摘要
目的:优化铁皮石斛黄酮苷类成分HPLC特征图谱分析方法,明确不同种源的共性成分与特异性成分。方法:色谱条件采用Kromasil 100-5 C18色谱柱;流动相四氢呋喃-乙腈-甲醇(10∶22∶5)-0.05%磷酸水溶液梯度洗脱;检测波长340nm;柱温30℃;流速1.0mL·min^-1。结果:27批铁皮石斛共标示出13个黄酮类特征峰,鉴别了6个黄酮碳苷(新西兰牡荆苷Ⅱ,新西兰牡荆苷Ⅰ,夏佛塔苷,异夏佛塔苷,佛莱心苷,异佛莱心苷)与1个黄酮氧苷(芦丁)共7个特征峰。不同批次样品检出7~11个特征峰,其中新西兰牡荆苷Ⅱ为不同来源样品相对稳定的共有峰,芦丁、佛莱心苷与异佛莱心苷特征峰差异较大。根据特征峰的相对丰度,可将样品分为3大类,其中第Ⅰ类10批样品主要来源于广东、江西、福建、浙江(武义)等丹霞地貌(即"丹霞种"),以检出明显的芦丁强峰为特征;第Ⅱ类11批样品主要来源于云南、广西等地(包括了"云南广南种""广西铁皮兰种"),以能检出佛莱心苷与异佛莱心苷特征峰为特征;而第Ⅲ类6批样品主要来源于浙江(即"浙江本地种"),以难以检出佛莱心苷与异佛莱心苷特征峰,检出不同程度的芦丁峰为特征。比较"丹霞种"与"广西铁皮兰种"的仿野生和大棚栽培铁皮石斛HPLC特征图谱,结果仿野生铁皮石斛中的特征峰在大棚栽培石斛中也能稳定检出,验证了铁皮石斛种源的可靠性。结论:所建立的分析方法能对铁皮石斛黄酮类特征峰达到较好的分离效果,重复性好,基本明确了不同种源铁皮石斛的共性及特异性黄酮苷类成分。提示新西兰牡荆苷Ⅱ适合作为特征图谱的参照物峰;可根据芦丁相对丰度或佛莱心苷与异佛莱心苷特征峰的检出及相对丰度作为铁皮石斛"丹霞种""云南广南与广西铁皮兰种"与"浙江本地种"的判断依据。
Objective: To optimize the HPLC characteristic spectrum of flavonoid glycosides of Dendrobium officinale, and identify the common and specific components of different provenances. Method:Kromasil 100-5 C18 column was adopted, with tetrahydrofuran-acetonitrile-methanol(10 ∶ 22 ∶ 5)-0. 05%phosphoric acid as mobile phase(gradient elution). The detection wavelength was 340 nm, the column temperature was 30 ℃,and the flow rate was 1. 0 mL·min-1. Result: 13 flavonoid characteristic peaks were marked in 27 batches of D. officinale,and 7 characteristic peaks of 6 flavonoid C-glycosides(vicenin Ⅱ,viceninⅠ,schaftoside,isoschaftoside,violanthin and isoviolanthin) and one flavonoid O-glycosides(rutin) was identified. 7-11 characteristic peaks were detected in different batches of samples. Among them,vicenin Ⅱ was a relatively stable common peak in different source samples,and the characteristic peaks of rutin,schaftoside and isoschaftoside were quite different. According to the relative abundance of the characteristic peaks,the samples could be divided into three categories. Among them,the first category had 10 batches of samples,which mainly came from Danxia landforms of Guangdong, Jiangxi, Fujian and Zhejiang(Wuyi) Province(which called " Danxia landform species") and characterized by detection of obvious peak of rutin. The second category had 11 batches of samples,which mainly came from Yunnan and Guangxi Province(which included " Yunnan Guangnan species" and " Guangxi Tiepilan species") and characterized by detection of violanthin and isoviolanthin. And the third category had 6 batches of samples,which were mainly derived from Zhejiang Province(which called " native species from Zhejiang") and characterized by detection of different degrees of rutin peak,but it was difficult to detect the characteristic peaks of violanthin and isoviolanthin. HPLC characteristic chromatograms of D. officinale in bionics wild cultivation and greenhouse of " Danxia landform species" and " Guangxi Tiepilan species" were compared. The results showed that the characteristic peaks in D. officinale planted in greenhouse could be detected stably,which verified the reliability of the source in D. officinale. Conclusion: The analytical method has a better separation effect on flavonoids of D. officinale,with a good reproducibility. The commonness and specificities of flavonoid glycosides components of D. officinale from different categories have basically confirmed. This suggests that Vicenin Ⅱ is suitable to be a reference peak for characteristic chromatogram. Both the relative abundance of rutin and the detection or relative abundance of violanthin and isoviolanthin peaks could be used as a reference to judge the categories of D. officinale in " Danxia landform species " or " Tiepilan species from Yunnan,South Guangdong and Guangxi" or " native species from Zhejiang".
作者
梁芷韵
谢镇山
黄月纯
袁媛
周楚娟
王雅文
魏刚
LIANG Zhi-yun;XIE Zhen-shan;HUANG Yue-chun;YUAN Yuan;ZHOU Chu-juan;WANG Ya-wen;WEI Gang(School of the First Clinical Medical,Guangzhou University of Chinese Medicine,Guangzhou 510005,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China;National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;School of Pharmaeentical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Shaoguan Research Institute of Dendrobium Officinal ,Shaoguan 512005,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2019年第1期22-28,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
中央本级重大增减支项目(2060302)
广州中医药大学高水平大学基金项目(2050205)
关键词
铁皮石斛
特征图谱
黄酮苷类成分
丹霞种
云南广南种
广西铁皮兰种
浙江本地种
Dendrobium officinale
characteristic chromatogram
flavonoid glycosides
Danxia landform species
Yunnan Guangnan species
Guangxi Tiepilan species
native species from Zhejiang
