Simultaneous Determination of Six Flavonoids in Astragali Radix Directional Processed with Four Enzymes by UPLC

Peng-peng LIU; Ji SHI; Fan ZHANG; Guo-shun SHAN; Tian-zhu JIA

doi:10.13422/j.cnki.syfjx.20200654

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Simultaneous Determination of Six Flavonoids in Astragali Radix Directional Processed with Four Enzymes by UPLC

Pharmacy Fundamentals | 更新时间：2021-02-09

- Simultaneous Determination of Six Flavonoids in Astragali Radix Directional Processed with Four Enzymes by UPLC
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 10, Pages: 94-99(2020)
- 作者机构：
  
  辽宁中医药大学　药学院，国家中医药管理局中药炮制原理解析重点实验室，辽宁省中药炮制工程技术研究中心，辽宁大连　 116600
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20200654
  CLC： R22; R943.1; R28; TQ92; O657.7
- Received：12 September 2019，
  
  Published Online：29 November 2019，
  
  Published：20 May 2020
- 稿件说明：
移动端阅览
Peng-peng LIU, Ji SHI, Fan ZHANG, et al. Simultaneous Determination of Six Flavonoids in Astragali Radix Directional Processed with Four Enzymes by UPLC[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(10): 94-99.
DOI：

Peng-peng LIU, Ji SHI, Fan ZHANG, et al. Simultaneous Determination of Six Flavonoids in Astragali Radix Directional Processed with Four Enzymes by UPLC[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(10): 94-99. DOI： 10.13422/j.cnki.syfjx.20200654.

摘要

目的：

建立黄芪中6种黄酮类成分的含量测定方法，研究4种酶(复合酶、植物纤维素酶、蜗牛酶及

-葡萄糖苷酶)定向炮制对黄芪中黄酮苷类成分及其苷元含量的影响。

方法：

采用ACQUITY UPLC HSS T

色谱柱(2.1 mm×100 mm，1.8 μm)，流动相0.1%甲酸水溶液-0.1%甲酸乙腈溶液梯度洗脱，检测波长260 nm，流速0.3 mL·min

-1

，柱温30 ℃，进样量2 μL。

结果：

6种成分毛蕊异黄酮苷、毛蕊异黄酮、芒柄花苷、芒柄花素、黄芪紫檀烷苷、美迪紫檀素的分离度良好，在各自线性范围内线性关系良好(

≥0.998 5)，精密度、稳定性、重复性试验的RSD均

5.0%；平均加样回收率97.62%～101.13%，RSD 1.4%～2.7%。在0.5 g·L

-1

酶溶液水平，毛蕊异黄酮苷、毛蕊异黄酮、芒柄花苷、芒柄花素、黄芪紫檀烷苷、美迪紫檀素在复合酶制黄芪中的质量分数分别为0.082 0，0.335 9，0.055 9，0.104 9，0.015 0，0.009 7 mg·g

-1

，在纤维素酶制黄芪中的质量分数分别为0.105 7，0.364 2，0.070 2，0.117 4，0.020 8，0.012 5 mg·g

-1

，在蜗牛酶制黄芪中的质量分数分别为0.031 4，0.510 0，0.043 5，0.210 9，0.013 0，0.013 0 mg·g

-1

，在

-葡萄糖苷酶制黄芪中的质量分数分别为0.085 3，0.312 4，0.061 5，0.110 8，0.005 8，0.009 6 mg·g

-1

。

结论：

黄芪经4种酶炮制后，除黄芪紫檀烷苷外，毛蕊异黄酮苷、芒柄花苷的含量均降低，但这3种黄酮苷类成分所对应苷元的含量均显著增加。该方法操作简便、准确、重复性好，适用于黄芪中6种黄酮类成分的含量测定，可为黄芪的酶定向炮制研究提供参考。

Abstract

Objective:

To establish an UPLC method for the simultaneous determination of 6 flavonoids

and to research for the effect of Astragali Radix directional processed with four enzymes (complex enzyme

plant cellulase

snail enzyme

and

-glucosidase) on the contents of flavonoid glycosides and their aglycones in this herb.

Method:

Chromatographic separation was carried out on ACQUITY UPLC HSS T

column (2.1 mm×100 mm

1.8 μm) with the mobile phase of 0.1 mol·L

-1

formic acid solution-0.1 mol·L

-1

formic acid acetonitrile solution for gradient elution. The detection wavelength was set at 260 nm

the flow rate was 0.3 mL·min

-1

the column temperature was 30 ℃

and the injection volume was 2 μL.

Result:

Calycosin-glucoside

calycosin

ononin

formononetin

10-dimethoxy-pterocarpan-3-

-glucoside and 3-hydroxy-9

10-dimethoxy-pterocarpan showed good linear relationships within their own ranges (

≥0.998 5)

the relative standard deviations (RSDs) of precision

stability and repeatability were all

5.0%

and the average recovery was 97.62%-101.13% with RSDs of 1.4%-2.7%. In 0.5 g·L

-1

level of enzyme solution

the contents of calycosin-glucoside

calycosin

ononin

formononetin

10-dimethoxy-pterocarpan-3-

-glucoside and 3-hydroxy-9

10-dimethoxy-pterocarpan in Astragali Radix processed with complex enzyme were 0.082 0

0.335 9

0.055 9

0.104 9

0.015 0

0.009 7 mg·g

-1

the contents of them in Astragali Radix processed with plant cellulase were 0.105 7

0.364 2

0.070 2

0.117 4

0.020 8

0.012 5 mg·g

-1

their contents in Astragali Radix processed with snail enzyme were 0.031 4

0.510 0

0.043 5

0.210 9

0.013 0

0.013 0 mg·g

-1

and their contents in Astragali Radix processed with

-glucosidase were 0.085 3

0.312 4

0.061 5

0.110 8

0.005 8

0.009 6 mg·g

-1

respectively.

Conclusion:

After the processing of Astragali Radix by four enzymes

in addition to 9

10-dimethoxy-pterocarpan-3-

-glucoside

the contents of calycosin-glucoside and ononin are reduced

but the contents of their three corresponding aglycones are significantly increased. The established method is simple

accurate and reproducible

and is suitable for the simultaneous determination of 6 flavonoids in Astragali Radix

which can provide a reference for this herb directional processed with enzymes.

关键词

Keywords

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