摘要
目的：建立基于固相萃取和含内标HPLC特征图谱的大黄质量评价方法。方法：采用水回流提取,固相萃取净化的方法,获取大黄药材中的总蒽醌;用HPLC法建立大黄总蒽醌的特征图谱;DAD结合MSn对色谱峰进行定性,各个特征峰与内标峰的相对峰面积进行定量,从而对大黄质量进行评价。结果：大黄提取的最佳条件为料液比1∶150,回流提取60 min;提取液采用Accu BONDⅡSPE ODS-C18Cartridges固相萃取柱,上样量1 m L,3 m L 0.3%甲酸溶液分2次淋洗,然后3 m L 25%甲醇溶液（含0.3%甲酸）淋洗,3 m L甲醇洗脱,指标成分回收率为96.3%-103%;以5-羟甲基糠醛为内标物,建立了大黄总蒽醌的HPLC特征图谱,标定了12个特征峰;考察了10批大黄特征峰的相对保留时间,其RSD〈3%,计算了各特征峰与内标峰的相对峰面积,通过与2015年版中国药典一部含量测定方法进行对比、主成分分析和聚类分析,表明各个特征峰与内标峰的相对峰面积可以基本反映大黄中蒽醌类成分的含量,并可以将唐古特大黄、药用大黄和掌叶大黄进行区分。结论：该方法操作简便、快捷、专属性强,通过外加内标物达到无对照品也能进行含量测定的目的,减少外界条件对特征图谱的影响,提高测定结果的准确性,可以为大黄的质量评价研究提供参考。
Objective： To establish a quality evaluation method for Rhei Radix et Rhizoma based on SPE and internal standard HPLC characteristic chromatogram. Methods： Using water reflux extraction and SPE methods,the total Rhei Radix et Rhizoma anthraquinones were obtained. By HPLC method,the characteristic chromatogram of Rhei Radix et Rhizoma was established,and the qualitative research was performed with DAD and MSn. Relative area of the characteristic peaks in the samples was calculated by the addition of an internal standard to evaluate the quality of Rhei Radix et Rhizoma. Results： The results showed that the optimum condition were as follows,the solid-liquid ratio was 1∶ 150; the extraction time was 60 min. SPE optimum condition for the introduction of Accu BOND Ⅱ SPE ODS-C18 Cartridges SPE were as follows,the loaded concentration was 1 m L,the SPE column was rinsed twice with 3 m L 0. 3% formic acid,then rinsed with 3 m L 25% methanol（ containing 0. 3% formic acid） and finally eluted with 3 m L of methanol. The recovery rate was 96. 3% - 103%. With 5-hydroxymethylfurfural as an internal standard,the characteristic chromatogram was established,and twelve peaks were selected as characteristic peaks. In ten batches of samples,the RSD of relative retention time was 3%,and the relative area of the characteristic peaks were calculated. The principal component analysis and cluster analysis with the results of the Chinese Pharmacopoeia（ 2015 edition）,which indicated that the characteristic chromatogram could reflect the anthraquinones in Rhei Radix et Rhizoma,and distinguish the Rheum palmatum,Rheum tanguticum and Rheum officinale. Conclusion： It is concluded that the established method is simple,fast and specific,which can provide a reference for the evaluation of the quality of Rhei Radix et Rhizoma.
作者
王亮
郭威
周倩
张军
于宗渊
WANG Liang GUO Wei ZHOU Qian ZHANG Jun YU Zong-yuan(Shandong Academy of Traditional Chinese Medicine, Jinan 250014, China School of Pharmaceutical Sciences, Shandong Univer- sity of Traditional Chinese Medicine,Jinan 250355 ,China)
出处
《中药材》
CAS
CSCD
北大核心
2016年第6期1327-1332,共6页
Journal of Chinese Medicinal Materials
基金
山东省科技发展计划(2011GSF11901)
关键词
大黄
高效液相色谱
特征图谱
固相萃取
内标
5-羟甲基糠醛
Rhei Radix et Rhizoma
HPLC
Characteristic chromatogram
SPE
Internal standard
5-hydroxymethylfurfural
