HPLC法测定藏药独一味及其伪品糙苏中7个成分含量

Simultaneous determination of seven components in Tibetan medicine Lamiophlomis Herba and its counterfeit Phlomis Radix

目的:采用高效液相色谱法同时测定藏药独一味及其伪品糙苏中胡麻属苷、山栀苷甲酯、绿原酸、8-O-乙酰山栀苷甲酯、连翘酯苷B、木犀草苷和毛蕊花糖苷7个成分的含量,并比较两者成分的差异。方法:采用HSS T3 C18色谱柱(4.6 mm×250 mm,5μm),以乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱(0~11 min,9%A;11~35 min,9%A→18%A;35~50 min,18%A),流速1.0 mL·min^(-1),检测波长为235 nm,柱温30℃。结果:50 min内独一味和糙苏的主要色谱峰能够达到完全分离,胡麻属苷、山栀苷甲酯、绿原酸、8-O-乙酰山栀苷甲酯、连翘酯苷B、木犀草苷和毛蕊花糖苷质量浓度分别在2.026~81.04μg·mL^(-1)(r=0.999 9,n=6)、2.064~82.56μg·mL^(-1)(r=0.999 9,n=6)、1.924~96.20μg·mL^(-1)(r=0.999 8,n=6)、1.912~76.48μg·mL^(-1)(r=0.999 9,n=6)、1.874~74.96μg·mL^(-1)(r=0.999 8,n=6)、1.948~77.92μg·mL^(-1)(r=0.999 9,n=6)和1.890~75.60μg·mL^(-1)(r=0.999 9,n=6)范围内线性关系良好。11批独一味中胡麻属苷、山栀苷甲酯、绿原酸、8-O-乙酰山栀苷甲酯、连翘酯苷B、木犀草苷和毛蕊花糖苷含量范围范围分别为0.265%~0.872%、0.354%~1.058%、0.124%~0.641%、0.182%~1.426%、0.121%~0.719%、0.138%~1.301%和0.091%~1.173%;12批糙苏中胡麻属苷、山栀苷甲酯、8-O-乙酰山栀苷甲酯、连翘酯苷B和毛蕊花糖苷含量范围分别为0.196%~0.751%、0.358%~0.652%、0.204%~0.822%、0.101%~0.761%和0.024%~0.682%,均未检出绿原酸和木犀草苷。结论:所建立方法操作简单,具有较好的重复性和稳定性,结合伪品的同时测定,可用于独一味药材的质量控制。

Objective：To establish a new method for simultaneous determination of sesamoside,shanzhiside methyl ester,chlorogenic acid,8-O-acetyl shanzhiside methylester,forsythoside B,luteoloside and acteoside inTibetan medicine Lamiophlomis Herba and its counterfeit Phlomis Radix.Methods：HPLC analysis was performed on a HSS T3 C18 column（4.6 mm×250 mm,5 μm）.The mobile phase consisted of acetonitrile-0.1% phosphate acid solution as a linear gradient elution（0-11 min,9%A;11-35 min,9%A → 18%A;35-50 min,18%A）and the flow rate was 1 mL·min-1.The detection wavelength was 235 nm and the column temperature was 30 ℃. Results：Seven characteristic peaks were simultaneously determined by HPLC within 50 min.The linear ranges of sesamoside,shanzhiside methyl ester,chlorogenic acid,8-O-acetyl shanzhiside methylester,forsythoside B,luteoloside and acteoside were 2.026-81.04 μg·mL-1（r=0.999 9,n=6）,2.064-82.56 μg·mL-1（r=0.999 9, n=6）,1.924-96.20 μg·mL-1（r=0.999 8,n=6）,1.912-76.48 μg·mL-1（r=0.999 9,n=6）,1.874-74.96μg·mL-1（r=0.999 8, n=6）,1.948-77.92 μg·mL-1（r=0.999 9,n=6）and 1.890-75.60 μg·mL-1（r=0.999 9,n=6）,respectively.The contents of the above seven components in 11 samples of Lamiophlomis Herba were 0.265%-0.872%,0.354%-1.058%,0.124%-0.641%,0.182%-1.426%,0.121%-0.719%,0.138%-1.301 % and 0.091%-1.173%.The contents of sesamoside,shanzhiside methyl ester,8-O-acetyl shanzhiside methylester,forsythoside B,and acteoside of 12 samples of Phlomis Radix were 0.196%-0.751%,0.358%-0.652%,0.204%-0.822%,0.101%-0.761% and 0.024%-0.682%,respectively;chlorogenic acid and luteoloside were not detected in 12 samples of Phlomis Radix.Conclusion：The developed HPLC method could be used to distinguish between Lamiophlomis Herba and Phlomis Radix and could be used for the quality control of the crude drug.