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G1006858

LC/MS Analysis of Nucleosides on Ascentis® Express F5

application for LC-MS

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technique(s)

LC/MS: suitable

test parameters

column: Ascentis Express F5, 15 cm x 2.1 mm I.D., 2.7 μm particles, 90Å (53571-U)
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
column temp.: 15 °C
detector: MS, ESI (-) QTOF, EIC
injection: 5 μL
sample: 100 μg/mL of 12 nucleosides except N6-methyladenosine in 99:1 10mM ammonium acetate pH 5.5 in water : methanol

suitability

application for LC-MS

application(s)

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.

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.

Other Notes

1-methyladenosine undergoes a Dimroth rearrangement to yield N6-methyladenosine in the standard solution mix.

Legal Information

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

Analyte

Description
    Pseudouridine
    Cytidine, BioReagent, suitable for cell culture, powder, ≥99%
    Uridine, ≥99%
    2-Thiocytidine
    5-Methylcytidine, ≥99%
    Ribothymidine
    2′-O-Methylcytidine
    Inosine, ≥99% (HPLC)
    Guanosine, ≥98%
    3-Methylcytidine methosulfate
    1-Methyladenosine
    7-Methylguanosine, ≥90%
    N6-Methyladenosine
    See All (13)

analytical column

Product No.
Description
Pricing

mobile phase component

Product No.
Description
Pricing

standard

Product No.
Description
Pricing

M4254

5-Methylcytidine, ≥99%

M0627

7-Methylguanosine, ≥90%

U3750

Uridine, ≥99%

C4654

Cytidine, BioReagent, suitable for cell culture, powder, ≥99%

I4125

Inosine, ≥99% (HPLC)

G6752

Guanosine, ≥98%

47310-U

Nucleosides Test Mix, in 1% sodium formate (varied conc.), analytical standard

PHR1138

Adenosine, Pharmaceutical Secondary Standard; Certified Reference Material

PHR1514

Disodium Guanylate, Pharmaceutical Secondary Standard; Certified Reference Material

PHR2244

Uridine, Pharmaceutical Secondary Standard; Certified Reference Material

PHR2411

Guanosine, Pharmaceutical Secondary Standard; Certified Reference Material

PHR2340

5′-Adenylic Acid, Pharmaceutical Secondary Standard; Certified Reference Material

PHR3163

Zidovudine Related Compound D, pharmaceutical secondary standard

PHR3802

Cytidine, certified reference material, pharmaceutical secondary standard

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