Citation:
LIU Ya, ZHANG Bo. Microscale Sample Transfer Technology for Capillary Two-Dimensional Liquid Chromatography[J]. Chinese Journal of Analytical Chemistry,
;2019, 47(11): 1803-1808.
doi:
10.19756/j.issn.0253-3820.191490
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Hydrophilic interaction chromatography (HILIC) coupled with reverse phase liquid chromatography (RPLC) is the most commonly used two-dimensional separation mode. However, it is suffered from the incompatibility of mobile phase. To overcome this issue, a microscale sample transfer technology was developed for capillary HILIC-RPLC two-dimensional liquid chromatography in this work. The sample transfer technology was consisted of weak elution make-up liquid and trap column, so that the strong elution solvent from the first dimension could be diluted effectively. Through optimization of make-up flow rate, the organic concentration of the first dimension effluent was reduced to less than 5%, and up to 95% sample transfer efficiency was achieved. This technology was applied to the two-dimensional separation of five standard protein digest. When 8 sample fractions from the first dimension separation were transferred to the second dimension separation, a theoretical peak capacity of 680 could be obtained. In addition, the peak capacity could be improved with the increasing sampling frequency, suggesting that this technology could be effectively applied to high-resolution separation of complex mixtures.
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