Improving the reproducibility in capillary electrophoresis by incorporating current drift in mobility and peak area calculations
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Improving the reproducibility in capillary electrophoresis by incorporating current drift in mobility and peak area calculations. / Petersen, Nickolaj J.; Hansen, Steen H.
In: Electrophoresis, Vol. 33, No. 6, 2012, p. 1021-31.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Improving the reproducibility in capillary electrophoresis by incorporating current drift in mobility and peak area calculations
AU - Petersen, Nickolaj J.
AU - Hansen, Steen H
N1 - © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
PY - 2012
Y1 - 2012
N2 - The traditional way of calculating mobility and peak areas in capillary electrophoresis does not take into account the changes in the buffer viscosity at different thermostatic control and that the analytes may accelerate during the individual runs due to Joule heating effects. We present a method for accounting for these changes based on the monitored changes in current during the separation. The calculation method requires measuring the initial resistance of the buffer filled capillary, performed using a 0.2 min voltage ramping at the start of a separation. The mobility calculation corrected for current drift allowed identification of the tested analytes independent from capillary dimensions, electric field strengths and temperature control. Furthermore, the peak areas become less influenced by the experimental conditions, since the velocities of the analytes passing the detector are corrected for the acceleration during the run. The short voltage ramping could be further used to evaluate the heat transfer of the capillary to the surroundings and to estimate the temperature changes during the separation. The temperature was shown to change the ionization of 2-phenylethylamine in accordance to a pKa dependency of primary amines reported in literature.
AB - The traditional way of calculating mobility and peak areas in capillary electrophoresis does not take into account the changes in the buffer viscosity at different thermostatic control and that the analytes may accelerate during the individual runs due to Joule heating effects. We present a method for accounting for these changes based on the monitored changes in current during the separation. The calculation method requires measuring the initial resistance of the buffer filled capillary, performed using a 0.2 min voltage ramping at the start of a separation. The mobility calculation corrected for current drift allowed identification of the tested analytes independent from capillary dimensions, electric field strengths and temperature control. Furthermore, the peak areas become less influenced by the experimental conditions, since the velocities of the analytes passing the detector are corrected for the acceleration during the run. The short voltage ramping could be further used to evaluate the heat transfer of the capillary to the surroundings and to estimate the temperature changes during the separation. The temperature was shown to change the ionization of 2-phenylethylamine in accordance to a pKa dependency of primary amines reported in literature.
KW - Electromagnetic Fields
KW - Electrophoresis, Capillary
KW - Models, Chemical
KW - Phenethylamines
KW - Reproducibility of Results
KW - Temperature
KW - Viscosity
U2 - 10.1002/elps.201100551
DO - 10.1002/elps.201100551
M3 - Journal article
C2 - 22528422
VL - 33
SP - 1021
EP - 1031
JO - Electrophoresis
JF - Electrophoresis
SN - 0173-0835
IS - 6
ER -
ID: 38519012