二妙丸HPLC指纹图谱及UPLC-LTQ-Orbitrap成分分析  

HPLC fingerprint of Ermiao Pills and chemical constituents analysis by UPLC-LTQ-Orbitrap

作　　者：雷蓉 周亚楠 刘永利 冯丽 

机构地区：[1]河北省药品检验研究院,河北石家庄050011

出　　处：《中草药》2017年第17期3511-3516,共6页Chinese Traditional and Herbal Drugs

基　　金：国家"重大新药创制"子课题:中成药标准研究平台建设(2012ZX09304005005)

摘　　要：目的建立二妙丸的HPLC指纹图谱质量评价方法,并对二妙丸中的化学成分进行UPLC-LTQ-Orbitrap定性分析。方法指纹图谱采用HPLC法,色谱柱为Shiseido C18柱(250 mm×4.6 mm,5μm),乙腈-0.1%甲酸水溶液为流动相进行梯度洗脱,体积流量为1.0 m L/min,检测波长为330 nm,以黄柏、苍术混合对照药材生成的色谱图为标准对照图谱,采用"中国药典委员会相似度评价软件(2012A)"对样品进行相似度评价;采用UPLC-LTQ-Orbitrap技术对二妙丸中化学成分进行定性分析,色谱柱为Thermo Scientific Syncronis C18柱(100 mm×2.1 mm,1.9μm),乙腈-0.1%甲酸水溶液梯度洗脱,经DAD检测器后采用LTQ-Orbitrap Elite质谱正离子模式采集,各成分的一级全扫描离子在离子阱内进行CID多级质谱、FT检测高分辨质谱数据分析。结果所建立的指纹图谱有10个共有色谱峰,20批样品的相似度在0.869~0.992。通过对照品指认、文献比对以及高分辨质谱数据解析,鉴定了21个化学成分,分别为neo-chlorogenic acid、木兰箭毒碱、竹叶椒碱、木兰花碱、3-O-feruloylquinic acid、蝙蝠葛碱、demethyleneberberine、oxyberberine、非洲防己碱、药根碱、小檗红碱、巴马汀、小檗碱、丁香酸、咖啡酸、(E)-4-(3-hydroxyprop-1-en1yl)-2-methoxyphenol、白术内酯II、acetosyringone、白术内酯I、selina-4(14),7(11)-dien-8-one、苍术素。结论建立的二妙丸HPLC指纹图谱专属性强,重现性好,结合液质联用技术的定性分析,可全面评价二妙丸的内在质量。Objective To establish a quality assessment method for Ermiao Pills（EP） based on HPLC fingerprint and qualitatively analyze the chemical constituents. Methods The chromatographic column Shiseido C18（250 mm × 4.6 mm, 5 μm） was used, acetonitrile-0.1% formic acid as mobile phase with gradient elution at the flow rate of 1.0 m L/min, and the detection wavelength was 330 nm. The standard chromatographic fingerprint was synthesized from chromatogram of the mixed standard herbs of Phellodendron Rupr. and Atractylodes DC, and the similarity evaluation of Ermiao Pills samples was carried out by the Similarity Evaluation System for Chromatographic Fingerprints of TCM（Chinese Pharmacopoeia Commission, version 2012A）. Ultra high-performance liquid chromatography coupled with linear ion trap-Orbitrap Elite mass spectrometer（UPLC-LTQ-Orbitrap） was used to characterize the chemical constituents of Ermiao Pills. A Thermo Scientific Syncronis C18（100 mm × 2.1 mm, 1.9 μm） column and a gradient elution of acetonitrile-0.1% formic acid were used for UPLC separation. The combination of ESI-LTQ-Orbitrap mass analyzer with a linear ion trap was applied for high resolution mass spectrometry and collision-induced dissociation（CID）. Results The chromatographic fingerprints were generated with 10 common peaks. The similarity scores of 20 samples between each material batch and the reference fingerprint ranged from 0.869—0.992. Twenty-one components were identified via referring to reference components and literatures and analyzing MS data, they were neo-chlorogenic acid, magnocurarine, xanthoplanine, magnoflorine, 3-O-feruloylquinic acid, menisperine, demethyleneberberine, oxyberberine, columbamine, jatrorrhizine, berberubine, palmatine, berberine, syringic acid, caffeic acid,（E）-4-（3-hydroxyprop-1-en1yl）-2-methoxyphenol, atractylenolide II, acetosyringone, atractylenolide I, selina-4（14）, 7（11）-dien-8-one, and atractylodin. Conclusion The established method of fingerprint is specific, combine

关 键 词：二妙丸 指纹图谱 HPLC UPLC LTQ-Orbitrap 质量评价 成分分析 相似度评价 neo-chlorogenic ACID 木兰箭毒碱 竹叶椒碱 木兰花碱 3-O-feruloylquinic ACID 蝙蝠葛碱 demethyleneberberine oxyberberine 非洲防己碱 药根碱 小檗红碱 巴马汀 小檗碱 丁香酸 咖啡酸 (E)-4-(3-hydroxyprop-1-en1yl)-2-methoxyphenol 白术内酯Ⅱ ACETOSYRINGONE 白术内酯Ⅰ selina-4(14) 7(11)-dien-8-one 苍术素 

分 类 号：R286.02[医药卫生—中药学]