasma has been developed by using diazepam as an internal standard. Chromatographic separation was performed on a Zorbax SB-C18 column (30 mm × 2.1 mm, 3.5 μm) with a mobile phase consisting of methanol and aqueous 25 mM ammonium acetate using gradient elution. A diethyl ether extraction method was used for the extraction of five anti-epilepsy drugs. The final extract was injected for analysis by LC-MS/MS. The method was validated within the concentration range of 50–5000 ng mL−1 for five anti-epilepsy drugs. The precision of the assay (RSD%) was less than 10% at all concentration levels within the tested range. The method recoveries for all samples were more than 90%. The results indicate that the method is specific, sensitive and accurate, and suitable to study the pharmacokinetics, to adjust the dosage for individual administration, and to monitor the drug-concentration and drug abuse of the five anti-epilepsy drugs.

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References

  1. Schmidt D. Drug treatment of epilepsy: Options and limitations. Epilepsy Behav, 2009, 15: 56–65

    Article  Google Scholar 

  2. Sánchez MB, Herranz JL, Leno C, Arteaga R, Oterino A, Valdizán EM, Nicolás JM, Adín J, Armijo JA. Genetic factors associated with drug-resistance of epilepsy: Relevance of stratification by patient age and aetiology of epilepsy. Seizure, 2010, 19: 93–101

    Article  Google Scholar 

  3. Jabbaribar F, Mortazavi SA, Jouyban A. Determination of phenytoin and its major metabolite in human serum by MEKC. J Chromatogr Sci, 2009, 47: 877–880

    CAS  Google Scholar 

  4. Ali AAP, Kamiar AP. Antinuclear antibodies in children with epilepsy treated by carbamazepine. Epilepsy Res, 2008, 80: 229–230

    Article  Google Scholar 

  5. Badiu CL. Sensitivity of thalamic GABAergic currents to clonazepam does not differ between control and genetic absence epilepsy rats. Brain Res, 2004, 1026: 261–266

    Article  CAS  Google Scholar 

  6. Lane SD, Gowin JL. GABAergic modulation of human social interaction in a prisoner’s dilemma model by acute administration of alprazolam. Behav Pharmacol, 2009, 20: 657–661

    Article  CAS  Google Scholar 

  7. Miura M, Otani K, Ohkubo T. Identification of human cytochrome P450 enzymes involved in the formation of 4-hydroxyestazolam from estazolam. Xenobiotica, 2005, 35: 455–465

    Article  CAS  Google Scholar 

  8. Allqvist A, Wennerholm A, Svensson JO, Rajaa AM. Simultaneous quantification of alprazolam, 4- and α-hydroxyalprazolam in plasma samples using liquid chromatography mass spectrometry. J Chromatogr B, 2005, 814: 127–131

    Article  CAS  Google Scholar 

  9. van Rooyen GF, Badenhorst D, Swart KJ, Hundt HKL, Scanes T, Hundt AF. Determination of carbamazepine and carbamazepine 10,11-epoxide in human plasma by tandem liquid chromatographymass spectrometry with electro spray ionization. J Chromatogr B, 2002, 769: 1–7

    Article  Google Scholar 

  10. Quintela O, Lopez P, Bermejo AM, Lopez-Rivadulla M, Determination of methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine and alprazolam in human plasma by liquid chromatography-electro spray ionization mass spectrometry. J Chromatogr B, 2006, 834: 188–194

    CAS  Google Scholar 

  11. Zhu Y, Chiang H, Wulster-Radcliffe H, Hilt R, Wong P, Kissinger CB, Kissinger PT. Liquid chromatography/tandem mass spectrometry for the determination of carbamazepine and its main metabolite in rat plasma utilizing an automated blood sampling system. J Pharm Biomed Anal, 2005, 38: 119–125

    Google Scholar 

  12. Bares IF, Pehourcq F, Jarry C. Development of a rapid RP-HPLC method for the determination of clonazepam in human plasma. J Pharmaceut Biomed Anal, 2004, 36: 865–869

    Article  CAS  Google Scholar 

  13. Kelly JF, Greenblatt DJ. Rapid and sensitive gas chromatographic determination of estazolam. J Chromatogr B, 1993, 621: 102–104

    Article  CAS  Google Scholar 

  14. Liu YZ, Zhou SX, Zeng L. Simultaneous determination of concentration of 5 anti-epilepsy drugs in plasma by solid phase extraction RP-HPLC method. Central South Pharmacy, 2008, 6: 686–688

    CAS  Google Scholar 

  15. Xu RN, Fan L, Rieser MJ. Recent advances in high throughput quantitative bioanalysis by LC-MS/MS. J Pharm Biom Anal, 2007, 44: 342

    Article  CAS  Google Scholar 

  16. Ma SG, Zhu MS. Recent advances in applications of liquid chromatography-tandem mass spectrometry to the analysis of reactive drug metabolites. Chem-Biol Inter, 2009, 179: 25–37

    Article  CAS  Google Scholar 

  17. Vanhoutte K, Dongen WV, Esmans EL. On-line nanoscale liquid chromatography nano-electrospray mass spectrometry: Effect of the mobile phase composition and the electrospray tip design on the performance of a nanoflow TM electrospray probe. Rapid Commun Mass Spect, 1998, 12: 15–24

    Article  CAS  Google Scholar 

  18. Garcia MC. The effect of the mobile phase additives on sensitivity in the analysis of peptides and proteins by high-performance liquid chromatography-electrospray mass spectrometry. J Chromatogr B, 2005, 825: 111–123

    Article  CAS  Google Scholar 

  19. Rochfort SJ, Trenerry CV, Imsic M, Panozzo J, Jones R Class targeted metabolomics: ESI ion trap screening methods for glucosinolates based on MSn fragmentation. Phytochemistry, 2008, 69: 1671–1679

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Institute of Pharmaceutical Analysis, College of Pharmacy, Wuhan University, Wuhan, 430072, China

    QinHua Chen, Weißig Stefanie, ShangWen Luo & ZiLin Chen

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  1. QinHua Chen
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  2. Weißig Stefanie
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  3. ShangWen Luo
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  4. ZiLin Chen
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Correspondence to ZiLin Chen.

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Chen, Q., Stefanie, W., Luo, S. et al. Simultaneous determination of five anti-epilepsy drugs in human plasma using liquid chromatography-mass spectrometry. Sci. China Chem. 53, 2373–2378 (2010). https://doi.org/10.1007/s11426-010-4009-9

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  • DOI: https://doi.org/10.1007/s11426-010-4009-9

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