On-line capillary isoelectric focusing-mass spectrometry for quantitative analysis of peptides and proteins
On-line capillary isoelectric focusing-mass spectrometry for quantitative analysis of peptides and proteins
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DOI：
10.1016/j.jpba.2004.11.025
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发表时间：
2005-03-09
影响因子：
3.4
通讯作者：
Matsuzaki, K
中科院分区：
文献类型：
--
作者：
Kuroda, Y;Yukinaga, H;Matsuzaki, K
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 mu M each, Delta pl = 0.2) at an Rs value of 2.29. The determined concentration of angiotensin II (0.1-5 mu M) well correlated (R = 0.999) with that obtained by the conventional RP-HPLC method. The limit of detection was 0.22 mu M, which was about 10 times lower than that by UV detection (2 mu M). The repeatability and accuracy were < 15 and < 11%, respectively. cIEF-MS was also applied to determine human tetrasialo-transferrin concentration. The good linearity (R-2 = 0.998) was also observed between the transferrin concentration (0.5-1.2 g/L) and peak area ratio IS; (beta-lactoglobulin B) with acceptable accuracy (< 1.9%) and repeatability (similar to 10% at 1 g/L). (c) 2004 Elsevier B.V. All rights reserved.
