UFLC法测定当归芍药散中6个活性成分的含量

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，2.2 μm），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 mL·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5 μL。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10~500 μg·mL^-1（r=0.999 9）、2~100 μg·mL^-1（r=0.999 9）、0.2~10 μg·mL^-1（r=0.999 9）、2~100 μg·mL^-1（r=0.999 7）、2~100 μg·mL^-1（r=0.999 3）和0.1~5 μg·mL^-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Ⅰ 0.002 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 atractylenolideⅠ in Danggui Shaoyao San. Methods:Chromatographic separation was performed on a Shim-Pack XR-ODS column (75 mm×3.0 mm,2.2 μm). The mobile phase consisted of 0.1% phosphoric acid(A) and acetonitrile(B) at a flow rate of 0.4 mL·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 232 nm(paeoniflorin,albiflorin and ferulic acid) and 208 nm(24-acetyl alisol A and 23-acetyl alisol B),and the equilibration time was 2 min. AtractylenolideⅠ was determined with isocratic elution(0-6 min,38%B) at 232 nm. The column temperature was maintained at 35 and the injection volume was 5 μL. Results:The linear ranges were 10-500 μg·mL^-1(r=0.999 9) for paeoniflorin,2-100 μg·mL^-1(r=0.999 9) for albiflorin,0.2-10 μg·mL^-1(r=0.999 9) for ferulic acid,2-100 μg·mL^-1(r=0.999 7) for 24-acetyl alisol A,2-100 μg·mL^-1(r=0.999 3) for 23-acetyl alisol B and 0.1-5 μg·mL^-1(r=0.999 8) for atractylenolide Ⅰ. 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,and 0.002 6%-0.003 0% for atractylenolideⅠ. Conclusion:The method can provide scientific basis for the quality control of Danggui Shaoyao San.