HPLC-CAD法同时测定风湿定片中7种活性成分的含量

Simultaneous Determination of 7 Kinds of Active Components in Fengshiding Tablet by HPLC-CAD

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 cid,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 μ L. 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 0.11,0.06,0.06,0.10,0.11,0.08,0.07 μg/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.