TY - JOUR
T1 - Fast, selective, and sensitive analysis of low-abundance peptides in human plasma by electromembrane exctraction
AU - Balchen, Marte
AU - Lund, Hanne
AU - Reubsaet, Léon
AU - Pedersen-Bjergaard, Stig
PY - 2012/2/24
Y1 - 2012/2/24
N2 - A totally new concept based on electrokinetic migration was evaluated for the extraction of three biologically active peptides from human plasma. Angiotensin 2, leu-enkephalin, and endomorphin 1 migrated from a diluted human plasma sample (2mL, positive electrode), through a supported liquid membrane (SLM) of 1-octanol, di-isobutylketon, and di-(2-ethylhexyl) phosphate (DEHP) (55:35:10, w/w/w), and into an acidified acceptor solution (25μL 50mM HCl, negative electrode) by the application of an electrical potential (20V) across the SLM. After only five min of extraction, the acceptor solution was injected and analyzed directly by liquid chromatography. The three peptides were quantified by tandem mass spectrometry, with acceptable linearity ranging from 100.0 to 1000.0pgmL -1 (r 2 in the range 0.9736-0.9988), and repeatability (RSD) ranging between 15% and 24% (n=5), using plasma spiked with the three peptides in 100pgmL -1 concentration. The estimated detection limits (S/N ratio of 3:1) for angiotensin 2, leu-enkephalin, and endomorphin 1, were 60, 24, and 24pgmL -1, respectively. With this novel approach based on electromembrane extraction (EME) coupled to LC-MS/MS, endogenous concentrations of the peptides were detected in non-spiked human plasma samples, with a total analysis time less than 50min. These experimental findings were highly interesting, and showed the opportunities for EME with regard to future peptide extractions.
AB - A totally new concept based on electrokinetic migration was evaluated for the extraction of three biologically active peptides from human plasma. Angiotensin 2, leu-enkephalin, and endomorphin 1 migrated from a diluted human plasma sample (2mL, positive electrode), through a supported liquid membrane (SLM) of 1-octanol, di-isobutylketon, and di-(2-ethylhexyl) phosphate (DEHP) (55:35:10, w/w/w), and into an acidified acceptor solution (25μL 50mM HCl, negative electrode) by the application of an electrical potential (20V) across the SLM. After only five min of extraction, the acceptor solution was injected and analyzed directly by liquid chromatography. The three peptides were quantified by tandem mass spectrometry, with acceptable linearity ranging from 100.0 to 1000.0pgmL -1 (r 2 in the range 0.9736-0.9988), and repeatability (RSD) ranging between 15% and 24% (n=5), using plasma spiked with the three peptides in 100pgmL -1 concentration. The estimated detection limits (S/N ratio of 3:1) for angiotensin 2, leu-enkephalin, and endomorphin 1, were 60, 24, and 24pgmL -1, respectively. With this novel approach based on electromembrane extraction (EME) coupled to LC-MS/MS, endogenous concentrations of the peptides were detected in non-spiked human plasma samples, with a total analysis time less than 50min. These experimental findings were highly interesting, and showed the opportunities for EME with regard to future peptide extractions.
KW - Former Faculty of Pharmaceutical Sciences
U2 - 10.1016/j.aca.2011.02.058
DO - 10.1016/j.aca.2011.02.058
M3 - Journal article
C2 - 22284872
SN - 0003-2670
VL - 716
SP - 16
EP - 23
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
ER -