大叶千斤拔根、茎的HPLC指纹图谱及成分差异研究

目的： 为了研究大叶千斤拔根、茎的化学成分差异，首先建立大叶千斤拔根的HPLC指纹图谱，对根、茎的成分差异进行定性分析；其次，针对二者共有成分染料木苷、6″-O-丙二酰染料木苷、染料木素建立含量测定方法，进行定量评价。方法： 采用Agilent Zorbax SB-phenyl（4.6 mm×250 mm，5 μm）色谱柱，以乙腈-0.5%乙酸水溶液为流动相，梯度洗脱，流速1.0 mL·min^-1，检测波长260 nm；采用中药指纹图谱相似度评价系统（2.0版）进行相似度评价；对3个成分的含量测定结果，采用SPSS（16.0）软件t检验分析，确定区分根、茎的指标成分。结果： 11批根的图谱相似度为0.919~0.984，7批茎的图谱相似度为0.638~0.848；染料木苷、6″-O-丙二酰染料木苷、染料木素3个成分的含量在根中分别为0.64~1.30、0.62~1.21、0.13~0.47 mg·g^-1，在茎中分别为0.07~0.31、0.14~1.00、0.10~0.28 mg·g^-1。结论： 大叶千斤拔根、茎的化学成分存在一定差异，染料木苷、6″-O-丙二酰染料木苷、染料木素3个成分总量可用于大叶千斤拔根的质量控制，染料木苷含量或染料木苷与染料木素的含量比值可作为区分根、茎的指标。

Objective: To evaluate chemical constituent differences of root and stem of Moghania macrophylla,HPLC fingerprint of the root was established,and constituent differences of root and stem samples were qualitatively analyzed. Meanwhile,determination method for three main constituents(genistin,6″-O-malonylgenistin and genistein)found in both root and stem samples was established for quantitative analysis. Methods: HPLC analysis was performed on Agilent Zorbax SB-phenyl column(4.6 mm×250 mm,5 μm)with a mixture of 0.5% acetic acid solution and acetonitrile as the mobile phase in gradient elution at a flow rate of 1.0 mL·min^-1. The detection wavelength was set at 260 nm. The fingerprint similarity was analyzed with Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica(version 2.0). The differential compounds were found by t test with SPSS software(version 16.0). Results: The fingerprint similarities of eleven root samples ranged from 0.919 to 0.984,while those of seven stem samples ranged from 0.638 to 0.848. The contents of genistin,6″-O-malonylgenistin and genistein were 0.64-1.30 mg·g^-1,0.62-1.21 mg·g^-1 and 0.13-0.47 mg·g^-1 in 11 root samples,and 0.07-0.31 mg·g^-1,0.14-1.00 mg·g^-1 and 0.10-0.28 mg·g^-1 in 7 stem samples. Conclusion: The chemical constituents in root and stem of Moghania macrophylla were different. The total contents of genistin,6″-O-malonylgenistin and genistein can be used as marker for quality evaluation of root. The content of genistin or the ratio of genistin to genistein can be used as differential marker for root and stem.

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