Pharmaceutics and Pharmacokinetic Division, CSIR-Central Drug Research Institute, Lucknow, India.
Pharmaceutics and Pharmacokinetics Division of CSIR-Central Drug Research Institute, Jawaharlal Nehru University, New Delhi, India.
J Mass Spectrom. 2024 Oct;59(10):e5088. doi: 10.1002/jms.5088.
Bacterial keratitis (BK) is an infection that causes inflammation of the cornea and, if severe, can result in blindness. Topical fluoroquinolones combined with corticosteroids have been shown to be useful in the treatment of BK. A rapid, selective, and sensitive bioanalytical method for simultaneous quantification of Gatifloxacin (GAT) and Dexamethasone (DEX) has been developed and validated using tandem mass spectrometry (LC-MS/MS). Optimal separation was accomplished in under 5 min using an Agilent Zorbax C18 column (100 mm × 4.6 mm, 3.5 μm). The mobile phase was composed of a blend of 0.2% formic acid in triple distilled water and methanol with a flow rate of 0.65 mL/min in isocratic mode. GAT and DEX were detected in positive electrospray ionization multiple reaction monitoring mode (MRM), and the retention time was found to be at 1.64 and 2.93 min, respectively. The linearity of GAT and DEX was found to be in the range of 1.56-400 ng mL with good precision and accuracy. The method was validated according to USFDA regulatory guidelines. The validated method was effectively utilized for preclinical pharmacokinetic analysis of GAT and DEX in rabbit tear fluid following the topical application of a commercial formulation.
细菌性角膜炎 (BK) 是一种感染，可引起角膜炎症，如果严重，可能导致失明。局部应用氟喹诺酮类药物联合皮质类固醇已被证明对 BK 的治疗有效。本研究建立并验证了一种使用串联质谱（LC-MS/MS）同时定量检测加替沙星（GAT）和地塞米松（DEX）的快速、选择性和灵敏的生物分析方法。采用安捷伦 Zorbax C18 柱（100mm×4.6mm，3.5μm），在 5min 内即可实现最佳分离。流动相由三重蒸馏水和甲醇中的 0.2%甲酸混合而成，以 0.65mL/min 的流速在等度模式下运行。GAT 和 DEX 采用正电喷雾电离多反应监测模式（MRM）进行检测，保留时间分别为 1.64 和 2.93min。GAT 和 DEX 的线性范围为 1.56-400ng/mL，具有良好的精密度和准确度。该方法符合美国 FDA 监管指南的要求。该方法已成功应用于商业化制剂局部给药后兔泪液中 GAT 和 DEX 的临床前药代动力学分析。
