, Volume 71, Issue 5–6, pp 419–422 | Cite as

Development and Validation of a Novel Gradient LC Method for Simultaneous Determination of Isoniazid and Acetylisoniazid in Human Plasma

  • Reza Mahjub
  • Hossein Khalili
  • Mohsen Amini


The goal of this study was to develop and validate a new gradient high-performance liquid chromatography method for the simultaneous determination of isoniazid (INH) and acetylisoniazid (Ac-INH) in human plasma samples. A C18 reversed-phase column was employed for separation followed by UV detection at 266 nm. The calibration involved the use of five concentration levels ranging from 1 to 20 μg mL−1 for both analytes. The developed method was validated using ICH guidelines. The calibration curve was found to be linear with correlation coefficient values (r 2) above 0.9991 and the highest RSD% values for intra-day assays were found to be 6.34 and 2.57% for INH and Ac-INH, respectively. The highest RSD% values for inter-day assays were 9.31 and 10.17% for INH and Ac-INH, respectively. LOD was calculated to be 0.1 and 0.15 μg mL−1 for INH and Ac-INH, respectively. LOQ was calculated to be 0.33 and 0.5 μg mL−1 for INH and Ac-INH, respectively.


Column liquid chromatography Plasma Isoniazid Acetyl isoniazid 



This study was made possible with financial supports from the deputy of research, Tehran University of Medical Sciences and Pharmaceutical Research Center.


  1. 1.
    Schaaf HS, Parkin DP, Seifart HI, Werely CJ, Hesseling PB, Van Helden PBMaritz JS, Donald PR (2005) Arch Dis Child 90:614–618CrossRefGoogle Scholar
  2. 2.
    Drug Information Handbook International (2006) Monograph of isoniazid, Ohio, Lexi-comp Inc, The American Pharmacists Association (APhA)Google Scholar
  3. 3.
    Schoeman JF, Morkel A, Seifart HI, Parkin DP, van Helden PD, Hewlett RH, Donald PR (1998) Pediatr Neurosurg 29:64–68CrossRefGoogle Scholar
  4. 4.
    Steichen O, Martinez-Almoyna L, de Broucker T (2006) Rev Mal Respir 23:157–160Google Scholar
  5. 5.
    Mohan B, Sharda N, Singh S (2003) J Pharm Biomed Anal 31:607–612CrossRefGoogle Scholar
  6. 6.
    Unsalan S, Sancar M, Bekce B, Clark PM, Karagoz T, Izzettin FV, Rollas S (2005) Chromatographia 61:595–598CrossRefGoogle Scholar
  7. 7.
    Huang L, Marzan F, Jayewardene AL, Lizac PS, Xiaohua L, Aweeka FT (2009) J Chromatogr B 877:285–290CrossRefGoogle Scholar
  8. 8.
    Prasad B, Singh S (2009) J Pharm Biopharm Anal. doi: 10.1016/j.jpba.2009.07.014
  9. 9.
    Um SW, Lee SW, Kwon SY, Yoon HI, Park KU, Song J, Lee CT, Lee JH (2007) Int J Tuberc Lung Dis 11:972–978Google Scholar
  10. 10.
    Seifart HI, Gent WL, Parkin DP, van Jaarsveld PP, Donald PR (1995) J Chromatogr B 674:269–275CrossRefGoogle Scholar
  11. 11.
    Mattar KM, Mayet AY, Ayoola EA, Bawazir SA, Al-Faleh FZ, Al Wazzan A (2004) J Clin Pharm Ther 29:443–447CrossRefGoogle Scholar
  12. 12.
    Rey E, Gendrel D, Treluyer JM, Tran A, Pariente-Khayat A, Athis P, Pons G (2001) Fundam Clin Pharmacol 15:355–359CrossRefGoogle Scholar
  13. 13.
    Fox HH, Gibas JT (1953) J Org Chem 18:1375–1379CrossRefGoogle Scholar
  14. 14.
    Fox HH, Gibas JT (1953) J Org Chem 18:983–989CrossRefGoogle Scholar
  15. 15.
    Fox HH, Gibas JT (1953) J Org Chem 18:994–1002CrossRefGoogle Scholar

Copyright information

© Vieweg+Teubner Verlag | Springer Fachmedien Wiesbaden GmbH 2010

Authors and Affiliations

  1. 1.Department of Pharmaceutics, Faculty of PharmacyTehran University of Medical SciencesTehranIran
  2. 2.Department of Clinical Pharmacy (Pharmacotherapy), Faculty of PharmacyTehran University of Medical SciencesTehranIran
  3. 3.Department of Medicinal Chemistry, Faculty of PharmacyTehran University of Medical SciencesTehranIran
  4. 4.Drug Design and Development Research CenterTehran University of Medical SciencesTehranIran

Personalised recommendations