HPLC-CAD法同时测定风湿定片中7种活性成分的含量
|
摘要
目的:建立同时测定风湿定片中甘草酸、水杨苷、甘草次酸、毒藜碱、丹皮酚、欧前胡素及异欧前胡素含量的方法。方法:采用高效液相色谱-电雾式检测器(HPLC-CAD)法。色谱柱为Thermo Hypersil BDS C18,流动相为乙腈-10 mmol/L甲酸铵(甲酸调pH至4.5,梯度洗脱),流速为1.0 m L/min,柱温为35℃,CAD雾化器温度为35℃,气压为(59.6±0.1)psi,检测频率为10 Hz,进样量为20μL。结果:甘草酸、水杨苷、甘草次酸、毒藜碱、丹皮酚、欧前胡素及异欧前胡素检测质量浓度线性范围为0.42～16.80、0.28～11.20、0.16～6.20、0.98～39.40、1.02～41.00、0.11～4.40、0.10～3.80μg/mL(r≥0.999 1);检测限分别为0.03、0.02、0.02、0.03、0.03、0.02、0.02μg/mL,定量限分别为0.11、0.06、0.06、0.10、0.11、0.08、0.07μg/mL;精密度、稳定性(48 h)、重复性试验的RSD均<2.0%(n=6或n=7);平均回收率为98.61%～99.76%,RSD为1.11%～1.51%(n=6)。结论:建立的方法操作简便、灵敏度高、结果准确,可用于风湿定片中甘草酸、水杨苷、甘草次酸、毒藜碱、丹皮酚、欧前胡素及异欧前胡素含量的同时测定。
OBJECTIVE:To establish the method for simultaneous determination of glycyrrhizic acid,salicin,glycyrrhetinic acid,chenopodium,paeonol,imperatorin and isoimperatorin in Fengshiding tablet. METHODS:HPLC-CAD method was adopted.The determination was performed on Thermo Hypersil BDS C18 column with mobile phase consisted of acetonitrile-10 mmol/L ammonium formate(pH value adjusted to 4.5 using formic acid,gradient elution) at the flow rate of 1.0 mL/min. The column temperature was set at 35 ℃. CAD atomizer temperature was 35 ℃,and air pressure was(59.6±0.1)psi. The detection frequency was 10 Hz, and sample size was 20 μL. RESULTS: The linear range of glycyrrhizic acid, salicin, glycyrrhetinic acid,chenopodium,paeonol,imperatorin and isoimperatorin were 0.42-16.80,0.28-11.20,0.16-6.20,0.98-39.40,1.02-41.00,0.11-4.40,0.10-3.80 μg/mL(r≥0.999 1),respectively. The detection limits were 0.03,0.02,0.02,0.03,0.03,0.02,0.02 μg/mL,respectively;quantitation limits were 0.11,0.06,0.06,0.10,0.11,0.08,0.07μg/mL,respectively. RSDs of precision,stability(48 h)and reproducibility tests were all lower than 2.0%(n=6 or n=7). Average recoveries rate were 98.61%-99.76%,and RSDs were 1.11%-1.51%(n=6). CONCLUSIONS:The established method is simple,sensitive,accurate,and can be used for simultaneous determination of glycyrrhizic acid, salicin, glycyrrhetinic acid, chenopodium, paeonol, imperatorin and isoimperatorin in Fengshiding tablet.
OBJECTIVE:To establish the method for simultaneous determination of glycyrrhizic acid,salicin,glycyrrhetinic acid,chenopodium,paeonol,imperatorin and isoimperatorin in Fengshiding tablet. METHODS:HPLC-CAD method was adopted.The determination was performed on Thermo Hypersil BDS C18 column with mobile phase consisted of acetonitrile-10 mmol/L ammonium formate(pH value adjusted to 4.5 using formic acid,gradient elution) at the flow rate of 1.0 mL/min. The column temperature was set at 35 ℃. CAD atomizer temperature was 35 ℃,and air pressure was(59.6±0.1)psi. The detection frequency was 10 Hz, and sample size was 20 μL. RESULTS: The linear range of glycyrrhizic acid, salicin, glycyrrhetinic acid,chenopodium,paeonol,imperatorin and isoimperatorin were 0.42-16.80,0.28-11.20,0.16-6.20,0.98-39.40,1.02-41.00,0.11-4.40,0.10-3.80 μg/mL(r≥0.999 1),respectively. The detection limits were 0.03,0.02,0.02,0.03,0.03,0.02,0.02 μg/mL,respectively;quantitation limits were 0.11,0.06,0.06,0.10,0.11,0.08,0.07μg/mL,respectively. RSDs of precision,stability(48 h)and reproducibility tests were all lower than 2.0%(n=6 or n=7). Average recoveries rate were 98.61%-99.76%,and RSDs were 1.11%-1.51%(n=6). CONCLUSIONS:The established method is simple,sensitive,accurate,and can be used for simultaneous determination of glycyrrhizic acid, salicin, glycyrrhetinic acid, chenopodium, paeonol, imperatorin and isoimperatorin in Fengshiding tablet.
引文
[1]国家药典委员会.中华人民共和国药典:一部[S]. 2015年版.北京:中国医药科技出版社,2015:686-687.
[2]王其勇,许亚玲. HPLC法测定八角枫药材不同药用部位中L(-)-八角枫碱的含量[J].中国药房,2016,27(27):3877-3879.
[3] AKAO T,YOSHINO T,KOBASHI K,et al. Evaluation of salicin as an antipyretic prodrug that does not cause gastric injury[J]. Planta Med,2002,68(8):714-718.
[4]金贤兰.徐长卿药理作用及临床应用研究进展[J].现代医药卫生,2010,26(19):2947-2948.
[5]朱艺欣,李宝莉,马宏胜,等.白芷的有效成分提取、药理作用及临床应用研究进展[J].中国医药导报,2014,11(31):159-162、166.
[6]高雪岩,王文全,魏胜利,等.甘草及其活性成分的药理活性研究进展[J].中国中药杂志,2009,34(21):2695-2700.
[7]梁从庆,李书渊,房志坚. HPLC法测定风湿定片中毒藜碱的含量[J].中药新药与临床药理,2004,15(4):268-270.
[8]叶伟峰,姚建标.高效液相色谱法测定风湿定片中丹皮酚的含量[J].儿科药学杂志,2008,14(2):41-43.
[9]曾上敏,秦斯民,曹华玲. HPLC法同时测定风湿定片中甘草酸和欧前胡素的含量[J].山东化工,2017,46(11):98-100、103.
[10]刘路,高旋,杨永健. HPLC-电雾式检测器的应用[J].中国医药工业杂志,2012,43(3):227-231.
[2]王其勇,许亚玲. HPLC法测定八角枫药材不同药用部位中L(-)-八角枫碱的含量[J].中国药房,2016,27(27):3877-3879.
[3] AKAO T,YOSHINO T,KOBASHI K,et al. Evaluation of salicin as an antipyretic prodrug that does not cause gastric injury[J]. Planta Med,2002,68(8):714-718.
[4]金贤兰.徐长卿药理作用及临床应用研究进展[J].现代医药卫生,2010,26(19):2947-2948.
[5]朱艺欣,李宝莉,马宏胜,等.白芷的有效成分提取、药理作用及临床应用研究进展[J].中国医药导报,2014,11(31):159-162、166.
[6]高雪岩,王文全,魏胜利,等.甘草及其活性成分的药理活性研究进展[J].中国中药杂志,2009,34(21):2695-2700.
[7]梁从庆,李书渊,房志坚. HPLC法测定风湿定片中毒藜碱的含量[J].中药新药与临床药理,2004,15(4):268-270.
[8]叶伟峰,姚建标.高效液相色谱法测定风湿定片中丹皮酚的含量[J].儿科药学杂志,2008,14(2):41-43.
[9]曾上敏,秦斯民,曹华玲. HPLC法同时测定风湿定片中甘草酸和欧前胡素的含量[J].山东化工,2017,46(11):98-100、103.
[10]刘路,高旋,杨永健. HPLC-电雾式检测器的应用[J].中国医药工业杂志,2012,43(3):227-231.