超高效二维液相色谱法测定复方南星止痛膏中双酯型乌头生物碱的含量

目的：采用在线中心切割超高效二维液相色谱（2D-UPLC）法，建立同时测定复方南星止痛膏中3个双酯型乌头生物碱（新乌头碱、乌头碱和次乌头碱）的含量测定方法。方法：一维UPLC采用Waters XBridgeBEH C₁₈色谱柱（2.1 mm×50 mm 1.7μm），以甲醇-0.008%二乙胺为流动相，梯度洗脱，流速0.1 mL·min^-1，柱温18℃；二维UPLC采用Waters XBridge BEH C₁₈色谱柱（3.0 mm×100 mm 1.7μm），以乙腈-0.01%磷酸为流动相，梯度洗脱，流速0.7 mL·min^-1，柱温40℃。进样量1μL，检测波长235 nm。结果：新乌头碱、乌头碱和次乌头碱的线性范围分别为0.66~9.90、0.55~8.20和0.91~13.72μg·mL^-1，R²分别为0.999 5、0.999 9和0.999 6，线性关系良好；3个成分的平均加样回收率在82.5%~88.8%之间，方法的准确度和稳定性均较好；复方南星止痛膏中新乌头碱、乌头碱和次乌头碱含量分别在3.2~5.0、0.9~1.3、8.5~10.4 μg·g^-1范围内。结论：该方法有效改善了复方南星止痛膏中待测组分的分离度，极大提高了检测灵敏度，对制剂中的3个双酯型乌头生物碱进行准确定量分析，提高产品的质量控制水平，并为临床用药提供理论支持。

Objective: To establish an on-line center-cutting two-dimensional ultra performance liquid chromategraphy (2D-UPLC) method for the determination of three double ester alkaloids (mesaconitine,aconitine and hypaconitine) in compound Nanxing Zhitong plaster. Methods: The first dimensional column was Waters XBridge BEH C₁₈ (2.1 mm×50 mm 1.7 μm),with methanol and 0.008% diethylamine as the mobile phase in gradient elution at a flow rate of 0.1 mL·min^-1,and the column temperature was 18℃. The second dimensional column was Waters XBridge BEH C₁₈ (3.0 mm×100 mm 1.7μm),with acetonitrile-0.01% phosphoric acid as the mobile phase in gradient elution at a flow rate of 0.7 mL·min^-1,and the column temperature was 40℃. The injection volume was 1μL and the detection wavelength was 235 nm. Results: The linear ranges of mesaconitine, aconitine and hypaconitine were 0.66-9.90 μg·mL^-1,0.55-8.20μg·mL^-1 and 0.91-13.72μg·mL^-1 with R² of 0.999 5,0.999 9 and 0.999 6,respectively,which indicated good linear relationship. The average recoveries of the three compounds were 82.5%-88.8%. The contents of mesaconitine, aconitine and hypaconitine in the compound Nanxing Zhitong plaster were within the range of 3.2-5.0 μg·g^-1, 0.9-1.3 μg·g^-1 and 8.5-10.4 μg·g^-1.Conclusion: It was proved that the method could effectively improve separation and sensitivity,and could determine the three diester alkaloids in an accurate way. The investigation provided theoretical basis for enhancement of quality control of compound Nanxing Zhitong plaster and for clinic application.

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