Liu Xiao, Liu Jingjing, Cai Hao, Li Songlin, Ma Xiaoqing, Lou Yajing, Qin Kunming, Guan Hongyue, Cai Baochang
Department of Chinese Materia Medica, College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, People's Republic of China ; Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing, People's Republic of China ; National First-Class Key Discipline for Science of Chinese Materia Medica, Nanjing University of Chinese Medicine, Nanjing, People's Republic of China ; Nanjing Haichang Chinese Medicine Group Corporation, Nanjing, People's Republic of China.
Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, People's Republic of China.
Pharmacogn Mag. 2014 Jul;10(39):338-45. doi: 10.4103/0973-1296.137376.
Harmful sulfur-fumigation processing method is abused during Radix Angelicae Dahuricae preparation. However, the analytical technique characterizing Radix Angelicae Dahuricae before and after the sulfur-fumigation process is absent.
The high performance liquid chromatography (HPLC) technique was adopted to develop methods combining finger-print analysis and multi-ingredients simultaneous determination for quality evaluation of Radix Angelicae Dahuricae before and after the sulfur-fumigation process. The chromatographic fingerprint method was established for qualitative analysis coupled with statistical cluster analysis basing on Euclidean distance. Additionally, a determination method was developed for quantitative analysis, which was able to assay the concentrations of the major coumarins including imperatorin, isoimperatorin, xanthotoxin, xanthotoxol, isoimpinellin, oxypeucedanin, and bergapten in Radix Angelicae Dahuricae simultaneously. The separations of the two methods were both achieved on a Hypersil octadecylsilyl C18 column (250 mm × 4.6 mm, 5 μm) at 35°C under different strategic gradient elution programs. The detection wavelength was set at 254 nm all the time. Method validation data indicated that the methods were both reliable and applicable. They were then used to assay different Radix Angelicae Dahuricae samples collected from good agricultural practice (GAP) bases and local herbal markets.
The successful application demonstrated that the combination of HPLC fingerprint and simultaneous quantification of multi-ingredients offers an efficient approach for quality evaluation of Radix Angelicae Dahuricae before and after the sulfur-fumigation process.
In order to discriminate Radix Angelicae Dahuricae before and after the sulfur-fumigation process, oxypeucedanin, and xanthotoxol were the most sensitive biomarkers and should be determined.
白芷炮制过程中存在滥用有害的硫熏加工方法的现象。然而,目前缺乏表征硫熏前后白芷的分析技术。
采用高效液相色谱(HPLC)技术,建立指纹图谱分析与多成分同时测定相结合的方法,用于硫熏前后白芷的质量评价。建立色谱指纹图谱法进行定性分析,并基于欧氏距离进行统计聚类分析。此外,开发了一种定量分析方法,能够同时测定白芷中主要香豆素类成分的含量,包括欧前胡素、异欧前胡素、花椒毒素、氧化前胡素、异茴芹内酯、氧化前胡内酯和佛手柑内酯。两种方法均在Hypersil十八烷基硅烷C18柱(250 mm×4.6 mm,5μm)上,于35℃下采用不同的梯度洗脱程序进行分离。检测波长始终设置为254 nm。方法验证数据表明,这两种方法均可靠且适用。然后用它们对白芷的不同样品进行测定,这些样品分别采集自良好农业规范(GAP)基地和当地中药材市场。
成功的应用表明,HPLC指纹图谱与多成分同时定量相结合为硫熏前后白芷的质量评价提供了一种有效的方法。
为了鉴别硫熏前后的白芷,氧化前胡内酯和氧化前胡素是最敏感的生物标志物,应予以测定。