Validasi Metode Analisis Ofloksasin dalam Plasma In Vitro secara Kromatografi Cair Kinerja Tinggi-Fluoresensi Mengacu pada European Medicines Agency Guideline
Abstract: Ofloxacin is an
antibiotic from second generation of fluoroquinolone's group. Concentration of
ofloxacin in human plasma is low, so it requires a selective, accurate,
sensitive method for analysis it. In this study, the optimization and
validation of ofloxacin analysis in human plasma using high performance liquid
chromatography-fluorescence with ciprofloxacin-HCl as an internal standard were
carried out. Separation of ofloxacin was performed using C18 (Waters, Sunfire
TM 5 µm; 250 x 4.6 mm) column with an isocratic mobile phase consisted of
triethylamine 1% on water pH 3.0–acetonitrile (84:16) an in the flow rate of
1.0 mL/min, and on column temperature
40oC whereas the detection was carried
out at excitation of 300 nm and emission of 500 nm. Plasma extraction was done
by deproteination using methanol, through the process of vortex and
centrifugation (10000 rpm) for 2 minutes and 10 minutes consecutively. The
method was valid and linear within the concentration ranged from 21,4 ng/mL to
4280 ng/mL with LLOQ of 21,4 ng/mL. Intra-day and inter-day accuracy and
presicion was not more than + 20% for LLOQ and not more than + 15% for QCL,
QCM, and QCH samples in both % diff and coefficient of variation. Ofloxacin was
stable in human plasma at least three freeze and thaw cycle, for at least 24
hours in room temperature and 28 days at -20oC. This bioanalytical method
fulfilled the acceptance criteria following EMEA guideline.
Keywords: siprofloksasin HCL;
fluoresensi; KCKT; ofloksasin; validasi; ciprofloxacin HCl, fluoresence, HPLC,
ofloxacin, validation
Penulis: Letitia Tania , Eme
Stepani Sitepu , Yahdiana Harahap
Kode Jurnal: jpfarmasidd160721
