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Quote : 刘应蛟,楚世峰,胡耀梅,裴刚,周小江,陈乃宏.玄参中阿魏酸和肉桂酸的含量测定及神经保护活性研究[J].湖南中医药大学学报英文版,2018,38(12):1393-1397.[Click to copy ]
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玄参中阿魏酸和肉桂酸的含量测定及神经保护活性研究
刘应蛟,楚世峰,胡耀梅,裴刚,周小江,陈乃宏
(湖南中医药大学药学院, 湖南 长沙 410208;湖南省中药饮片标准化及功能工程技术研究中心, 湖南 长沙 410208;中国医学科学院北京协和医学院药物研究所, 北京 100050)
摘要:
    目的 建立中药玄参中阿魏酸和肉桂酸的HPLC含量测定方法,观察其水解前后的变化,以及对鱼藤酮和去血清模型的神经保护作用。方法 采用Agilent 5 TC-C18柱(4.6 mm×250 mm,5 μm),以乙腈-0.05%磷酸溶液为流动相,梯度洗脱,检测波长为290 nm,柱温30℃;建立体外鱼藤酮及去血清模型,用不同浓度阿魏酸和肉桂酸预处理,以MTT法测定细胞存活率。结果 阿魏酸和肉桂酸水解前后回收率分别为98.20%和91.41%、97.22%和90.96%,RSD均未超过2.0%,6批不同产地药材中阿魏酸和肉桂酸水解前后的含量分别为0.028%~0.041%和0.073%~0.102%、0.035%~0.142%和0.163%~0.336%。阿魏酸(10 μmol/L)可显著增加去血清和鱼藤酮处理细胞的存活率(P<0.01);肉桂酸(0.1 μmol/L)能明显增加鱼藤酮处理细胞的存活率(P<0.05)。结论 本方法简便、精密度高、重现性好,为玄参中具神经保护作用的阿魏酸和肉桂酸动态变化提供科学依据。
关键词:  玄参  阿魏酸  肉桂酸  高效液相色谱  神经保护
DOI:10.3969/j.issn.1674-070X.2018.12.009
Received:July 25, 2018  
基金项目:国家中医药标准化项目(ZYBZH-Y-HUN-24,ZYBZH-Y-HEB-16);湖南省教育厅创新平台开放基金项目(15K091);湖南省重点研究计划项目(2017NK2261)。
Determination of Ferulic Acid and Cinnamic Acid in Scrophularia ningpoensis and Their Neuroprotective Activity
LIU Yingjiao,CHU Shifeng,HU Yaomei,PEI Gang,ZHOU Xiaojiang,CHEN Naihong
(School of Pharmacy, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan Engineering Technology Center of Standardization and Function of Chinese Herbal Decoction Pieces, Changsha, Hunan 410208, China;Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China)
Abstract:
    Objective To develop a high-performance liquid chromatography-based method for determination of ferulic acid and cinnamic acid in Scrophularia ningpoensis, to evaluate their changes after hydrolysis, and to analyze their neuroprotective effects on rotenone-treated and serum-free cell culture models.Methods The content of ferulic acid and cinnamic acid was determined using an Agilent 5TC-C18 column (250 mm×4.6 mm, 5 μm) with a mobile phase of acetonitrile-0.05% phosphoric acid for gradient elution at a detection wavelength of 290 nm and a column temperature of 25℃. Pretreated with different concentrations of ferulic acid and cinnamic acid, rotenone-treated and serum-free cell culture models were established in vitro. Cell viability was determined by MTT assay.Results The recovery rates before and after hydrolysis were 98.20% and 91.41% for ferulic acid, and 97.22% and 90.96% for cinnamic acid, respectively. The relative standard deviations were both less than 2.0%. In six batches of medicinal materials originating from different places, the content before and after hydrolysis was 0.028%-0.041% and 0.073%-0.102% for ferulic acid, and 0.035%-0.142% and 0.163%-0.336% for cinnamic acid, respectively. Ferulic acid (10 μmol/L) significantly increased the viability of cells treated by rotenone and serum deprivation (P<0.01). Cinnamic acid (0.1 μmol/L) significantly increased the viability of cells treated by rotenone (P<0.05).Conclusion This method is simple, highly accurate, and reproducible, which provides a scientific basis for the dynamic changes in two components with neuroprotective activity, ferulic acid and cinnamic acid, in Scrophularia ningpoensis.
Key words:  Scrophularia ningpoensis  ferulic acid  cinnamic acid  high-performance liquid chromatography  neuroprotective
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