Development and Validation of Stability Indicating HPLC Method for Simultaneous Estimation of Amoxicillin and Clavulanic Acid in Injection

Durga Mallikarjuna Rao Tippa, N. Singh

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DOI: 10.4236/ajac.2010.13013                

Abstract

A simple, fast, precise, accurate and rugged stability indicating high performance liquid chromatography (HPLC) method has been developed for simultaneous estimation of Amoxicillin and Clavulanic acid from injectable dosage form. The stability indicating capability of the method was proven by subjecting the drugs to stress conditions as per ICH recommended test conditions such as alkaline and acid hydrolysis, oxidation, photolysis, thermal degradation and resolution of the degradation products formed therein. The separation was obtained using a mobile phase composition at a ratio of 95:5 (v/v) of pH 5.0 buffer and methanol on Inertsil C18 column (250 × 4.0 mm, 4 μm) with UV detection at 220 nm at a flow rate of 1 ml/minute. The photodiode array detector was used for stress studies. The order of elution of peaks was Clavulanic acid followed by Amoxicillin. The linear calibration range was found to be 79.51 to 315.32 μg/ml for Amoxicillin and 17.82 to 67.90 μg/ml for Clavulanic acid. The Amoxicillin and Clavulanic acid were found to be stable in solution up to 24 hours. The method validation data showed excellent results for precision, linearity, specificity, limit of detection, limit of quantification and robustness. The present method can be successfully used for routine quality control and stability studies.

Keywords

Amoxicillin, Clavulanic Acid, HPLC, Stability Indicating

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	C. Reading and M. Cole, “Clavulanic Acid: A Beta Lactamase-Inhibiting Beta-Lactam from Streptomyces Clavuligerus,” Antimicrobial Agents and Chemotherapy, Vol. 11, No. 5, 1977, pp. 852-857.
[2]	M. Foulstone and C. Reading, “Assay of Amoxicillin and Clavulanic Acid, the Component of Augmentin, in Biological Fluids with High Performance Liquid Chromatography,” Antimicrobial Agents and Chemotherapy, Vol. 22, No. 5, November 1982, pp. 753-762.
[3]	P. A. Hunter, K. Coleman, J. Fisher, D. Taylor and E. Taylor, “Clavulanic Acid, a Novel B-Lactam with Broad Spectrum B-Lactamase Inhibitory Properties. Synergistic Activity with Ampicillin and Amoxicillin,” Drugs Under Experimental and Clinical Research, Vol. 5, 1979, pp. 1-6.
[4]	K. R. Reddy, P. Brillant and E. R. Schiff, “Amoxicillin-Clavulanate Potassium Associated Cholestasis,” Gastroenterology, Vol. 96, No. 4, April 1989, pp. 135-141.
[5]	C. Hall, F. Curran, D. W. Burdon and M. R. B. Keighley, “A Randomized Trial to Compare Amoxicillin and Clavulanate with Metonidazole Plus Gentamycin in Prophylaxis in Elective Colarectl Suregery,” Journal of Antimicrobial Chemotherapy, Vol. 24, No. (Suppl)B, 1989, pp. 195-208.
[6]	A. P. R. Willson, S. Shrimpton and M. Jaderberg, “A Meta Analysis of the Use of Amoxicillin-Clavulanic Acid in Surgical Prophylaxis,” Journal of Hospital Infection, Vol. 22, No. (Suppl)A, 1992, pp. 9-21.
[7]	The Department of Health, “British Pharmacopoeia,” DH, London, Vol. I, 2009, pp. 353-361.
[8]	“Physicians’ Desk Reference,” 60th Edition, Thomson PDR Company, Montvale, 2006, p. 1331.
[9]	J. B. Veer, Y. A. Hekster, A. M. Baars and E. Van Der Klerjn, “Rapid Determination of Amoxicillin and Ampicillin in Body Fluids by Means of High Performance Liquid Chromatography,” Journal of Chromatography, Vol. 1145, 1978, pp. 496-501.
[10]	M. J. Lebella, W. L. Wilson and G. Lawriault, “High Performance Liquid Chromatographic Determination of Amoxicillin in Pharmaceutical Dosage Forms,” Journal of Chromatography, Vol. 202, No. 1, 1980, pp. 144-147.
[11]	De Pourcq, J. Hoebus, E. Roets, J. Hoogmartens and H. Vanderhaeghe, “Quantiative Determination of Amoxicillin and Its Decomposition Products by High Performance Chromatography,” Journal of Chromatography, Vol. 321, 1985, pp. 441-443.
[12]	A. S. Low, R. B. Taylor and J. M. Gould, “Determination of Clavulanic Acid by a Sensitive HPLC Method,” Journal of Antimicrobial Chemotherapy, Vol. 24, No. (Suppl)B, 1989, pp. 83-86.
[13]	J. Haginaka, T. Nakagawa, Y. Nishind and T. Uno, “High Performance Liquid Chromatography Determination of Clavulanic Acid in Human Urine,” Journal of Antibiotics, Vol. 34, 1981, pp. 1189-1194.
[14]	ICH, “Validation of Analytical Procedures: Text and Methodology,” International Conference on Harmonisation Guidance Documents, Q2(R1), 2005.
[15]	ICH, “Photo Stability Testing of New Drug Substances and Products,” International Conference on Harmonisation Guidance Documents, Q1B, 2005.
[16]	ICH, “Stability Testing of New Drug Substances and Products,” International Conference on Harmonisation Guidance Documents, Q1A(R2), 2005.