UFLC法测定当归芍药散中6个活性成分的含量

Determination of six active components in Danggui Shaoyao San by UFLC

目的:建立测定当归芍药散中芍药苷、芍药内酯苷、阿魏酸、24-乙酰泽泻醇A、23-乙酰泽泻醇B和白术内酯Ⅰ含量的超快速液相色谱法。方法:采用Shim-Pack XR-ODS色谱柱(75 mm×3.0 mm,2.2μm),以0.1%磷酸水溶液(A)-乙腈(B)为流动相,测定芍药苷、芍药内酯苷、阿魏酸、24-乙酰泽泻醇A和23-乙酰泽泻醇B时采用梯度洗脱(0~10 min,14%B;10~11 min,14%B→80%B;11~20 min,80%B),平衡时间为2 min,测定白术内酯Ⅰ时采用等度洗脱(0~6 min,38%B),流速0.4 m L·min^(-1),柱温为35℃,检测波长为232 nm(芍药苷、芍药内酯苷、阿魏酸、白术内酯Ⅰ)和208 nm(24-乙酰泽泻醇A和23-乙酰泽泻醇B),进样量为5μL。结果:芍药苷、芍药内酯苷、阿魏酸、24-乙酰泽泻醇A、23-乙酰泽泻醇B和白术内酯Ⅰ质量浓度分别在10~500μg·m L^(-1)(r=0.999 9)、2~100μg·m L^(-1)(r=0.999 9)、0.2~10μg·m L^(-1)(r=0.999 9)、2~100μg·m L^(-1)(r=0.999 7)、2~100μg·m L^(-1)(r=0.999 3)和0.1~5μg·m L^(-1)(r=0.999 8)范围内与峰面积呈良好的线性关系;平均回收率(n=9)分别为98.6%,96.8%,97.7%,96.9%,97.0%和97.6%。6批当归芍药散样品测定结果:芍药苷0.91%~0.98%,芍药内酯苷0.20%~0.23%,阿魏酸0.034%~0.052%,24-乙酰泽泻醇A 0.041%~0.043%,23-乙酰泽泻醇B 0.020 1%~0.024 9%,白术内酯Ⅰ0.002 6%~0.003 0%。结论:该方法可为当归芍药散的质量控制研究提供科学依据。

Objective：To develop an ultra-fast liquid chromatography（UFLC）method for determination of paeoniflorin,albiflorin,ferulic acid,24-acetyl alisol A,23-acetyl alisol B and atractylenolideⅠin Danggui Shaoyao San.Methods：Chromatographic separation was performed on a Shim-Pack XR-ODS column（75 mm×3.0 mm,2.2μm）.The mobile phase consisted of 0.1%phosphoric acid（A）and acetonitrile（B）at a flow rate of 0.4 m L·min^（-1）.Paeoniflorin,albiflorin,ferulic acid,24-acetyl alisol A and 23-acetyl alisol B were determined with gradient elution（0-10 min,14%B;10-11 min,14%B→80%B;11-20 min,80%B）at 232nm（paeoniflorin,albiflorin and ferulic acid）and 208 nm（24-acetyl alisol A and 23-acetyl alisol B）,and the equilibration time was 2 min.AtractylenolideⅠwas determined with isocratic elution（0-6 min,38%B）at 232nm.The column temperature was maintained at 35℃and the injection volume was 5μL.Results：The linear ranges were 10-500μg·m L^（-1）（r=0.999 9）for paeoniflorin,2-100μg·m L^（-1）（r=0.999 9）for albiflorin,0.2-10μg·m L^（-1）（r=0.999 9）for ferulic acid,2-100μg·m L^（-1）（r=0.999 7）for 24-acetyl alisol A,2-100μg·m L^（-1）（r=0.999 3）for 23-acetyl alisol B and 0.1-5μg·m L^（-1）（r=0.999 8）for atractylenolideⅠ.The average recovery（n=9）of the six components were 98.6%,96.8%,97.7%,96.9%,97.0%and 97.6%,respectively.The content ranges of six components in six batches were 0.91%-0.98%for paeoniflorin,0.20%-0.23%for albiflorin,0.034%-0.052%for ferulic acid,0.041%-0.043%for 24-acetyl alisol A,0.020 1%-0.024 9%for 23-acetyl alisol B,and0.002 6%-0.003 0%for atractylenolideⅠ.Conclusion：The method can provide scientific basis for the quality control of Danggui Shaoyao San.