盐酸消旋山莨菪碱注射液有关物质的研究

目的：采用液质联用和核磁共振技术确定盐酸消旋山莨菪碱注射液中未知杂质的结构，并采用高效液相色谱法对该杂质进行控制。方法：采用Agilent 1200-6320 IontrapXCT液相色谱/质谱联用仪及核磁共振仪对未知杂质分析，确定结构后，采用外标法进行该杂质的测定。使用C₁₈色谱柱（250 mm×4.6 mm，5 μm），以0.01 mol·L^-1磷酸二氢钾（含0.15%三乙胺溶液，用磷酸调pH 6.5）-甲醇（70：30）为流动相；流速1.0 m·L min^-1，检测波长为220 nm。结果：液质联用和核磁共振的结果显示，未知杂质为盐酸消旋山莨菪碱降解产物托品酸。外标法测定过程中，HPLC各杂质峰与主峰均分离良好，方法的精密度良好，杂质托品酸在3.77~11.31 μg·mL^-1范围内与峰面积呈良好的线性关系，回归方程为Y=28.68X-2.632， r=0.9999，定量限为0.0377 μg·mL^-1。结论：研究确定了杂质托品酸的结构，并建立了HPLC检测方法，经方法学验证，该法适用于有关物质的测定。

Objective: To do a structure analysis of unknown impurities in raceanisodamine hydrochloride injection by LC-MS and NMR and determinate the related substance by external standard method. Methods: Agilent 1200-6320 IontrapXCT LC-MS,¹H-NMR and ¹³C-NMR were used to determine structures of unknown impurities.After the structure was identified,the determination of the impurity was carried out by external standard method.The separation was performed on a C₁₈ column(250 mm×4.6 mm,5 μm);the mobile phase was 0.01 mol·L^-1 potassium dihydrogen phosphate solution (including 0.15% triethylamine,adjusted to pH 6.5 with phosphoric acid)-methanol (70:30);the flow rate was 1.0 mL·min^-1 and the detection wavelength was 220 nm. Results: The unknown impurity was tropic acids,which were the degradation product of raceanisodamine.Under the external standard method,all impurity peaks in the HPLC chromatogram were separated well from the main peak;the reproducibility and precision of the method were good;the linear range of tropic acid was 3.77-11.31 μg·mL^-1,and the linear equation was Y=28.68X-2.632,r=0.9999.The limit of quantification was 0.0377 μg·mL^-1 Conclusion: The impurity was tropic acid,whose structure was ascertained through study.An HPLC method was established,and methodology verification proved that the method was suitable for determination of related substances in raceanisodamine hydrochloride injection.

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