苦瓜皂苷的HPLC-ELSD指纹图谱研究

目的：建立苦瓜皂苷类成分的高效液相色谱-蒸发光散射（HPLC-ELSD）指纹图谱。方法：新鲜苦瓜果肉匀浆，冷冻干燥后经50%甲醇超声提取，HP-20大孔树脂固相萃取纯化得到苦瓜总皂苷。采用HPLC-ELSD法对14批次不同品种的苦瓜总皂苷进行检测，色谱柱为YMC-Pack ODS-A（5 μm，250 mm×4.6 mm）；以乙腈-水梯度洗脱（0 min，25%乙腈；40 min，35%乙腈；80 min，50%乙腈），流速1.0 mL·min^-1；漂移管温度：108.0℃；气体：空气；气流速度：3.0 L·min^-1。所得指纹图谱采用中药色谱指纹图谱相似度评价软件进行分析。结果：确定了12个共有峰作为苦瓜皂苷的特征指纹峰，并指认了其中4个色谱峰的化合物结构；14批次不同品种苦瓜皂苷指纹图谱与对照指纹图谱的相似度介于0.493~0.880之间。结论：本法经方法学验证，可为苦瓜的品种鉴定、品质评价和质量控制提供参考依据。

Objective: To establish an HPLC-ELSD chromatographic fingerprint analysis method for the total triterpenoid saponins in the fruits of Momordica charantia L.. Methods: A total of 14 batches of freeze-dried fruits of M.charantia L.varieties were selected for the study.The total triterpenoid saponins were extracted using 50% methanol,purified by solid phase extraction over HP-20 macroporous resin,and then analyzed by HPLC-ELSD.The samples were separated on YMC-Pack ODS-A column (250 mm×4.6 mm,5 μm),and eluted with a gradient of acetonitrile and water (0 min,25% acetonitrile;40 min,35% acetonitrile;80 min,50% acetonitrile) at a flow rate of 1.0 mL·min^-1. The drift tube temperature was 108 ℃;the gas was air;and the flow rate was 3.0 L·min^-1.The obtained HPLC fingerprints were then analyzed by the similarity evaluation software. Results: A total of 12 peaks were designated as characteristic common peaks,and 4 of them were identified by comparing with authentic compounds.Similarity index of 14 batches of samples varied from 0.493 to 0.880. Conclusion: The methodology validation shows that the established method could be used for species identification,quality evaluation and control of the fresh fruits of M.charantia.