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Standard Peptide Mixture
Precise and Fast Equilibration Time
Method Conditions
| Column |
Cogent Bidentate C8 300™, 5µm, 300A |
| Catalog No. |
40008-75P-3M |
| Dimensions |
4.6 x 75 mm |
| Solvents |
A: DI water + 0.1% trifluoroacetic acid (TFA)
B: acetonitrile + 0.1% TFA |
| Gradient |
| Time (min.) |
%B |
| 0.0 |
9 |
| 5.0 |
21 |
| 20.0 |
27 |
| 21.0 |
9 |
|
| Post time |
5 min |
| Flow rate |
1.0 mL/min. |
| Sample Prep |
1. Gly-Tyr
2. Val-Tyr-Val
3. Methionine enkephalin
4. Angiotensin II
5. Leucine enkephalin
|
| Detection |
UV 214 nm. |
Discussion
HPLC in various modes is a main technique for the characterization of peptides. Reverse Phase HPLC is employed for the initial analysis
and the final large scale purification of peptides. The first step of the production of synthetic peptides usually involves an initial
separation of the peptides in the mixture on an analytical scale. Next the purification and collection of the target peptide follows.
The Figure shown here presents the use of RP-HPLC with a Cogent Bidentate C8 300™ column in the separation of a five peptide
mixture. The 300A pore size of the sorbent is ideal for separation of small peptides chosen for the presented chromatogram. This column
is a great choice for the gradient analysis of these due to the very fast equilibration time between injections (less than 5
minutes).
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