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catalog description
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CelerityCE™ Application Sheet
Protein Mixture on C18
- Short Analysis Time
- Simple, Easy, Fast
- C18 Bonded Capillaries with High Surface Area
Method Conditions:
Voltage: 30kV
Capillary: CelerityCE™ C18 Bonded Capillary,
20µm x 45cm
Injection: 5 Seconds @ 5” Vacuum (12.5 cm Hg Vacuum)
Run Buffer: 30mM Acetic Acid/37.5mM Aminobutyric Acid
pH: 4.41
Detection: UV 211nm
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Method:
A solution of Proteins was prepared and injected onto a CelerityCE™ C18 High Surface Area Capillary using the
ABI 270A-HT CE system. A voltage of 30kV was applied across the capillary resulting in 15µA. The mobile
phase 30mM Acetic Acid/ 37.5mM Gamma Amino Butyric Acid was prepared using Good Laboratory Practice. The
Capillary was conditioned following the procedure suggested by the capillary manufacturer. Injection was
Hydrodynamic for 5 seconds under 12.5cm of Hg
PEAKS:
1.Lysozyme (turkey)
2.Cytochrome-C
3.Ribonuclease A
4.Myoglobin
5.Unknown Impurity
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Discussion and Rationale:
These proteins are series of compounds that are well separated during a short analysis time. Basic compounds
that are as hydrophobic as these compounds are usually require a more hydrophilic phase to be separated
with such high efficiency and high symmetry. These proteins are usually problematic for uncoated
capillaries.
UV detection of 211nm was selected and sharp, well resolved peaks result. Each of these proteins are
separated isocratically.
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