A stability-indicating reversed-phase HPLC method for assay of milbemycin oxime (MO) and estimation of its related compounds has been developed and validated as per VICH and ICH method validation guidelines. The stability-indicating capability of this method has been demonstrated by adequate separation of all process-related impurities including all potential degradation products of milbemycin oxime (MO) that were generated using stress degradation conditions that are prescribed in ICH guideline. The stability-indicating capabilities of this method were further challenged by analyzing aged stability samples of MO active pharmaceutical ingredient (API). Chromatographic separation of MO and its related compounds was achieved using an isocratic elution at a flow rate of 0.5 mL/min on a Supelco Ascentis Express C18 column (100 mm × 3.0 mm, 2.7 µm particle size, 90 Å pore size) maintained at 50 °C. The isocratic mobile phase was composed of 30% v/v of 0.05% phosphoric acid in aqueous solution and 70% v/v of a mixture of methanol and acetonitrile (6:4, v/v). UV detection at 244 nm was employed to monitor the analytes.

已经开发出了一种稳定性指示型反相HPLC方法，用于测定米尔贝霉素肟（MO）并估算其相关化合物，并已按照VICH和ICH方法验证指南进行了验证。通过充分分离所有工艺相关的杂质，包括使用ICH指南中规定的应力降解条件产生的米尔贝霉素肟（MO）的所有潜在降解产物，已证明了该方法的稳定性。通过分析MO活性药物成分（API）的老化稳定性样品，进一步挑战了该方法的稳定性指示能力。在Supelco Ascentis Express C18色谱柱（100 mm×3.0 mm，2。保持在50°C的7 µm粒径（90Å孔径）。等度流动相由水溶液中30％v / v的0.05％磷酸和70％v / v的甲醇和乙腈的混合物（6：4，v / v）组成。采用244 nm的紫外检测来监测分析物。