HPLC法测定神农丸中士的宁与马钱子碱的含量

目的：采用氨基键合硅胶为固定相的正相高效液相色谱法，研究建立神农丸中士的宁与马钱子碱的含量测定方法，并进行方法学研究。方法：以ZORBAX NH2氨基键合硅胶柱为色谱柱（4.6 mm×250 mm，5 μm），以乙腈-异丙醇-3%磷酸溶液（80：10：10）为流动相，以260 nm为检测波长，流速1.0 mL·min^-1，柱温：40℃。结果：士的宁进样量在0.051 2~0.512 μg范围内线性关系良好，回归方程：Y=1 989.6X+3.610，r=0.999 9；马钱子碱进样量在0.057 6~0.576 μg范围内线性关系良好，回归方程：Y=1 932.8X-2.569，r=1.000，士的宁与马钱子碱的平均回收率分别为98.6%、96.7%。结论：本方法操作简便，供试品色谱图基线稳定，主成分峰峰形对称，分离度高，方法的重复性好、准确度高，可用于神农丸中士的宁与马钱子碱的含量测定。

Objective:To establish a method for the determination of strychnine and brucine in Shennong pills by NP-HPLC of amino-bonded silica gel column,and study its methodology.Method:The stationary phase was amino-bonded silica gel column(ZORBAX NH₂,4.6 mm×250 mm,5 μm).The mobile phase consisted of acetonitrile-isopropanol-3% sulphuric acid(80:10:10)at a flow rate of 1.0 mL·min^-1.The detecting wavelength was 260 nm.The column temperature was 40℃.Result:The linear range of strychnine was 0.051 2-0.512 μg and the linear calibration curve was Y=1 989.6X+3.610(r=0.999 9);The linear range of brucine was 0.057 6-0.576 μg and the linear calibration curve was Y=1 932.8X-2.569(r=1.000).The average recovery rates of strychnine and brucine were 98.6% and 96.7% respectively.Conclusion:This method is simple.The base line is straight.The main peaks' shape are symmetric and the degree of separation between the main peaks is high in the chromatogram.The method is repeatable and accurate,which can be applied to the determination of the contents of strychnine and brucine in Shennong pills.