Separating UDP and CDP Sugars with ANP and Increased Sensitivity

This Method can be used to analyze UDP and CDP Sugars using UDP-Hexanolamine (a metabolite) as an Internal standard. The Sugar Nucleotides used in this Application Note are a mixture of compounds that occur in plants and their structure is proprietary.

A potentially powerful tool for profiling Sugar Nucleotides in Metabolomic studies, this Method uses an Inverse Gradient (HILIC like); the Mobile Phase uses high organic component which enhances Mass Spec response and assures lower detection limits.

Method Conditions
Column: Cogent Diamond Hydride™, 4μm, 100Å
Catalog No.: 70000-15P-2
Dimensions: 2.1 x 150mm
Mobile Phase:
A: DI Water / 0.1% Ammonium Formate (pH 7.2)
B: 90% Acetonitrile / 10% DI Water / 0.1% Ammonium Formate (pH 6)
Gradient:

Time (minutes) %B
0 95
10 75
12 75
12.1 95
15 95

Post Time: 5 minutes
Flow rate: 0.3mL / minute
Detection: ESI – neg – Agilent 6410 Triple Quadrupole Mass Spectrometer
Mass Data:
1. Compound 1 – the monitored MRM transitions were m/z 535 to m/z 323
2. Compound 2 – the monitored MRM transitions were m/z 564 to m/z 322
3. UDP Hexanolamine (internal standard) – the monitored MRM transitions were m/z 502 to m/z 258
—–b(MRM = multiple reaction monitoring in LC/MS/MS)

Notes: Sugar Nucleotides among other metabolites are an important group of compounds to be analyzed when one is trying to understand cellular response to genetic or environmental perturbations.

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