HPLC法测定藏药独一味及其伪品糙苏中7个成分含量

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，5 μm），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2.026~81.04 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）、1.912~76.48μg·mL^-1（r=0.999 9，n=6）、1.874~74.96 μg·mL^-1（r=0.999 8，n=6）、1.948~77.92 μg·mL^-1（r=0.999 9，n=6）和1.890~75.60 μg·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 in Tibetan 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.