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On-line large-volume electroextraction coupled to liquid chromatography-mass spectrometry to improve detection of peptides

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Author: Lindenburg, P.W. · Tempels, F.W.A. · Tjaden, U.R. · Greef, J. van der · Hankemeier, T.
Type:article
Date:2012
Source:Journal of Chromatography A, 1249, 17-24
Identifier: 462859
doi: doi:10.1016/j.chroma.2012.06.016
Keywords: Biology · Bioanalysis · Electroextraction · Liquid chromatography · Mass spectrometry · Peptides · Sample preconcentration · Analytical procedure · Bioanalysis · Calibration curves · Dye crystal violet · Electroextraction · Feasibility studies · Liquid chromatography-Mass spectrometry · Liquid-liquid interfaces · Liquid-liquid systems · Model peptides · Organic phase · Plasma analysis · Sample handling · Sample preconcentration · Sample volume · Electric fields · Liquid chromatography · Mass spectrometry · Plasmas · Peptides · angiotensin I · angiotensin II · angiotensin II [3-8] · crystal violet · peptide · article · automation · blood analysis · electroextraction · extraction · feasibility study · limit of detection · liquid chromatography · liquid liquid extraction · mass spectrometry · peptide analysis · priority journal · process optimization · Healthy for Life · Healthy Living · Life · MSB - Microbiology and Systems Biology · EELS - Earth, Environmental and Life Sciences

Abstract

Electroextraction (EE) takes place in a two-phase liquid-liquid system, consisting of an aqueous and an organic phase, where an applied electric field causes ions to be extracted from one phase into the other, to be concentrated close after the liquid-liquid interface. The extraction takes place in a wide-bore capillary that is connected to a 2-way 10-port switching valve, which serves to couple capillary EE (cEE) with LC-MS. In this set-up, volumes as high as 100 μL can be extracted, which is a ten times larger volume than has been reported, earlier. After a feasibility study using the cationic purple dye crystal violet, the method was coupled to LC-MS and large volume cEE of several model peptides was optimized. The cEE-LC-MS method had good repeatability, good linearity and LODs between 0.5 and 10. nM. The whole procedure was automated and could be used routinely. Finally, the method was applied to plasma analysis and calibration curves of the relevant plasma peptides angiotensin 1 and 2 as well as the fragment angiotensin 2 (3-8) showed good linearity and repeatability; LOD values were 10-50. nM. Analysis of unspiked plasma resulted in 60 putative endogenous peptides, underlining the great potential of EE as on-line sample concentrating technique. On-line large volume cEE-LC-MS allows for enrichment, separation and detection of plasma peptides from large sample volumes, minimizes sample handling and can be an important step in full automation of analytical procedures. © 2012 Elsevier B.V. Chemicals/CAS: angiotensin I, 9041-90-1; angiotensin II, 11128-99-7; crystal violet, 467-63-0, 548-62-9