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目的:建立不同产地灯台叶HPLC指纹图谱,为灯台叶产地鉴别、质量控制和评价提供科学依据,推动傣药现代化发展进程。方法:采用正交试验设计优化灯台叶超声提取方法,建立HPLC指纹图谱。色谱条件:采用Agilent ZORBAX Eclipse XDB C18色谱柱(250 mm×4.6 mm,5μm),以0.1%甲酸水(A)-乙腈(B)为流动相,梯度洗脱(0~5 min,5%B;5~35 min,5%B→42%B;35~40 min,42%B→80%B;40~50 min,80%B→95%B;50~65 min,95%B),流速1.0 m L·min~(-1),柱温30℃,检测波长287 nm,进样量5μL。结合相似度评价及聚类分析对不同产地灯台叶指纹图谱进行比较。结果:正交实验结果表明,70%甲醇水溶液超声50 min,料液比1∶10为最佳提取条件。建立了灯台叶HPLC指纹图谱,确定了10个共有峰。海南中和、广西南宁北湖村、海南海口高新区等3个产地样品相似度小于0.8,系统聚类中被逐一分开;海南万宁、广东广州和海南海口海府大道3个产地样品相似度介于0.8~0.9之间;海南文昌、广东东莞、海南乐东、海南琼海、广西南宁六安村、西双版纳勐海、西双版纳勐腊7个产地样品相似度均大于0.9,系统聚类分析能聚为一类。结论:不同产地灯台叶化学组分较相似,但含量差异明显。海南文昌、广东东莞、海南乐东、海南琼海、广西南宁六安村5个产地样品与传统药用产地西双版纳灯台叶样品质量相似,可以作为傣药灯台叶药材来源。
OBJECTIVE: To establish HPLC fingerprints of lamp holder leaves in different areas to provide a scientific basis for the identification, quality control and evaluation of lamp holder leaves and to promote the modernization of Dai drug. Methods: Orthogonal design was used to optimize the ultrasonic extraction of lampshade and HPLC fingerprint was established. Chromatographic conditions: gradient elution (0-5 min, 5% B on a Agilent ZORBAX Eclipse XDB C18 column (250 mm × 4.6 mm, 5 μm) using 0.1% formic acid in water ; 5 to 35 min, 5% B to 42% B; 35 to 40 min, 42% B to 80% B; 40 to 50 min, 80% B to 95% B; The flow rate was 1.0 m L · min -1, the column temperature was 30 ℃, the detection wavelength was 287 nm, and the injection volume was 5 μL. Combining the similarity evaluation and cluster analysis, the fingerprints of lampshade from different places were compared. Results: The results of orthogonal experiment showed that 70% methanol aqueous solution was sonicated for 50 min and the ratio of solid to liquid 1:10 was the best extraction condition. The HPLC fingerprint of the lamp holder was established and 10 common peaks were identified. Similarities of Hainan Zhonghe, Beihu Village, Nanning, Guangxi and Hainan Haikou High-tech Zone were less than 0.8, and the similarity in the samples was separated one by one in the system cluster. Sample similarity of samples from three producing areas in Hainan Wanning, Guangzhou Guangdong and Haikou Haikou The similarity of samples from Hainan Wenchang, Guangdong Dongguan, Hainan Ledong, Hainan Qionghai, Nan’an Guangxi Lu’an, Menghai Xishuangbanna and Mengla Xishuangbanna were both greater than 0.9, and the cluster analysis could be clustered into one class. Conclusion: The chemical components of leaves from different places are similar, but the content is obviously different. Hainan Wenchang, Dongguan, Hainan Ledong, Hainan Qionghai, Nan’an, Guangxi Lu’an five samples of origin and traditional medicinal Xishuangbanna lampstand sample quality is similar, can be used as a source of Dai medicine dai’ai.