基于茜素红S-苯硼酸光化学比色传感器的附子配方颗粒强心活性评价方法的建立  

Establishment of cardiotonic activity evaluation method of Fuzi formula granules based on alizarin red S-phenylboronic acid photochemical colorimetric sensor

作　　者：陈露梦 刘惠 贺亚男[1] 罗传红 贾明艳 周永峰 杨景屏 谭鹏 杨明 张定堃[1] CHEN Lu-meng;LIU Hui;HE Ya-nan;LUO Chuan-hong;JIA Ming-yan;ZHOU Yong-feng;YANG Jing-ping;TAN Peng;YANG Ming;ZHANG Ding-kun(State Key Laboratory of Southwestern Chinese Medicine Resources,School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Sichuan Xingshengyuan Pharmaceutical Co.,Ltd.,Chengdu 621700,China;State Key Laboratory of Biological Evaluation of TCM Quality,National Administration of TCM,Sichuan Academy of Chinese Medicine Sciences,Chengdu 610041,China;Key Laboratory of Modern Preparation of TCM,Ministry of Education,Jiangxi University of Chinese Medicine,Nanchang 330004,China)

机构地区：[1]成都中医药大学药学院,西南特色中药资源国家重点实验室,四川成都611137 [2]四川新盛源药业有限公司,四川江油621700 [3]四川省中医药科学院,国家中医药管理局中药质量生物评价重点研究室,四川成都610041 [4]江西中医药大学,现代中药制剂教育部重点实验室,江西南昌330004

出　　处：《中草药》2022年第6期1761-1767,共7页Chinese Traditional and Herbal Drugs

基　　金：四川省中医药管理局科学技术研究专项(2021MS016);四川省中医药管理局中医药重点学科建设项目(川中医药函[2020]84号);国家自然科学基金青年科学基金项目(82003907);四川省重点研发项目(2022YFS0429)。

摘　　要：目的 基于附子强心组分去甲乌药碱、去甲猪毛菜碱和棍掌碱共有的肾上腺素类似结构,构建了以邻二酚羟基基团与茜素红S-苯硼酸体系置换反应为核心的光化学比色传感器,用于整体评价附子配方颗粒的强心作用。方法 以传感器的响应强度为评价指标,对超声提取时间、料液比、提取液浓度和乙二胺四乙酸二钠(EDTA-2Na)浓度等供试品的制备条件进行优化;并对该方法的精密度、重复性和稳定性进行考察;用平板扫描仪获取23批附子配方颗粒的|ΔG|值(绿色G,|ΔG|=|G;-G;|),同时采用LC-MS/MS测定附子配方颗粒中3种生物碱的含量,将|ΔG|与强心成分总含量进行相关性分析。结果 供试品制备方法优选为取附子配方颗粒粉末0.5 g,加10 mL甲醇,300 W、60 kHz超声提取15 min,将上清液全部转移至蒸发皿中,80℃挥干,4 mL 0.13 g/mL的EDTA-2Na溶液洗脱蒸发皿上物质,离心,取上清液,调pH至8.0左右,缓冲溶液稀释3倍。方法学考察表明该方法精密度、重复性和稳定性良好。23批样品的强心成分总含量的差异较大,为127.83~6 928.27μg/g;|ΔG|的范围为0~12.5,两者的相关系数(r)高达0.96,表明强心成分总含量与|ΔG|高度相关。结论 该方法具有一定的科学性与准确性,可整体性评价附子配方颗粒强心药效。Objective Based on the similar structure of adrenaline shared by higenamine,salsolinol and coryneine,a photochemical colorimetric sensor based on the displacement reaction of o-diphenol hydroxyl group and alizarin red S-phenylboric acid system was proposed to evaluate the cardiotonic effect of Fuzi(Aconiti Lateralis Radix Praeparata) formula granules as a whole.Methods Taking the response intensity of the sensor as the evaluation index,the preparation conditions of test samples such as ultrasonic extraction time,solid-liquid ratio,extraction solution concentration and EDTA-2 Na concentration were optimized;then the precision,repeatability and stability of the method were investigated;The |ΔG| value(G is green,|ΔG| = |G;-G;|) of each sample was obtained by response values determination of 23 batches of Fuzi formula granules;at the same time,the contents of three alkaloids in Fuzi formula granules were determined by LC-MS/MS.The correlation between |ΔG| and the total content of cardiac components was analyzed.Results The preparation method of the test article was preferably as follows:take 0.5 g of Fuzi formula granular powder,add 10 mL of methanol,and ultrasonic at 300 W and 60 kHz for 15 min,put the supernatant in an evaporating dish,volatilize it at 80 ℃,elute the substance on the evaporating dish with 4 mL of 0.13 g/mL EDTA-2 Na solution,centrifuge,take the supernatant,adjust the pH to 8.0,and dilute the buffer solution three times;The methodological investigation showed that the method had good precision,repeatability and stability.The total content of cardiac components in 23 batches of samples varied greatly,127.83—6 928.27 μg/g;The range of |ΔG| was 0—12.5,and the correlation coefficient(r) was as high as 0.96,indicating that the total content of cardiac components was highly correlated with |ΔG|.Conclusion The method is scientific and accurate,and can evaluate the cardiotonic effect of Fuzi formula granules as a whole.

关 键 词：附子 强心作用 配方颗粒 光化学比色传感器 质量评价 茜素红S 苯硼酸 去甲乌药碱 去甲猪毛菜碱 棍掌碱 肾上腺素 

分 类 号：R283.6[医药卫生—中药学]