A Rapid and Simple High-Performance Liquid Chromatographic Method for Determination of Levofloxacin in Human Plasma
ABSTRACT: To conduct a
bioequivalence study for a copy product of levofloxacin (LEV), a simple and
validated analytical method was needed, but the previous developed methods were
still too complicated. For this reason, a simple and rapid high performance
liquid chromatography method was developed and validated for LEV quantification
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 that
adjusted at pH 3.0 (13:7:80 v/v/v) and pumped at a flow rate of 1.5 mL/min.
Detection was performed under UV detector at wavelength of 280 nm. Samples were
prepared by adding acetonitrile and followed by centrifugation to precipitate
plasma protein. Then followed successively by evaporation and reconstitution
step. The optimized method meets the requirements of validation parameters
which included linearity (r = 0.995), sensitivity (LLOQ and LOD was 1.77 and
0.57 µg/mL respectively), accuracy (%error above LLOQ ≤ 12% and LLOQ ≤ 20%),
precision (RSD ≤ 9%), and robustness in the ranges of 1.77-28.83 µg/mL.
Therefore, the method can be used as a routine analysis of LEV in human plasma
as well as in bioequivalence study of LEV.
Author: Dion Notario, Sudibyo
Martono, Zullies Ikawati, Arief Rahman Hakim, Fathul Jannah, Endang
Lukitaningsih
Journal Code: jpkimiagg170018
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