A Rapid and Simple High-Performance Liquid Chromatographic Method for Determination of Levofloxacin in Human Plasma

Dion Notario; Sudibyo Martono; Zullies Ikawati; Arief Rahman Hakim; Fathul Jannah; Endang Lukitaningsih

doi:10.22146/ijc.23552

A Rapid and Simple High-Performance Liquid Chromatographic Method for Determination of Levofloxacin in Human Plasma

https://doi.org/10.22146/ijc.23552

Dion Notario⁽¹⁾, Sudibyo Martono⁽²⁾, Zullies Ikawati⁽³⁾, Arief Rahman Hakim⁽⁴⁾, Fathul Jannah⁽⁵⁾, Endang Lukitaningsih^(6*)

(1) Department of Pharmacy, Faculty of Science and Technology, Ma Chung University, Villa Puncak Bukit Tidar, Malang 65151, East Java
(2) Faculty of Pharmacy, Universitas Gadjah Mada, Sekip Utara, Yogyakarta 55281
(3) Faculty of Pharmacy, Universitas Gadjah Mada, Sekip Utara, Yogyakarta 55281
(4) Faculty of Pharmacy, Universitas Gadjah Mada, Sekip Utara, Yogyakarta 55281
(5) Faculty of Pharmacy, Universitas Gadjah Mada, Sekip Utara, Yogyakarta 55281
(6) Faculty of Pharmacy, Universitas Gadjah Mada, Sekip Utara, Yogyakarta 55281
(*) Corresponding Author

Abstract

A simple and rapid high-performance liquid chromatography method was developed and validated for quantifying LEV in human plasma. Chromatographic separation was performed under isocratic elution on a Luna Phenomenex^® C18 (150 × 4.6 mm, 5 µm) column. The mobile phase was comprised of acetonitrile, methanol, and phosphate buffer 25 mM pH 3.0 (13:7:80 v/v/v) and pumped at a flow rate of 1.5 mL/min. Detection was performed by UV detector at a wavelength of 280 nm. Samples were pre-treated with acetonitrile followed by centrifugation, evaporation, and reconstitution step. The method proved linear (r = 0.995), sensitive (LLOQ and LOD was 1.8 and 0.6 µg/mL respectively), accurate (% error above LLOQ ≤ 12% and LLOQ ≤ 20%), precise (RSD ≤ 9%), robust in the ranges of 1.8-28.8 µg/mL, rapid (separation time not more than 10 min), and simple (use no organic additive in mobile phase). The method was showed reliable for quantifying LEV in human plasma.

Keywords

levofloxacin; HPLC-UV; validation; human plasma

Full Text:

Full Text Pdf

References

[1] Naber, K.G., Roscher, K., Botto, H., and Schaefer, V., 2008, Oral levofloxacin 500 mg once daily in the treatment of chronic bacterial prostatitis, Int. J. Antimicrob. Agents, 32 (2), 145–53.

[2] Zhang, Y., Huang, H., Ren, Z., Zheng, H., Yu, Y., Lü, X., Xiao, Z.K., Yang, H.F., Xiu Q.Y., Chen, B.Y., Yue H.M., Hao Q.L., Huang, J.A., Ma, H., Xiao, W., Guo D.Y., Si B., Sun, S.H., Zhang W., Li, Q.H., Shen H.H., Duan, J., Li, H.Y., Yao W.Z., Gu J.M., Xia, Q.M., Ying, K.J., Liu, A., Yang, H.P., Shi, M.H., Sun, T.Y., Ding, G.H., and Wu, G.M., 2009, Clinical evaluation of oral levofloxacin 500 mg once-daily dosage for treatment of lower respiratory tract infections and urinary tract infections: a prospective multicenter study in China, J. Infect. Chemother., 15 (5), 301–311.

[3] Peedikayil, M.C., Alsohaibani, F.I., and Alkhenizan, A.H., 2014, Levofloxacin-based first-line therapy versus standard first-line therapy for Helicobacter pylori eradication: meta-analysis of randomized controlled trials, PloS One, 9 (1), e85620.

[4] Medscape, 2011, FDA Approves First Generic Versions of Levofloxacin. Available from: http://www.medscape.com/viewarticle/745007.

[5] Furlanut, M., Brollo, L., Lugatti, E., Di Qual, E., Dolcet, F., Talmassons, G., and Pea, F., 2003, Pharmacokinetic aspects of levofloxacin 500 mg once daily during sequential intravenous/oral therapy in patients with lower respiratory tract infections, J. Antimicrob. Chemother., 51(1), 101–106.

[6] Caufield, W.V., and Stewart, J.T., 2002, Determination of zidovudine and levofloxacin in human plasma by reversed phase HPLC and solid phase extraction, J. Liq. Chromatogr. Related Technol., 25 (12), 1791–1805.

[7] Djabarouti, S., Boselli, E., Allaouchiche, B., Ba, B., Nguyen, A.T., Gordien, J.B., Bernadou, J.M., Saux, M.C., and Breilh, D., 2004, Determination of levofloxacin in plasma, bronchoalveolar lavage and bone tissues by high-performance liquid chromatography with ultraviolet detection using a fully automated extraction method, J. Chromatogr. B, 799 (1), 165–172.

[8] Nemutlu, E., Kır, S., Özyüncü, Ö., and Beksaç, M.S., 2007, Simultaneous separation and determination of seven quinolones using HPLC: analysis of levofloxacin and moxifloxacin in plasma and amniotic fluid, Chromatographia, 66 (Suppl. 1), 15–24.

[9] Qin, G.D., Xiao, M.Z., Zhou, Y.D., Yang, J., He, H.X., He, Y, and Zeng, Y., 2013, Tamsulosin alters levofloxacin pharmacokinetics in prostates derived from rats with acute bacterial prostatitis, Asian J. Androl.,15 (2), 254–260.

[10] Kontou, P., Manika, K., Chatzika, K., Papaioannou, M., Sionidou, M., Pitsiou, G., and Kioumis, I., 2013, Pharmacokinetics of moxifloxacin and high-dose levofloxacin in severe lower respiratory tract infections, Int. J. Antimicrob. Agents, 42 (3), 262–267.

[11] Nguyen, H.A., Grellet, J., Ba, B.B., Quentin, C., and Saux, M-C., 2004, Simultaneous determination of levofloxacin, gatifloxacin and moxifloxacin in serum by liquid chromatography with column switching, J. Chromatogr. B, 810 (1), 77–83.

[12] Sousa, J., Alves, G., Campos, G., Fortuna, A., and Falcão, A., 2013, First liquid chromatography method for the simultaneous determination of levofloxacin, pazufloxacin, gatifloxacin, moxifloxacin and trovafloxacin in human plasma, J. Chromatogr. B, 930,104–111.

[13] Tasso, L., de Andrade, C., and Costa, T.D., 2011, Pharmacokinetic/pharmacodynamic modelling of the bactericidal activity of free lung concentrations of levofloxacin and gatifloxacin against Streptococcus pneumoniae, Int. J. Antimicrob. Agents, 38 (4), 307–313.

[14] Watabe, S., Yokoyama, Y., Nakazawa, K., Shinozaki, K., Hiraoka, R., Takeshita, K., and Suzuki, Y., 2010, Simultaneous measurement of pazufloxacin, ciprofloxacin, and levofloxacin in human serum by high-performance liquid chromatography with fluorescence detection, J. Chromatogr. B, 878 (19), 1555–1561.

[15] Conte, J.E., Golden, J.A., McIver, M., and Zurlinden, E., 2006, Intrapulmonary pharmacokinetics and pharmacodynamics of high-dose levofloxacin in healthy volunteer subjects, Int. J. Antimicrob. Agents, 28 (2), 114–121.

[16] Ji, H.Y., Jeong, D.W., Kim, Y.H., Kim, H-.., Sohn, D.R., and Lee, H.S., 2006, Hydrophilic interaction liquid chromatography–tandem mass spectrometry for the determination of levofloxacin in human plasma, J. Pharm. Biomed. Anal., 41 (2), 622–627.

[17] Fang, P.F., Cai, H.L., Li, H.D., Zhu, R.H., Tan, Q.Y., Gao, W., Liu, Y.P., Zhang, W.Y., Chen, Y.C., and Zhang, F., 2010, Simultaneous determination of isoniazid, rifampicin, levofloxacin in mouse tissues and plasma by high performance liquid chromatography–tandem mass spectrometry, J. Chromatogr. B, 878 (24), 2286–2291.

[18] Snyder, L.R., Kirkland, J.J., and Dolan, J.W., 2011, Introduction to Modern Liquid Chromatography, Wiley & Sons, New Jersey.

[19] Völgyi, G., Ruiz, R., Box, K., Comer, J., Bosch, E., and Takács-Novák, K., 2007, Potentiometric and spectrophotometric pK_a determination of water-insoluble compounds: Validation study in a new cosolvent system, Anal. Chim. Acta, 583 (2), 418–428.

[20] Völgyi, G., Vizseralek, G., Takacs-Novak, K., Avdeef, A., and Tam, K.Y., 2012, Predicting the exposure and antibacterial activity of fluoroquinolones based on physicochemical properties, Eur. J. Pharm. Sci., 47 (1), 21–27.

[21] Badwaik, R.T., Dashputra, A.V., and Gupta, M., 2012, Determination of levofloxacin in some commercial oral formulations by using spectrophotometer and HPLC, IJMPS, 3 (3), 14–19.

[22] Cazedey, E.C.L., and Salgado, H.R.N., 2012, Spectrophotometric determination of ciprofloxacin hydrochloride in ophthalmic solution, Adv. Anal. Chem., 2 (6), 74–79.

[23] FDA, 2001, Guidance for Industry: Bioanalytical Method Validation, US Department of Health and Human Services, Food and Drug Administration Center for Drug Evaluation Research (CDER), Center for Veterinary Medicine (CVM)

[24] FDA, 1994, Reviewer Guidance, Validation of Chromatographic Method, Analytical Methods Technical Committee of the Chemistry Manufacturing Controls Coordinating Committee (CMCCC) of the Center for Drug Evaluation and Research at the Food and Drug Administration Rockville.

[25] Almeida, A.M., Castel-Branco, M.M., and Falcão, A.C., 2002, Linear regression for calibration lines revisited: weighting schemes for bioanalytical methods, J. Chromatogr. B, 774 (2), 215–222.

DOI: https://doi.org/10.22146/ijc.23552