FLOROKİNOLONLARIN HPLC-DAD İLE ANALİZİ İÇİN YENİ YÖNTEM GELİŞTİRİLMESİ

Öz

Amaç: Bu çalışmanın amacı, farklı nesil florokinolonlardan; siprofloksasin, levofloksasin, enrofloksasin ve moksifloksasinin ayrılması ve eş zamanlı analizine olanak sağlayan yeni, kolay, hızlı ve hassas bir HPLC-DAD yöntemi geliştirmektir. Gereç ve Yöntem: Literatürlerde, etken maddede florokinolonların tek başına veya ikili karışımlarının ayrılmaları, analizi ve miktar tayinleri ile ilgili çeşitli yöntemler mevcuttur. Bu çalışmada siprofloksasin, levofloksasin, enrofloksasin ve moksifloksasin için en etkili ayırımı sağlayacak yeni bir HPLC-DAD yönteminin oluşturulması hedeflenmiştir. Farklı asidik ve bazik hareketli fazlar, tampon çözeltiler ve ayırım tipleri denenmiştir. En etkili ve seçici yöntemin XTerra, C18 (100 x 4.6 mm, tanecik boyutu 3.5 µm) analitik kolon ve metanol:borat tamponu (pH=9.1, 100 mM) içeren hareketli faz ile gradient elüsyonla 0.6 ml/dak akış hızında gerçekleştirilmiştir. Genel olarak florokinolonların kromatografik tekniklerle analizinde floresan dedektör kullanıldığı gözlenmiştir. Yaptığımız çalışmada ise ayırım 280 nm'de DAD dedektörü kullanılarak başarılmış ve siprofloksasin, levofloksasin, enrofloksasin ve moksifloksasinin eş zamanlı tayinleri gerçekleştirilmiştir. Kalibrasyon eğrileri çalışılan florokinolonların herbiri için 0.5-10 µg/ml konsantrasyon aralığında doğrusaldır. Geliştirilen yöntem için validasyon çalışmaları da yapılmıştır. Sonuç ve Tartışma: Siprofloksasin, levofloksasin, enrofloksasin ve moksifloksasinin eş zamanlı tayinine izin veren basit, hızlı, hassas ve valide bir HPLC-DAD yöntemi geliştirilmiştir.

Anahtar Kelimeler

• Enrofloksasin
• HPLC-DAD
• levofloksasin
• moksifloksasin
• siprofloksasin

THE NEW METHOD DEVELOPMENT FOR THE DETERMINATION OF FLUOROQUINOLONES BY HPLC-DAD

Öz

Objective: The aim of this study was to develop a new, simple, rapid and sensitive HPLC-DAD method for the separation and simultaneous analysis of ciprofloxacin, levofloxacin, enrofloxacin and moxifloxacin from different generation of fluoroquinolones. Material and Method: In the literature, there are various methods for the separation, analysis and quantification of fluoroquinolones alone or in binary mixtures in active substance. In this study, it was aimed to develop a new HPLC-DAD method that would provide the most effective separation for ciprofloxacin, ofloxacin, enrofloxacin and moxifloxacin. Different acidic and basic mobile phases, buffer solutions and separation types were tested. The most efficient and selective method XTerra, C18 (100 x 4.6 mm, particle size 3.5 µm) analytical column and mobile phase containing methanol:borate buffer (pH=9.1, 100 mM) were used for gradient elution at a flow rate of 0.6 ml/min. In general, fluorescent detector was used in the analysis of fluoroquinolones by chromatographic techniques. In our study, separation was achieved by using DAD detector at 280 nm and simultaneous determinations of ciprofloxacin, levofloxacin, enrofloxacin and moxifloxacin were performed. The calibration curves were linear in the concentration range of 0.5-10 µg/ml for each of the fluoroquinolones studied. Validation studies were also performed for the developed method. Result and Discussion: A simple, rapid, sensitive and validated HPLC-DAD method allowing simultaneous determination of ciprofloxacin, levofloxacin, enrofloxacin and moxifloxacin was developed.

Anahtar Kelimeler

• Ciprofloxacin
• enrofloxacin
• HPLC-DAD
• levofloxacin
• moxifloxacin

Kaynakça

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