Development and Validation of a RP-HPLC Method for Simultaneous Determination of Levofloxacin and Moxifloxacin in Pharmaceutical Dosage Forms


HPLC, method development, validation, levofloxacin, moxifloxacin.

How to Cite

Farjahan Nur, Asma Rahman, Md. Zakir Sultan, Md. Gias Uddin, Mohammad A. Rashid, Amir Hassan, & Mansoor Ahmed. (2021). Development and Validation of a RP-HPLC Method for Simultaneous Determination of Levofloxacin and Moxifloxacin in Pharmaceutical Dosage Forms. Journal of Basic & Applied Sciences, 9, 633–638.


A simple, fast and economic reversed phase high performance liquid chromatographic (HPLC) method has been successfully developed and validated for simultaneous determination of fluoroquinolone analogs namely levofloxacin and moxifloxacin in both pure form (as API) and in pharmaceutical dosage forms. The method was validated according to the guidelines of ICH, FDA and USP with respect to accuracy, precision and linearity. For method development a C-18 bonded silica column (250 x 4.6 mm, 5μ, Phenomenex, Inc) was used with a mobile phase comprising of 10% aqueous solution of acetic acid and acetonitrile in a ratio of 80:20 v/v. The flow rate was 0.5 mL/min and effluents were monitored at 300 nm and the retention times were found to be at 7.0±0.1 min and 10.59±0.1 min for levofloxacin and moxifloxacin, respectively. The recovery was found to be more than 99% for each spiked samples of levofloxacin and moxifloxacin, demonstrating the accuracy of the protocol. Intra-day and inter-day precisions of the new method were less than the maximum allowable limit (RSD% £ 2.0) according to FDA. The method showed linear response with correlation coefficient value of 0.9975 in both the cases.

Therefore, the developed method was found to be simpler, accurate, reproducible, efficient and less time consuming and can be successfully applied for the simultaneous assay of levofloxacin and moxifloxacin formulations.HPLC, method development, validation, levofloxacin, moxifloxacin.


Tanaka M, Kurata T, Fujisawa C, Oshima Y, Aoki H, Okazaki O, et al. Mechanistic study of inhibition of levofloxacin absorption by aluminum hydroxide. Antimicrob Agents Chemother 1993; 37(10): 2173-8.

Eliopoulos GM, Eliopoulos CT, Hoope DC, Wolfson JS. In: Quinolone antibacterial agents, American Society for Microbiology, Washington 1993; pp. 161-93. (accessed on June 2011).

Ronald AR, Low DE. Fluoroquinolone antibiotics: Milestones in drug: Therapy, Birkhauser Verlag, Basel, Switzerland 2003; pp. 107-19.

Ball P. Adverse drug reactions: implications for the development of fluoroquinolones. J Antimicrob Chemother 2003; 51(Suppl. S1): 21-7.

Appelbaum PC, Hunter PA. The fluoroquinolone antibacterials: past, present and future perspectives. Int J Antimicrob Agents 2002; 16(1): 5-15.

Sultan MZ, Lee KM, Moon SS. Antibacterial effect of naturally occurring unsaturated fatty acids from Prunus japonica against Propionibacterium acnes. Orient Pharm Exp Med 2009; 9: 90-6.

Kumar PS, Krishnan SN, Kumar VN, Anilkumar G, Kumar GK. HPLC method development of levofloxacin by RP-HPLC in its bulk dosage forms. Int J Res Ayurv Pharm 2011; 2: 1790-2.

Tejakumar R, Chitra A, Amrithraj RV, Kumar NS. New RP-HPLC method development and validation for estimation of levofloxacin in tablet dosage form. J Glob Trends Pharm Sci 2011; 2(3): 264-76.

Subbaiah PR, Kumudhavalli MV, Saravanan C, Kumar M, Chandira RM. Method development and validation for estimation of moxifloxacin HCl in tablet dosage form by RP-HPLC method. Pharma Anal Acta 2010; 1(2): 1-2.

Kumar SA, Mangamma K, Anusha M, Priyadarsini JV, Kumar VR. A validated RP-HPLC method for the analysis of moxifloxacin hydrochloride in pharmaceutical dosage forms. Pharmanest 2010; 1(2): 347-52.

United States Pharmacopoeia 30 - National Formulary 25 (USP 30 - NF 25), United States Pharmacopeial Convention, Rockville, MD 2007.

International Conference on Harmonization: ICH Harmonized Tripartite Guideline- Validation of Analytical Procedures: Text and Methodology Q2 (R1): 2005.

USA Food and Drug Administration, Methods, method verification and validation; Document No.: ORA-Lab. 5.4.5, version No.: 1.5; 2009.

British Pharmacopoeia (BP)-2009, The Stationary Office, London 2002.

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.