铁皮石斛黄酮苷类成分HPLC特征图谱优化及不同种源特征性分析

梁芷韵; 谢镇山; 黄月纯; 袁媛; 周楚娟; 王雅文; 魏刚

doi:10.13422/j.cnki.syfjx.20190112

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铁皮石斛黄酮苷类成分HPLC特征图谱优化及不同种源特征性分析

铁皮石斛种质资源专题 | 更新时间：2021-02-09

- 铁皮石斛黄酮苷类成分HPLC特征图谱优化及不同种源特征性分析
- HPLC Characteristic Spectrum Optimization of Flavonoid Glycosides on Dendrobium officinale and Characteristics Analysis of Different Provenances
- 中国实验方剂学杂志 2019年25卷第1期页码：22-28
- 作者机构：
  
  1.广州中医药大学　第一临床医学院，广州　 510006
  2.广州中医药大学　第一附属医院，广州　 510405
  3.中国中医科学院　中药资源中心，北京　 100700
  4.广州中医药大学　中药学院，广州　 510006
  5.韶关市铁皮石斛研究院，广东　韶关　 512005
- 作者简介：
  
  梁芷韵，在读硕士，从事中药质量标准研究与指纹图谱分析工作，E-mail：976788804@qq.com
  黄月纯，主任中药师，硕士生导师，从事中药质量标准、指纹图谱及活性成分研究，E-mail:huangyuechun@163.com；
  袁媛，研究员，从事中药鉴定与分子生药学研究，Tel：010-64087649，E-mail:y_yuan0732@163.com
- 基金信息：
  
  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)
- DOI：10.13422/j.cnki.syfjx.20190112
  中图分类号： R284.1;R22;R2-031
- 收稿日期：2018-08-24，
  
  网络出版日期：2018-10-19，
  
  纸质出版日期：2019-01-05
- 稿件说明：
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梁芷韵, 谢镇山, 黄月纯, 等. 铁皮石斛黄酮苷类成分HPLC特征图谱优化及不同种源特征性分析[J]. 中国实验方剂学杂志, 2019,25(1):22-28.

Zhi-yun LIANG, Zhen-shan XIE, Yue-chun HUANG, et al. HPLC Characteristic Spectrum Optimization of Flavonoid Glycosides on Dendrobium officinale and Characteristics Analysis of Different Provenances[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(1): 22-28.
梁芷韵, 谢镇山, 黄月纯, 等. 铁皮石斛黄酮苷类成分HPLC特征图谱优化及不同种源特征性分析[J]. 中国实验方剂学杂志, 2019,25(1):22-28. DOI： 10.13422/j.cnki.syfjx.20190112.

Zhi-yun LIANG, Zhen-shan XIE, Yue-chun HUANG, et al. HPLC Characteristic Spectrum Optimization of Flavonoid Glycosides on Dendrobium officinale and Characteristics Analysis of Different Provenances[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(1): 22-28. DOI： 10.13422/j.cnki.syfjx.20190112.

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 C

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340 nm；柱温30 ℃；流速1.0 mL·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。

Abstract

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 C

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

officinale

and 7 characteristic peaks of 6 flavonoid

-glycosides (vicenin Ⅱ

vicenin Ⅰ

schaftoside

isoschaftoside

violanthin and isoviolanthin) and one flavonoid

-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

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

officinale

planted in greenhouse could be detected stably

which verified the reliability of the source in

officinale

Conclusion:

The analytical method has a better separation effect on flavonoids of

officinale

with a good reproducibility. The commonness and specificities of flavonoid glycosides components of

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

officinale

in " Danxia landform species" or " Tiepilan species from Yunnan

South Guangdong and Guangxi" or " native species from Zhejiang" .

关键词

Keywords

references

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