YMC i-Mail June 2023

Application Note

Advantages of bioinert hardware for the analysis of phosphopeptides

When working with biomolecules, loss of analytes due to absorption by the metal hardware of the column is a well-known issue. Biomolecules with phosphate groups such as phosphorylated peptides tend to adhere strongly to the column surface. The use of bioinert column hardware is a great advantage and enables:

  • higher recovery
  • better peak shapes
  • greater reproducibility

In this Application Note the analysis of phosphorylated peptides using a bioinert YMC-Accura Triart column is compared to a standard stainless steel column.



Figure: Separation of a phosphopeptide mixture using a bioinert YMC-Accura Triart C18 column compared to a stainless steel column.

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New Technical Note

The best choice for poly(dT) oligonucleotide analysis – YMC-Triart Bio C18 columns

IP-RP is commonly used for the analysis of oligonucleotides because even oligonucleotides with minimal size differences such as poly(dT)s can be separated due to the high resolution. Poly(dT) oligonucleotides are single stranded nucleotide chains solely consisting of thymine as bases.

In this Technical Note poly(dT) oligonucleotides of different lengths (10 - 120mer) are analysed with a YMC-Triart Bio C18 column and two other columns especially designed for oligonucleotides. An elevated temperature of 65 °C and a TEA/HFIP buffer system was used. This comparison shows that YMC-Triart Bio C18 provides superior peak shapes, recovery and resolution across all lengths and is therefore the best choice for the analysis of poly(dT) oligonucleotides.

For full method details you can download the Technical Note.
 



Figure: Separation of poly(dT) oligonucleotides by IP-RP using YMC-Triart Bio C18 and two columns dedicated for oligonucleotides.

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How to choose the correct pore size for oligonucleotide analysis by SEC

SEC is a suitable method for the analysis of long chain oligonucleotides with length up to 200mer. The choice of the optimal pore size is crucial for the success of the method.

In this Application Note, analyses of oligonucleotides of various lengths are described and a selection guide for columns for different DNA is discussed - a practical addition to your local application database.

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