知识与财富的链接
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20μL。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2.125～34.000μg/mL(r=0.999 9)、28.700～459.200μg/m L(r=0.999 7)、3.675～58.800μg/mL(r=0.999 7)、1.235～19.760μg/m L(r=0.999 8)、2.300～36.800μg/m L(r=0.999 8)、0.955～15.280μg/mL(r=0.999 7)、36.000～576.000μg/m L(r=0.999 7)、1.500～24.000μg/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、0.198μg/mL,检测限分别为0.041、0.031、0.029、0.010、0.004、0.007、0.201、0.059μg/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 20μL.RESULTS:The linear range of albiflorin,paeoniflorin,liquiritin,liquiritigenin,cinnamic acid,cinnamaldehyde,ammonium glycyrrhetate and 6-ginger phenol were 2.125-34.000μg/mL(r=0.999 9),28.700-459.200μg/mL(r=0.999 7),3.675-58.800μg/mL(r=0.999 7),1.235-19.760μg/mL(r=0.999 8),2.300-36.800μg/mL(r=0.999 8),0.955-15.280μg/mL(r=0.999 7),36.000-576.000μg/mL(r=0.999 7)and 1.500-24.000μg/mL(r=0.999 7),respectively.The quantitative limits were 0.135,0.102,0.096,0.033,0.013,0.023,0.663,0.198μg/mL;the detection limits were 0.041,0.031,0.029,0.010,0.004,0.007,0.201,0.059μg/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.
