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"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. Waktu retensi ofloksasin dan siprofloksasin HCl adalah 7,476 menit dan 8,446 menit dengan total waktu analisis 10 menit. 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. Hasil penelitian menunjukkan bahwa spesifisitas dan selektivitas metode ekstraksi ditunjukkan dengan tidak adanya puncak pengganggu pada saat retensi analit dan baku dalam. 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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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. Retention time of ofloxacin and ciprofloxacin-HCl were 7.476 and 8.446 minutes respectively with total analytical run time was 10 minutes. Plasma extraction was done by eproteination using methanol, through the process of vortex and centrifugation (10000 rpm) for 2 minutes and 10 minutes consecutively. The results showed that the extraction method was specific and selective as there was no interfering peaks of human plasma at the retention times of the ofloxacin and the internal standard. 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 guidelines."

"Infeksi saluran kemih (ISK) merupakan penyakit infeksi yang cukup sering terjadi di masyarakat. Dari berbagai penelitian di Indonesia dan di luar negeri, telah menunjukkan penurunan kepekaan bakteri penyebab ISK terhadap antibiotik golongan fluorokuinolon. Hal ini dikhawatirkan menjadi kendala dalam penanggulangan ISK di Indonesia. Penelitian ini bertujuan untuk mengetahui pola kepekaan bakteri Gram negatif yaitu Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeuroginosa Enterobacter aerogenes, dan Proteus mirabilis dari penderita infeksi saluran kemih terhadap siprofloksasin, gatifloksasin, ofloksasin, dan moksifloksasin. Penelitian ini dilakukan dengan cara menganalisis data sekunder sebanyak 3268 isolat urin dengan kultur positif dari Laboratorium Mikrobiologi Klinik FKUI pada Januari 2001 sampai Desember 2005 dan telah dilakukan uji resistensi sesuai dengan NCCLS. Dari hasil analisis didapatkan angka kepekaan Escherichia coli terhadap siprofloksasin, ofloksasin, gatifloksasin dan moksifloksasin adalah 54.5%, 59.4%, 54.5%, dan 38.0%; kepekaan Klebsiella pneumoniae terhadap siprofloksasin, ofloksasin, gatifloksasin dan moksifloksasin adalah 46.0%, 54.2%, 48.1%, dan 34.9%; kepekaan Pseudomonas aeruginosa terhadap siprofloksasin, ofloksasin, gatifloksasin dan moksifloksasin adalah 43.9%, 43.9%, 44.9%, dan 38.1%; kepekaan Enterobacter aerogenes terhadap siprofloksasin, ofloksasin, dan gatifloksasin adalah 58.7%, 63.8%, dan 65.5%; kepekaan Proteus mirabilis terhadap siprofloksasin, ofloksasin, dan gatifloksasin adalah 80.5%, 83.9%, dan 70.0%.

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Urinary tract infection (UTI) is a common infectious disease in the community practice. Studies in Indonesia and overseas showed the decrease of sensitivity of bacteria causing UTI to fluoroquinolone. This problem is potentially leading to difficulty in the treatment of UTI in Indonesia.

This study objective was to investigate the sensitivity pattern of Gram negative bacteria such as Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeuroginosa, Enterobacter aerogenes, and Proteus mirabilis taken from UTI patient to ciprofloxacin, gatifloxacin, ofloxacin, and moxifloxacin.

This study was conducted by analyzing secondary data of 3268 isolated urine with positive culture from Clinical Microbiology Laboratory of FMUI since January 2001 to December 2005. Resistance test had been performed in guidance of NCCLS.

Results of the analysis indicate that sensitivity patterns of Escherichia coli to ciprofloxacin, ofloxacin, gatifloxacin, moxifloxacin were 54.5%, 59.4%, 54.5%, and 38.0%, respectively; Klebsiella pneumoniae to ciprofloxacin, ofloxacin, gatifloxacin, and moxifloxacin were 46.0%, 54.2%, 48.1%, and 34.9%; Pseudomonas aeruginosa to ciprofloxacin, ofloxacin, gatifloxacin, and moxifloxacin were 43.9%, 43.9%, 44.9%, and 38.1%; Enterobacter aerogenes to ciprofloxacin, ofloxacin, and gatifloxacin were 58.7%, 63.8%, and 65.5%; Proteus mirabilis to ciprofloxacin, ofloxacin, and gatifloxacin were 80.5%, 83.9%, and 70.0%."