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

Guanine (G) is one of the four nucleobases used to form DNA and is a derivative of purine. It pairs with Cytosine (C) in the double helix of DNA (in RNA it is replaced by Uracil, U). Adding a ribose ring to guanine transforms it into the nucleoside Guanosine. Removing a hydroxyl group transforms Guanosine into Deoxyguanosine which is a key building block of DNA. Using SIELC’s newly introduced BIST™ method, Deoxyguanosine, Guanine and Guanosine 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 Phosphoric acid (H3PO4), which act 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. The effect each ionic modifier has on the retention and separation characteristics of these compounds is shown here. Using this new and unique analysis method, Deoxyguanosine, Guanine and Guanosine can be separated, retained, and UV detected at 260 nm.

High Performance Liquid Chromatography (HPLC) Method for Analyses of Deoxyguanosine, Guanine and Guanosine

Condition

Column	BIST™ B+, 4.6×150 mm, 5µm, 100A
Mobile Phase	MeCN
Buffer	H3PO4, H2SO4, HFGA (Hexafluoroglutaric acid) – 0.2%,
Flow Rate	1.0 ml/min
Detection	UV 260 nm
Retention time

Description

Class of Compounds	Nucleosides
Analyzing Compounds	Deoxyguanosine, Guanine and Guanosine