高效液相色谱法同时测定苎麻根中5-咖啡酰奎宁酸与绿原酸和咖啡酸的含量

Determination of 5-caffeoylquinic acid,chlorogenic acid and caffeic acid in Boehmeria nivea by HPLC

目的建立高效液相色谱法同时测定苎麻根中5-咖啡酰奎宁酸、绿原酸和咖啡酸的含量。方法用高效液相色谱法对苎麻根中的5-咖啡奎宁酸、绿原酸和咖啡酸进行测定。色谱柱:Agilent ZORBAX SB-C_(18)(4.6 mm×250 mm,5μm);流动相为乙腈-0.4%磷酸溶液梯度洗脱;检测波长:326 nm。考察该方法的专属性、标准曲线和定量下限、精密度与回收率、稳定性及重现性。结果 5-咖啡酰奎宁酸、绿原酸、咖啡酸的分别在26.32~263.20 ng、103.13~1031.26ng、43.32~433.20 ng线性关系良好,回归方程分别为Y=17567.60X+205.61(r=0.999 1),Y=25927.14X+246.23(r=0.999 9),Y=16839.22X+71.06(r=0.999 4);平均加样回收率分别为101.83%,98.77%,99.77%。结论本研究建立的方法检测苎麻根中5-咖啡酰奎宁酸、绿原酸、咖啡酸的含量,方法重现性好,结果准确、可靠,为苎麻根药材的质量控制研究奠定了良好的基础。

Objective To establish a high performance liquid chroma- tograghy（HPLC） method to determine the content of 5 -caffeoylquinic acid, chlorogenic acid and caffeic acid in Boehmeria nivea. Methods The product was analyzed by HPLC, using Agilent ZORBAX SB - Cs column （4.6 mm 250 mm, 5 μm） , with gradient elution acetonitrile - 0. 4% phosphate as the mobile phase. The flow rate was 1.0 mL 2 min-1 and the wavelength was 326 nm. The specificity, standard curve, limit of quantification, precision and recovery, stability and repeatability were measured. Results There was a good linear relationship of 5 -caf- feoylquinic acid, chlorogenic acid and caffeic acid in 26. 32 -263.20 ng, 103.13 - 1031.26 ng and 43.32 -433.20 ng respectively. The re- gression equations were Y = 17567.60X ＋ 205.61 （ r --- 0. 999 1 ）, Y = 25927. 14X ＋ 246. 23 （ r = 0. 999 9 ）, Y = 16839.22X ＋ 71.06 （ r = 0.999 4） . The average recoveries were 101.83%, 98.77% and 99. 77%, respectively. Conclusion The method established in this study for detecting the content of 5 -caffeoylquinic, chlorogenic acid and caffeic acid in Boehmeria nivea has good reproducibility, accurate and reliable results. The method can be used for the quality control of Boeh- meria nivea.