三七不同部位及总皂苷提取物的HPLC指纹图谱研究.doc

三七不同部位及总皂苷提取物的HPLC指纹图谱研究 崔翰明，张秋燕，彭 亮，路文龙，王 阶* 中国中医科学院广安门医院（北京市宣武区北线阁5号，100053） 摘要：目的 建立三七不同部位及总皂苷提取物的HPLC指纹图谱分析方法。方法 三七不同药用部位（主根、筋条、剪口、叶、花）和不同规格（20-100头）及总皂苷提取物采用甲醇提取，RP-HPLC/UV法测定，Kromasil C18（250mm×4.6mm，5μm）色谱柱；乙腈和水（v/v）线性梯度洗脱；流速：1mL·min-1；柱温：30℃；检测波长：203nm；进样量：20μL。结果 三七在80min的色谱图中共确定17个特征峰，主要峰分离良好，经对照品和HPLC/MS判定峰4为三七皂苷R1、峰5为人参皂苷Rg1和Re、峰6为Rf、峰7为Rb1、峰8为Rc、峰9为Rb2、峰10为Rb3、峰11为Rd。以峰4为参照峰（S），计算17个特征峰的相对保留时间和相对峰面积比，各峰精密度、稳定性和测定重现性RSD均小于6.47％。通过相似度软件评价，不同规格三七根（20-100头）和剪口、筋条的指纹图谱近似，相似度大于0.971，但三七根和根茎与花和叶差异很大，相似度分别为0.596和0.564。三七总皂苷提取物因药用部位及生产工艺不同，其主要特征峰的峰面积及比例存在较大差异。结论 本方法简便、准确，重现性良好，可用于准确分辨三七不同药用部位及其总皂苷提取物的质量。 关键词：三七；高效液相色谱；指纹图谱；三七总皂苷 Study on the fingerprint of different part of Panax Notoginseng and total saponins by HPLC CUI Han-ming, ZHANG Qiu-yan, PENG Liang, LU Wen-long, Wang Jie * Guang’anmen Hospital, China Academy of Chinese Medical Science; Beijing 100053,China Abstract: Objective: To establish an analysis method of HPLC fingerprint of the different part of Panax Notoginseng and its total saponins. Method: The different part (including root, rhizome, branch root, leaf, flower) and different specifications of (20-100 pieces per 500g) Panax Notoginseng and its total saponins were extracted with methanol. The HPLC conditions were as following: Kromasil C18 column (250×4.6mm，5μm), acetonitrile and water linearity gradient elution, flow rate at 1.0mL·min-1, column temperature at 30℃, wavelength 203nm, injection 20μL. Result: The primary peaks in 80min HPLC chromatogram of the different part of Panax Notoginseng were well separated. The 4th，5th，6th，7th，8th，9th，10th，11th peak were identified as Notoginsenoside R1, ginsenoside Rg1 and Re, Rf, Rb1, Rc, Rb2, Rb3, Rd by control sample and HPLC/MS technique, respectively. 17 character peaks were selected as the fingerprint peaks of Panax Notoginseng. The relative retain time and relative area of 17 peaks were calculated by taking Notoginsenoside R1 peak(4th) as the referential peak. The RSDs of precise and stability a