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目的:建立同时测定桂枝加芍药汤中芍药内酯苷、芍药苷、甘草苷、甘草素、肉桂酸、桂皮醛、甘草酸铵、6-姜酚含量的方法。方法:采用高效液相色谱法。色谱柱为Kromasil C18,流动相为乙腈-0.1%磷酸溶液(梯度洗脱),流速为1.0 mL/min,检测波长为235 nm(0~35 min,芍药内酯苷、芍药苷、甘草苷)、280 nm(35~65 min,甘草素、肉桂酸、桂皮醛、甘草酸铵、6-姜酚),柱温为25 ℃,进样量为20 μL。结果:芍药内酯苷、芍药苷、甘草苷、甘草素、肉桂酸、桂皮醛、甘草酸铵、6-姜酚检测质量浓度线性范围分别为2.125~34.000 μg/mL(r=0.999 9)、28.700~459.200 μg/mL(r=0.999 7)、3.675~58.800 μg/mL(r=0.999 7)、1.235~19.760 μg/mL(r=0.999 8)、2.300~36.800 μg/mL(r=0.999 8)、0.955~15.280 μg/mL(r=0.999 7)、36.000~576.000 μg/mL(r=0.999 7)、1.500~24.000 μg/mL(r=0.999 7);定量限分别为0.135、0.102、0.096、0.033、0.013、0.023、0.663、0.198 μg/mL,检测限分别为0.041、0.031、0.029、0.010、0.004、0.007、0.201、0.059 μg/mL;精密度、稳定性、重复性试验的RSD均小于3%(n=6);加样回收率分别为97.47%~100.76%(RSD=1.33%,n=6)、98.15%~103.50%(RSD=1.82%,n=6)、95.65%~100.84%(RSD=2.38%,n=6)、96.75%~100.32%(RSD=1.31%,n=6)、95.88%~102.75%(RSD=2.52%,n=6)、95.63%~100.63%(RSD=2.00%,n=6)、96.78%~100.45%(RSD=1.35%,n=6)、95.71%~100.48%(RSD=1.80%,n=6)。结论:该方法准确、可靠,专属性好,可用于同时测定桂枝加芍药汤中8种成分的含量。
OBJECTIVE: To establish a method for simultaneous determination of albiflorin, paeoniflorin, liquiritin, liquiritigenin, cinnamic acid, cinnamaldehyde, ammonium glycyrrhetate and 6-ginger phenol in Guizhijiashaoyao decoction. METHODS: HPLC method was adopted. The determination was performed on Kromasil C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelengths were 235 nm(0-35 min,albiflorin, paeoniflorin, liquiritin)、280 nm(35-65 min,liquiritigenin, cinnamic acid, cinnamaldehyde, ammonium glycyrrhetate and 6-ginger phenol). The column temperature was 25 ℃, and the sample size was 20 μL. RESULTS: The linear range of albiflorin, paeoniflorin, liquiritin, liquiritigenin, cinnamic acid, cinnamaldehyde, ammonium glycyrrhetate and 6-ginger phenol were 2.125-34.000 μg/mL(r=0.999 9), 28.700-459.200 μg/mL(r=0.999 7), 3.675-58.800 μg/mL(r=0.999 7), 1.235-19.760 μg/mL(r=0.999 8), 2.300-36.800 μg/mL(r=0.999 8), 0.955-15.280 μg/mL(r=0.999 7), 36.000-576.000 μg/mL(r=0.999 7) and 1.500-24.000 μg/mL(r=0.999 7), respectively. The quantitative limits were 0.135, 0.102, 0.096, 0.033, 0.013, 0.023, 0.663, 0.198 μg/mL; the detection limits were 0.041, 0.031, 0.029, 0.010, 0.004, 0.007, 0.201, 0.059 μg/mL. RSDs of precision, stability and reproducibility tests were all lower than 3% (n=6). The recovery rates were 97.47%-100.76%(RSD=1.33%,n=6), 98.15%-103.50%(RSD=1.82%,n=6), 95.65%-100.84%(RSD=2.38%,n=6), 96.75%-100.32%(RSD=1.31%,n=6), 95.88%-102.75%(RSD=2.52%,n=6), 95.63%-100.63%(RSD=2.00%,n=6), 96.78%-100.45%(RSD=1.35%,n=6), 95.71%-100.48%(RSD=1.80%,n=6). CONCLUSIONS: The method is accurate, reliable and exclusive, and suitable for simultaneous determination of 8 ingredients in Guizhijiashaoyao decoction.
桂枝加芍药汤高效液相色谱法含量测定芍药内酯苷芍药苷甘草苷甘草素肉桂酸桂皮醛甘草酸铵6-姜酚
Guizhijiashaoyao decoctionHPLCContent determinationAlbiflorinPaeoniflorinLiquiritinLiquiritigeninCinnamic acidCinnamaldehydeAmmonium glycyrrhetate6-ginger phenol
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