目的：建立同时测定乌头碱等6 种单酯和双酯型生物碱含量的高效液相色谱法，对生附子、黑顺片和白附片及生附子与大黄配伍后生物碱的含量进行测定，从化学成分角度为阐释附子炮制及配伍减毒提供参考。方法：采用Phenomenex Gemini C18 色谱柱（4.6 mmx250 mm，5 μm），以乙腈-40 mmol·L-1 醋酸铵缓冲液（氨水调pH9.8）为流动相，梯度洗脱，流速1.0 mL·min-1，检测波长235 nm。结果：附子中6 种酯型生物碱分离良好，各成分的质量浓度与峰面积在测定范围内均呈现良好的线性关系（r＞0.999），平均加样回收率为96.9%~102.4%；与生附子相比，黑顺片、白附片及生附子大黄配伍中双酯型生物碱的含量均明显降低。结论：该方法稳定可靠、简便易行，可用于附子类药材中6 种酯型生物碱的同时定量分析；炮制或配伍可明显降低附子中生物碱类成分的含量，从而降低其毒性。

This study was aimed to establish an HPLC method to simultaneous determine 6 kinds of monoester and diester aconitum alkaloids. The content of alkaloids in aconite roots, black and white prepared lateral root of aconite and the compatibility of aconite roots with rhubarb were determined. This study provided reference for the interpretation of the attenuation of processing and compatibility of medicines from chemical component aspect. The HPLC analysis was performed on a Phenomenex Gemini C18 (4.6 mm x250 mm, 5 μm) with 40 mmol·L-1 ammonium acetate (adjusted to pH 9.8 with ammonia water) and acetonitrile as mobile phase, with a gradient elution at the flow rate of 1.0 mL·min-1 and a detection wavelength of 235 nm. The results showed that 6 alkaloids in aconite roots achieved favorable separation and a good linearity relationship (r > 0.999) over the studied concentration range. The extraction recoveries were ranged from 96.9% to 102.4% for the 6 alkaloids. The content of diester alkaloids decreased markedly in processed products and the compatibility of aconite roots with rhubarb compared with the raw aconite roots. It was concluded that this method was stable, reliable, simple and practical. It can be used for the simultaneous determination of 6 kinds of monoester and diester aconitum alkaloids in aconite roots. This processing and compatibility can significantly reduce the content of alkaloids in aconite roots in order to reduce its toxicity.

沈阳药科大学大学生创新训练计划项目：附子配伍减毒作用的代谢组学研究，负责人：熊志立。