1Build your own CE workshop, Brno Oct 15-16, 2018

BUILD YOUR OWN CAPILLARY ELECTROPHORESISINSTRUMENT

Petr Kubáň

Department of Bioanalytical InstrumentationCentral Institute of Technology (CEITEC) – Masaryk University

Institute of Analytical Chemistry of the Czech Academy of SciencesVeveří 97, 602 00, Brno, Czech Republic

Email: petr.kuban@gmail.com

2Build your own CE workshop, Brno Oct 15-16, 2018

LITERATURE (open source)

Tutorial: Capillary electrophoresishttp://www.esi.umontreal.ca/~badiaa/CE%20basics.pdf

Basic Principles and Modes of Capillary Electrophoresishttps://www.springer.com/cda/content/document/cda_downloaddocument/9780896036451-c2.pdf?SGWID=0-0-45-454204-p173727706

HP primer: Capillary electrophoresis: An Introductionhttp://www.colby.edu/chemistry/CH332/laboratory/Agilent%20CE%20Primer.pdf

Introduction to capillary electrophoresis (Beckman coulter)https://ls.beckmancoulter.co.jp/files/appli_note/CEPrimer1.pdf

Basics of Capillary Electrophoresis (Ronald E, Majors, HP)matematicas.udea.edu.co/~carlopez/cnq533/CE_Basics.pdf

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CAPILLARY ELECTROPHORESIS

separation technique (liquid phase)

narrow-bore fused silica capillaries (10–75 μm id)

small sample volume required (1 to 50 nl injected)

high voltages (10 to 30 kV) and high electric fields (100 to 1000 V/cm) are applied across the capillary

high resistance of the capillary limits current generation and internal heating (currents typically µA)

high efficiency (N>10.000 to 100.000)

short analysis time

4Build your own CE workshop, Brno Oct 15-16, 2018

sample sample

high voltage

separation electrolyte

capillary

+

+

++_

___

+ _

detector

CAPILLARY ELECTROPHORESIS SYSTEM

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PHOTO OF A SIMPLEST CE SYSTEM

HV supply

DAQ

capillary

VIAL + electrode

VIAL + electrode

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B

HV C4D

T

CPDAS

INJ + G

Agilent

Prince

Wynsep

EH systems

Home build CE systems

7Build your own CE workshop, Brno Oct 15-16, 2018

Basics of Capillary Electrophoresis(Ronald E, Majors, HP)matematicas.udea.edu.co/~carlopez/cnq533/CE_Basics.pdf

ELECTROOSMOTIC FLOW

Negatively charged

fused silica surface

(Si-O ¯ )

Hydrated cations

accumulate near

the surface

Bulk flow is towards

the cathode upon

application of the

electric field

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ELECTROPHORETIC MOBILITY IN CE, µA

A

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https://www.youtube.com/watch?v=wStV1rFjHOo

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EOF (CE) vs. Pressure flow (HPLC)

Basics of Capillary Electrophoresis(Ronald E, Majors, HP)matematicas.udea.edu.co/~carlopez/cnq533/CE_Basics.pdf

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INJECTION IN CAPILLARY ELECTROPHORESIS

Basics of Capillary Electrophoresis (Ronald E, Majors, HP) matematicas.udea.edu.co/~carlopez/cnq533/CE_Basics.pdf

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BUFFER (BACKGROUND ELECTROLYTE) CONSIDERATIONS

Always need to consider co- and counter- ion

Good buffering capacity (pKa), pH influence separation

Co-ion mobility matching the analytes

Suitable solubility, no interaction with analytes/wall

Suitable optical properties/conductivity

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BGE

Asr

CONCENTRATION PROFILE ELECTROPHEROGRAM

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direct UV detection

indirect UV detection

µA > µBGE µA = µBGE µA < µBGE

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Conductivity detection

µA> µBGE µA = µBGE µA < µBGE

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FUSED SILICA CAPILLARIES

ID 10-75 um

Fused silica, PTFE, PEEK

Shape (circular, rectangular)

PRODUCERS

Polymicrotechnologies MOLEX

Microquartz

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HIGH VOLTAGE POWER SUPPLIES

Ready to use Modules

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PHOTO OF OUR IN-HOUSE BUILT HVPS

Based on EMCO DX250 module

Based on Spellmann UM8*4 module

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DETECTORS

TYPE Commercially available

DIY

UV-VIS absorbance YES YES

Contactless conductometric (C4D, CCD etc.) YES YES

Fluorescence (LIF, LED-LIF) YES YES

Amperometric NO YES

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DATA ACQUISITION DEVICES

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CapillaryCE SYSTEM CONSISTS OF 3 MAJOR PARTS

CONCLUSIONS

HV power suppplyDetector

CE is easy to build from ready modules ~10 minutes

CE is cheap – total cost: 500-3000 EUR

CE is easily made portable

CE is the best separation technique for the OPEN SOURCE HW