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The Ultra Performance Liquid Chromatography (UPLC) was developed and validated for determination of Norfloxacin raw material. The chromatographic conditions using Acquity UPLC BEH C18 column, 2.1 × 100 mm, 1.7 μm, column maintained at 30 °C, using 0.1% trifluoroacetic acid solution and acetonitrile mixed by gradient system as a mobile phase, flow rate 0.4 mL/min, runtime 14 minutes per injection and PDA detector at 265 nm. The chromatographic system suitability meets the suitability requirements of the United States Pharmacopoeia 41th revision (USP41). The specificity and selectivity were proved by good separation of Norfloxacin from other related compounds including Norfloxacin related compound A, Norfloxacin related compound E, Norfloxacin related compound H, Norfloxacin related compound K and degradation products. The relationship between concentration and area under curve was linear over the range of 5 to 25 μg of Norfloxacin per mL with the correlation coefficient of 0.99998. The accuracy of the results assessed by percentage recoveries of three concentration levels i.e. 5, 15 and 25 μg/mL were 100.05, 100.83 and 101.12, respectively. The system precision, performed by 3 standard solutions gave % RSD of 0.10, 0.16 and 0.15. The intermediate precision was no statistical difference of the results when the analysts, instruments and days were changed. The variations of different batches of column, column temperature, injection volume, concentration of acid in mobile phase, and flow rate did not affect the chromatographic system suitability. Limit of detection and limit of quantitation were 0.06 and 0.18 μg/mL, respectively. The UPLC method for determination of Norfloxacin raw material is simple, specific, accurate, precise, and robust. It reduces working time and reagent expense. In addition, the advantage of using not extremely high column temperature is to prolong the column life time. Therefore, the proposed method is suitable for use as a standard method for the determination of Norfloxacin raw material.

Key words: Norfloxacin, UPLC