HPLC Method for Analysis of Oligonucleotides dA 5 mer on BIST A Column

https://sielc.com/hplc-method-for-analysis-of-oligo5a
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Chromatogram

Description

· Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies · HPLC Method for Analysis of DNA Oligo, 5 bases (AAAAA) on BIST A Column by SIELC Technologies · Synthesis of DNA Oligonucleotide (AAAAA) · The application of a DNA oligo with a sequence “AAAAA” can vary based on the context:

Using SIELC’s newly introduced BIST™ method, this oligonucleotide can be retained on a negatively-charged, cation-exchange BIST™ A column. There are two keys to this retention method: 1) a multi-charged, positive buffer, such as TMEDA formate, which acts as a bridge, linking the negatively charged dye to the negatively-charged column surface and 2) a mobile phase consisting mostly of organic solvent (such as MeCN) to minimize the formation of a solvation layer around the charged analytes. Using this new and unique analysis method, oligonucleotide can be separated, retained, and detected at 260 nm.

Method Parameters

Mobile PhaseMeCN – 60%
BufferTMEDA Formate pH 4.0 – 20 mM
Flow Rate1.0 ml/min
DetectionUV 260 nm
Sample0.021 mg/ml in EtOH/H2O – 50/50%
Injection volume1 µl
LOD*15 ppb
Class of CompoundsOligonucleotides
Analyzing CompoundsOligonucleotides

HPLC Column Used

BIST A, 4.6 x 100 mm, 5 µm, 100 A, surface coated