On-line capillary isoelectric focusing–mass spectrometry for quantitative analysis of peptides and proteins

On-line capillary isoelectric focusing–mass spectrometry (cIEF–MS) was applied to determine concentrations of peptides and proteins using angiotensin II and human tetrasialo-transferrin as the model samples. The concentration of the carrier ampholyte was optimized for both resolution and ion intensi...

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Bibliographic Details
Published in:Journal of pharmaceutical and biomedical analysis 2005-03, Vol.37 (3), p.423-428
Main Authors: Kuroda, Yukihiro, Yukinaga, Hideo, Kitano, Masahiro, Noguchi, Taeko, Nemati, Mahboob, Shibukawa, Akimasa, Nakagawa, Terumichi, Matsuzaki, Katsumi
Format: Article
Language:English
Subjects:
Analysis
Analytical, structural and metabolic biochemistry
Angiotensin
Animals
Biological and medical sciences
Capillary isoelectric focusing
Cattle
Electrophoresis, Capillary - methods
Fundamental and applied biological sciences. Psychology
General pharmacology
Humans
Isoelectric Focusing - methods
Mass spectrometry
Mass Spectrometry - methods
Medical sciences
Online Systems
Peptides - analysis
Pharmacology. Drug treatments
Proteins - analysis
Quantitative analysis
Sheep
Transferrin
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Summary:On-line capillary isoelectric focusing–mass spectrometry (cIEF–MS) was applied to determine concentrations of peptides and proteins using angiotensin II and human tetrasialo-transferrin as the model samples. The concentration of the carrier ampholyte was optimized for both resolution and ion intensity. cIEF–MS employing 1% Pharmalyte 3–10 and a sheath liquid containing water/methanol/acetic acid (50/49/1) resolved angiotensin I and II (5 μM each, Δp I = 0.2) at an Rs value of 2.29. The determined concentration of angiotensin II (0.1–5 μM) well correlated ( R = 0.999) with that obtained by the conventional RP–HPLC method. The limit of detection was 0.22 μM, which was about 10 times lower than that by UV detection (2 μM). The repeatability and accuracy were
ISSN:0731-7085
1873-264X
DOI:10.1016/j.jpba.2004.11.025