HPLC法同时测定杠板归不同部位中5种成分的含量

Simultaneous determination of five components in different parts of Polygonum Perfoliatum L.by HPLC

目的建立高效液相色谱法同时测定杠板归不同部位(茎、叶、根)中5种成分的含量。方法采用Luna?-C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈-0.1%磷酸水为流动相梯度洗脱,流速为1.0 mL/min,检测波长为320 nm,柱温为30℃,测定杠板归不同部位中绿原酸、咖啡酸、芦丁、阿魏酸、槲皮素的含量。结果绿原酸、咖啡酸、芦丁、阿魏酸、槲皮素分别在0.046~4.6μg/mL,0.212~21.2μg/mL,0.055~5.5μg/mL,0.084~8.4μg/mL,0.114~11.4μg/mL范围内质量浓度与峰面积呈线性关系良好(r>0.9990),平均加样回收率为97.65%~102.09%,RSD为1.76%~2.44%。不同部位中5种成分含量差异较大,绿原酸和咖啡酸2个成分在叶中含量高,且远远高于根和茎中的含量;芦丁和阿魏酸主要存在于茎和根中,叶中未检测到;槲皮素在茎中含量最高,叶中次之,根中最低。结论建立的HPLC方法准确可靠、重复性好、专属性强,可用于杠板归不同部位中5个成分含量的同时测定。

Objective To establish an HPLC method for the simultaneous determination of the contents of five compounds in stems,leaves,and roots of Polygonum Perfoliatum L..Methods The quantification of chlorogenic acid,caffeic acid,rutin,ferulic acid,and quercetin was conducted on a Luna-C 18 column(4.6 mm×250 mm,5μm).The mobile phase was consisted of acetonitrile and 0.1% phosphoric acid solution for gradient elution with the flow rate of 1.0 mL/min.The column temperature was kept at 30℃ and the detection wavelength was set at 320 nm.Results Chlorogenic acid,caffeic acid,rutin,ferulic acid,and quercetin showed good linear relationships in the range of 0.046-4.6μg/mL,0.212-21.2μg/mL,0.055-5.5μg/mL,0.084-8.4μg/mL,0.114-11.4μg/mL(all r>0.9990),and the average recoveries were 97.65%-102.09%with the RSD of 1.76%-2.44%.The contents of these five components in different parts were significantly different.The contents of chlorogenic acid and caffeic acid were higher in leaves than those in roots and stems.Rutin and ferulic acid were mainly found in stems and roots,but not detected in leaves.The content of quercetin was the highest in the stem,and the lowest in the roots.Conclusion The established method is accurate,reliable,reproducible,and highly specific,which can be used for the simultaneous determination of the contents of five components in different parts of Polygonum Perfoliatum L..