Content Comparison of Mannose, Glucose and Narinhenin in <italic style="font-style: italic">Dendrobium officinale</italic> and <italic style="font-style: italic">Dendrobium huoshanense</italic>

Ya-wen WANG; Zhi-yun LIANG; Zhen-shan XIE; Jia-hong YE; Si-jian OU; Chu-juan ZHOU; Yuan YUAN; Yue-chun HUANG; Gang WEI

doi:10.13422/j.cnki.syfjx.20182418

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Content Comparison of Mannose, Glucose and Narinhenin in Dendrobium officinale and Dendrobium huoshanense

Topic on Germplasm Resources of Dendrobii Officinalis Caulis | 更新时间：2021-02-09

- Content Comparison of Mannose, Glucose and Narinhenin in Dendrobium officinale and Dendrobium huoshanense
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 1, Pages: 35-42(2019)
- 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　中药学院，广州　 510006
  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　第一附属医院，广州　 510405
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20182418
  CLC： R284.1;R289;R22;R927.2;R931.6
- Received：22 June 2018，
  
  Published Online：28 September 2018，
  
  Published：05 January 2019
- 稿件说明：
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Ya-wen WANG, Zhi-yun LIANG, Zhen-shan XIE, et al. Content Comparison of Mannose, Glucose and Narinhenin in Dendrobium officinale and Dendrobium huoshanense[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(1): 35-42.
DOI：

Ya-wen WANG, Zhi-yun LIANG, Zhen-shan XIE, et al. Content Comparison of Mannose, Glucose and Narinhenin in Dendrobium officinale and Dendrobium huoshanense[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(1): 35-42. DOI： 10.13422/j.cnki.syfjx.20182418.

摘要

目的：

优化铁皮石斛中甘露糖与葡萄糖的柱前衍生HPLC含量测定方法以及柚皮素HPLC含量测定方法，比较铁皮石斛与霍山石斛中这3种成分的含量差异。

方法：

在2015年版《中国药典》铁皮石斛甘露糖柱前衍生HPLC含量测定项下色谱条件基础上，选择乙腈-0.02 mol·L

－1

乙酸铵溶液为流动相系统梯度洗脱，同时测定甘露糖与葡萄糖的含量，并分析甘露糖与葡萄糖的峰面积比值；采用Kromasil 100-5 C

色谱柱（4.6 mm×250 mm，5 μm）；检测波长250 nm；流速1.0 mL·min

－1

；柱温30 ℃。柚皮素HPLC含量测定采用Kromasil 100-5 C

色谱柱（4.6 mm×250 mm，5 μm）；流动相乙腈-甲醇-0.4%磷酸溶液，梯度洗脱；检测波长290 nm；流速0.8 mL·min

－1

；柱温40 ℃。

结果：

甘露糖与葡萄糖在0.15～3.0，0.075～2.25 μg线性关系良好（

=0.999 9），平均加样回收率分别为99.01%（RSD 2.1%），101.69%（RSD 2.0%），重复性、耐用性等其他方法学研究符合要求。43批不同产区铁皮石斛中甘露糖、葡萄糖以及两者的含量之和分别在12.75%～36.40%，2.93%～18.39%，19.23%～54.58%，除极少数样品外，基本符合2015年版《中国药典》甘露糖含量限度要求，甘露糖与葡萄糖含量之和也接近总多糖含量限度要求；含量与产区相关性不显著。12批霍山石斛的总糖则分别在14.33%～29.47%，6.64%～15.20%，25.73%～44.37%，其含量以及峰面积比值基本落在铁皮石斛范围期间，多批次的平均含量也基本与铁皮石斛一致（约33%左右）。柚皮素在0.020 8～0.832 0 μg线性关系良好（

=0.999 9），平均加样回收率为101.96%（RSD 1.8%）。11批铁皮石斛与7批霍山石斛的柚皮素含量分别为0.053 2～0.122 4 mg·g

－1

（均值为0.081 0 mg·g

－1

），0.040 3～0.090 0 mg·g

－1

（均值为 0.068 3 mg·g

－1

），铁皮石斛含量稍高于霍山石斛，但含量均亦未达到0.02%的质量标准下限的常规要求。

结论：

铁皮石斛中甘露糖与葡萄糖HPLC含量测定方法重复性较好，用两者含量之和替代具有较大误差的总多糖含量作为测定指标具有可行性；单糖含量测定可应用于霍山石斛的定量质控指标；但依据2种石斛的总多糖含量、水解后的单糖含量与峰面积比值以及柚皮素含量，无法区分铁皮石斛与霍山石斛，需结合其他专属性方法方能对两种石斛进行区别。

Abstract

Objective:

To optimize the pre-column derivation high performance liquid chromatography (HPLC) content determination method of

-mannose and

-glucose as well as the content determination method of narinhenin in

Dendrobium officinale

and

huoshanense

and compare the contents of

-mannose

-glucose and narinhenin between

officinale

and

huoshanense

Method:

A pre-column derivation HPLC method modified by

Chinese Pharmacopoeia

(Ch.P) 2015 was used to simultaneously determine the contents of

-mannose and

-glucose

with acetonitrile-0.02 mol·L

-1

ammonium acetate solution as mobile phase for gradient elution. Kromasil 100-5 C

was performed with the wavelength set at 250 nm

and the flow rate was 1 mL·min

-1

; column temperature was 30 ℃. HPLC content determination of narinhenin was performed on Kromasil 100-5 C

with the acetonitrile-methanol-0.4% phosphoric acid solution as mobile phase for gradient elution

and the wavelength was set at 290 nm; the flow rate was 0.8 mL·min

-1

and column temperature was 40 ℃.

Result:

-mannose and

-glucose showed a good linear relationship within the range of 0.15-3.0 μg and 0.075-2.25 μg (

=0.999 9); and their average recoveries were 99.01% (RSD 2.1%) and 101.69% (RSD 2.0%) respectively. In addition

the other methodological researches such as repeatability and durability all met the requirements. The contents of

-mannose(

)

-glucose(

) and sum of them (

) were 12.75%-36.40%

2.93%-18.39% and 19.23%-54.58% in 43 batch of

officinale

. Almost all of the results except very few samples reached the

-mannose standard in Ch.P 2015

and the total content of

-mannose and

-glucose was also up to the total polysccharide standard in Ch.P. The correlation between content and origin was not significant. The contents of

-mannose(

)

-glucose(

) and sum of them (

) were 14.33%-29.47%

6.64%-15.20%

and 25.73%-44.37% in 12 batch of

huoshanense

. These contents and ratio of peak areas of

-mannose to

-glucose (

) were within the scope of

officinale

in addition

their average contents were basically the same with those in

officinale

(about 33%). Next

naringenin showed a good linear relationship within the range of 0.020 8-0.832 0 μg (

=0.999 9)

and its average recovery was 101.96% (RSD 1.8%). The content of naringenin was 0.053 2-0.122 4 mg·g

-1

(average value of 0.081 0 mg·g

-1

) in 11 batch of

officinale

slightly higher than 0.040 3-0.090 0 mg·g

-1

(average value of 0.068 3 mg·g

-1

) in 7 batch of

huoshanense

. All of these results of narinfenin did not reach the content lower limit in Ch.P.

Conclusion:

The method used to determinate the content of

-mannose and

-glucose is reproducible

and their sum content is possible to substitute the total polysccaride determination (with higher errors) in

officinale;

monosaccharide content determination can be used for quantitative quality control of

huoshanense

. However

it could not distinguish

officinale

and

huoshanense

by determining the contents of polysccharide

-glucose

-mannose and narinhenin

and shall be combined with other specificity methods for further identification.

关键词

Keywords

references

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Content Comparison of Mannose, Glucose and Narinhenin in Dendrobium officinale and Dendrobium huoshanense

DOI：10.13422/j.cnki.syfjx.20182418

摘要

Abstract

关键词

Keywords

references