Method Conditions:
Voltage: 28 kV
Capillary: MicroSolvCE™ Bare Fused Silica 25µm 27cm long
Injection: Hydrodynamic
Run Buffer: CElixir™ Accelerator™ Solution, pH 6.2, 7.2, 8.2 and 9.2
Organic Additive: None
Detection: 190nm UV
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Using a Beckman™ P/ACE 5000 capillary electrophoresis instrument and a bare fused silica capillary, the separation of a protein mixture was found. A solution of Bovine Albumin, Lactalbulin, Trypsin Inhibitor and Carbonic Anhydrase was prepared using good laboratory practice. Four different CElixir pH’s were used to “scout” for the method that showed the most separation. Ramping the voltage for one minute before beginning the separation was employed after inserting a 1 sec injection of a “water plug”. This aided in the precision of this method.

Discussion and Rationale:
The first run that was performed was done in the “borate” CElixir™ at pH 9.2 without sufficient. Since CElixir™ does not suffer from pH hysteresis when changing pH, the same capillary was used without any conditioning steps. CElixir™ pH 8.2 was then run without sufficient resolution. The third run was performed by using CElixir™ pH 6.2 and resolution was achieved.

A fourth run was preformed by adding together CElixir™ Accelerator solutions of pH 6.2 and pH 8.2 (according the CElixir™ Manual) that resulted in an CElixir of pH 7.2. This run did not produce any better resolution but indicates that with more pH optimization, the first two peaks may show baseline resolution.

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