Development and Validation of Stability Indicating HPTLC Assay for Determination of Gemifloxacin Mesylate in Dosage Forms


Simple and sensitive stability-indicating high performance thin layer chromatography (HPTLC) assay was developed and validated for quantitative determination of the antibacterial drug, gemifloxacin mesylate (GFX) in presence of its degradation products and ambroxol hydrochloride. The chromatographic separation was performed on HPTLC precoated silica gel plate 60F254 as stationary phase. The mobile phase consisted of a mixture of ethyl acetate: methanol: 25% ammonia, (8:4.5:3, v/v/v). The detection was performed using fluorescence mode and the emission intensity was measured using optical filter K400 after excitation at 342 nm. The Rf value for GFX was 0.47 ± 0.03. Good correlation coefficient was obtained over the concentration range of 1.5 - 180 ng/band. The LOD and LOQ of the proposed method were 0.28 and 0.86 ng/band, respectively. The proposed method was successfully applied for the analysis of GFX in its single and combined dosage forms. Moreover, it was utilized to investigate the kinetics of acidic, alkaline, neutral, oxidative and photolytic degradation of the drug. The apparent kinetic-order rate constants and half-life times of the degradation process were calculated. Furthermore, the proposed method was successfully applied for investigating the factors affecting the storage of GFX.

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Mahmoud, A. , Atia, N. , El-Shabouri, S. and El-Koussi, W. (2015) Development and Validation of Stability Indicating HPTLC Assay for Determination of Gemifloxacin Mesylate in Dosage Forms. American Journal of Analytical Chemistry, 6, 85-97. doi: 10.4236/ajac.2015.62008.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] ICH Guidance for Industry (Q1A) (R2) (2003) Stability Testing of New Drug Substances and Products in Federal Register. ICH Guidance for Industry, Geneva.
[2] El Zeany, B.A., Moustafa, A.A. and Farid, N.F. (2003) Determination of Zolpidem Hemitartrate by Quantitative HPTLC and LC. Journal of Pharmaceutical and Biomedical Analysis, 33, 393-401.
[3] Guermouche, M.H., Habel, D. and Guermouche, S. (1999) Assay of Tinidazole in Human Serum by High-Performance Thin-Layer Chromatography: Comparison with High-Performance Liquid Chromatography. Journal of AOAC International, 82, 244-247.
[4] Kaale, E., Risha, P. and Layloff, T. (2011) TLC for Pharmaceutical Analysis in Resource Limited Countries. Journal of Chromatography A, 1218, 2732-2736.
[5] Le, T.P. and Xiong, Y.Q. (2001) Gemifloxacin. Drugs of Today, 37, 401-410.
[6] Yoo, B.K., Triller, D.M., Yong, C.S. and Lodise, T.P. (2004) Gemifloxacin: A New Fluoroquinolone Approved for Treatment of Respiratory Infections. The Annals of Pharmacotherapy, 38, 1226-1235.
[7] Hong, C.Y. (2001) Discovery of Gemifloxacin (Factive, LB20304a): A Quinolone of New a Generation. Il Farmaco, 56, 41-44.
[8] Atia, N.N., Mahmoud, A.M., El-Shabouri, S.R. and El-Koussi, W.M. (2013) Two Validated Spectrofluorometric Methods for Determination of Gemifloxacin Mesylate in Tablets and Human Plasma. International Journal of Analytical Chemistry, 2013, 1-11.
[9] Doyle, E., Fowles, S.E., McDonnell, D.F., McCarthy, R. and White, S.A. (2000) Rapid Determination of Gemifloxacin in Human Plasma by High-Performance Liquid Chromatography-Tandem Mass Spectrometry. Journal of Chromatography B: Biomedical Sciences and Applications, 746, 191-198.
[10] Al-Hadiya, B.M.H., Khady, A.A. and Mostafa, G.A.E. (2010) Validated Liquid Chromatographic-Fluorescence Method for the Quantitation of Gemifloxacin in Human Plasma. Talanta, 83, 110-116.
[11] Rote, A.R. and Pingle, S.P. (2009) Reverse Phase-HPLC and HPTLC Methods for Determination of Gemifloxacin Mesylate in Human Plasma. Journal of Chromatography B, 877, 3719-3723.
[12] El-Koussi, W.M., Atia, N.N., Mahmoud, A.M. and El-Shabouri, S.R. (2014) HPTLC Method for Direct Determination of Gemifloxacin Mesylate in Human Plasma. Journal of Chromatography B, 967, 98-101.
[13] Raja, T. and Rao, A.L. (2012) Development and Validation of HPTLC Method for the Simultaneous Estimation of Gemifloxacin Mesylate and Ambroxol Hydrochloride in Bulk and Tablet Dosage Form. Analytical Chemistry Letters, 2, 152-158.
[14] Elbashir, A., Saad, B., Ali, A.S.M., Al-Azzam, K. and Aboul-Enein, H. (2008) Validated Stability Indicating Assay of Gemifloxacin and Lomefloxacin in Tablet Formulations by Capillary Electrophoresis. Journal of Liquid Chromatography & Related Technologies, 31, 1465-1477.
[15] Ranjane, P.N., Gandhi, S.V., Kadukar, S.S. and Bothara, K.G. (2010) Stability Indicating RP-LC Method for the Determination of Gemifloxacin Mesylate. Chromatographia, 71, 1113-1117.
[16] Gandhimathi, N., Ravi, T.K. and Bharathi, N. (2010) Study of Hydrolytic and Oxidative Behavior of Gemifloxacin Mesylate in Aqueous Solution by LC-MS. Journal of Global Pharma Technology, 2, 81-85.
[17] Paim, C.S., Führ, F., Barth, A.B., Goncalves, C.E.I., Nardi, N., Steppe, M. and Schapoval, E.E.S. (2011) Gemifloxacin Mesylate Stability Evaluation Applying a Validated Bioassay Method and in Vitro Cytotoxic Study. Talanta, 83, 1774-1779.
[18] Nageswara Rao, R., Naidu, C.G., Prasad, K.G. and Narasimha, R. (2011) Development and Validation of a RP-HPLC Method for Stability-Indicating Assay of Gemifloxacin Mesylate Including Identification of Related Substances by LC-ESI-MS/MS, 1H and 13C NMR Spectroscopy. Biomedical Chromatography, 25, 1222-1229.
[19] Paim, S., Fuhr, F., Miron, S., Pereira, L., Malesuik, D., Steppe, M. and Schapoval, E.S. (2010) Stability-Indicating LC Assay and Determination of System Suitability Limits with a Robustness Test of Gemifloxacin Mesylate in Tablets. Current Analytical Chemistry, 6, 269-276.
[20] Fried, B. and Sherma, J. (1999) Thin-Layer Chromatography. Vol. 81, CRC Press, Boca Raton.
[21] Fried, B. and Sherma, J. (1982) Thin Layer Chromatography. Vol. 17, Marcel Dekker, Inc., New York, 308.
[22] ICH Guidance for Industry, Q2 (R1) (2005) Validation of Analytical Procedures: Text and Methodology. Federal Register, Geneva, 1-13.
[23] Wainwright, N., Collins, P. and Ferguson, J. (1993) Photosensitivity Associated with Antibacterial Agents. Drug Safety, 9, 437-440.
[24] Vousden, M., Ferguson, J., Richards, J., Bird, N. and Allen, A. (1999) Evaluation of Phototoxic Potential of Gemifloxacin in Healthy Volunteers Compared with Ciprofloxacin. Chemotherapy, 45, 512-520.
[25] Hidalgo, M.E., Pessoa, C., Fernandez, E. and Cardenas, A.M. (1993) Comparative Determination of Photodegradation Kinetics of Quinolones. Journal of Photochemistry and Photobiology A: Chemistry, 73, 135-138.
[26] Endruweit, A., Johnson, M.S. and Long, A.C. (2006) Curing of Composite Components by Ultraviolet Radiation: A Review. Polymer Composites, 27, 119-128.

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