Electrophoresis is a technique used to separate molecules according to their size and charge by applying an electric field to move them through a medium. Capillary electrophoresis improves on traditional electrophoresis by using a narrow capillary tube instead of a slab gel. Molecules migrate through the capillary at different rates depending on their charge and size. The electric field causes positively and negatively charged molecules to move in opposite directions. Capillary electrophoresis has applications in genetic analysis, pharmaceutical analysis, and protein characterization due to its high efficiency, small sample size requirements, and ability to automate.

2. ELECTROPHORESIS
MIGRATION OF IONS BY ATTRACTION OR REPULSION IN AN ELECTRIC FIELD
OR
MIGRATION OF CHARGED PARTICLES UNDER THE INFLUENCE OF ELECTRIC FIELD

3. CAPILLARY ELECTROPHORESIS
• IT IS A SEPERATION TECHNIQUE THAT SEPERATES MOLECULES IN AN ELECTRIC FIELD ACCORDING TO SIZE
AND CHARGE
• THE MIGRATION RATE OF THE PARTICLE AND THE APPLIED ELECTRIC FIELD ARE DIRECTLY PROPORTIONAL
THAT IS GREATER THE FIELD STRENGTH, FASTER WILL BE THE MOBILITYOF PARTICLES.

4. THEORY
• ELECTROPHORETIC FLOW:
o IT IS THE PROCESS IN WHICH SAMPLE IONS MOVE UNDER THE INFLUENCE OF AN APPLIED VOLTAGE.
oTHE ION UNDERGOES A FORCE THAT IS EQUAL TO THE SAMPLE OF THE ELECTROPHORETIC MOBILITY AND THE ELECTRIC FIELD
STRENGTH.
oTHE FLOW OF IONS IS TOWARDS THE OPPOSITE CHARGED ELECTRODE.
𝜇𝐸𝑃 = 𝑞 ∕ 6𝜂𝜋𝑟
𝜇𝐸𝑃 = ELECTROPHORETIC MOBILITY
𝑞 = CHARGE ON IONS
𝜂 = VISCOSITY
𝑟 = RADIUS
𝜇𝐸𝑃 = 𝑞 ∕ 6𝜂𝜋𝑟

5. THEORY
• ELECROSMOTIC FLOW
oOSMOSIS UNDER THE INFLUENCE OF AN ELECTRIC FIELD
oTHE SPEED OF EOF CAN BE ADJUSTED BY CHANGING THE BUFFER PH
oBULK MOVEMENT OF SOLUTES IS CAUSED BY EOF
oEOF IS USUALLY SUFFICIENT TO SWEEP ALL +VE, NEUTRAL, -VE, SPECIES TOWARDS THE SAME END
𝜇𝐸𝑂𝐹 = 𝐶. 𝜁 ∕ 4𝜋𝜂
𝜇𝐸𝑂𝐹 = ELECTRO OSMOTICMOBILITY
𝐶 = DIELECTRIC CONSTANT
𝜁 = ZETA POTENTIAL
𝜂 = VISCOSITY
𝜇𝐸𝑂𝐹 = 𝐶. 𝜁 ∕ 4𝜋𝜂

6. INSTRUMENTATION

7. INSTRUMENTATION
• HIGH VOLTAGE POWER SUPPLY – 5 TO 30 KV
• BUFFER SOLUTION – SODIUM DIHYDROGEN PHOSPHATE
• CAPILLARY TUBE –
i. INTERNAL DIAMETER 10 – 100MM & 20 – 100CM LENGTH
ii. MADE OF FUSED SILICA COATED WITH POLYIMIDE
• SAMPLE INJECTION –
i. HYDRODYNAMIC INJECTION – BY APPLYING PRESSURE, VACUUM & GRAVITATION
ii. ELECTROKINETIC INJECTION – BY USING ELECTRIC SUPPLY
• DETECTORS –
i. SIMILAR TO THOSE IN GC & HPLC

8. MODES OF CAPILLARY ELECTROPHORESIS
• CAPILLARY ZONE ELECTROPHORESIS
• CAPILLARY GEL ELECTROPHORESIS
• CAPILLARY ISOELECTRIC FOCUSING
• CAPILLARY ISOTACHOPHORESIS

9. CAPILLARY ZONE ELECTROPHORESIS
• ANALYTES MOVE IN THE EOF BUT SEPARATE INTO BANDS BECAUSE OF DIFFERENCES IN THEIR
ELECTROPHORETIC MOBILITIES, 𝜇
• DIFFERENCE IN 𝜇 MAKE EACH ANALYTIC’S OVERALL MIGRATION VELOCITY SLIGHTLY DIFFERENT &
DIFFERENCE IN MIGRATION

10. CAPILLARY GEL ELECTROPHORESIS
• IT USES SEPARATION BASED ON THE DIFFERENCE IN SOLUTE SIZE AS A PARTICLE MIGRATE THROUGH GEL
• GELS PREVENT THE CAPILLARY WALLS FROM ABSORBING THEN SOLUTE

11. CAPILLARY ISOELECTRIC FOCUSING
• IT IS A TECHNIQUE COMMONLY USED TO SEPARATE PEPTIDES AND PROTEINS
• IT RUNS IN A PH GRADIENT WHERE PH IS LOW AT ANODE AND HIGH AT CATHODE

12. CAPILLARY ISOTACHOPHORESIS
• IT IS A TECHNIQUE BASED ON THE MIGRATION OF SAMPLE COMPONENTS BETWEEN LEADING AND
TERMINATING ELECTROLYTES

13. APPLICATIONS
• GENETIC ANALYSIS
• ANALYSIS OF PHARMACEUTICALS
• PROTEIN CHARACTERIZATION
• COUNTER ION ANALYSIS IN DRUG DISCOVERY

14. ADVANTAGES:
• EASY& PREDICTABLE SELECTIVITY
• CAN BE AUTOMATED
• SMALL SAMPLE REQUIRED (1 TO 10 ML)
• FAST SEPARATION ( 1 T0 45 MIN)
• HIGH SEPERATION EFFICIENCY
DISADVANTAGES
• PREPERATIVE SCALE PREPERATIONS CAN NOT BE DONE
• DIFFICULT FOR LOW CONCENTRATIONS AND LARGE VOLUMES

15. THANK YOU