P1254503571aMGbA

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Pherogram Normalization for CE and MEKC
analyses of bio-matrices
Jetse Reijenga1, Han Martens2, Andrea Giuliani3
and Marcella Chiari3
1. Department of Chemical Engineering and
Chemistry, Eindhoven University of Technology,
The Netherlands 2. Martens Engineering BV,
Heerlen, The Netherlands 3. Institute of
Biocatalysis and Molecular Recognition, C.N.R.,
Milano, Italy
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• migration time reproducibility, limitations due
  to
• Different injection volumes
• Stacking differences
• Non-uniform fieldstrengths
• EOF variations
• Coating deterioration

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reference migration time
tiR LdLt / V(ui ueof)
sample migration time
ti LdLt / ? V(ui ueof ?)

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calculation of ?, ? (?)
Peak 1. LdLt /Vt1 - ueof LdLt / ?Vt1R - ueof -
? Peak 2. LdLt /Vt2 - ueof LdLt / ?Vt2R - ueof
- ?

• (1/t1 - 1/t2)/(1/t1R - 1/t2R)
• 2?V/LdLt ? (1/t1 1/t2)/? - (1/t1R 1/t2R)

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migration time correction
tiR 1/(1/?ti - ?/2)
FIELD shift factor 1/?
EOF shift factor ?ti/tiR
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CEqualizer
Freeware, download from http//www.ceyork.f2s.com
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• what CEqualizer does
• Load data files (ref new)
• Select identification peaks
• Calculate correction factors ?, ?
• Display FIELD and EOF shift factors
• Normalize with respect to reference
• Save normalized new data file

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Example DNA ladder
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Example Steroids with sweeping MEKC
J.-B. Kim, J.P. Quirino, K. Otsuka, S. Terabe, J.
Chromatogr. A 916 (2001) 123
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Example Opioids in urine
A.B.Wey, W. Thormann, J. Chromatogr. A 916 (2001)
225
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Example inorganic anions with twin CE detection
system
R. Bodor, D. Kaniansky, M. Masár, J. Chromatogr.
A 916 (2001) 31
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Example ß-blockers in large volume stacking CE
J.P. Quirino, S. Terabe, Electrophoresis 21
(2000) 355
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Example aromatic amines in large volume stacking
CE
J.P. Quirino, S. Terabe, Electrophoresis 21
(2000) 355
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• Conclusions
• Diagnose peak shifts samples vs standards
• Distinguish Fieldstrength and EOF effects
• Normalise sample Electropherograms

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Acknowledgements data files of published material
were provided by D. Kaniansky (Bratislava),
J.-B. Kim and S. Terabe (Hyogo), J.P. Quirino
(Sunnyvale) and W. Thormann (Berne)
CEqualizer on http//www.ceyork.f2s.com