Peptide purity, measured by analytical reversed phase HPLC, is the percentage of the peptide compared to impurities that absorb at 210-220 nm(peptide bond absorption wavelength). Peptide purity does not take into account water and salts usually present in the sample. Sometimes alternative methods such as ion exchange, dialysis, precipitation or gel filtration chromatography are also used. Purity of peptide is a critical factor that affects the success and meaningful outcome of a research project, thus peptide purity analysis, detection of contamination and residual content are widely performed in various industries. Common contaminates of peptide product are resulted from chemical synthesis, culturing medium, or incorporated from extraction and purification procedures. Impurities consist of deletion sequences, incompletely deprotected sequences, truncated sequences and side products of the synthesis process. Residual salt and water that do not absorb at this wavelength are not quantified. In experimental planning, it is important to consider that some peptide or contamination may be toxic to cells. This kind of toxicity o cells can generally be avoided by purifying the peptide with proper methods, which will result in only trace amounts of toxic impurities.
Recommended Purity Based on Application of Peptide:
|95% to 99%|
Recommended for quantitative assays and structural studies.
|85% to 94%|
Suitable for most qualitative and semi-quantitative biochemical assays.
|70% to 84%|
Suitable for a number of screening and research applications as shown right.
Figure 1. Peptides with different purities (Analyzed by HPLC absorbance at 214 nm)
Creative proteomics has already developed highly sensitive, reversed phase HPLC platform to analyze peptide purity.
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