黑顺片中6种酯型生物碱提取方法对比研究

目的：对比不同溶剂提取对黑顺片中6种酯型生物碱含量的影响，建立HPLC同时测定其含量的方法。方法：采用4种不同溶剂提取制备供试品溶液，以6种成分含量归一化法处理后综合评分为评价指标，比较4种提取溶剂对含量的影响。色谱条件：采用Zorbax SB-C₁₈（4.6 mm×250 mm，5 μm）色谱柱，以乙腈-40 mmol·L^-1醋酸铵为流动相，梯度洗脱，流速1.0 mL·min^-1，检测波长235 nm，柱温30℃。结果：6种成分含量综合评分显示，氨试液-乙醚法依次优于水、氨试液-异丙醇-乙酸乙酯和酸性甲醇法，确定氨试液-乙醚为最佳提取溶剂。苯甲酰新乌头碱、苯甲酰乌头原碱、苯甲酰次乌头碱、新乌头碱、乌头碱、次乌头碱进样量分别在0.437 5～8.750 μg（r=0.999 8），0.101～2.020 μg（r=0.999 8），0.062 5～1.250 μg（r=0.999 9），0.010 4～0.208 μg（r=0.999 8），0.014 2～0.284 μg（r=0.999 8），0.052 8～1.056 μg（r=0.999 8）范围与色谱峰面积呈良好的线性关系；加样回收率（n=6）均在95.99%～104.6%，RSD均小于3%。结论：氨试液-乙醚为6种生物碱最佳提取溶剂，建立的含量测定方法简单、准确、重复性好，为黑顺片的质量评价奠定了基础。

Objective: To compare the effect of different solvents on the content extraction of six ester-type alkaloids in Heishun Pian(Radix Aconiti Lateralis Praeparata),and establish an HPLC method for simultaneously determination of the six ester-type alkaloids.Methods: To compare the extraction efficiency of six ester-type alkaloids in Radix Aconiti Lateralis Praeparata by different extracted solvent with evaluation index of normalization to six contents.The separation was performed on a Zorbax SB-C₁₈(4.6 mm×250 mm,5 μm) column with the gradient elution of acetonitrile-40 mmol·L^-1 ammonium acetate.The flow rate was 1 mL·min^-1,and the detection wavelength was set at 235 nm.The column temperature was maintained at 30℃.Results: The solvent of ammonia solution-diethyl ether was the best among water,ammonia solution-ethyl acetate and acid methanol. Benzoylmesaconine,benzoylaconine,benzoylhypaconine,mesaconine,aconine and hypaconine had good linearity in the ranges of 0.437 5-8.750 μg(r=0.999 8),0.101-2.020 μg(r=0.999 8),0.062 5-1.250 μg(r=0.999 9), 0.010 4-0.208 μg(r=0.999 8),0.0142-0.284 μg(r=0.999 8),and 0.052 8-1.056 μg(r=0.999 8), respectively.The RSDs of recoveries(95.99%-104.6%) were all below 3%.Conclusion: This method,based on the solvent of ammonia solution-diethyl ether,is simple,reliable and accurate,which can provide a basis for Radix Aconiti Lateralis Praeparata quality evaluation and quality control.