【作者】 黄琳娜；

【导师】 莫卫民； 曹小吉；

【作者基本信息】 浙江工业大学 ， 应用化学， 2010， 硕士

【摘要】 超高效液相色谱-质谱法（UPLC-MS）是21世纪初发展起来的一种分析技术,它是结合了UPLC的高效分离能力与MS的高灵敏度和极强的专属性的分离检测技术。它具有应用范围广、分离能力强、灵敏度高、分析速度快和自动化程度高等特点,目前已成为有机物分析,特别是药物分析的重要方法之一。本论文开展了应用新型串联色谱柱分离头孢菌素类药物的研究;超高效液相色谱-多级质谱串联技术对仙鹤草提取物中仙鹤草酚的研究;以及超高效液相色谱-多级质谱串联柱后添加过渡金属在线区分黑鳗藤提取物中C21甾体糖苷同分异构体的研究。此外,论文详细综述了色谱质谱联用技术的工作原理以及其在药物分析中的应用。首先,对多孔壳层Halo-C₁₈柱、Zorbax Eclipse XDB-C₁₈柱、Zorbax SB-C₁₈柱、ACQUIT UPLC BEH-C₁₈柱以及多孔壳层Halo-C₁₈与Zorbax SB-C₁₈的串联柱分离分析头孢菌素类药物进行了比较研究。结果表明:Halo-C₁₈柱的分离效果明显优于Zorbax Eclipse XDB-C₁₈柱和Zorbax SB-C₁₈柱。串联柱的柱效、分离度几乎达到ACQUIT UPLC BEH-C₁₈柱的水平,而其柱压却只有UPLC柱的一半。多孔壳层Halo-C₁₈串联柱在一定程度上可以取代超高压色谱柱在常规液相色谱仪上实现高通量快速分离。其次,应用液质联用技术（UPLC/ESI-IT-MSⁿ）结合在线的高分辨HPLC/ESI-TOF-MS对仙鹤草藤茎中的仙鹤草酚进行快速筛选。实验还对仙鹤草酚的[M-H]-离子的质谱裂解机理进行探讨,得到表征其三个环单元取代基的特征碎片离子,进而成功鉴定了仙鹤草藤茎中19个具有抗肿瘤活性的仙鹤草酚结构,其中有12个是新化合物。最后,由于金属离子常常能诱导化合物的质谱裂解模式发生改变这为鉴定化合物结构提供了新的契机。在以前研究的基础上,本实验对在线的过渡金属离子诱导的C₂₁甾体糖苷的质谱裂解机理进行了详细系统的研究。并依据C₂₁甾体糖苷-过渡金属离子复合物的质谱裂解机理成功区分了两对由非还原末端单糖异构引起的同分异构体。更多还原

【Abstract】 Ultra performance liquid chromatography and mass spectrometry （UPLC-MS） has grown to become a mature and routinely used technique in analytical chemistry since 2004. UPLC-MS is an online coupling system of UPLC, a separation technique that can be applied to the extremely complex mixtures, and MS, the most sensitive method of molecular analysis that has the potential to yield information of the molecular weight as well as the structure of the analyte. It provides a low limit of detection and as an identifier for structural information.UPLC-MS has become a powerful and important technique and method in analysis of medicines in the complex mixtures.In this thesis, we comparative study on the separation of cephalosporins by the new rapid liquid chromatography column. We successfully developed electrospray ionization tandem mass spectrometry coupled with ultra performance liquid chromatography to screen for agrimols with potent activity against Staphylococcus aureus from Rhinacanthus nasutus （ L.） Kurz and C₂₁ steroidal glycosides with immunological activities from Stephanotis mucronata. In order to differentiate isomers in extracts, we also investigated C₂₁ steroidal glycosides by ultra performance liquid chromatography tandem mass spectrometry using ferrous chloride as a postcolumn reagent based on previous investigations of offline regioisomer characterization of C21 steroidal glycosides. In addition, the principle of LC-MS technique and its extensive application in pharmaceutical analysis were reviewed in detail.First, the characteristics and performance of five different C₁₈-bonded silica columns including Halo-C₁₈ column, Zorbax Eclipse XDB-C₁₈ column, Zorbax SB-C₁₈ column, ACQUIT UPLC BEH-C₁₈ column and the coupled column of Halo-C₁₈ column and Zorbax SB-C₁₈ column to separate the cephalosporins in the gradient elution mode were investigated. The results indicate that the Halo-C₁₈ column provides a higher performance than that of Zorbax Eclipse XDB-C₁₈ column and Zorbax SB-C₁₈ column. Furthermore, the efficiency and resolution of the coupled column were nearly same as that of UPLC column. However, the back pressure of the coupled column was 50 % lower than that of the UPLC column. It is obvious that the coupled column can offer rapid, high performance separation of compounds instead of UPLC column with a conventional chromatography instrument.Second, Ultra performance liquid chromatography tandem mass spectrometry was developed for screening of grimols which were homologous series in the roots of R.nasutus by means of online high–resolution HPLC/TOF-MS. Fragmentation pathways and major fragment ions were determined. As a result, nineteen phloroglucinol derivatives including five groups of isomeric compounds are rapidly analyzed and identified, and twelve of these are observed here for the first time in this plant.At last, The fragmentation patterns of compound induced by metal ion can be significantly altered, allowing new opportunities for structural determination. We have systemically investigated fragmentation patterns of C₂₁ steroidal glycosides induced using the ferrous chloride as a postcolumn reagent, the non-reducing end glycoside stereoisomers were differentiated.更多还原