Flow variation as a factor determining repeatability of the internal standard-based qualitative and quantitative analyses by capillary electrophoresis.
The use of migration times and peak areas referred to another sample component - internal standard, brings many benefits in improving reliability of capillary electrophoresis. However, it is quite commonly overlooked that despite relative migration time and peak area ratio are more stable than the absolute values upon alteration in the flow rate, some shift should always be expected. The present work offers a new look at this analytically-important issue. We have derived a simple model allowing to estimate the magnitude of error for the selected pair of molecules of known mobilities upon the given flow alteration. Then, we have confronted the theoretical predictions with the experimental results obtained for the model sample separated in various flow conditions reached by the external pressure manipulation, including several internal standards of different mobilities. A good agreement has been obtained, pointing out that the magnitude of error may be large even for the seemingly "good" internal standards. Several potentially useful means have been tested to address this issue: the use of electrophoretic mobilities and electrophoretic mobility ratios instead migration times in the qualitative analysis, and performing time-correction of peak area ratios, or alternatively, transformation of electropherograms from the time-related scale into the electrophoretic mobility-related scale in the quantitative analysis. We have also considered some additional factors. The results may be of interest for all users dealing with the development and optimization of analytical methods using capillary electrophoresis.
['Nowak|Paweł Mateusz|PM|', 'Woźniakiewicz|Michał|M|', 'Kościelniak|Paweł|P|']
D019075:Electrophoresis, Capillary D011312:Pressure D012015:Reference Standards D015203:Reproducibility of Results D012212:Rheology D013997:Time Factors
Capillary electrophoresis Internal standard Electrophoretic mobility Electroosmotic flow Voltage ramping