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目的:建立同时测定桂枝芍药知母汤提取物中异甘草素、6-姜辣素、白术内酯Ⅲ、白术内酯Ⅰ、白术内酯Ⅱ含量的方法。方法:桂枝芍药知母汤采用70%乙醇提取得到提取物,采用HPLC法测定含量。色谱条件:采用Zorbax SB-C18(4.6 mm×250mm,5μm)色谱柱,以乙腈-0.01%甲酸水为流动相,梯度洗脱,流速1.0 m L·min-1,检测波长220 nm(6-姜辣素、白术内酯Ⅲ、白术内酯Ⅰ),275 nm(白术内酯Ⅱ),370 nm(异甘草素),柱温30℃。结果:异甘草素、6-姜辣素、白术内酯Ⅲ、白术内酯Ⅰ、白术内酯Ⅱ进样量分别在0.013 2~0.264μg(r=0.999 8),0.164 0~3.280μg(r=0.999 9),0.051 1~1.022μg(r=0.999 9),0.051 6~1.032μg(r=0.999 9),0.029 6~0.592μg(r=0.999 9)范围内与色谱峰面积呈良好的线性关系;加样回收率(n=6)均在96.2%~103.3%,RSD均小于3%。异甘草素、6-姜辣素、白术内酯Ⅲ、白术内酯Ⅰ和白术内酯Ⅱ含量分别为0.064 4、1.158、0.319 1、0.247 2、0.160 5 mg·g-1。结论:本方法简单、准确,重复性好,为桂枝芍药知母汤物质基础研究提供了基础。
Objective: To establish a method for the simultaneous determination of isoliquiritigenin, 6 - gingerol, atractyloside Ⅲ, atractylodes rhizome Ⅰ and atractyloside Ⅱ in Guizhi Shaoyao Zhimu Tang extract. Methods: Guizhi Shaoyao Zhimu Tang was extracted with 70% ethanol to get the extract, and the content was determined by HPLC. Chromatographic conditions: The mobile phase was eluted with a gradient of acetonitrile-0.01% formic acid in water on a Zorbax SB-C18 (4.6 mm × 250 mm, 5 μm) column with a flow rate of 1.0 mL · min- Gingerol, atractylodes rhizome Ⅲ, atractylenolide Ⅰ), 275 nm (atractyloside Ⅱ), 370 nm (isoliquiritigenin) at 30 ℃. Results: The injection rates of isoliquiritigenin, 6-gingerol, atractylenolide III, atractylenolide I and atractylenolide II were 0.013 2-0.264 μg (r = 0.999 8), 0.164 0-3.280 μg = 0.999 9), 0.051 1 ~ 1.022μg (r = 0.999 9), 0.051 6 ~ 1.032μg (r = 0.999 9) and 0.029 6 ~ 0.592μg (r = 0.999 9) (N = 6) were 96.2% ~ 103.3%, RSD were less than 3%. Isoliquiritigenin, 6-gingerol, atractylenolide III, atractylen I and atractylenolide II were 0.064 4,1.158,0.319 1,0.247 2,0.160 5 mg · g -1, respectively. Conclusion: The method is simple, accurate and reproducible. It provides the basis for the material basic research of Guizhi Shaoyao Zhimu Tang.