title: Development and validation of a reversed phase liquid chromatographic method for analysis of oxytetracycline and related impurities.
abstract: A simple, robust and fast high-performance liquid chromatographic method is described for the analysis of oxytetracycline and its related impurities. The principal peak and impurities are all baseline separated in 20 min using an Inertsil C₈ (150 mm × 4.6 mm, 5 μm) column kept at 50 °C. The mobile phase consists of a gradient mixture of mobile phases A (0.05% trifluoroacetic acid in water) and B (acetonitrile-methanol-tetrahydrofuran, 80:15:5, v/v/v) pumped at a flow rate of 1.3 ml/min. UV detection was performed at 254 nm. The developed method was validated for its robustness, sensitivity, precision and linearity in the range from limit of quantification (LOQ) to 120%. The limits of detection (LOD) and LOQ were found to be 0.08 μg/ml and 0.32 μg/ml, respectively. This method allows the separation of oxytetracycline from all known and 5 unknown impurities, which is better than previously reported in the literature. Moreover, the simple mobile phase composition devoid of non-volatile buffers made the method suitable to interface with mass spectrometry for further characterization of unknown impurities. The developed method has been applied for determination of related substances in oxytetracycline bulk samples available from four manufacturers. The validation results demonstrate that the method is reliable for quantification of oxytetracycline and its impurities.
authors.affiliation: Laboratorium voor Farmaceutische Analyse, Faculteit Farmaceutische Wetenschappen, Katholieke Universiteit Leuven, O&N2, PB-923, Herestraat 49, B-3000 Leuven, Belgium.
authors.forename: Getu
authors.initials: G
authors.lastname: Kahsay
authors.forename: Fairouz
authors.initials: F
authors.lastname: Shraim
authors.forename: Philippe
authors.initials: P
authors.lastname: Villatte
authors.forename: Jacques
authors.initials: J
authors.lastname: Rotger
authors.forename: Céline
authors.initials: C
authors.lastname: Cassus-Coussère
authors.forename: Ann
authors.initials: A
authors.lastname: Van Schepdael
authors.forename: Jos
authors.initials: J
authors.lastname: Hoogmartens
authors.forename: Erwin
authors.initials: E
authors.lastname: Adams
mesh_terms: D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000737:chemistry / Q000191:economics; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D004340:Drug Contamination; D005060:Europe; D017408:Guidelines as Topic; D057230:Limit of Detection; D015394:Molecular Structure; D008970:Molecular Weight; D010118:Oxytetracycline / Q000032:analysis* / Q000737:chemistry / Q000191:economics; D011786:Quality Control; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D013056:Spectrophotometry, Ultraviolet; D013754:Tetracyclines / Q000032:analysis / Q000737:chemistry
chemical_list: D000900:Anti-Bacterial Agents; D013754:Tetracyclines; C107318:2-acetyl-2-decarboxamidooxytetracycline; D010118:Oxytetracycline