UPLC-MS/MS联合细胞膜固相色谱法分析补阳还五汤作用于神经元样PC12细胞的效应成分

郑华珠; 王利胜; 吴银爱; 范有明; 孟英姣; 沈晓

doi:10.13422/j.cnki.syfjx.2018040014

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UPLC-MS/MS联合细胞膜固相色谱法分析补阳还五汤作用于神经元样PC12细胞的效应成分

药剂与炮制 | 更新时间：2024-01-04

- UPLC-MS/MS联合细胞膜固相色谱法分析补阳还五汤作用于神经元样PC12细胞的效应成分
- Effective Components in Buyang Huanwu Tang on Neuron-like PC12 Cells by UPLC-MS/MS Combined with Cell Membrane Solid-phase Chromatography
- 中国实验方剂学杂志 2018年24卷第4期页码：14-20
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金项目（81373973，81573872）;广东省省级科技计划项目（2013B032500015）
- DOI：10.13422/j.cnki.syfjx.2018040014
  中图分类号： R284.1
- 纸质出版日期：2018
- 稿件说明：
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郑华珠, 王利胜, 吴银爱, 等. UPLC-MS/MS联合细胞膜固相色谱法分析补阳还五汤作用于神经元样PC12细胞的效应成分[J]. 中国实验方剂学杂志, 2018,24(4):14-20.

ZHENG Hua-zhu, WANG Li-sheng, WU Yin-ai, et al. Effective Components in Buyang Huanwu Tang on Neuron-like PC12 Cells by UPLC-MS/MS Combined with Cell Membrane Solid-phase Chromatography[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(4): 14-20.
郑华珠, 王利胜, 吴银爱, 等. UPLC-MS/MS联合细胞膜固相色谱法分析补阳还五汤作用于神经元样PC12细胞的效应成分[J]. 中国实验方剂学杂志, 2018,24(4):14-20. DOI： 10.13422/j.cnki.syfjx.2018040014.

ZHENG Hua-zhu, WANG Li-sheng, WU Yin-ai, et al. Effective Components in Buyang Huanwu Tang on Neuron-like PC12 Cells by UPLC-MS/MS Combined with Cell Membrane Solid-phase Chromatography[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(4): 14-20. DOI： 10.13422/j.cnki.syfjx.2018040014.

摘要

目的：筛选补阳还五汤（BYHWT）中特异作用于神经元样PC12细胞的化学成分，为该复方的后续研究与应用提供实验依据。方法：采用固相萃取、神经元样PC12细胞细胞膜固相色谱法富集补阳还五汤特异结合神经元样PC12的效应成分，采用超高效液相色谱联用电喷雾飞行时间质谱（UPLC-MS/MS）鉴定特异结合成分，色谱条件为流动相甲醇-0.1%甲酸水溶液梯度洗脱，进样量3 μL，流速0.3 mL·min-1；质谱条件为雾化器压力和辅助气压力均为379.2 kPa，气帘气压力241.3 kPa，离子源温度500 ℃，喷雾电压－4.5 kV。结果：补阳还五汤特异结合神经元PC12细胞的化学成分可能为6-羟基山柰酚-3，6-二-O-葡萄糖苷，6-羟基山柰酚-3，6，7-三-O-葡萄糖苷和毛蕊异黄酮苷。结论：6-羟基山柰酚-3，6-二-O-葡萄糖苷，6-羟基山柰酚-3，6，7-三-O-葡萄糖苷和毛蕊异黄酮苷可特异结合神经元样PC12细胞，可能是补阳还五汤中发挥神经保护的效应成分。

Abstract

Objective: To screen the specific binding components in Buyang Huanwu Tang(BYHWT) on neuron-like PC12 cells

and to provide experimental basis for further study and application of this compound. Method: A method on analysis neuron-like PC12 cells specificity binding components in BYHWT by solid-phase extraction(SPE)

neuron-like PC12 cells binding assay combined with SPE was employed

and the samples were analyzed by UPLC-MS/MS

mobile phase was methanol-0.1% formic acid aqueous solution under gradient elution

the injection volume was 3 μL and the flow rate was set as 0.3 mL·min-1.The mass spectrometric conditions were set as following:atomizer pressure and auxiliary gas pressure of 379.2 kPa

air curtain gas pressure of 241.3 kPa

ion source temperature at 500 ℃

spray voltage of －4.5 kV. Result: The neuron-like PC12 cells specificity binding components in BYHWT were found as follows:6-hydroxykaempferol-3

7-tri-O-glucoside

6-hydroxykaempferol-3

6-di-O-glucoside and calycosin-7-O-β-D-glucoside. Conclusion: These three components may be the important compositions for neuroprotective function in BYHWT.

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