Abstract：
A reverse phase high-performance liquid chromatography (HPLC) method has been developed for the quantification of a typical drug Dasatinib (DST) and its related impurities in pharmaceuticals. Kinetex C18 (4.6 × 150 mm, 5 μm) column was used in the chromatographic separations, using buffer (1.36 g of KH2PO4 in 1000 mL of water, pH = 7.8; adjusted with diluted KOH solution) with solvent as acetonitrile and mode of elution as the gradient. The flow rate is 0.9 mL/min, column oven temperature as 45°C and the overall gradient run time as 65 min. The developed method was found to produce symmetric and good separation between the process-related and degradation impurities. Method optimization is achieved with photodiode array at 305 nm over the concentration range of 0.5 mg/mL and degradation studies were carried out under acidic, alkaline, oxidative, photolytic and thermal conditions to demonstrate the stability indicating capability of the method. Two major impurities were found in forced degradation studies in the HPLC analysis, the unknown, acid degradants were enriched and isolated by preparative HPLC, then characterized through high-resolution mass spectrometry, nuclear magnetic resonance spectroscopy and Fourier transform infrared spectroscopy. The unknown acid degradation impurity was showing Exact Mass of 521.11, molecular formula C22H25Cl2N7O2S and its chemical name as 2-(5-chloro-6-(4-(2-hydroxyethyl) piperazin-1-yl)-2-methylpyrimidin-4-ylamino)-N-(2-chloro-6-methylphenyl) thiazole-5-carboxamide. Another impurity (oxidative degradant) found as known DST N-oxide Impurity-L and its chemical name as 4-(6-((5-((2-chloro-6-methylphenyl) carbamoyl) thiazol-2-yl) amino)-2-methylpyrimidin-4-yl)-1-(2-hydroxyethyl) piperazine 1-oxide. The analytical HPLC method was further validated as per ICH guidelines.
摘要：
已开发出一种反相高效液相色谱（HPLC）方法，用于定量药物中的典型药物达沙替尼（DST）及其相关杂质。KinetexC18(4.6×150mm,5μm)柱用于色谱分离，使用缓冲液（1.36克KH2PO4在1000毫升水中，pH=7.8;用稀释的KOH溶液调节),溶剂为乙腈,洗脱模式为梯度。流速为0.9mL/min,柱烘箱温度为45°C，总梯度运行时间为65分钟。发现所开发的方法在与过程相关的杂质和降解杂质之间产生对称且良好的分离。在0.5mg/mL的浓度范围内，使用305nm的光电二极管阵列实现了方法优化，并在酸性条件下进行了降解研究，碱性,氧化，光解和热条件，以证明该方法的稳定性指示能力。在HPLC分析的强制降解研究中发现了两种主要杂质，未知,通过制备型HPLC富集和分离酸降解物，然后通过高分辨率质谱表征，核磁共振光谱和傅里叶变换红外光谱。未知的酸降解杂质显示出521.11的确切质量，分子式为C22H25Cl2N7O2S，其化学名称为2-（5-氯-6-（4-（2-羟乙基）哌嗪-1-基）-2-甲基嘧啶-4-基氨基）-N-（2-氯-6-甲基苯基）噻唑-5-甲酰胺。发现的另一种杂质(氧化降解剂)为已知的DSTN-氧化物杂质-L,其化学名称为4-(6-(5-((2-氯-6-甲基苯基)氨基甲酰基)噻唑-2-基)氨基)-2-甲基嘧啶-4-基)-1-(2-羟乙基)哌嗪1-氧化物。根据ICH指南进一步验证分析型HPLC方法。
