一测多评法同时测定莨菪浸膏片中3个生物碱的含量

目的：建立同时测定莨菪浸膏片中阿托品、东莨菪碱、山莨菪碱含量的一测多评法。方法：采用Phenomenex C₁₈色谱柱（4.6 mm×250 mm，5 μm），流动相为甲醇（A）-0.05%磷酸水溶液（B），梯度洗脱，流速为1.0 mL·min^-1，柱温为30℃，检测波长为218 nm。以硫酸阿托品为内参物，计算阿托品、东莨菪碱、山莨菪碱的相对校正因子，采用一测多评法和外标法分别测定3个生物碱含量，并比较两者测定结果的差异。结果：经方法学验证，分离度、精密度、重复性、稳定性良好，4批莨菪浸膏片中阿托品、东莨菪碱、山莨菪碱质量浓度分别在分别在22.00~66.01、9.48~28.45、6.32~18.97 μg·mL^-1范围内线性关系良好。阿托品、东莨菪碱、山莨菪碱的相对校正因子分别为0.855 1、0.950 3、0.760 5，各相对校正因子重现性良好。一测多评法测得批号为1509559、151004、151005、151006的样品中阿托品含量分别为15.788、26.008、26.254、26.518μg·片^-1，东莨菪碱含量分别为7.765、1.117、1.121、1.009 μg·片^-1，山莨菪碱含量分别为0.605、0.823、0.654、0.482 μg·片^-1，其结果与外标法测定结果基本一致。结论：本法可用于莨菪浸膏片中莨菪烷类生物碱的质量控制。

Objective: To establish a quantitative analysis of multi-components by single-marker(QAMS)method for simultaneous determination of atropine,scopolamine and anisodamine in Hyoscyamus extract tablets.Methods: The separation was carried out on a Phenomenex C₁₈ column(4.6 mm×250 mm,5 μm)with methanol(A)and 0.05% phosphoric acid solution(B)as mobile phase in gradient elution program.The flow rate was 1.0 mL·min^-1;the column temperature was 30℃;the detection wavelength was set at 218 nm.Atropine sulfate was selected as the internal reference substance.The relative correction factors(RCFs)of atropine,scopolamine and anisodamine were calculated.The contents of three alkaloids were determined by both external standard method(ESM)and QAMS method and the results were compared.Results: The results of resolution,accuracy,precision and stability were satisfactory in method validation.Good linear correlations were found in the ranges of 22.00-66.01 μg·mL^-1,9.48-28.45 μg·mL^-1 and 6.32-18.97 μg·mL^-1 for atropine,scopolamine and anisodamine,respectively.RCFs of atropine,scopolamine and anisodamine were 0.855 1,0.950 3 and 0.760 5,respectively with good reproducibility.Through QAMS quantitative method,the contents of atropine in samples No.1509559,151004,151005,151006 were 15.788,26.008,26.254,26.518 μg per tablet,respectively;the contents of scopolamine were 7.765,1.117,1.121,1.009 μg per tablet,respectively;the contents of anisodamine were 0.605,0.823,0.654,0.482 μg per tablet,respectively.The quantitative results of external standard method and QAMS method were basically identical.Conclusion: The QAMS method can be used for the quality control of tropane alkaloids in Hyoscyamus extract tablets.