畲药柳叶蜡梅质量评价和谱效关系研究

Study on Quality Assessment and Fingerprint-effect Relationship of She Medicine?Chimonanthus Salicifolius S. Y. Hu

关键词:  畲药  柳叶蜡梅  液相指纹图谱  抗氧化活性  谱效关系

Key words:  She medicine  Chimonanthus salicifolius S. Y. Hu  HPLC fingerprint  antioxidant activity  fingerprint-effect relationship

摘要:

目的 评价不同产地及不同采收时间的柳叶蜡梅药材质量,建立谱效关系模型,挖掘与抗氧化活性有显著相关的色谱特征峰。方法 建立可同时定量多种成分(芦丁、异槲皮苷、山柰酚-3-O-芸香糖苷、槲皮苷、槲皮素、山柰酚)的液相指纹图谱方法,测定不同来源柳叶蜡梅中总黄酮含量,并采用DPPH法测定柳叶蜡梅样本的清除率SR及IC₅₀值,建立基于灰色关联度和相似度算法的谱效关系模型,辨别与抗氧化活性有显著相关的色谱特征峰。结果 不同产地柳叶蜡梅样本的总黄酮及其单体成分含量、抗氧化活性等指标存在较大差异,但同一基地来源的质量差异较小,松阳标准化基地样本IC50值<4.5 mg·mL^-1,RSD值<20%,显示整体抗氧化能力较强。7-8月份的柳叶蜡梅样本黄酮类物质累积最多且抗氧化能力最强,尤其是山柰酚-3-O-芸香糖苷含量最高,最高可达0.87%。此外,基于色谱数据的三维主成分分析结果显示,松阳和开化标准化种植基地的柳叶蜡梅质量属性较为接近,而与来自莲都地区的野生样本差距较大。通过灰色关联度和相关系数分析算法构建的谱效关系结果显示,山柰酚-3-O-芸香糖苷、槲皮苷、异槲皮苷、芦丁等黄酮成分均与抗氧化活性显著相关。结论 本研究明确了柳叶蜡梅最佳采摘期以及产地对药材质量的影响,初步明确山柰酚-3-O-芸香糖苷、槲皮苷、异槲皮苷、芦丁与抗氧化活性关联度较强,为柳叶蜡梅质量综合评价和药效物质基础研究提供参考。

Abstract:

OBJECTIVE To evaluate the quality of Chimonanthus salicifolius S. Y. Hu from different habitats and harvest time, establishing fingerprint-effect relationship models and assessing characteristic peaks significantly related to antioxidant activity. METHODS The HPLC fingerprint method was developed to quantitative determine rutin, isoquercitrin, kaempferol-3-O-rutinoside, quercitrin, quercetin and kaempferol, and the total flavonoid contents of Chimonanthus salicifolius S. Y. Hu leaves from different habitats were measured. The SR and IC₅₀ values of the Chimonanthus salicifolius S. Y. Hu samples were determined by DPPH assay. The fingerprint-effect relationship model based on grey relationship analysis and correlation coefficient method was investigated, to identify the characteristic chromatographic peaks which were significantly associated with antioxidant activity.RESULTS The contents of total flavonoid and its monomer components and antioxidant activities of Chimonanthus salicifolius S. Y. Hu from different habitats displayed marked differences. However, the quality differences of the samples from the same cultivation base had little difference. The IC₅₀ and RSD values of the samples from Songyang standardized cultivation base was <4.5 mg·mL^-1 and 20% respectively, indicating overall strong antioxidant capacities. For different harvest time, raw materials harvested in July and August exhibited highest values of flavonoids and antioxidant capacity, especially kaempferol-3-O-rutinoside, the highest content attainable was 0.87%. In addition, the results of three-dimensional principal component analysis based on chromatographic data showed that, Chimonanthus salicifolius S. Y. Hu samples from Songyang and Kaihua standardized cultivation bases had similar quality attributes, however, there was a big gap between the quality of wild samples from Liandu and the samples harvested from Songyang and Kaihua standardized cultivation bases. The results of spectral efficiency relationship constructed by grey correlation degree and correlation coefficient analysis algorithm showed that, kaempferol-3-O-rutinoside, quercitrin, isoquercitrin and rutin displayed a substantial correlation with antioxidant activities. CONCLUSION This study reveal the appropriate harvest time and the influence of habitats on the quality of Chimonanthus salicifolius S. Y. Hu. It is preliminarily confirmed that kaempferol-3-O-rutinoside, quercitrin, isoquercitrin and rutin display a substantial correlation with antioxidant activities. The results provide certain scientific basis for quality evaluation and effective substances ofChimonanthus salicifolius S. Y. Hu.