高效液相色谱-质谱分析指导下制备黄芩中系列黄酮成分对照品
郭鹤男;杨学东;刘军;郑文凤
【摘 要】旅游英语作文在高效液相色谱-质谱分析指导下,针对性地分离制备了黄芩药材中系列黄酮成分对照品.首先对黄芩药材乙醇提取物进行液相色谱-质谱分析,获得各色谱峰的保留时间、紫外光谱和质谱特征.经波谱数据解析结合文献对比,鉴定了黄芩药材中的19种黄酮类成分.然后根据液相色谱-质谱分析结果和文献,设计了目标成分对照品的制备流程,采用低压制备柱色谱法依次制备了黄芩苷、汉黄芩苷、黄芩素、汉黄芩素和千层纸素A共5种黄酮类成分的对照品.结果表明这5种黄酮类成分对照品的纯度均大于98％.该方法可用于针对性地快速分离制备中药中的化学成分.%Flavonoid reference standards were targeted-prepared from Scutellariae Radix under the guidance of high performance liquid chromatography-mass spectrometry (HPLC-MS) analysis. With HPLC-MS analysis of Scutellariae Radix, 19 flavonoid components were identified by analyzing and comparing their retention times, ultraviolet spectra, and mass spectrometry data with literature. The paration and purification protocols of all targeted flavonoid reference standards were optimally designed according t
o the results of HPLC-MS analysis and related literature. The ethanol extract of Scutellariae Radix was suspended in water and extracted with petroleum ether, ethyl acetate, and n-butanol successively. The ethyl acetate extract and n-butanol extract were parately subjected to primary paration by low pressure rever pha preparative chromatography. Then the fractions containing targeted compounds were further purified by low pressure rever and normal phas preparative chromatography. Finally, baicalin and wogonoside reference standards were obtained from n-butanol extract;baicaelin, wogonin, and oroxylin A reference standards were obtained from ethyl acetate extract. The structures of the 5 reference standards were identified by mass spectrometry (MS) and ' H nuclear magnetic resonance ('H NMR) spectroscopy. The HPLC analytical results showed that the purities of the 5 reference standards were all above 98%. It is demonstrated that the rapid targeted-preparation method under the guidance of the HPLC-MS analysis is applicable for the isolation and preparation of chemical components in traditional Chine medicines.
【期刊名称】《色谱》
shaggy>last name七律长征翻译【年(卷),期】2012(030)007
李阳口语宝典【总页数】3年级上册英语6页(P690-695)
【关键词】高效液相色谱-质谱联用;黄芩;黄酮;对照品铍青铜英文
【作 者】郭鹤男;杨学东;刘军;郑文凤
【作者单位】天津大学药物科学与技术学院,天津300072;天津大学药物科学与技术学院,天津300072;中国计量科学研究院化学计量与分析科学研究所,北京100013;天津大学药物科学与技术学院,天津300072
【正文语种】中 文
正好英文【中图分类】O658
Abstract:Flavonoid reference standards were targeted-prepared from Scutellariae Radix under the guidance of high performance liquid chromatography-mass spectrometry(HPLC-MS)analysis.With HPLCMS analysis of Scutellariae Radix，19 flavo
noid components were identified by analyzing and comparing their retention times，ultraviolet spectra，and mass spectrometry data with literature.The paration and purification protocols ofall targeted flavonoid reference standards were optimally designed according to the results of HPLC-MS analysis and related literature.The ethanol extract of Scutellariae Radix was suspended in water and extracted with petroleum ether，ethylacetate，and n-butanolsuccessively.The ethyl acetate extract and n-butanolextract were parately subjected to primary paration by low pressure rever pha preparative chromatography.Then the fractions containing targeted compounds were further purified by low pressure rever and normal phas preparative chromatography.Finally，baicalin and wogonoside reference standards were obtained from n-butanol extract;baicaelin，wogonin，and oroxylin A reference standards were obtained from ethyl acetate extract.The structures of the 5 reference standards were identified by mass spectrometry(MS)and1H nuclear magnetic resonance(1H NMR)spectroscopy.The HPLC analytical results showed that the purities of the 5 reference standards were all above 98%.It is demonstrated that the rapid targeted-prepar
ation method under the guidance of the HPLC-MS analysis is applicable for the isolation and preparation of chemical components in traditional Chine medicines.
Key words:high performance liquid chromatography-mass spectrometry(HPLC-MS);Scutellariae Radix;flavonoid;reference standard
黄芩(Scutellariae Radix)为唇形科植物黄芩(Scutellaria baicalensis Georgi)的干燥根，具有清热燥湿、泻火解毒、止血、安胎等作用［1］。黄酮类化合物是黄芩中的主要活性成分，文献［2］报道已从黄芩中分离得到约70种黄酮类成分，其中黄芩苷、汉黄芩苷、黄芩素和汉黄芩素等是其主要特征性成分。2010年版《中国药典》中仅将黄芩药材中黄芩苷的含量不低于9%作为其质量控制的标准，这不足以有效控制黄芩药材的质量水平。近年来，采用多种成分测定的方法对黄芩药材的质量进行控制已成为一种趋势［3，4］，所以经常需要黄芩中存在的系列黄酮类化学对照品用于黄芩类药材的质量控制。
传统的中药化学成分提取分离方法耗时、费力，并且具有一定的盲目性，在未对分离得到的化合物进行结构鉴定前往往对化合物的信息了解很少。高效液相色谱-二极管阵列检测器-电喷雾-质谱(high performance liquid chromatography-diode array detector-electrospra
y ionization-mass spectrometryn，HPLCDAD-ESI-MSn)联用技术具有分析速度快、分辨率高、灵敏度高的特点，能在线提供每个色谱峰的紫外光谱、相对分子质量和结构信息，可以快速鉴定出分析样品的成分，已广泛应用于中药中化学成分的快速鉴定［5－7］。Han等［8］和Liu等［9］利用HPLC-DADESI-MSn对黄芩药材中的多种黄酮类成分进行了较为系统的研究。采用HPLC-DAD-ESI-MSn方法可以快速鉴定出黄芩药材中的化学成分，从而指导目标化学成分的快速分离制备。
本研究先采用HPLC-DAD-ESI-MSn联用法对黄芩药材乙醇提取物中的化学成分进行鉴定，确定目标成分。之后，依据其色谱行为制定分离方案，针对性地制备黄芩苷(baicalin)、汉黄芩苷(wogonoside)、黄芩素(baicaelin)、汉黄芩素(wogonin)以及千层纸素A(oroxylin A)5种黄酮类化学成分(化学结构如图1所示)的对照品，为中药中化学成分的快速分离制备提供一种新思路。
1.1 仪器、试剂与材料
Agilent1200/6310液相色谱-质谱联用仪(美国安捷伦公司);Dionex Ultimate 3000高效液相色谱仪(美国戴安公司);XS105分析天平(瑞士Mettler Toledo公司)。
硅胶(青岛海洋化工有限公司);乙腈、甲醇(色谱纯，天津康科德公司)，水为Milli-Q超纯水，其他试剂均为分析纯。黄芩药材经承德职业学院李世教授收集并鉴定为河北承德产黄芩道地药材。
样品溶液的制备:称取1.0 g黄芩药材粉末，用40 mL的70%(如无特殊说明均为体积分数)乙醇超声提取30 min，提取液过0.22μm微孔滤膜。
1.2 液相色谱-质谱定性分析
1.2.1 色谱条件
尚艺美发
保护柱:Cosmosil MS-ⅡC18，色谱柱:Cosmosil AR-ⅡC18(250 mm×4.6 mm，5μm);流动相A为0.06%的三氟乙酸水溶液，流动相B为甲醇-乙腈混合溶液(90∶10，v/v)。梯度洗脱程序:0～15 min，30%B～35%B;15～20 min，35%B～39%B;20～64 min，39%B;64～79 min，39%B～55%B;79～105 min，55%B。柱温:30℃;流速:1.0 mL/min;进样体积:10μL;DAD扫描范围:200～400 nm。
1.2.2 质谱条件
英国东伦敦大学
电离源:电喷雾离子源(ESI);检测方式:正、负离子检测模式;扫描范围:m/z 100～1 000;离子源温度:350℃;雾化压力:206.8 Pa(30.0 psi);干燥氮气流速:8.0 L/min;毛细管电压:3.5 kV。
