论文部分内容阅读
讨论了影响复杂游离地卟啉混合物高效液相色谱(HPLC)分析效果的主要因素,指出二溶剂梯度洗脱体系和三溶剂梯度洗脱体系有着实际的差异,不能简单地等同看待;流动相的溶剂配比是控制分析效果的决定性因素,同时,只有溶剂体系与洗脱梯度的有效配合才能取得良好的分析效果。探索了一套游离地卟啉二溶剂梯度HPLC分析条件。采用在流动相中同时加入醋酸和吡啶的方法,当溶剂A配比为正已烷∶二氯甲烷∶丙酮∶吡啶=500∶200∶50∶1.5,溶剂B配比为正已烷∶醋酸=105∶1时,采用使流动相起始极性较低的梯度洗脱方式,对于较低柱效的色谱柱(硅胶柱)亦能获得良好的分析效果。通过典型样品的质谱(MS)和HPLC的对比分析,给该分析条件下HPLC峰群进行了初步定性,认为对大多数地卟啉样品能实现ETIO,DPEP和Di-DPEP三种类型卟啉的较有效的分离。图5表3参4
The main factors influencing the HPLC analysis of complex free porphyrin mixtures are discussed. It is pointed out that there are practical differences between the two-solvent gradient elution system and the three-solvent gradient elution system and can not be simply treated as equivalent. The mobile phase Solvent ratio is the decisive factor in the control of the analytical results, and only the effective combination of the solvent system and the elution gradient can achieve a good analytical result. A set of free porphyrin di-solvent gradient HPLC analysis conditions was explored. Adopting the method of adding acetic acid and pyridine in the mobile phase at the same time, when the ratio of solvent A is n-hexane: dichloromethane: acetone: pyridine = 500: 200: 105: 1, the mobile phase with a lower initial gradient of the gradient elution method, for the lower efficiency of the column (silica gel column) also get good results. Through the comparative analysis of typical samples by mass spectrometry (MS) and HPLC, the HPLC peak population under this analytical condition was preliminary qualitative. It is considered that most porphyrin samples can achieve the three types of porphyrins of ETIO, DPEP and Di-DPEP More effective separation. Figure 5 Table 3 Reference 4