Abstract
Ofloxacin is an antibiotic from second generation of fluoroquinolones group. Concentration of ofloxacin in human plasma is in low level, and due to that fact, it requires a selective, accurate, sensitive method of analysis. 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, SunfireTM 5 µm; 250 x 4.6 mm) column with an isocratic mobile phase consisted of triethylamine 1% in water pH 3.0–acetonitrile (84:16) in the flow rate of 1.0 mL/min, 40oC column temperature 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.
Bahasa Abstract
Ofloksasin merupakan salah satu antibiotika golongan fluorokuinolon generasi kedua. Konsentrasi ofloksasin dalam plasma kecil, sehingga diperlukan metode analisis yang selektif, akurat, dan sensitif. Pada penelitian ini, dilakukan optimasi dan validasi metode analisis ofloksasin dalam plasma in vitro menggunakan kromatografi cair kinerja tinggi deteksi fluoresensi dengan siprofloksasin HCl sebagai baku dalam. Pemisahan dilakukan dengan menggunakan kolom C18 (Waters, SunfireTM 5 µm; 250 x 4,6 mm) dengan fase gerak isokratik yang terdiri dari trietilamin 1% dalam air pH 3,0–asetonitril (84:16), laju alir 1,0 mL/menit, suhu kolom 40oC, dan deteksi ofloksasin dilakukan pada panjang gelombang eksitasi 300 nm dan emisi 500 nm. Proses ekstraksi plasma dilakukan dengan metode pengendapan protein menggunakan metanol melalui proses vorteks selama 2 menit dan sentrifugasi pada kecepatan 10000 rpm selama 10 menit. Validitas dan linearitas metode analisis berada pada rentang konsentrasi 21,4 ng/mL – 4280 ng/mL dengan LLOQ 21,4 ng/mL. Nilai % diff dan koefisien variasi untuk akurasi dan presisi intra hari dan antar hari tidak lebih dari + 20% untuk konsentrasi LLOQ dan + 15% untuk konsentrasi rendah, sedang, dan tinggi. Ofloksasin dalam plasma dinyatakan stabil selama 3 siklus beku dan cair, stabil minimal 24 jam pada suhu kamar dan selama minimal 28 hari pada suhu -20oC. Metode analisis ofloksasin dalam plasma ini sudah memenuhi kriteria penerimaan sesuai persyaratan validasi pedoman EMEA.
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Recommended Citation
Tania, Letitia; Sitepu, Eme Stepani; and Harahap, Yahdiana
(2016)
"Validasi Metode Analisis Ofloksasin dalam Plasma In Vitro secara Kromatografi Cair Kinerja Tinggi-Fluoresensi Mengacu pada European Medicines Agency Guideline,"
Pharmaceutical Sciences and Research: Vol. 3:
No.
2, Article 1.
DOI: 10.7454/psr.v3i2.3518
Available at:
https://scholarhub.ui.ac.id/psr/vol3/iss2/1
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Natural Products Chemistry and Pharmacognosy Commons, Other Pharmacy and Pharmaceutical Sciences Commons, Pharmaceutics and Drug Design Commons
