Validation and Application of a New Optimized RP-HPLC-Fluorescent Detection Method for Norfloxacin
A new reverse phase-high performance liquid chromatography (RP-HPLC) method with fluorescent detector (FLD) was developed and optimized for Norfloxacin determination in human plasma. Mobile phase specifications, extraction method and excitation and emission wavelengths were varied for optimization. HPLC system contained a reverse phase C18 (5 μm, 4.6 mm×150 mm) column with FLD operated at excitation 330 nm and emission 440 nm. The optimized mobile phase consisted of 14% acetonitrile in buffer solution. The aqueous phase was prepared by mixing 2g of citric acid, 2g sodium acetate and 1 ml of triethylamine in 1 L of Milli-Q water was run at a flow rate of 1.2 mL/min. The standard curve was linear for the range tested (0.156–20 μg/mL) and the coefficient of determination was 0.9978. Aceclofenac sodium was used as internal standard. A detection limit of 0.078 μg/mL was achieved. Run time was set at 10 minutes because retention time of norfloxacin was 0.99 min. which shows the rapidness of this method of analysis. The present assay showed good accuracy, precision and sensitivity for Norfloxacin determination in human plasma with a new internal standard and can be applied pharmacokinetic evaluation of Norfloxacin tablets after oral administration in human.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1084960Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF
 A. Delon, F. Huguet, P. Courtois, J.M. Vierfond, S. Bouquet and W. Couet. (1997, February). Pharmacokinetic-pharmacodynamic contributions to the convulsant activity of Pefloxacin and Norfloxacin in rats. J. Pharmacol. Exp. Ther. (Online). 280(2): 983-987. Available: http://www.springerlink.com/index/T81R444M 32M30185.pdf
 H. Salem. (2005, February). Spectrofluorimetric, Atomic absorption spectrometric and spectrophotometric determination of some fluoroquinolones. Am. J. Appl Sci. (Online). 2(3): 719-729. Available: http://www.scipub.org/fulltext/ajas/ajas23 719-729.pdf
 K.A. Al-Rashood, K.I. Al-Khamis, Y.M. El-Sayed, S. Al-Bella, M.A. Al-Yaman, S.M. Alam and R. Dha. (2000, July). Bioequivalence evaluation of Norfloxacin 400 mg tablets (Uroxin and Noroxin) in healthy human volunteers. J. Biopharm. Drug Dispos. (Online). 21(5): 177-179. Available: http://www. repository.ksu.edu.sa
 V.K. Boppana and B.N. Swanson. (1982, May). Detrmination of Norfloxacin, a new nalidixic acid analog, in human serum and urine by HPLC. J. Antimicrob. Chemother. (Online). 21(5): 808-810. Available: http:// www.aac.asm.org/cgi/reprint/21/5/808.pdf
 Merck & Company. Product information. Noroxin (R), Norfloxacin. Merck & Company. (2001, August). Available: http:// www.merck.com/product/home.html
 L.T. Pauliukonis, D.G. Musson and W.F. Bayne. (1984, January). Quantitation of Norfloxacin, a new antibacterial agent in human plasma and urine by ion-pair reverse -phase chromatography. J. pharm. Sci. (Online). 73(1): 99-102. http://www.ncbi.nlm.nih.gov/pubmed/622 9625.
 J.M. Hermann and K. Christian. (1998, July). Determination of norfloxacin in human plasma and urine by high-performance liquid chromatography and fluorescence detection. J. Chromatogr. A. (Online). 812 (1-2): 381-385. Available: http://www.linkinghub.elsevier.com/retrieve/pii/S0021967398004014.
 V.F. Samanidou, E.A. Christodoulou and I.N. Papadoyannis. (2003, March). Direct determination of four quinolones, Enoxacin, Norfloxacin, Ofloxacin & Ciprofloxacin, in pharmaceutical & blood serum by HPLC. Anal. Bioanal. Chem. (Online). 375(5): 623-9. http://www.springerlink.com/index/MR6HG7H0WN7QGY21.pdf
 S.J. Morton, V.H. Shull and J.D. Dick. (1986, August). Determination of Norfloxacin and Ciprofloxacin in serum & urine by HPLC. J. antimicrobial. chemother. (Online). 30(2): 325-332. Available: http:// www.ncbi.nlm.nih.gov