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(5μm,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.