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Note: During the preparation of the OTCEC capillaries, about 95% of the silanol groups on the capillary inner wall are
removed by our production process, therefore undesirable adsorption effects between the positively charged analytes and
the ionised silanol groups are significantly reduced. As a result, very efficient and highly reproducible peptide
separations are achieved, as shown in presented figures.
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Cytochrome-c Tryptic Digest
Method Conditions:
Voltage: 20 kV
Capillary: CelerityCE™ Cholesterol Bonded Capillary, i.d. 50µm, total length 70cm.
Catalog No.: 04925-50
Injection: 3 seconds @ 50 mbar
Separation Buffer: 20 nM NH4Ac + 20% acetonitrile, pH 6.8
Sample: Cytochrome-c Tryptic Digest, 0.2 µg/mL.
Detection: ESI MS
Instrument: Agilent 3D CE capillary electrophoresis system coupled to an Agilent 1100 series LC/MSD-SL ion
trap mass spectrometer through an Agilent G1607A orthogonal electrospray interface (Agilent Technologies,
Waldbronn, Germany).
Electrical contact at the electrospray needle tip was established via a liquid sheath flow
delivered by an Agilent 1100 series isocratic LC pump. All system control and data acquisition were conducted
with Agilent ChemStation and MSD Trap Control software. |
Discussion:
The efficient separation of a cytochrome-c tryptic digest, as illustrated in Figures A and B demonstrates the potential of
CelerityCE™ capillaries for proteomic applications. In Figure (A) Total Ion Chromatogram of the tryptic digest of the
peptide is shown. Figure (B) shows single ion chromatogram of ions with a m/z of 261.2 that corresponds to the isobaric
tryptic peptides T6 and T11. Separation of the two ions with the same m/z ratio verifies that they have a different sequence.
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