Incorporating high-pressure electroosmotic pump and a nano-flow gradient generator into a miniaturized liquid chromatographic system for peptide analysis
- Univ. of Oklahoma, Norman, OK (United States). Dept. of Chemistry and Biochemistry
- Univ. of Oklahoma, Norman, OK (United States). Dept. of Chemistry and Biochemistry; Veritas Laboratories, LLC, Tifton, GA (United States)
Here we integrate a high-pressure electroosmotic pump (EOP), a nanoflow gradient generator, and a capillary column into a miniaturized liquid chromatographic system that can be directly coupled with a mass spectrometer for proteomic analysis. We have recently developed a low-cost high-pressure EOP capable of generating pressure of tens of thousands psi, ideal for uses in miniaturized HPLC. The pump worked smoothly when it was used for isocratic elutions. When it was used for gradient elutions, generating reproducible gradient profiles was challenging; because the pump rate fluctuated when the pump was used to pump high-content organic solvents. This presents an issue for separating proteins/peptides since high-content organic solvents are often utilized. In this work, we solve this problem by incorporating our high-pressure EOP with a nano-flow gradient generator so that the EOP needs only to pump an aqueous solution. With this combination, we develop a capillary-based nano-HPLC system capable of performing nano-flow gradient elution; the pump rate is stable, and the gradient profiles are reproducible and can be conveniently tuned. Finally, to demonstrate its utility, we couple it with either a UV absorbance detector or a mass spectrometer for peptide separations.
- Research Organization:
- MicroChem Solutions, Norman, OK (United States)
- Sponsoring Organization:
- USDOE
- Contributing Organization:
- University of Oklahoma, Norman, Ok (United States)
- Grant/Contract Number:
- SC0006351; AR11-003
- OSTI ID:
- 1337176
- Journal Information:
- Analytica Chimica Acta, Vol. 844, Issue C; ISSN 0003-2670
- Publisher:
- ElsevierCopyright Statement
- Country of Publication:
- United States
- Language:
- English
Web of Science
Review: Electric field driven pumping in microfluidic device
|
journal | December 2017 |
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