Simultaneous Determination of Eight Active Components in Material Benchmark of Shentong Zhuyutang by HPLC-DAD Wavelength Switching

Yan-fen CHENG; Sheng-ju WANG; Yi-han WU; Xiao WANG; Ting WANG; Jin PEI; Jin-ming ZHANG; Chao-mei FU

doi:10.13422/j.cnki.syfjx.20191954

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Simultaneous Determination of Eight Active Components in Material Benchmark of Shentong Zhuyutang by HPLC-DAD Wavelength Switching

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- Simultaneous Determination of Eight Active Components in Material Benchmark of Shentong Zhuyutang by HPLC-DAD Wavelength Switching
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 4, Pages: 16-22(2020)
- 作者机构：
  
  成都中医药大学　药学院，中药材标准化教育部重点实验室，四川省中药资源系统研究与开发利用重点实验室——省部共建国家重点实验室培育基地，成都　 611137
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191954
  CLC： R22; R28; R94; R927.2; O657.7
- Received：12 May 2019，
  
  Published Online：19 June 2019，
  
  Published：20 February 2020
- 稿件说明：
移动端阅览
Yan-fen CHENG, Sheng-ju WANG, Yi-han WU, et al. Simultaneous Determination of Eight Active Components in Material Benchmark of Shentong Zhuyutang by HPLC-DAD Wavelength Switching[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(4): 16-22.
DOI：

Yan-fen CHENG, Sheng-ju WANG, Yi-han WU, et al. Simultaneous Determination of Eight Active Components in Material Benchmark of Shentong Zhuyutang by HPLC-DAD Wavelength Switching[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(4): 16-22. DOI： 10.13422/j.cnki.syfjx.20191954.

摘要

目的：

采用高效液相色谱-二极管阵列检测器(HPLC-DAD)波长切换法同时测定15批身痛逐瘀汤物质基准中肌苷，马钱苷酸，绿原酸，苦杏仁苷，羟基红花黄色素A，龙胆苦苷，阿魏酸，甘草苷8种有效成分的含量。

方法：

采用Thermo Hypersil GOLD C

色谱柱(4.6 mm×250 mm，5 μm)，以乙腈-0.1%磷酸水溶液为流动相梯度洗脱，检测波长分别为248 nm(0～11 min，检测肌苷)，235 nm(11～14 min，检测马钱苷酸)，324 nm(14～16 min，检测绿原酸)，220 nm(16～19 min，检测苦杏仁苷、羟基红花黄色素A)，274 nm(19～26 min，检测龙胆苦苷)，247 nm(26～54 min，检测阿魏酸、甘草苷)，流速设定1.0 mL·min

-1

，柱温25 ℃。根据样品中8个成分的含量，采用SIMCA 14.1软件对15批身痛逐瘀汤物质基准样品进行正交偏最小二乘法-判别分析(OPLS-DA)，以评价各批样品之间的质量差异。

结果：

各待测组分分离度良好，肌苷，马钱苷酸，绿原酸，苦杏仁苷，羟基红花黄色素A，龙胆苦苷，阿魏酸，甘草苷的质量浓度分别在2.1～67.2，1.812 5～58，1.937 5～62，5.212 5～166.8，8.45～270.4，7.075～226.4，1.775～56.8，3.875～124 mg·L

-1

(

均≥0.999 6)与峰面积呈良好线性关系；平均加样回收率分别为99.23%，100.09%，99.33%，98.85%，99.15%，98.75%，99.42%，98.96%；RSD均

2%。15批供试品中上述8种成分的质量分数依次为0.183 5～0.250 3，0.173 1～0.265 3，0.069 5～0.169 8，0.959 2～1.458 2，1.905 4～2.553 3，0.933 3～1.997 5，0.084 6～0.143 4，0.212 5～0.704 3 mg·g

-1

。通过OPLS-DA发现甘草苷、阿魏酸、龙胆苦苷、羟基红花黄色素A是影响不同批次身痛逐瘀汤物质基准质量贡献较大的4种成分。

结论：

建立的多成分含量测定的方法结果可靠、操作简便且符合方法学验证要求，适用于身痛逐瘀汤复方制剂研发的质量控制。

Abstract

Objective:

To develop high performance liquid chromatography-diode array detector (HPLC-DAD) wavelength switching for simultaneously determining the contents of inosine

loganic acid

chlorogenic acid

amygdalin

hydroxysafflor yellow A

gentiopicroside

ferulic acid and liquiritin in 15 batches of material benchmarks of Shentong Zhuyutang.

Method:

The quantitative analysis was carried out on a Thermo Hypersil GOLD C

column (4.6 mm×250 mm

5 μm) with mobile phase of acetonitrile-0.1%phosphoric acid aqueous solution for gradient elution

the flow rate was 1.0 mL·min

-1

the detection wavelengths were set as 248 nm (0-11 min

inosine)

235 nm (11-14 min

loganic acid)

324 nm (14-16 min

chlorogenic acid)

220 nm (16-19 min

amygdalin and hydroxysafflor yellow A)

274 nm (19-26 min

gentiopicroside)

247 nm (26-54 min

ferulic acid and liquiritin)

the column temperature was maintained at 25 ℃. According to the contents of eight active components in 15 batches of material benchmarks

orthogonal partial least squares discriminant analysis (OPLS-DA) in SIMCA 14.1 was used to evaluate the quality difference of each batch of samples.

Result:

Each component had good separations

the linear ranges of the above 8 components were 2.1-67.2

1.812 5-58

1.937 5-62

5.212 5-166.8

8.45-270.4

7.075-226.4

1.775-56.8

3.875-124 mg·L

-1

respectively (

≥0.999 6). The average recoveries of them were 99.23%

100.09%

99.33%

98.85%

99.15%

98.75%

99.42%

98.96%

respectively (RSD

2%). The contents of the above eight components in 15 batches of material benchmarks were 0.183 5-0.250 3

0.173 1-0.265 3

0.069 5-0.169 8

0.959 2-1.458 2

1.905 4-2.553 3

0.933 3-1.997 5

0.084 6-0.143 4

0.212 5-0.704 3 mg·g

-1

respectively. Liquiritin

ferulic acid

gentiopicroside and hydroxysafflor yellow A were determined to have significant impact on the quality of different batches of material benchmarks of Shentong Zhuyutang through OPLS-DA.

Conclusion:

The established method for simultaneous determination of multi-components is reliable

simple and in line with the requirements of methodological verification. It is suitable for the quality control of research and development of compound preparations of Shentong Zhuyutang.

关键词

Keywords

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