不同醋制方法对白芍6种活性成分影响的定量质控

Quantitative Quality Control for Effects of Different Vinegar-Processing on Six Active Components Contained in Baishao(Paeoniae Radix Alba)

目的建立HPLC法探讨不同醋制方法对白芍6种活性成分的影响。方法采用Waters Symmetry C色谱柱(4.6 mm×250 mm,5μm);流动相:乙腈-0.1%磷酸水,梯度洗脱;检测波长:230 nm;流速:1.0 mL/min;柱温:35℃。结果白芍中的没食子酸、氧化芍药苷、芍药内酯苷、芍药苷、1,2,3,4,6-O-五没食子酰葡萄糖、苯甲酰芍药苷分别在0.77~99.14、0.38~97.52、1.62~208.6、9.66~1236.19、1.17~298.86、0.40~51.52μg/mL浓度范围内线性关系良好。白芍中的没食子酸、氧化芍药苷、芍药内酯苷、芍药苷、1,2,3,4,6-O-五没食子酰葡萄糖、苯甲酰芍药苷的平均加样回收率为99.64%~106.16%,相对标准偏差为0.77%~2.59%。白芍经醋炙后,6种成分的含量均有显著性提升;而白芍经醋浸后,除芍药苷含量无显著性差异之外,没食子酸、芍药内酯苷、1,2,3,4,6-O-五没食子酰葡萄糖的含量均有显著性降低,氧化芍药苷、苯甲酰芍药苷的含量均有显著性提升。醋炙白芍与醋浸白芍相比,其6种成分的含量均有显著性提升。结论该研究所建分析方法准确可靠,专属性强,可初步揭示炮制过程中“加热”操作的必要性,为白芍临床炮制品的合理使用和质量控制提供参考。

Objective To establish an HPLC method for investigating the effects of different vinegar-processing(vinegar-stir-frying and vinegar-immersing)on six active components contained in Baishao(Paeoniae Radix Alba).Methods The analyses of methanol extracts of crude and vinegar-processed Baishao(Paeoniae Radix Alba)were performed on a Waters Symmetry Ccolumn(4.6 mm×250 mm,5μm).The mobile phase was composed of acetonitrile and 0.1%aqueous phosphoric acid for gradient elution.The detection wavelength was set at 230 nm.The flow rate was 1.0 mL/min and the column temperature was set at 35℃.Results There were good linear relationships among gallic acid,oxypaeoniflorin,albiflorin,paeoniflorin,1,2,3,4,6-O-pentagalloyl glucose and benzoylpaeoniflorin contained in Baishao(Paeoniae Radix Alba)within ranges of 0.77~99.14,0.38~97.52,1.62~208.6,9.66~1236.19,1.17~298.86 and 0.40~51.52μg/mL,respectively.The average recoveries of these components were 99.64%~106.16%,and the RSD values of them were 0.77%~2.59%.The contents of six components contained in Baishao(Paeoniae Radix Alba)were significantly increased after vinegar-stir-frying of Baishao(Paeoniae Radix Alba).The contents of gallic acid,albiflorin and 1,2,3,4,6-O-pentagalloyl glucose contained in Baishao(Paeoniae Radix Alba)were significantly decreased,while the contents of oxypaeoniflorin and benzoylpaeoniflorin contained in Baishao(Paeoniae Radix Alba)were significantly increased after vinegar-immersing of Baishao(Paeoniae Radix Alba)except for no significant difference in the content of paeoniflorin.Compared to vinegar-immersed Baishao(Paeoniae Radix Alba),the contents of six components contained in vinegar-stir-fried Baishao(Paeoniae Radix Alba)were significantly higher than those contained in vinegar-immersed one.Conclusion With strong specificity,the accurate and reliable analytical method established in this study can preliminarily reveal the necessity of‘higher temperature operation’in the processing process,and provides reference for the rational use and quality control of the processed products of Baishao(Paeoniae Radix Alba)in clinic.