桂枝汤类方化学成分含量研究

Content change in categorized formulas about Guizhi Decoction

目的:建立HPLC法同时测定桂枝汤(桂枝、白芍、生姜、甘草和大枣)类方中原儿茶酸、香豆素、桂皮酸、桂皮醛、芍药苷和甘草酸含量的方法,并探索配伍对各成分含量的影响。方法:用SUPELCO Discovery C18柱,0.05%磷酸-乙腈为流动相,梯度洗脱;流速:1.0mL/min;柱温:30℃;检测波长为230nm和254nm;测定桂枝汤、桂枝加桂汤和桂枝加芍药汤中原儿茶酸、香豆素、桂皮酸、桂皮醛、芍药苷和甘草酸的含量,不同配伍之间的差异采用SPSS软件进行方差分析。结果:建立了同时测定原儿茶酸、香豆素、桂皮酸、桂皮醛、芍药苷和甘草酸含量的高效液相色谱法和相应的回归方程,与单味桂枝相比,三个类方中原儿茶酸含量增加、桂皮酸含量减少,桂枝汤和桂枝加桂汤中香豆素含量增加;与单味芍药相比,三个类方中芍药苷含量减少;与单味甘草相比,三个类方中甘草酸含量增加。结论:本方法专属、重复性好,可用于桂枝汤类方中原儿茶酸、香豆素、桂皮酸、桂皮醛、芍药苷和甘草酸的含量测定。不同配伍对各成分含量有影响,其影响非常复杂。

AIM： To determine the contents of protocatechuic acid,peoniflorin,coumarin,cinnamic acid,cinnamaldehyde and glycyrrhizic acid in categorized formulas about Guizhi Decoction （Ramulus cinnamoni,Radix paeoniae alba,Rhizoma zingiberis recens,Radix et Rhizoma glycyrrhizae and Fructus jujubae） by HPLC.METHODS： The gradient elution mode was applied in chromatographic separation.The C18 column was used with the mobile phase of 0.05% phosphoric acid-acetonitrile,flow rate was at 1.0 mL/min,detection wavelength at 230 nm and 254 nm,and the column temperature was at 30 ℃.The contents of the above-mentioned six constituents were determined in Guizhi Decoction,Guizhi Decoction plus Ramulus cinnamoni and Guizhi Decoction plus Radix paeoniae alba,respectively.The differences among all combinations were tested by one-way analysis of variance using SPSS software.RESULTS： The concentrations of the above-mentioned six constituents in different decoctions were simultaneously determined by HPLC and the linear equations of six constituents were established.Compared with Ramulus Cinnamomi alone,cinnamic acid content decreased and protocatechuic acid content increased in categorized formulas about Guizhi Decoctions significantly,but then coumarin content increased in Guizhi Decoction and Guizhi Decoction plus Radix paeoniae alba.Compared with Radix Paeoniae Alba alone,peoniflorin content decreased in categorized formulas about Guizhi Decoctions significantly.Compared with Radix et Rhizoma Glycyrrhiza alone,glycyrrhizic acid content increased in categorized formulas about Guizhi Decoctions significantly.CONCLUSION：The methods are accurate,reproducible and suitable for determineation of the contents of six constituents in categorized formulas about Guizhi Decoctions,Results show that the contents change with different decoctions.