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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.
|Column||BIST™ B+, 4.6×150 mm, 5µm, 100A|
|Buffer||H3PO4, H2SO4, HFGA (Hexafluoroglutaric acid) – 0.2%,|
|Flow Rate||1.0 ml/min|
|Detection||UV 260 nm|
|Class of Compounds||Nucleosides|
|Analyzing Compounds||Deoxyguanosine, Guanine and Guanosine|