芫花根HPLC色谱指纹图谱研究  

HPLC Fingerprint of the Roots of Daphne Genkwa

作　　者：郑维发[1] 陈才法[1] 石枫[1] 

机构地区：[1]徐州师范大学江苏省药用植物生物技术重点实验室,江苏徐州221116

出　　处：《解放军药学学报》2005年第1期4-9,共6页Pharmaceutical Journal of Chinese People's Liberation Army

基　　金：教育部科学技术研究重点项目 ;No .0 3 0 49;江苏省高校自然科学基金重点项目 ;No.0 2KJA3 60 0 0 2 ;江苏省药用植物生物技术重点实室开放基金;No.KJSO2 114

摘　　要：目的　建立芫花根HPLC色谱指纹图谱 ,为芫花根有效成分筛选、药理毒理学、药物代谢动力学以及脱毒提取工艺研究提供可靠的检测方法。方法　利用柱层析 (硅胶 ,SephadexLH 2 0 )从芫花根乙醇提取物中分离代表性化合物 ;以HPLC方法 ,梯度洗脱 ,分别测定不同地区芫花根各样品 10批。色谱条件 :流动相A为甲醇 ;流动相B为水 (乙酸调pH为 3.0 )。梯度洗脱A/B体积比 :0～ 2 0min ,35 / 6 5 ;2 1～ 4 0min ,4 5 / 5 5 ;4 1～ 6 0min ,5 0 / 5 0 ;6 1～ 85min ,6 5 / 35 ,分析时间 12 0min。流速 :1ml/min ,柱温 :35℃。结果　 10批芫花根乙醇提取物得到的色谱指纹图谱有 2 3个共有峰 ,可分为 6个区段。保留时间 0～ 11min出现 2小峰 ;保留时间 11～ 2 3min出现 2个峰 ;保留时间 2 3～ 35min出现 1峰 ;35～ 6 6min出现 14个峰 ,主要特征峰 6 ,7,11,12 ,13,18,19号峰在此区域 ,峰 13为最强峰 ;6 6～ 72min出现 1个峰 ,72～ 85min出现 3个峰。不同产地的芫花根指纹图谱有一定差异。结论　芫花根乙醇提取物HPLC指纹图谱具有很强的特征性及专属性 ,可作为检测方法用于芫花根黄酮、香豆素类化合物有效成分筛选、协同作用、药代动力学、毒理学以及芫花根脱毒提取工艺研究。Aim To establish a sensitive and specific HPLC method for biomedical assay on pharmacology,toxicology,pharmacokinetics of active constituents in crude extract of the root of Daphne genkwa (ERD) as well as for the assay of setting up extracting techniques with minimum content of toxic constituents.Methods 10 Batches of samples from different growing regions were assayed respectively by HPLC in optimized gradient elution for obtaining HPLC profile of ERD.The HPLC system consisted of a Waters 600 pump and a 2487 ultraviolet detector.Data were acquired and processed with the Millennium 2010 workstation.HPLC analysis was performed with the mixture of methanol (A) and H 2O (acidified to pH 3.0 with acetic acid) (B) as mobile phase.The gradient elution was programmed as the following A/B ratio in volume:0～20min:35/65;21～40min:45/55;41～60min:50/50;61～85min:65/35.The column temperature was setup at 35℃ and the flow rate was 1ml/min.Results The HPLC chromatographic fingerprint of 10 batches of ERD displayed 23 characteristic peaks.These peaks distributed in 6 regions in the following patterns: from 0～11min,there appeared 2 low peaks;11～23min,appeared 2 peaks;23～35min,appeared 1 peak;35～66min,appeared 14 peaks including main diagnostic peak 6,7,11,13,18 and 19;66～72min appeared 1 peak;72～85min,appeared 3 peaks.The fingerprint of the roots of Daphne genkwa from different growing regions indicated differences in the composition of the secondary metabolites.Conclusion HPLC fingerprint of ERD indicates high specificity,which can be used for studies on pharmacology and toxicology as well as pharmacokinetics of flavonoids and coumarins in the extract.The fingerprint can also be used in establishing technics for extracting the active constituents from the species with minimum content of toxic diterpenoids.

关 键 词：芫花 色谱指纹图谱 乙醇提取物 梯度洗脱 保留时间 有效成分 提取工艺 脱毒 区段 筛选 

分 类 号：R282.71[医药卫生—中药学] R284[医药卫生—中医学]