高效、快速液相色谱系统的构建及评价

【作者】 李笃信；

【导师】 雷武； 李彤；

【作者基本信息】 南京理工大学 ， 应用化学， 2007， 硕士

【摘要】 论文总结了Flash色谱法、整体柱色谱法、超高压色谱法和高温色谱法的特征，系统探讨了与高压和高温相关的问题及解决方法。研究了流量和温度对色谱保留参数的影响规律。提高压力(流量)可以极大加快分离速度。而升高柱温，在改善最佳流速范围的同时，不仅使色谱柱背压降低，也可能对分离选择性起到一定的调节作用。采用亚2μm色谱柱构建了一套超高压液相色谱系统。所建立的系统最高耐压60MPa，系统稳定性良好，可以满足液相色谱定量和定性分析的需要。使用1.9μm填料色谱柱与常规5μm填料色谱柱比较，相同流速下的分析速度提高了8倍，而流动相消耗仅为1／14，节约了时间和溶剂。通过对常规仪器的改进，构建了一套高温液相色谱系统，在不同分离模式下通过对六味地黄丸的分析评价系统。系统具有分离速度快、柱效高的特点。四支串联色谱柱的绝对柱效达到7.6万，分离六味地黄丸时色谱峰数可增加一倍以上。以正相色谱柱和反相色谱柱串联构建的连续二维色谱系统，结合了正相色谱与反相色谱的特点；将紫外检测器和ELSD串联使用，可以得到更加详尽的检测信息。更多还原

【Abstract】 This paper summarized the Flash LC, MLC, UHPLC and HTLC, and focused on thehigh-pressure and high-temperature-related problems and solutions. The effect of flow rateand temperature on retention was studied. The increase in pressure (flow rate) couldgreatly accelerate separation speed. Elevation of temperature could raised the optimumvelocity, decreased column back pressure, enhanced resolution, adjusted selective andreduced the asymmetry to improve the peaks.Based on sub-2μm column, an UHPLC system was constructed. This system couldgenerate maximum pressure at 60MPa; system stability was good enough to meet thequantitative and qualitative analysis. In the analysis of the standard sample, compared with5μm, columns packed with 1.9μm particles was more faster, and it’s 8 times faster whenseparated at the same velocity and only 1/14 of solvent consumption.The conventional HPLC was modified to HTLC system. Separations of LiuweiDihuang Pill under different mode were used to evaluate the HTLC system. Separation ofLiuwei Dihuang Pill used HTLC achieved faster and high through-put analysis, and easy tocreate sophisticated fingerprint. If 4 columns used in series, the absolute plate numberreached 7.6M, peaks of Liuwei Dihuang Pill timed, especially the antecedent peak of theless retained components got enough separation and the fingerprint of sample was morecomprehensive. Continues two-dimensional chromatography system constituted of normaland reverse columns speeded up the separation speed. The mode of UV and ELSD in serialhad feature of high sensitivity and universal which gave more detailed information.更多还原