Zero dead volume capillary electrophoresis and laser induced fluorescence detection online derivatization device

Abstract

The invention relates to a zero dead volume capillary electrophoresis and laser induced fluorescence detection online derivatization device, in particular, a zero dead volume hollow fiber film reactor is connected behind a capillary electrophoresis separation column, a fluorescence derivatization agent enters a system through a film and directly carries out online derivatization on samples such as protein, polypeptide and the like, and a product is detected through a laser induced fluorescence detector behind the reactor. The zero dead volume capillary electrophoresis and laser induced fluorescence detection online derivatization device has the characteristics that online fluorescence derivatization is carried out on samples such as micro (even trace) proteins, polypeptide and the like by using the hollow fiber film reactor, and the defects of large sample loss, complexity in operation, long time consumption and the like in an offline derivatization process are overcome. Compared with other online reactors, the zero dead volume capillary electrophoresis and laser induced fluorescence detection online derivatization device has the advantages of small dead volume, less consumption of reagents, controllable derivatization temperature and time, low cost, multiple functions, convenience and stability for operation, and wide application prospect.

Description

【Technical field】 [0001] The invention relates to the field of capillary electrophoresis separation technology and fluorescence detection technology, in particular to the manufacture and use method of a zero-dead volume capillary electrophoresis-laser-induced fluorescence detection (CE-LIF) online derivative device. 【Background technique】 [0002] As an important class of life-active substances, the specific identification and detection of proteins, peptides and other samples is of great significance to the study of the origin and evolution of life, metabolism and genetics of organisms. Capillary electrophoresis (CE) has become one of the most important and effective analysis methods for protein and peptide samples due to its various separation modes, high efficiency, and rapidity. The introduction of the laser-induced fluorescence detector (LIF) is a milestone in the development history of capillary electrophoresis detection. It overcomes the defect of low detection sensiti...

AI Technical Summary

Problems solved by technology

However, pre-column derivatization requires a certain amount of sample, and low-concentration samples are easily lost or introduce unnecessary impurities during off-line operation; recently, some research groups have reported using in-column reactions, that is, using derivatization reagents and target analytes The migration rate is different, so that the two form an intersecting interface during the migration process in the capillary to complete the derivatization, but the limitation of this method is that the reaction contact time is extremely short, the derivation efficiency is low, and it is not suitable for complex samples.

Conventional post-column derivatization devices (such as sheath flow cells, switching valves, etc.) not only require high micro-processing of the instrument, but also the sample is diluted by a high factor, and the short contact time also requires the derivatization reaction to be completed in an instant, and there is a bottleneck in development.

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