Agilent PLRP-S Ion-Pair Reversed-Phase Purification Application Note

About this manual

The document discusses ion-exchange and ion-pair reversed-phase (IP-RP) chromatography techniques for analyzing and purifying synthetic oligonucleotides. It highlights that IP-RP uses organic solvents with volatile eluents, resulting in smaller volumes compared to the aqueous eluents required for ion-exchange.

The note focuses on Agilent PLRP-S reversed-phase materials, specifically evaluating different pore sizes (100A, 300A, 1000A, and 4000A). It presents data on dynamic binding capacity and mass transfer characteristics at ambient and elevated temperatures using TEAA and TBuABr ion-pairing agents. The text concludes that the PLRP-S 100A material is recommended for typical 15 to 20mer oligonucleotides due to its high surface area and capacity.

What's inside

• Introduction
• Materials and Methods
• Separation of Phosphodiester in a Thiolated Oligonucleotide
• Determination of Binding Capacity
• Results and Discussion
• Conclusions

Frequently asked questions

Which PLRP-S pore size is recommended for purifying typical 15 to 20mer oligonucleotides?

The small pore high surface area PLRP-S 100A is recommended.

Which ion-pairing agent is preferred for resolving residual phosphodiesters from phosphorothioate oligonucleotides?

The TEAA ion-pairing agent is preferred.