Analysis on Sugar Spectrum Difference Between Wild-simulated and Transplanted Astragali Radix

LI Hufeng; LI Ke; LYU Wanwan; FENG Shihong; WEN Yuwei; LI Zhenyu; QIN Xuemei; DU Yuguang

doi:10.13422/j.cnki.syfjx.20230663

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Analysis on Sugar Spectrum Difference Between Wild-simulated and Transplanted Astragali Radix

更新时间：2023-09-21

- Analysis on Sugar Spectrum Difference Between Wild-simulated and Transplanted Astragali Radix
  增强出版
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 20, Pages: 141-148(2023)
- 作者机构：
  
  1.山西大学中医药现代研究中心，化学生物学与分子工程教育部重点实验室，地产中药功效物质研究与利用山西省重点实验室，太原 030006
  2.中国科学院过程工程研究所，北京 100190
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20230663
  CLC： R22;R28;R931;O657
- Published：20 October 2023，
  
  Published Online：17 February 2023，
  
  Received：06 December 2022，
- 稿件说明：
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李虎峰,李科,吕弯弯等.仿野生黄芪与移栽黄芪的糖谱差异分析[J].中国实验方剂学杂志,2023,29(20):141-148.

LI Hufeng,LI Ke,LYU Wanwan,et al.Analysis on Sugar Spectrum Difference Between Wild-simulated and Transplanted Astragali Radix[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(20):141-148.
李虎峰,李科,吕弯弯等.仿野生黄芪与移栽黄芪的糖谱差异分析[J].中国实验方剂学杂志,2023,29(20):141-148. DOI： 10.13422/j.cnki.syfjx.20230663.

LI Hufeng,LI Ke,LYU Wanwan,et al.Analysis on Sugar Spectrum Difference Between Wild-simulated and Transplanted Astragali Radix[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(20):141-148. DOI： 10.13422/j.cnki.syfjx.20230663.

摘要

目的

建立仿野生黄芪和移栽黄芪多糖、寡糖、单糖特征糖谱，分析二者的糖谱差异，为黄芪品质评价提供依据。

方法

通过高效液相色谱-蒸发光散射检测法（HPLC-ELSD）对18批仿野生黄芪和12批移栽黄芪的多糖进行相对分子质量分布表征，建立2种黄芪多糖的特征图谱，对相对分子质量为10 kDa的多糖组分APS-Ⅱ的峰面积占比进行差异分析，并通过受试者工作特征（ROC）曲线确定APS-Ⅱ峰面积占比临界值。采用三氟乙酸（TFA）将APS-Ⅱ进行部分酸水解，基于HPLC-ELSD建立2种黄芪寡糖特征图谱，并通过主成分分析（PCA）及正交偏最小二乘法-判别分析（OPLS-DA）寻找差异寡糖特征。采用TFA将2种黄芪的APS-Ⅱ完全酸水解并衍生化处理，基于HPLC建立2种黄芪单糖特征图谱，对2种黄芪单糖峰面积占比进行PCA及OPLS-DA处理，分析二者APS-Ⅱ单糖组成的差异。

结果

黄芪多糖特征糖谱表明，APS-Ⅱ的峰面积占比为主要差异，仿野生黄芪和移栽黄芪APS-Ⅱ峰面积占比分别为89.17%~97.17%和80.14%~91.96%。ROC曲线确定2种黄芪APS-Ⅱ峰面积占比差异的临界值为92.28%；对APS-Ⅱ寡糖组分进行多元统计分析发现，聚合度≥10的寡糖峰面积占比为主要差异，仿野生黄芪为11.835%~19.092%，移栽黄芪为2.778%~7.017%；单糖特征糖谱分析结果表明，2种黄芪均由6种单糖组成，葡萄糖和阿拉伯糖为差异单糖组分，仿野生黄芪中葡萄糖和阿拉伯糖峰面积占比分别为85%~93.9%、2.7%~5.8%，移栽黄芪为74.3%~87.3%、5.3%~10.7%，提示2种黄芪多糖组分APS-Ⅱ的结构可能有所不同。

结论

仿野生黄芪和移栽黄芪间的糖谱差异可能与APS-Ⅱ的含量及结构相关，本研究可为黄芪糖类物质研究及药材品质评价提供参考依据。

Abstract

Objective

To establish the characteristic sugar spectrum of polysaccharides， oligosaccharides and monosaccharides of wild-simulated and transplanted Astragali Radix， and find out the difference of the sugar spectrum between the two， so as to provide a basis for quality evaluation of Astragali Radix.

Method

The relative molecular weight distribution of polysaccharides from 18 batches of wild-simulated Astragali Radix and 12 batches of transplanted Astragali Radix were characterized by high performance liquid chromatography-evaporative light scattering detection（HPLC-ELSD） to establish the characteristic chromatograms of two kinds of polysaccharides. The difference in the peak area ratio of APS-Ⅱ， a polysaccharide component with a relative molecular weight of 10 kDa， in two kinds of Astragali Radix was analyzed， and the critical value of peak area ratio of APS-Ⅱ was determined by receiver operating characteristic（ROC） curve. At the same time， APS-Ⅱ was partially acid-hydrolyzed by trifluoroacetic acid（TFA） to establish characteristic spectra of two kinds of oligosaccharides from Astragali Radix based on HPLC-ELSD， and the characteristics of differential oligosaccharides were found by principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA）. Two kinds of APS-Ⅱ were completely acid-hydrolyzed by TFA and derivatized to establish characteristic spectra of two kinds of monosaccharides from Astragali Radix based on HPLC， PCA and OPLS-DA were performed on the peak area ratio of two kinds of monosaccharides to explore the differences in the composition of two kinds of APS-Ⅱ monosaccharides.

Result

The characteristic sugar spectrum of polysaccharides from Astragali Radix showed that the peak area ratio of APS-Ⅱ was the main difference， and the peak area of APS-Ⅱ of wild-simulated and transplanted Astragali Radix were 89.17%-97.17% and 80.14%-91.96%， respectively. The ROC curve determined the critical value of 92.28% for the difference of APS-Ⅱ peak area ratio of the two kinds of Astragali Radix. The multivariate analysis of APS-Ⅱ oligosaccharides revealed that the peak area ratio of oligosaccharides with polymerization degree≥10 was the main difference， which ranged from 11.835%-19.092% for wild-simulated products and 2.778%-7.017% for transplanted products. The results of monosaccharide characteristic sugar spectrum analysis showed that both Astragali Radix species consisted of six monosaccharides， and glucose and arabinose were the differential monosaccharide fractions. The peak area ratios of glucose and arabinose in wild-simulated products were 85%-93.9% and 2.7%-5.8%， respectively， while those of transplanted products were 74.3%-87.3% and 5.3%-10.7%， suggesting that the structures of the two polysaccharide fractions APS-Ⅱ of Astragali Radix may be different.

Conclusion

The difference of sugar spectrum between two kinds of Astragali Radix may be related to the content and structure of APS-Ⅱ， and this study may provide a reference for the study of carbohydrates in Astragali Radix and the quality evaluation of medicinal materials.

关键词

黄芪多糖糖谱寡糖单糖品质评价高效液相色谱法（HPLC）

Keywords

Astragali Radixpolysaccharidessugar spectrumoligosaccharidesmonosaccharidesquality evaluationhigh performance liquid chromatography（HPLC）

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