Evaluation of phytic acid as a buffer additive for the separation of proteins in capillary electrophoresis.
The use of phytic acid to improve protein analysis by capillary electrophoresis (CE) is becoming more and more popular. Due to its size and number of negative charges (up to 12) it provides a high ionic strength combined with a low conductance resulting in an efficient decrease of wall adsorption for proteins. Because of its twelve acidic groups, phytic acid can be used as a buffer over a wide pH range (pH 2-11). The limited wall adsorption of proteins using phytic acid-containing buffers is observed for buffers with a pH of 5.5 and higher. With a monoprotic buffer, most of the investigated proteins show wall adsorption at the pH values studied. In case of a phytic acid buffer, wall adsorption is reduced by a factor of 2-4. The use of phytic acid both as a modifier and as a pH buffer results in more pronounced differences between the various protein mobilities compared with the use of monoprotic buffers. As a result this feature can be used to improve resolution in protein separations.
Veraart
J R
JR
Free University, Department of General and Analytical Chemistry, Amsterdam, The Netherlands.
Schouten
Y
Y
Gooijer
C
C
Lingeman
H
H
D002021:Buffers; D019075:Electrophoresis, Capillary; D006863:Hydrogen-Ion Concentration; D010833:Phytic Acid / Q000737:chemistry*; D011506:Proteins / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet
D002021:Buffers; D011506:Proteins; D010833:Phytic Acid