波长切换法测定桂枝芍药知母汤提取物中5个脂溶性成分的含量

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×250 mm,5 μm)色谱柱,以乙腈-0.01%甲酸水为流动相,梯度洗脱,流速1.0 mL · 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 2~0.264 μg(r=0.999 8),0.164 0~3.280 μg(r=0.999 9),0.051 1~1.022 μg(r=0.999 9),0.051 6~1.032 μg(r=0.999 9),0.029 6~0.592 μg(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 an HPLC method with multi-wavelength for determination of isoliquiritigenin,6-gingerol,atractylenolide Ⅲ,atractylenolide I and atractylenolide Ⅱ in extract of Guizhi Shaoyao Zhimu decoction. Methods: The decoction extract was abstracted by 70% ethanol and quantified by HPLC.The separation was performed on a Zorbax SB-C₁₈(4.6 mm×150 mm,5 μm)column with the gradient elution of acetonitrile-0.01% formic acid water,at the flow rate of 1 mL · min^-1;the detection wavelength was set at 220 nm(6-gingerol,atractylenolide Ⅲ,atractylenolide Ⅰ),275 nm(atractylenolide Ⅱ),and 370 wnm(isoliquiritigenin);the column temperature was maintained at 30 ℃. Results: Isoliquiritigenin,6-gingerol,atractylenolide Ⅲ,atractylenolide Ⅰand atractylenolide Ⅱ had good linearity in the ranges of 0.013 2-0.264 μg(r=0.999 8),0.1640-3.280 μg(r=0.999 9),0.051 1-1.022 μg(r=0.999 9),0.051 6-1.032 μg(r=0.999 9)and 0.029 6-0.592 μg(r=0.999 9),respectively;the average recoveries(n=6)ranged from 96.2% to 103.3%,and the relative standard deviations were below 3%.The content of isoliquiritigenin,6-gingerol,atractylenolide Ⅲ,atractylenolide Ⅰand atractylenolide Ⅱ in extract were 0.0644,1.158,0.3191,0.2472,and 0.1605 mg · g^-1,respectively. Conclusion: This method is simple,reliable and accurate,and can provide basis for Guizhi Shaoyao Zhimu decoction basic research.