HPLC法同时测定桂枝加芍药汤中8种成分的含量
|
摘要
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 20 μ L. RESULTS:The linear range of albiflorin,paeoniflorin,liquiritin,liquiritigenin,cinnamic acid,cinnamaldehyde,ammonium glycyrrhetate and 6-ginger phenol were 2.125-34.000 μg/m L(r=0.999 9),28.700-459.200 μg/m L(r=0.999 7),3.675-58.800 μg/m L(r=0.999 7),1.235-19.760μg/m L(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 μ g/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 μ g/mL;the detection limits were 0.041,0.031,0.029,0.010,0.004,0.007,0.201,0.059 μ g/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.
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 μ L. RESULTS:The linear range of albiflorin,paeoniflorin,liquiritin,liquiritigenin,cinnamic acid,cinnamaldehyde,ammonium glycyrrhetate and 6-ginger phenol were 2.125-34.000 μg/m L(r=0.999 9),28.700-459.200 μg/m L(r=0.999 7),3.675-58.800 μg/m L(r=0.999 7),1.235-19.760μg/m L(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 μ g/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 μ g/mL;the detection limits were 0.041,0.031,0.029,0.010,0.004,0.007,0.201,0.059 μ g/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.
引文
[1]王海强.基于活性指数和液质联用技术的桂枝加芍药汤抗炎活性成分快速发现研究[D].杭州:浙江大学,2017.
[2]彭程,刘建和.刘建和运用桂枝加芍药汤加减治疗腹痛经验[J].湖南中医杂志,2016,32(5):28-29、35.
[3]危美红,张喜奎.张喜奎教授运用桂枝加芍药汤治疗脾阴虚腹痛的经验[J].国医论坛,2015,30(6):17-19.
[4]吴雪,段国相.桂枝加芍药汤治疗胃肠神经官能症临床观察[J].世界最新医学信息文摘,2014,14(7):205-206.
[5]许爱丽,苏晓兰,郭宇,等.桂枝汤类方在脾胃疾病中的应用[J].上海中医药杂志,2018,52(7):34-36.
[6]袁海建,李卫,金建明,等.桂枝汤化学成分、药理作用机制与临床应用研究进展[J].中国中药杂志,2017,42(23):4556-4564.
[7]冯博,房玉涛,徐瑞山.桂枝汤的现代临床应用及作用机制研究进展[J].中国中药杂志,2018,43(12):2442-2447.
[8]张利青,张占刚,付岩,等.桂皮醛药理作用的研究进展[J].中国中药杂志,2015,40(23):4568-4572.
[9]马骁.赤芍治疗淤胆型肝炎的物质基础与作用机制研究[D].成都:成都中医药大学,2016.
[10]王兵,王亚新,赵红燕,等.甘草的主要成分及其药理作用的研究进展[J].吉林医药学院学报,2013,34(3):215-218.
[11]赵文竹,张瑞雪,于志鹏,等.生姜的化学成分及生物活性研究进展[J].食品工业科技,2016,37(11):383-389.
[12]邵长森,张国青,韩真真,等.HPLC法同时测定温经汤中10种活性成分的含量[J].中国药房,2018,29(19):2640-2643.
[13]袁鹏飞,刘焕,尚明英,等.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[J].中国药房,2016,27(6):801-805.
[14]严优芍,钟伟健,郭丽冰.GC-MS分析桂枝汤不同配伍对桂枝挥发油成分的影响[J].中药材,2012,35(3):410-415.
[2]彭程,刘建和.刘建和运用桂枝加芍药汤加减治疗腹痛经验[J].湖南中医杂志,2016,32(5):28-29、35.
[3]危美红,张喜奎.张喜奎教授运用桂枝加芍药汤治疗脾阴虚腹痛的经验[J].国医论坛,2015,30(6):17-19.
[4]吴雪,段国相.桂枝加芍药汤治疗胃肠神经官能症临床观察[J].世界最新医学信息文摘,2014,14(7):205-206.
[5]许爱丽,苏晓兰,郭宇,等.桂枝汤类方在脾胃疾病中的应用[J].上海中医药杂志,2018,52(7):34-36.
[6]袁海建,李卫,金建明,等.桂枝汤化学成分、药理作用机制与临床应用研究进展[J].中国中药杂志,2017,42(23):4556-4564.
[7]冯博,房玉涛,徐瑞山.桂枝汤的现代临床应用及作用机制研究进展[J].中国中药杂志,2018,43(12):2442-2447.
[8]张利青,张占刚,付岩,等.桂皮醛药理作用的研究进展[J].中国中药杂志,2015,40(23):4568-4572.
[9]马骁.赤芍治疗淤胆型肝炎的物质基础与作用机制研究[D].成都:成都中医药大学,2016.
[10]王兵,王亚新,赵红燕,等.甘草的主要成分及其药理作用的研究进展[J].吉林医药学院学报,2013,34(3):215-218.
[11]赵文竹,张瑞雪,于志鹏,等.生姜的化学成分及生物活性研究进展[J].食品工业科技,2016,37(11):383-389.
[12]邵长森,张国青,韩真真,等.HPLC法同时测定温经汤中10种活性成分的含量[J].中国药房,2018,29(19):2640-2643.
[13]袁鹏飞,刘焕,尚明英,等.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[J].中国药房,2016,27(6):801-805.
[14]严优芍,钟伟健,郭丽冰.GC-MS分析桂枝汤不同配伍对桂枝挥发油成分的影响[J].中药材,2012,35(3):410-415.