FLOROKİNOLONLARIN HPLC-DAD İLE ANALİZİ İÇİN YENİ YÖNTEM GELİŞTİRİLMESİ

Yıl 2023, Cilt: 47 Sayı: 3, 987 - 993, 20.09.2023

Aysun Dinçel , Elif Damla Gök Topak , Feyyaz Onur

https://doi.org/10.33483/jfpau.1337313

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

• 1. Lesher, G.Y., Froelich EJ, Gruett, M.D., Bailey, J.H., Brundage, R.P. (1962). 1,8-Naphthyridine derivatives: A new class of chemotherapeutic agents. Journal of Medicinal and Pharmaceutical Chemistry, 5, 1063-1068. [CrossRef]
• 2. Andriole. V.T. (2005). The Quinolones: Past, present, and future. Clinical Infectious Diseases, 41, S113-S119. [CrossRef]
• 3. Owens, R.C. Jr, Ambrose, P.G. (2005). Amtimicrobial safety: Focus on fluoroquinolones. Clinical Infectious Disease, 40(Suppl 7), S144-S157. [CrossRef]
• 4. Bush, G.N., Diez-Santos, I., Abbott L.R., Maxwell A. (2020). Quinolones: Mechanism, Lethality and Their Contributions to Antibiotic Resistance. Molecules, 25, 5662. [CrossRef]
• 5. Mitscher, L.A. (2005). Bacterial topoisomerase inhibitors: Quinolone and pyridone antibacterial agents, Chemical Reviews, 105, 559-592. [CrossRef]
• 6. Çiftçi, E., Doğru, Ü. (2000). Florokinolonların pediatride kullanımı. Ankara Üniversitesi Tıp Fakültesi Mecmuası, 53(4), 281-291.
• 7. Pallo-Zimmerman L.M., Byron J.K., Graves T.K. (2010) Flouroquinolones: Then and now. Compendium Continuing Education for Veterinarians 32, 1-9.
• 8. Vinícius de Faria, L., Marques de Farias, D., Lisboa, T.P., Matos, M.A.C., Munoz, R.A.A., Matos, R.C. (2022). Batch injection analysis with amperometric detection for fluoroquinolone determination in urine, pharmaceutical formulations, and milk samples using a reduced graphene oxide–modified glassy carbon electrode, Analytical and Bioanalytical Chemistry 414, 5309-5318. [CrossRef]
• 9. Rambla-Alegre, M., Esteve-Romero, J., Carda-Broch, S. (2009). Validation of a MLC method with fluorescence detection for the determination of quinolones in urine samples by direct injection. Journal of Chromatography B Analytical Technologies in the Biomedical and Life Sciences, 877(31), 3975-3981. [CrossRef]
• 10. Maia, A.S., Ribeiro, A.R., Amorim, C.L., Barreiro, J.C., Cass, Q.B., Castro, P.M.L., Tiritan, M.E. (2014). Degradation of fluoroquinolone antibiotics and identification of metabolites/transformation products by liquid chromatography-tandem mass spectrometry. Journal of Chromatography A, 1333, 87-98. [CrossRef]
• 11. Watabe, S., Yokoyama, Y., Nakazawa, K., Shinozaki, K., Hiraoka, R., Takeshita, K., Suzuki, Y. (2010). Simultaneous measurement of pazufloxacin, ciprofloxacin, and levofloxacin in human serum by high-performance liquid chromatography with fluorescence detection. Journal of Chromatography B Analytical Technologies in the Biomedical and Life Sciences, 878, 1555-1561. [CrossRef]
• 12. Arroyo-Manzanares, N., Huertas-Pérez, J.F., Lombardo-Agüí, M., Gámiz-Gracia, L., GarcíaCampaña, A.M. (2015). A high-throughput method for the determination of quinolones in different matrices by ultra-high performance liquid chromatography with fluorescence detection. Analytical Methods, 7, 253-259. [CrossRef]
• 13. Kumar, A.K.H., Sudha, V., Srinivasan, R., Ramachandran, G. (2011). Simple and rapid liquid chromatography method for determination of moxifloxacin in saliva. Journal of Chromatography B Analytical Technologies in the Biomedical and Life Sciences, 879, 3663-3667. [CrossRef]
• 14. Yıldırım, S., Karakoç, H.N., Yaşar, A., Köksal, İ. (2020). Determination of levofloxacin, ciprofloxacin, moxifloxacin and gemifloxacin in urine and plasma by HPLC-FLD-DAD using pentafluorophenyl core-shell column: Application to drug monitoring. Biomedical Chromatography, 34(10), e4925. [CrossRef]
• 15. Xu, Y.H., Li, D., Liu, X.Y., Li, Y.Z., Lu, J. (2010). High performance liquid chromatography assay with ultraviolet detection for moxifloxacin: Validation and application to a pharmacokinetic study in chinese volunteers. Journal of Chromatography B Analytical Technologies in the Biomedical and Life Sciences, 878, 3437-3441. [CrossRef]
• 16. ICH, Harmonized Tripartite Guideline, Validation of analytical procedures: Text and Methodology Q2(R1), ICH Steering Commitee, 2014.
• 17. ICH, Harmonized Tripartite Guideline, on validation of analytical procedures Q2(R2), ICH Steering Commitee, 2022.
• 18. Jenke, D.R. (1996). Chromatographic method validation: A review of current practices and procedures. II. Guidelines for primary validation parameters. Journal of Liquid Chromatography and Related Technologies, 19(5), 737-757. [CrossRef]
• 19. Jenke, D.R. (1996). Chromatographic method validation: A review of current practices and procedures. III. Ruggedness, re-validation and system suitability, Journal of Liquid Chromatography and Related Technologies, 19(12), 1873-1891. [CrossRef]