Sumino Megumi, Saito Yuko, Ikegami Fumio, Namiki Takao
Department of Japanese Oriental (Kampo) Medicine, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan
Department of Japanese Oriental (Kampo) Medicine, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan.
J Chromatogr Sci. 2015 Feb;53(2):320-4. doi: 10.1093/chromsci/bmu062. Epub 2014 Jun 30.
We developed a simultaneous analysis method using high-performance liquid chromatography coupled with diode-array detector (HPLC-DAD) for six principal compounds (atractylenolide III, alisol A, alisol B, paeoniflorin, ferulic acid and (Z)-ligustilide) in a traditional Japanese (Kampo) medicine, tokishakuyakusan (TSS). The HPLC separation was conducted on a reversed-phase TSK-gel ODS-80TS column (4.6 i.d. × 250 mm, 5 µm) at 40°C with a 0.1% phosphoric acid-acetonitrile gradient system. The DAD detection wavelength was set at 205, 232 and 330 nm. Calibration curves for the compounds showed linear regressions with correlation coefficients of >0.999. The intra- and inter-day precision (i.e., the relative standard deviation) were in the range of 0.50-1.55 and 0.70-1.80%, respectively. The average recovery yields of the compounds ranged from 98.3 to 103%. The present results will contribute to shorter analysis times with less organic solvent compared with the individual analysis of each compound for the evaluation of TSS. The application of the established method to TSS will also provide helpful information for the further pharmacological and clinical studies.
我们开发了一种同时分析方法，使用高效液相色谱仪结合二极管阵列检测器（HPLC-DAD）来测定日本传统汉方药物芍药甘草汤（TSS）中的六种主要化合物（白术内酯III、泽泻醇A、泽泻醇B、芍药苷、阿魏酸和（Z）-藁本内酯）。HPLC分离在反相TSK-gel ODS-80TS柱（内径4.6×250 mm，5 µm）上于40°C进行，采用0.1%磷酸-乙腈梯度系统。DAD检测波长设定为205、232和330 nm。化合物的校准曲线显示线性回归，相关系数>0.999。日内和日间精密度（即相对标准偏差）分别在0.50 - 1.55%和0.70 - 1.80%范围内。化合物的平均回收率在98.3%至103%之间。与对每种化合物进行单独分析以评估TSS相比，本研究结果将有助于缩短分析时间并减少有机溶剂的使用。将所建立的方法应用于TSS也将为进一步的药理和临床研究提供有用信息。
