不同配伍配比对赤芍-当归药对中10个成分溶出量的影响研究

目的：建立赤芍-当归药对中没食子酸、氧化芍药苷、香草酸、芍药内酯苷、芍药苷、阿魏酸、1，2，3，4，6-O-五没食子酰葡萄糖、苯甲酰芍药苷、丹皮酚、藁本内酯10个成分的含量测定方法，考察不同配伍比例及配伍方式对赤芍-当归药对中10个主要活性成分溶出量的影响。方法：采用Wonda Cract ODS-2色谱柱（4.6 mm×250 mm，5 μm），以乙腈-0.1%磷酸水溶液为流动相，梯度洗脱，流速1.0 mL·min^-1，柱温25℃，变换波长检测赤芍-当归药对中10个成分，检测波长为210、230、260、274、275、325、350 nm。进一步测定赤芍-当归药对的单煎液、单煎合并液（1∶1）以及不同配比的合煎液（1∶1，1∶2，1∶3，2∶1，3∶1）中上述10个主要活性成分的含量。结果：没食子酸、氧化芍药苷、香草酸、芍药内酯苷、芍药苷、阿魏酸、1，2，3，4，6-O-五没食子酰葡萄糖、苯甲酰芍药苷、丹皮酚、藁本内酯质量浓度分别在1.49~74.50 μg·mL^-1（r=0.996 0）、2.88~144.00 μg·mL^-1（r=0.996 4）、2.96~147.99 μg·mL^-1（r=0.997 6）、2.94~147.00 μg·mL^-1（r=0.997 9）、2.99~149.50 μg·mL^-1（r=0.998 1）、2.24~111.99 μg·mL^-1（r=0.997 8）、3.01~150.50 μg·mL^-1（r=0.997 8）、2.98~148.99 μg·mL^-1（c0.998 4）、2.79~139.50（r=0.997 3）、6.00~300.00（r=0.998 2）μg·mL^-1范围内与峰面积呈良好的线性关系。精密度良好，RSD小于2.0%；回收率良好，3个加样浓度的平均回收率（n=9）为83.7%~100.4%，RSD为2.5%~7.2%。当赤芍-当归比例为2∶1时，各成分含量明显大于单煎液中含量，且多种成分在该比例中含量最高。当赤芍-当归比例为1∶1时，合煎液中含量较高的5个成分的溶出量明显多于单煎合并液。结论：本研究所建立的测定方法，结果可靠准确，操作简便，可用于赤芍-当归药对中10个成分的含量测定；赤芍-当归（2∶1）的配伍比例以及合煎的配伍方式有利于10个成分在水中的溶出。  

Objective:To establish a method for simultaneous determination of ten components (gallic acid,oxypaeoniflorin,vanillic acid, albiflorin,paeoniflorin, ferulic acid,1,2,3,4,6-penta-O-galloy-β-D-glucose,benzoylpaeoniflorin, paeonol, ligustilide)in Paeoniae Radix Rubra and Angelicae Sinensis Radix drug pair,and to study the influence of compatibility proportions on the ten components. Methods:A Wonda Cract ODS-2 column(250 mm×4.6 mm,5 μm)was used for the separation,with gradient elution of acetonitrile and 0.1% phosphoric acid as the mobile phase at a flow rate of 1.0 mL·min^-1.The detection wavelength was set at 210,230,260,274,275,325 and 350 nm. The column temperature was 25℃. And the ten components were determined in different compatibility proportions,including single decoction,mixed single decoctions(1:1)and mixed decoction(1:1,1:2,1:3,2:1 and 3:1). Results:The linear range was 1.49-74.50 μg·mL^-1 for gallic acid(r=0.996 0),2.88-144.00 μg·mL^-1 for oxypaeoniflorin(r=0.996 4),2.96-147.99 μg·mL^-1 for vanillic acid(r=0.997 6),2.94-147.00 μg·mL^-1 for albiflorin(r=0.997 9),2.99-149.50 μg·mL^-1 for peaoniflorin(r=0.998 1),2.24-111.99 μg·mL^-1 for ferulic acid(r=0.997 8),3.01-150.50 μg·mL^-1 for 1,2,3,4,6-penta-O-galloy-β-D-glucose(r=0.997 8),2.98-148.99 μg·mL^-1 for benzoylpaeoniflorin(r=0.998 4),2.79-139.50 μg·mL^-1 for paeonol(r=0.997 3),and 6.00-300.00 μg·mL^-1 for ligustilide(r=0.998 2). The results of the precision tests were satisfactory with relative standard deviations(RSDs)all below 2.0%. The average spike recoveries(n=9)at three concentration levels were 83.7%-100.4%,with RSDs within 2.5%-7.2%. Compared with the single decoction,when the proportion of Paeoniae Radix Rubra-Angelicae Sinensis Radix(m/m)was 2:1,the contents of the ten components increased obviously in mixed decoction. And the contents of most of ten components were the highest in the 2:1 drug pair. Compared with the mixed single decoctions,the contents of oxypaeoniflorin,vanillic acid,albiflorin,peaoniflorin and paeonol increased obviously in mixed decoction of Paeoniae Radix Rubra and Angelicae Sinensis Radix drug pair(1:1). Conclusion:The established method is simple and accurate,and can be applied to determinate ten components in Paeoniae Radix Rubra and Angelicae Sinensis Radix drug pair. The mixed decoction of Paeoniae Radix Rubra and Angelicae Sinensis Radix with proportion of 2:1 has an obvious effect in promoting the dissolution of the ten components. 

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