HPLC指纹图谱结合多指标含量测定的八角莲质量评价研究

Study on quality evaluation of Dysosma versipellis by HPLC fingerprint combined with multi-index content determination

建立八角莲HPLC指纹图谱,并同时测定5种化学成分的含量,为其质量评价提供依据。采用Thermo HyPURITY C 18色谱柱(250 mm×4.6 mm,5μm),以甲醇(A)-0.4%磷酸水溶液(B)为流动相,梯度洗脱,柱温30℃,体积流量1.0 mL/min,检测波长290 nm,进样量10μL,建立了八角莲的HPLC指纹图谱。16批样品指纹图谱中有28个共有峰,相似度较好,经与对照品比照确认了8个成分,对(+)-tanegool-7′-methylether、4′-去甲基鬼臼毒素、槲皮素、鬼臼毒素、山柰酚5种化学成分进行了定量分析,5个化学成分在各自范围内线性关系良好(r^2≥0.9990),精密度、重复性、稳定性均符合定量要求;加样回收率在98.7%~99.6%,RSD小于3%。所建立的方法简便、稳定、可靠,HPLC指纹图谱结合多指标成分含量测定可用于八角莲药材质量的评价。

To provide the scientific basis for the quality evaluation of Dysosma versipellis,its HPLC fingerprints was established,and the contents of five components were simultaneously determined.The analyses were performed on Thermo HyPURITY C 18 column(250 mm×4.6 mm,5μm),and the mobile phase comprised of MeOH(A)-0.4%phosphoric acid aqueous solution(B)with the gradient elution at a flow rate of 1.0 mL/min.The detection wavelength was set at 290 nm;the column temperature was maintained at 30℃and the injection volume was 10μL.The HPLC fingerprint with twenty-eight common peaks of D.versipellis in 16 batches of sample was established,and the similarities of samples were good.Eight components were identified by comparison with the reference substances.Five components,namely,(+)-tanegool-7′-methylether,4′-demethylpodophyllotoxin,quercetin,podophyllotoxin,kaempferol were analyzed quantitatively.The linear relationship of five components was good(r^2≥0.9990).The precision,repeatability and stability were all in line with quantitative requirements.The average recoveries(n=6)of five components were between 98.7%and 99.6%,and the RSD were less than 3%.The established method is simple,stable and reliable,and can be expected for the quality control of D.versipellis.