, Volume 69, Issue 3–4, pp 345–350 | Cite as

Development of Validated Chromatographic Methods for the Simultaneous Determination of Metronidazole and Spiramycin in Tablets

Full Short Communication


Two chromatographic methods have been described for the simultaneous determination of metronidazole (MET) and spiramycin (SPY) in their mixtures. The first method was based on a high performance thin layer chromatographic (HPTLC) separation of the two drugs followed by densitometric measurements of their spots at 240 nm. The separation was carried out on Merck TLC aluminum sheets of silica gel 60 F254 using methanol: chloroform (9:1, v/v) as a mobile phase. Analysis data was used for the linear regression line in the range of 1.0–2.0 and 0.8–2.0 μg band−1 for MET and SPY, respectively. The second method was based on a reversed-phase liquid chromatographic separation of the cited drugs on a C-18 column (5 μm, 250 × 4.6 mm, i.d.). The mobile phase consisted of a mixture of phosphate buffer of pH 2.4 and acetonitrile (70:30, v/v). The separation was carried out at ambient temperature with a flow rate of 1.0 mL min−1. Quantitation was achieved with UV detection at 232 nm based on peak area with linear calibration curves at concentration ranges 0.4–50.0 and 0.5–50.0 μg mL−1 for MET and SPY, respectively. The proposed chromatographic methods were successfully applied to the determination of the investigated drugs in pharmaceutical preparations. Both methods were validated in compliance with ICH guidelines; in terms of linearity, accuracy, precision, robustness, limits of detection and quantitation and other aspects of analytical validation.


High performance thin layer chromatography Reversed phase liquid chromatography Validation Metronidazole and spiramycin in tablets 


  1. 1.
    Hardman JG, Limbird LE (1996) Goodman & Gillman’s, The Pharmacological Basis of Therapeutics, 9th ed. McGraw, Hill, NewYorkGoogle Scholar
  2. 2.
    Zhong D, Shi X, Sun L, Chen X (2003) J Chromatogr B 791:45–53CrossRefGoogle Scholar
  3. 3.
    British Pharmacopoeia (2008) through internet communications,
  4. 4.
    Sagan C, Salvador A, Dubreuil D, Poulet PP, Duffaut D, Brumpt I (2005) J Pharm Biomed Anal 38:298–306CrossRefGoogle Scholar
  5. 5.
    Li F, Ye YY, Jia WP, Chen Q (2006) Yaowu Fenxi Zazhi 26:1311–1313Google Scholar
  6. 6.
    Menelaou A, Somogyi AA, Barclay ML, Bochner F (1999) J Chromatogr B 731:261–266CrossRefGoogle Scholar
  7. 7.
    Mishal A, Sober D (2005) J Pharm Biomed Anal 39:819–823CrossRefGoogle Scholar
  8. 8.
    Jin W, Li W, Xu Q, Dong Q (2000) Electrophoresis 21:1409–1414CrossRefGoogle Scholar
  9. 9.
    Agbaba D, Djurkovic M, Brboric J, Zivanov-stakic D (1998) J Planar Chromatogr Mod TLC 11:447–449Google Scholar
  10. 10.
    Bartlett PN, Ghoneim E, El-Hefnawy G, El-Hallag I (2005) Talanta 66:869–874CrossRefGoogle Scholar
  11. 11.
    Huet AC, Mortier L, Els Daeseleire, Fodey T, Elliott C, Delahaut P (2005) Anal Chim Acta 534:157–162CrossRefGoogle Scholar
  12. 12.
    Liu JF (1999) Yaowu Fenxi Zazhi 19:125–126Google Scholar
  13. 13.
    Kanfer I, Skinner MF, Walker RB (1998) J Chromatogr A 812:255–286CrossRefGoogle Scholar
  14. 14.
    Horie M, Saito K, Ishii R, Yoshida T, Haramaki Y, Nakazawa H (1998) J Chromatogr A 812:295–302CrossRefGoogle Scholar
  15. 15.
    Garcia-Mayor MA, Garcinuno RM, Fernandez-Hernado P, Durand-Alergia JS (2006) J Chromatogr A 1122:76–83CrossRefGoogle Scholar
  16. 16.
    Flurer CL (1996) Electrophoresis 17:359–366CrossRefGoogle Scholar
  17. 17.
    Sun C, Yu R, Yang Q, Sheng S, Zhao X (1987) Yaoxue Xuebao 22:515–519Google Scholar
  18. 18.
    Ozkan SA, Uslu B, Aboul-Enein HY (2003) Crit Rev Anal Chem 33:155–181CrossRefGoogle Scholar
  19. 19.
    Moffat AC, Osselton MD, Widdop B (2004) Clarke’s Analysis of Drugs and Poisons 3 rd ed., London, pp. 915–916, 1319–1320Google Scholar
  20. 20.
    Q2A, ICH, Q2A (R1), Validation of Analytical Procedures: Text and Methodology, International Conference on Harmonization, Geneva, (November 2005), (
  21. 21.
    Kaul N, Dhaneshwar SR, Agrawal H, Kakad A, Patil B (2005) J Pharm Biomed Anal 37:27–38CrossRefGoogle Scholar
  22. 22.
    Miller JN, Miller JC (2000) Statistics and Chemometrics for Analytical Chemistry, 4th ed edn. Prentice Hall, Harlow, England, pp 111–118Google Scholar
  23. 23.
    Armitage P, Berry G (1994) Statistical Methods in Medical Research, 3rd ed edn. Blackwell Scientific Publications, Oxford, England, pp 283–285Google Scholar
  24. 24.
    Sethi PD (1996) High Performance Thin Layer Chromatography: Quantitative Analysis of Pharmaceutical Formulations. CBS Publishers and Distributors, New Delhi, IndiaGoogle Scholar

Copyright information

© Vieweg+Teubner | GWV Fachverlage GmbH 2008

Authors and Affiliations

  1. 1.Department of Pharmaceutical Analytical Chemistry, Faculty of PharmacyUniversity of AlexandriaAlexandriaEgypt

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