基于UHPLC-Q-TOF-MS分析荭草花提取物在H9c2心肌细胞中效应物质

向文英; 梅朝叶; 孙慧园; 覃小丽; 谢玉敏; 巩仔鹏; 郑林; 李月婷; 黄勇

doi:10.13422/j.cnki.syfjx.2017120090

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基于UHPLC-Q-TOF-MS分析荭草花提取物在H9c2心肌细胞中效应物质

药物代谢 | 更新时间：2024-01-04

- 基于UHPLC-Q-TOF-MS分析荭草花提取物在H9c2心肌细胞中效应物质
- Identification of Effective Substances from Flower Extract in H9c2 Myocardial Cells by UHPLC-Q-TOF-MS
- 中国实验方剂学杂志 2017年23卷第12期页码：90-95
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金项目（81360664，81560630）;贵州省研究生卓越人才计划项目（黔教研合ZYRC字[2014]012）
- DOI：10.13422/j.cnki.syfjx.2017120090
  中图分类号： R285
- 纸质出版日期：2017
- 稿件说明：
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向文英, 梅朝叶, 孙慧园, 等. 基于UHPLC-Q-TOF-MS分析荭草花提取物在H9c2心肌细胞中效应物质[J]. 中国实验方剂学杂志, 2017,23(12):90-95.

XIANG Wen-ying, MEI Chao-ye, SUN Hui-yuan, et al. Identification of Effective Substances from Flower Extract in H9c2 Myocardial Cells by UHPLC-Q-TOF-MS[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(12): 90-95.
向文英, 梅朝叶, 孙慧园, 等. 基于UHPLC-Q-TOF-MS分析荭草花提取物在H9c2心肌细胞中效应物质[J]. 中国实验方剂学杂志, 2017,23(12):90-95. DOI： 10.13422/j.cnki.syfjx.2017120090.

XIANG Wen-ying, MEI Chao-ye, SUN Hui-yuan, et al. Identification of Effective Substances from Flower Extract in H9c2 Myocardial Cells by UHPLC-Q-TOF-MS[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(12): 90-95. DOI： 10.13422/j.cnki.syfjx.2017120090.

摘要

目的：利用H9c2心肌细胞和UHPLC-Q-TOF-MS技术筛选荭草花抗心肌缺血作用的活性成分。方法：用心肌细胞选择性地结合荭草花提取液中的活性成分，洗去未结合的其他成分后，使细胞靶点失活，被结合的活性成分从心肌细胞中释放出来，采用Eclipse Plus C18色谱柱（2.1 mm×100 mm，1.8 μm），流动相0.1%甲酸水溶液（A）-0.1%甲酸乙腈溶液（B）梯度洗脱（0~2 min，95%~79% A；2~5 min，79% A；5~6.5 min，79%~60% A；6.5~7.5 min，60%~55% A；7.5~9.5 min，55%~0% A；9.5~10 min，0%~95% A），电喷雾离子源（ESI）正、负离子模式下扫描采集数据，通过采用对照品比对及二级碎片离子对化学成分进行鉴定。结果：检测出荭草花提取物中可与心肌细胞结合的17个成分，采用对照品比对后，确定8个色谱峰所表征的化学成分，依次为原儿茶酸，儿茶素，山柰素-3-O-β-D-葡萄糖苷，槲皮素3-O-α-L-鼠李糖苷，山柰素-3-O-α-L-鼠李糖苷，N-p-香豆酰酪胺，槲皮素和山柰酚。结论：通过分析心肌细胞破碎液中与活性细胞相结合的成分，筛选出了荭草花提取物中能与心肌细胞有相互作用的成分，为明确荭草花的药效物质基础提供了科学依据。

Abstract

Objective: To screen effective substances from Polygonum orientale flower extract by UHPLC-Q-TOF-MS technology for H9c2 myocardial cells. Method: The active substances in P. orientale flower extract were combined with H9c2 myocardial cells under imitated physiological environments

then the unattached substances were washed off by phosphate buffer solution (PBS) with pH of 7.4.After that

the conjugated compounds were eluted by PBS with pH of 4.0.These compounds released from target sites were collected and handled through Oasis HLB solid phase extraction column to be condensed.The separation was performed on a Eclipse Plus C18 column

mass sepectrometric data were obtained in electrospray ionization (ESI) positive and negative modes.Meanwhile

the chemical components were identified by comparing the retention times with standard substance and secondary fragment ions. Result: Seventeen compounds from P. orientale flower extract were identified preliminarily

and eight compounds were identified as protocatechuic acid

cianidanol

kaempferol

kaempferol-3-O-β-D-glucoside

quercetin

quercetin-3-O-β-D-glucoside

kaempferol-3-O-α-L-rhamnoside

paprazine. Conclusion: Throngh analysis of the fracture fluid compounds from the H9c2 cells extraction combined with active cells

the bioactive constituents from P. orientale flower extract interacted with myocardial cells are screened

this research provides a scientific fundament for material basis of effectiveness of P. orientale flower.

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