Capillary Electrophoresis (CE)
Capillary Electrophoresis (CE)
Capillary Electrophoresis (CE)
Done by: Hussein Talal ID No.201117011 Supervised by: prof. Ashok Kumar
The rate at which the particle moves is directly proportional to the applied electric field--the greater the field strength, the fast the mobility.
If two ions are the same size, the one with greater charge will move the fastest.
For ions of the same charge, the smaller particle has less friction and overall faster migration rate.
For safety reasons one electrode is usually at ground and the other is biased positively or negatively
Anode
Cathode
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a detector
an output device, and
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These electrodes help to induce an electric field to initiate the migration of the sample from the anode to the cathode through the capillary tube
The capillary is made of fused silica and is sometimes coated with polyimide
Each side of the capillary tube is dipped in a vial containing the electrode and an electrolytic solution, or aqueous buffer.
Where is the electrophoretic mobility and E is the electric field strength. The electrophoretic mobility is proportional to the ionic charge of a sample and inversely proportional to any frictional forces present in the buffer.
The frictional forces experienced by an analyte ion depend on the viscosity () of the medium and the size and shape of the ion. Accordingly, the electrophoretic mobility of an analyte at a given pH is given by:
This flow occurs when the buffer running through the silica capillary has a pH greater than 3 and the SiOH groups lose a proton to become SiO- ions.
EOF = /4 *E
where is the dielectric constant of the solution, is the viscosity of the solution, E is the field strength, and is the zeta potential.
A fluorescence ion with the same sign of charge as the analytes is added to background electrolyte to provide a steady background signal.
The separation principle of MEKC is based on a differential partition between the micelle and the solvent (a pseudo-stationary phase). This principle can be employed with charged or neutral solutes and may involve stationary or mobile micelles.
MEKC has become valuable in the separation of very hydrophobic pharmaceuticals from their very polar metabolites.
Its most useful application appears to be in the form of on-line analyte concentration that can be used to concentrate a given sample prior to separation by CZE
3. HPLC is more thoroughly developed and has many mobile and stationary phases that can be implemented. 4. HPLC has more complex instrumentation, while CE is simpler for the operator. 5. HPLC has such a wide variety of column lengths and packing, whereas CE is limited to thin capillaries.
This technique is commonly employed in protein characterization as a mechanism to determine a protein's isoelectric point. Isoelectric Point: is the pH at which a particular molecule (protein) or surface carries no net electrical charge.
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Complex mix. of anions, cations or mixtures of anions with other analytes present e.g. in tissue culture media may serve as typical examples
Separation is performed using capillary ionic electrophoresis (CIE) Separations are carried out in fused silica columns Separation times are usually short (2-10) mins.
By raising the electrolyte pH, carbohydrates can acquire a negative charge and be easily characterized by CE under indirect detection.
Offers new selectivity, an alternative to HPLC High separation efficiency (105 to 106 theoretical plates) Small sample sizes (1-10 ul) Fast separations (1 to 45 min)
Disadvantages
Cannot do preparative scale separations Low concentrations and large volumes difficult Sticky compounds Species that are difficult to dissolve Reproducibility problems
Thank you
References:
1-http://www.scielo.br/scielo.php?pid=S010350532003000200016&script=sci_arttext 2http://chemwiki.ucdavis.edu/Analytical_Chemistry/Instrumental_Analysis/ Capillary_Electrophoresis 3-http://www.sciencedirect.com/science/article/pii/S0003269705005956 4-Journal of Chromatography B, 656 (1994) 3-27 Biomedical applications of capillary electrophoresis Z. Deyl* @, F. Tagliarob, I. MikSik Institute of Physiology, Academy of Sciences of the Czech Republic, Videiiskri 1083, 14220 Prague 4, Czech Republic Institute of Forensic Medicine, University of Verona, Policlinico Borgo Roma, 37134 Verona, Italy http://www.analyt.natureblink.com/publikace/biomed_rev.pdf