Capillary Electrophoretic Determination of 4-Hydroxyproline
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Capillary electrophoresis (CE) represents a relatively new separation technology that has gained acceptance in a wide variety
of applications. A discussion of the basic theory of CE separations is beyond the scope of this presentation, but is well
addressed in a number of recent texts (1–5). In principle, differential migration of analytes in a potential field is achieved as a result of individual differences
in mass/charge ratios. In CE, substantial advantage in separation speed, efficiency, and resolution is derived from the veryhigh
field strengths (e.g., 150–300 V/cm) typically used. In addition, high field strengths induce the effect of electroosmosis
in the column. The walls of the silica column have a negative charge and attract hydrated counter ions from the buffer. When
power is applied to the system, the positively charged ions with their associated water molecules, migrate toward the cathode
with substantial velocity. This results in the flow of water termed electroosmosis. The flow proceeds from the anode to the
cathode and serves as a pump. Because the electroosmotic water flow is much greater than the velocity of the analytes, all
components are swept to the cathode.
Affiliation(s): (2) Doheny Eye Institute, Los Angeles, CA
(3) Department of Food Science, University of Nebraska, Lincoln, NE
(3) Department of Food Science, University of Nebraska, Lincoln, NE
Book Title: Amino Acid Analysis Protocols
Series: Methods in Molecular Biology | Volume: 159 | Pub. Date: Sep-05-2000 | Page Range: 227-235 | DOI: 10.1385/1-59259-047-0:227
Subject: Biochemistry
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