Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies

Adenine (A) is one of the four nucleobases used to form DNA and RNA and is a derivative of purine. It pairs with Thyamine (T) in the double helix of DNA or Uracil (U) in RNA. Adenine is a nucleoside based on Adenine that is a key component of RNA and its derivatives AMP, ADP, and ATP are key compounds that provide energy to all living cells. Deoxyadenosine is similar to Adenine in that it has one less OH group, and is a key building block of DNA. Using SIELC’s newly introduced BIST™ method, Adenine, Deoxyadenosine and Adenosine can be retained and separated on a positively-charged anion-exchange BIST™ B+ column. 

There are two keys to this retention method: 1) a multi-charged, negative buffer, such as Sulfuric acid (H2SO4), which acts as a bridge, linking the positively-charged analytes to the positively-charged column surface and 2) a mobile phase consisting of a majority 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, Adenine, Deoxyadenosine and Adenosine can be separated, retained, and UV detected at 260 nm.

High Performance Liquid Chromatography (HPLC) Method for Analysis of Adenine, Deoxyadenosine and Adenosine

Condition

Column	BIST™ B+, 4.6×50 mm, 5µm, 100A
Mobile Phase	MeCN – 85%
Buffer	H2SO4 – 0.2%
Flow Rate	1.0 ml/min
Detection	UV 260 nm
Retention time	2.8, 3.2, 4.3 min

Description

Class of Compounds	Nucleosides
Analyzing Compounds	Adenine, Deoxyadenosine and Adenosine