盐酸替哌拉索杂质结构鉴定

Identification of a Degradation Impurity of Tipiracil Hydrochloride

目的:对盐酸替哌拉索碱性强制降解杂质进行结构鉴定,并建立盐酸替哌拉索及有关物质的HPLC测定方法。方法:以HPLC法对碱性强制降解杂质进行分析,并通过制备得到杂质单体,根据Q-TOF/MS以及~1H-NMR、^(13)C-NMR、~2D-NMR技术对其结构进行分析确证。采用Waters Atlantis T3柱(4.6 mm×150 mm,3μm)对盐酸替哌拉索的降解杂质和工艺杂质进行定量分析,以磷酸二氢钾、庚烷磺酸钠缓冲液(p H 3.0)-甲醇为流动相,梯度洗脱,检测波长为210 nm。结果:综合解析确证盐酸替哌拉索碱性强制降解杂质结构为4-(((5-氯-2,6-二酮-1,2,3,6-四氢嘧啶-4-基)甲基)氨基)丁酰胺。在该HPLC测定方法条件下主峰与各杂质峰间能达到基线分离,盐酸替哌拉索及其杂质检测限为0.05μg·m L-1,定量限为0.15μg·m L-1。结论:本研究鉴定了盐酸替哌拉索原药在剧烈条件下产生的杂质;此高效液相色谱法选择性高、重复性好,可作为盐酸替哌拉索原药质量控制的方法。

Objective: To identify the alkaline degradation product of tipiracil hydrochloride and establish an HPLC method for the determination of its related substances. Methods: The alkaline degradation product was analyzed by HPLC and its structural confirmation was unambiguously carried out by preparation followed by characterization using Q-TOF/MS and NMR. The test for degradation and process impurities of tipiracil hydrochloride is based on a Waters Atlantis T3 column(4.6 mm×150 mm, 3 μm); the mobile phase consisted of potassium dihydrogen phosphate and sodium heptane-sulphonate buffer solution(adjust to p H 3.0)- methanol with gradient elution; the detection wavelength was 210 nm. Results: Based on the spectral data, the alkaline degradation product of tipiracil hydrochloride was characterized as 4-(((5-chloro-2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)methyl)amino)butanamide. The main peak and every impurity peak were separated well. The LOD and LOQ of tipiracil hydrochloride and its impurities were 0.05 μg·m L-1and 0.15 μg·m L-1, respectively. Conclusion: The investigation identified the impurity of tipiracil hydrochloride produced under high p H conditions and the highly selective and reproducible HPLC offered an option in the quality control of tipiracil hydrochloride.