HPLC法同时测定腰痛片中芍药苷、川续断皂苷Ⅵ、补骨脂素和异补骨脂素含量

目的: 建立同时测定腰痛片(杜仲叶、赤芍、补骨脂、续断、狗脊等)中活性成分芍药苷、川续断皂苷Ⅵ、补骨脂素与异补骨脂素含量的方法。方法: 采用HPLC法,色谱柱为Inert Sustain C₁₈柱(250 mm×4.6 mm,5 μm),流动相为乙腈(A)-水(B),梯度洗脱(0 min,10%A;60 min,50%A),流速1 mL·min^-1,柱温25 ℃,检测波长为212 nm(川续断皂苷Ⅵ)、238 nm(芍药苷、补骨脂素和异补骨脂素),进样量10 μL。结果: 芍药苷、川续断皂苷Ⅵ、补骨脂素与异补骨脂素质量浓度分别在201.6~4 032 μg·mL^-1(r=0.999 5)、80~2 000 μg·mL^-1(r=0.999 9)、 11.04~220.8 μg·mL^-1(r=1.00 0)、8.7~174 μg·mL^-1(r=0.999 7)范围内与峰面积线性关系良好,平均回收率(n=9)分别为100.6%、100.6%、100.4%、99.93%,精密度试验RSD均小于0.80%。结论: 经方法学验证,该法能同时测定4种成分的含量,适用于腰痛片的质量控制。

Objective: To determine the content of four major components (paeoniflorin,asperosaponin Ⅵ,psoralen and isopsoralen) in Yaotong tablets by high performance liquid chromatography. Methods: The determination was conducted on the Insert Sustain-C₁₈ column (250 mm×4.6 mm,5 μm) with the mobile phase of acetonitrile(A)-water(B),gradient elution (0 min,10%A; 60 min,50%A) at the flow rate of 1 mL·min^-1; the column temperature was at 25 ℃; the detection wavelength was 212 nm for asperosaponin Ⅵ,and 238 nm for paeoniflorin,psoralen and isopsoralen; and the injection volume was 10 μL. Results: Good linear relationship of paeoniflorin,asperosaponin Ⅵ,psoralen and isopsoralen between peak area and concentrations were noted within 201.6 -4 032 μg·mL^-1(r=0.999 5),80-2 000 μg·mL^-1(r=0.999 9),11.04-220.8 μg·mL^-1(r=1.000),and 8.7-174 μg·mL^-1(r=0.999 7),respectively. The average recoveries (n=9) were 100.6%,100.6%,100.4%,99.93%,RSD of precision test was less than 0.8%. Conclusion: Methodology verification proved that the method could be applied to simultaneously determine four components and control the quality Yaotong tablets.

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