Department of Engineering Chemistry, A. U. College of Engineering (A), Andhra University, Visakhapatnam, Andhra Pradesh, 530003, India.
Research & Development, Aurobindo Pharma Ltd. Research Centre-II, Indrakaran, Sangareddy District, Hyderabad, Telangana, 502329, India.
Anal Sci. 2024 Nov;40(11):2005-2015. doi: 10.1007/s44211-024-00633-4. Epub 2024 Jul 30.
New stability indicating related substances and assay methods for trametinib acetic acid by RP-HPLC have been developed and validated through forced degradation and mass balance studies for the first time. In related substances (RS) method, trametinib acetic acid was successfully separated from its six related substances namely, cyclopropanamide impurity, desiodo trametinib, desacetyl trametinib, trione acetamide intermediate, trione intermediate, and trione PTSA intermediate using YMC-Triart C18 (150 × 4.6 mm, 3 µm) column. Orthophosphoric acid (0.15%) in water was used as buffer. Gradient elution was programmed using mobile phase-A (buffer and acetonitrile mixture in 80:20 v/v ratio) and mobile phase-B (buffer and acetonitrile mixture in 20:80 v/v ratio). Acetonitrile and methanol mixture (1:1 v/v) was used as diluent. Flow rate, injection volume, column temperature, and wavelengths were kept as 0.8 mL/min, 10 µL, 55 °C, and 240 nm, respectively. Desacetyl trametinib and cyclopropanamide impurity were identified as potential degradation impurities in acid and base degradation conditions, respectively. Assay method specific to above six related substances was also developed. Assay of forced degradation samples was also determined, and the mass balance was established by adding total impurities formed in RS method to assay of trametinib acetic acid.
首次通过强制降解和物料平衡研究，开发并验证了用于曲美替尼醋酸盐的有关物质和含量测定的反相高效液相色谱法（RP-HPLC）新方法，具有稳定性指示能力。在有关物质（RS）方法中，使用 YMC-Triart C18（150×4.6mm，3μm）柱，成功地将曲美替尼醋酸盐与其六种有关物质（环丙酰胺杂质、去碘曲美替尼、去乙酰曲美替尼、三酮乙酰胺中间产物、三酮中间产物和三酮 PTSA 中间产物）分离。以 0.15%正磷酸水溶液作为缓冲液。采用梯度洗脱程序，流动相 A（缓冲液和乙腈的 80:20 v/v 混合比）和流动相 B（缓冲液和乙腈的 20:80 v/v 混合比）。乙腈和甲醇混合液（1:1 v/v）用作稀释剂。流速、进样量、柱温、波长分别保持在 0.8mL/min、10μL、55°C 和 240nm。在酸和碱降解条件下，分别鉴定出去乙酰曲美替尼和环丙酰胺杂质为潜在的降解杂质。还开发了专门针对上述六种有关物质的含量测定方法。对强制降解样品进行了含量测定，并通过将 RS 方法中形成的总杂质加入到曲美替尼醋酸盐的含量测定中，建立了质量平衡。
