HPLC法同时测定温经汤中10种活性成分的含量

Simultaneous Determination of 10 Active Components in Wenjing Decoction by HPLC Method

目的:建立同时测定温经汤中10种活性成分(芍药内酯苷、芍药苷、β-蜕皮甾酮、甘草苷、阿魏酸、甘草素、肉桂酸、桂皮醛、丹皮酚和甘草酸铵)含量的方法。方法:采用高效液相色谱(HPLC)法。色谱柱为Kromasil C18,流动相为乙腈-0.1%磷酸水溶液(梯度洗脱),流速为1.0 mL/min,双波长切换检测(0~25 min为240 nm,检测芍药内酯苷、芍药苷、β-蜕皮甾酮、甘草苷、阿魏酸;25~65 min为275 nm,检测甘草素、肉桂酸、桂皮醛、丹皮酚、甘草酸铵),柱温为25℃,进样量为20μL。结果:芍药内酯苷、芍药苷、β-蜕皮甾酮、甘草苷、阿魏酸、甘草素、肉桂酸、桂皮醛、丹皮酚、甘草酸铵的检测质量浓度线性范围分别为4.04~80.80、19.32~368.40、2.62~52.40、17.52~350.40、2.07~41.40、4.02~80.40、0.56~11.20、1.69~33.80、2.18~43.60、72.10~1 442.00μg/m L(r均≥0.999 6),检测限分别为0.06、0.03、0.04、0.06、0.07、0.05、0.04、0.06、0.07、0.09μg/mL,定量限分别为0.17、0.09、0.13、0.19、0.20、0.18、0.11、0.15、0.18、0.25μg/mL,精密度、重复性、稳定性(24 h)试验的RSD均<3.5%(n=6~7),各种成分的平均加样回收率为97.72%~100.60%(RSD为0.80%~2.49%,n=6)。结论:建立的HPLC法准确可靠、专属性好,可用于温经汤中芍药内酯苷等10种活性成分含量的同时测定。

OBJECTIVE:To establish a method for simultaneous determination of 10 active components(albiflorin,paeoniflorin,β-ecdysterone,liquiritin,ferulic acid,liquiritigenin,cinnamic acid,cinnamaldehyde,paeonol and ammonium glycyrrhetate)in Wenjing decoction.METHODS:HPLC method was adopted.The determination was performed on Kromasil C18 column with mobile phase consisted of acetonitrile-0.1%phosphoric acid water solution(gradient elution)at the flow rate of 1.0 mL/min,in dual wavelength switching detection(0-25 min,240 nm for albiflorin,paeoniflorin,β-ecdysterone,liquiritin,ferulic acid;25-65 min,275 nm for liquiritigenin,cinnamic acid,cinnamaldehyde,paeonol,ammonium glycyrrhetate).The column temperature was set at 25℃,and sample size was 20μL.RESULTS:The linear range of albiflorin,paeoniflorin,β-ecdysterone,liquiritin,ferulic acid,liquiritigenin,cinnamic acid,cinnamaldehyde,paeonol and ammonium glycyrrhetate were 4.04-80.80,19.32-368.40,2.62-52.40,17.52-350.40,2.07-41.40,4.02-80.40,0.56-11.20,1.69-33.80,2.18-43.60 and 72.10-1 442.00μg/mL(all r≥0.999 6),respectively.The limits of detection were 0.06,0.03,0.04,0.06,0.07,0.05,0.04,0.06,0.07,0.09μg/mL,respectively.The limits of quantitation were 0.17,0.09,0.13,0.19,0.20,0.18,0.11,0.15,0.18,0.25μg/mL,respectively.RSDs of precision,reproducibility and stability tests(24 h)were all lower than 3.5%(n=6-7).The average recovery rates were 97.72%-100.60%with the RSDs of 0.80%-2.49%(n=6).CONCLUSIONS:The established HPLC method is accurate,reliable and specfic,and suitable for simultaneous determination of 10 active components as albiflorin in Wenjing decoction.