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G005644

HPLC Analysis of Nucleosides on Ascentis® Express OH5 (HILIC Mode)

application for HPLC

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technique(s)

HPLC: suitable

test parameters

column: Ascentis Express OH5, 10 cm x 2.1 mm I.D., 2.7 μm particles (53757-U)
mobile phase: [A] 5 mM ammonium acetate, pH 5.0 with acetic acid in 95:5, acetonitrile:water; [B] 5 mM ammonium acetate, pH 5.0 with acetic acid in 80:20, acetonitrile:water
gradient: 0% B held for 1 min; to 100% B in 10 min; held at 100% B for 1 min
flow rate: 0.3 mL/min
column temp.: 25 °C
detector: UV, 250 nm
injection: 2 μL
sample: 10 - 100 μg/mL in 95:5, acetonitrile:water (pH of buffer stock (in water) was adjusted before further dilution with water and/or acentonitrile)

suitability

application for HPLC

application(s)

life science and biopharma

Analysis Note

Nucleosides are involved in many aspects of metabolism. Analogs of nucleosides have medical application in antivirals or anticancer treatments. This application demonstrates the utility of hydrophilic interaction chromatography (HILIC) for the retention and resolution of nucleosides and analogs thereof. Elution is not solely explained by logD but by other ionic and solvophobic effects.

Legal Information

Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany

Analyte

Description

    5-Methyluridine
    DMT-2′O-TC-rU Phosphoramidite
    2-Thiocytidine dihydrate
    2′-O-Methylcytidine
    Inosine
    5-Methylcytidine
    Cytidine
    DMT-2′O-TC-rG(ib) Phosphoramidite
    1-Methyladenosine
    7-Methylguanosine
    See All (10)

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