LC/MS Analysis of Nucleosides on Ascentis® Express F5

LC/MS Analysis of Nucleosides on Ascentis® Express F5


column Ascentis Express F5, 15 cm x 2.1 mm I.D., 2.7 µm particles, 90Ã… (53571-U)
column temp. 15 °C
mobile phase [A] 5 mM ammonium acetate, pH 5.5 with acidic acid in water; [B] Methanol
gradient hold at 0% for 6 min; to 50% B in 4 min; hold at 50% B for 4 min; to 0% B in 0.1 min; to hold at 0% B for 5.9 min
flow rate 0.3 mL/min
pressure 5017-9367 psi
sample 100 µg/mL of 12 nucleosides except N6-methyladenosine in 99:1 10mM ammonium acetate pH 5.5 in water : methanol
injection 5 µL
detector MS, ESI (-) QTOF, EIC


Analysis Note Nucleosides are structural subunits of nucleic acids, the macromolecules that convey genetic information in living cells. They contain a molecule of sugar (pentose) bonded to a nitrogen-containing organic ring compound. The nitrogen-containing compound is either a pyrimidine (cytosine, thymine, or uracil) or a purine (adenine or guanine). Nucleosides, derivatives and / or analogues can be used in many different industries and applications as in agrochemistry (herbicides, fungicides and insecticides), therapeutic drugs including a range of antiviral (HIV, hepatitis C virus and herpes simplex) and even antibiotics. This application demonstrates reverse phase interactions with mobile phases that are shown to work well in mass spectrometry conditions.
Categories Analytical Chromatography, Nucleosides
Featured Industry Life Science and Biopharma
General description Detector: MS, ESI (-) QTOF, EIC, (Pseudouridine m/z 243 and 153; Cytidine m/z 242 and 109; Uridine m/z 243 and 111; 2-Thiocytidine m/z 258 and 126; Guanosine m/z 282 and 150; Inosine m/z 267 and 135; Ribothymidine m/z 257 and 125; 1-Methyladenosine m/z 280 and 148; 2′-O-Methyladenosine m/z 256, 213 and 108; 5-Methylcytidine m/z 256 and 124; 7-Methylguanosine m/z 124 and 164; 3-Methylcytidine methosulfate m/z 256 and 124; N6-Methyladenosine m/z 280 and 148.
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC
Other Notes 1-methyladenosine undergoes a Dimroth rearrangement to yield N6-methyladenosine in the standard solution mix.
suitability application for LC-MS


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