桂枝汤中9种主要成分含量的测定和不同煎煮方式的比较研究

Contents Determination of Nine Components in Guizhi Decoction and Comparison of Different Decoction Methods

目的：建立同时测定桂枝汤中9种主要成分含量的方法,比较传统煎煮和煎药机煎煮对桂枝汤中上述成分含量的影响。方法：采用高效液相色谱法。色谱柱为Agilent Zorbax SB C18,流动相为乙腈-0.1%磷酸（梯度洗脱）,流速为1.0 ml/min,检测波长为230 nm（芍药内酯苷、芍药苷、甘草苷）、254 nm（桂皮醛、甘草酸）、280 nm（没食子酸、香豆素、桂皮酸、2-甲氧基桂皮醛）,柱温为25℃,进样量为10μl。结果：没食子酸、芍药内酯苷、芍药苷、甘草苷、香豆素、桂皮酸、桂皮醛、2-甲氧基桂皮醛和甘草酸检测质量浓度线性范围分别为0.410 2~210.0（r=0.999 9）、0.994 0~254.5（r=0.999 9）、1.636 0~1 675.0（r=0.999 9）、0.988 3~506.0（r=0.999 6）、0.987 3~31.59（r=0.999 5）、0.486 8~124.6（r=0.999 5）、2.458 0~314.6（r=0.999 5）、0.034 3~1.096（r=0.999 8）、1.711 0~219.0μg/ml（r=0.999 7）;精密度、稳定性、重复性试验的RSD〈5%,加样回收率分别为93.56%~103.19%（RSD=4.00%,n=9）、101.51%~107.32%（RSD=2.21%,n=9）、95.08%~103.76%（RSD=2.87%,n=9）、100.82%~105.73%（RSD=1.85%,n=9）、85.08%~89.12%（RSD=1.40%,n=9）、92.31%~99.12%（RSD=2.71%,n=9）、99.17%~102.32%（RSD=1.24%,n=9）、100.15%~103.98%（RSD=1.18%,n=9）、99.93%~102.61%（RSD=1.03%,n=9）。煎药机煎煮所得总有效成分含量为4 565μg/g,传统煎煮所得总有效成分含量为2 742μg/g。结论：该方法操作简便、稳定、重复性好,可用于桂枝汤中9种主要成分含量的同时测定,其中没食子酸、芍药内酯苷和2-甲氧基桂皮醛的含量测定为桂枝汤中首次报道。煎药机煎煮所得总有效成分含量高于传统煎煮所得总有效成分。

OBJECTIVE：To establish a method for the contents determination of 9 components in Guizhi decoction,and compare the effects of traditional decoction method and the extracting machine decoction method on these contents in Guizhi decoction.METHODS：HPLC was performed on the column of Agilent Zorbax SB-C18 with mobile phaseof acetonitrile- 0.1% phosphoric acid（gradient elution）at a flow rate of 1.0 ml/min,the detection wavelength was 230 nm,254 nm and 280 nm,the column temperature was 25 ℃,and the injection volume was 10 μl. RESULTS：The linear range was 0.410 2-210.0 μg/ml for gallic acid（r=0.999 9）,0.994 0-254.5 μg/ml for albiflorin（r=0.999 9）,1.636 0-1 675.0 μg/ml for paeoniflorin（r=0.999 9）,0.988 3-506.0 μg/ml for liquiritin（r=0.999 6）,0.987 3-31.59 μg/ml for coumarin（r=0.999 5）,0.486 8-124.6 μg/ml for cinnamic acid（r=0.999 5）,2.458 0-314.6μg/ml for cinnamaldehyde（r=0.999 5）,0.034 3-1.096 μg/ml for 2-methoxy cinnamaldehyde（r=0.999 8）,and 1.711 0-219.0 μg/ml for glycyrrdhizic acid（r=0.999 7）;RSDs of precision,stability and reproducibility tests were lower than 5%,recoveries were93.56%-103.19%（RSD=4.00%,n=9）、101.51%-107.32%（RSD=2.21%,n=9）、95.08%-103.76%（RSD=2.87%,n=9）、100.82%-105.73%（RSD=1.85%,n=9）、85.08%-89.12%（RSD=1.40%,n=9）、92.31%-99.12%（RSD=2.71%,n=9）、99.17%-102.32%（RSD=1.24%,n=9）、100.15%-103.98%（RSD=1.18%,n=9）、99.93%-102.61%（RSD=1.03%,n=9）. The content of total effective components from the extracting machine decoction method was 4 565 μg/g,that from the traditional decoction method was 2 742 μg/g.CONCLUSIONS：The method is simple,stable and reproducible,and can be used for the simultaneous determination of 9 componentsin Guizhi decoction. The contents of gallic acid,albiflorin and 2-methoxy cinnamaldehyde are first reported. The total effective components from the extracting machine decoction method are higher than that from the traditional decoction method.