Peptide Production Confirmation Using the Cogent HPS© C18

An in-process Method for Qualifying a synthetic peptide is shown with 3 samples of tryptic “digests” were injected on the same Column. The Reproducibility from run to run on the samples is excellent giving the user a high level of confidence. The process and Method was further confirmed by coupling the HPLC Separation directory to Mass Spectrometry. The Mass data had a high degree of Accuracy and Reproducibility, verifying the peptide.

Method Conditions
Column: Cogent HPS C18™, 5μm, 120Å
Catalog No.: 75018-25P
Dimensions: 4.6 x 250mm
Mobile Phase:
A: DI Water with 0.1% Trifluoroacetic Acid (TFA)
B: Acetonitrile wtih 0.1% Trifluoroacetic Acid (TFA)
Gradient:

Time (minutes) %B
0 0
20 60
20.1 0

Temperature: 30°C
Injection vol.: 20μL
Flow rate: 1mL / minute
Detection: UV @ 254nm
Sample Preparation: 3 Different Samples from the same peptide batch were digested using Trypsin and injected onto the HPLC Column.

Notes: Cogent HPS C18 Columns are perfectly suited for peptide analysis. The pore diameter, the exhaustive end capping and the low metal content of the Silica make these Columns an excellent choice for Peptide Mapping after digestion with Trypsin also known as a Tryptic Digest. Unique Selectivity and low cost make these Columns the choice of many Protein laboratories worldwide when Reproducibility and Reliability are mandatory.

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