中药色谱分析06-色谱联用技术2分析.ppt

质谱条件： 检测器为Agilent 6200 TOF/MS装配ESI离子源。TOF/MS分析使用全扫描模式，质核比范围100-1100，负离子模式。ESI离子源条件如下：干燥气流速，8 L/min；干燥气温度，350℃；雾化器压力，35 psig；毛细管压力，4000V；碎片电压，130-325V动态调节；skimmer voltage, 60 V; octopole RF, 250 V。 化合物鉴别策略 样品处理 1、与对照品比对保留时间、碎片信息 与已知碎片骨架信息比对 LC-TOF/MS分析,获得色谱峰精确分子量 2、在化合物数据库中搜索匹配 化合物数据库建立 对照品处理 3、根据碎片信息推导可能的结构 比对质谱标准数据库，尝试鉴别非目标化合物，个别购买对照品确证 误差5 ppm，利用[M–H]-、[2M–H]- [M–2H+Na]-等分子离子峰信息匹配最有可能的化学结构式 通过查询注射液药材相关的植物化学和药理学相关文献获得，数据库包括化合物的化学式、精确分子量、名称、中文名、来源植物，CAS号，紫外信息，质谱信息，参考文献和注释。 与文献数据对比，明确鉴别目标化合物 目标化合物 非目标化合物 * C18柱TOFMS总离子流图 HILIC柱TOFMS总离子流图 34个化合物， 4组同分异构体 17个化合物， 1组同分异构体 * 非目标化合物的鉴别和确证 EIC:m/z=130.0609 碎片电压130V 128.0351的同位素分布离子 1 2 1、异亮氨酸 2、亮氨酸 * C6H13NO2 C12H22O11 非目标化合物的鉴别和确证 [2M-H]— [M+HCOO]— [M-H]— [M-H]— [2M-H]— -162 [Glucose -H]— 蔗糖 * 非目标化合物的鉴别和确证 [2M-2H+Na]— [M-H]— [M-H-NH2]— L-苯丙氨酸 C9H11NO2 * 示例：LC-DAD-MS/MS分离鉴定中药莲子心中三种生物碱 莲子心中主要有效成分－双苄基异喹啉类生物碱的药用价值日趋重视。其中，莲心碱（Ⅰ）、异莲心碱（Ⅱ）、甲基莲心碱（Ⅲ）为莲子心中双苄基异喹啉类生物碱的主要成分 J Pharm Biomed Anal. 43 (2007) 99-104. IF2007 2.761 Dried and powdered embryo of the seed of N. nucifera GAERTN. (10 kg) was soaked in 1% HCl for 1 week. The filtrate was then precipitated with 10% aqua ammonia until the pH of the solution reached 8.5 and 593 g of crude extract were collected as brown precipitates. The crude extract was extracted three times with chloroform, and then the organic phase was extracted two times with 3% NaOH. Finally, the extract solution was precipitated with diluted HCl and off-white precipitates were filtered and dried to obtain phenolic alkaloid sample (38.5 g). The primary stock solutions of crude extract (1 mg/ml) and phenolic alkaloid sample(1 mg/ml) were prepared by dissolving appropriate amount of substance in methanol. Working solutions were obtained by diluting the stock solutions with distilled water and 10μl acetic acid per ml solution was added. All the solutions were stored at 4 ?C. 282nmHPLC图 J Pharm Biomed Anal. 43 (2007) 99-104. IF2007 2.761 LC-DAD-MS/MS分离鉴定中药莲子心中三种生物碱 1.莲心碱 2.异莲心碱 3.甲基莲心碱 制备液相制得化学对照品 采用ESI-MS/M