title: Development and validation of stability indicating HPLC methods for related substances and assay analyses of amoxicillin and potassium clavulanate mixtures.
abstract: Antibacterial combinations consisting of the semisynthetic antibiotic amoxicillin (amox) and the β-lactamase inhibitor potassium clavulanate (clav) are commonly used and several chromatographic methods were reported for their quantification in mixtures. In the present work, single HPLC method for related substances analyses of amoxicillin and potassium clavulanate mixtures was developed and validated according to international conference on harmonization (ICH) guidelines. Eighteen amoxicillin and six potassium clavulanate impurities were successfully separated from each other by using triple gradient elution using a Thermo Hypersil Zorbax BDS C18 (250 mm×4.6mm, 3μm) column with 50μL injection volumes at a wavelength of 215nm. Commercially unavailable impurities were formed by degradation of amoxicillin and potassium clavulanate, identified by LC-MS studies and used during analytical method development and validation studies. Also, process related amoxicillin impurity-P was synthesized and characterized by using nuclear magnetic resonance (NMR) and mass spectroscopy (MS) for the first time. As complementary of this work, an assay method for amoxicillin and potassium clavulanate mixtures was developed and validated; stress-testing and stability studies of amox/clav mixtures was carried out under specified conditions according to ICH and analyzed by using validated stability-indicating assay and related substances methods.
authors.affiliation: DEVA Holding A.Ş., Çerkezköy-2 Production Plant, Karaağaç Mh. Fatih Blv. No: 26, Address No: 2278035833, Kapaklı, Tekirdağ, Turkey. Electronic address: ebellur@deva.com.tr.
authors.forename: Esen
authors.initials: E
authors.lastname: Bellur Atici
authors.affiliation: DEVA Holding A.Ş., Çerkezköy-2 Production Plant, Karaağaç Mh. Fatih Blv. No: 26, Address No: 2278035833, Kapaklı, Tekirdağ, Turkey.
authors.forename: Yücel
authors.initials: Y
authors.lastname: Yazar
authors.affiliation: DEVA Holding A.Ş., Çerkezköy-2 Production Plant, Karaağaç Mh. Fatih Blv. No: 26, Address No: 2278035833, Kapaklı, Tekirdağ, Turkey.
authors.forename: Çağan
authors.initials: Ç
authors.lastname: Ağtaş
authors.affiliation: DEVA Holding A.Ş., Çerkezköy-2 Production Plant, Karaağaç Mh. Fatih Blv. No: 26, Address No: 2278035833, Kapaklı, Tekirdağ, Turkey.
authors.forename: Nurten
authors.initials: N
authors.lastname: Ridvanoğlu
authors.affiliation: DEVA Holding A.Ş., Çerkezköy-2 Production Plant, Karaağaç Mh. Fatih Blv. No: 26, Address No: 2278035833, Kapaklı, Tekirdağ, Turkey.
authors.forename: Bekir
authors.initials: B
authors.lastname: Karlığa
mesh_terms: D019980:Amoxicillin-Potassium Clavulanate Combination / Q000032:analysis* / Q000737:chemistry / Q000592:standards; D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000737:chemistry / Q000592:standards; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004340:Drug Contamination / Q000517:prevention & control*; D004355:Drug Stability; D015394:Molecular Structure; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000900:Anti-Bacterial Agents; D019980:Amoxicillin-Potassium Clavulanate Combination
keywords: Amoxicillin; Assay; Clavulanic acid; Related substances; Stress-testing; Validation