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


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


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.


levofloxacin; HPLC-UV; validation; human plasma

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

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