Phosphorylated Sugar Nucleotides Analyzed with LC-MS - AppNote
May 11, 2012
/
/
/

Date: 11-MAY-2012   Last Updated: 22-AUGUST-2026

Nucleotide Sugars UDP, ADP, TDP and CDP

UDP Hexanolamine was used as the Internal Standard in the analysis of Nucleotide Sugars in this rapid analysis Method. The Mobile Phase was designed to maximize the detector response in LCMS for maximum Chromatographic Efficiency. The simple “Inverse Gradient” which produces a HILIC mode HPLC Method, was required for the results shown.

This Method can be used in measuring Metabolite Concentration.

Peaks:

1. ADP–Glucose, RT = 3.68 min, monitored MRM transitions were m/z 588 to m/z 346
2. Proprietary Sugar Nucleotide, RT = 6.03 min, monitored MRM transitions were m/z 563 to m/z 321
3. Proprietary Sugar Nucleotide, RT = 6.20 min, monitored MRM transitions were m/z 606 to m/z 385
4. CDP–Glucose, RT = 7.13 min, monitored MRM transitions were m/z 564 to m/z 322
5. UDP Hexanolamine (internal standard), RT = 8.40 min, monitored MRM transitions were m/z 502 to m/z 258
MRM = Multiple Reaction Monitoring in LCMS/MS

Method Conditions
Column: Cogent Diamond Hydride™, 4 μm, 100 Å
Catalog No.: 70000-15P-2
Dimensions: 2.1 x 150 mm

Mobile Phase
:

  • A: DI Water / 0.1% Ammonium Formate pH 7.2
  • B: 90:10 Acetonitrile / DI Water / 0.1% Ammonium Formate pH 6

Gradient:

Time (minutes)

%B

0

95

10.0

75

12.0

75

12.1

95

15.0

95


Flow rate
: 0.3 mL / minute

Sample Preparation
: 400 mL DI Water / 400 mL Acetonitrile / 20 mL of Stock Solution of each compound / 5 mL of 12% ammonia

Detection: ESI – NEG - Agilent 6210 MSD TOF Mass Spectrometer


Note: Please note the addition of small amount of Ammonia to the Sample Matrix. The alkaline environment of the Sample Matrix assured Efficient and Symmetrical Peaks for all analytes.



© Copyright 2026. MICROSOLV. All Rights Reserved. Website & Hosting by BlueTone Media