Analysis of Nucleotides – AppNote – MicroSolv Technology Corporation

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ARTICLES, APPNOTES, FAQ (9289)
- AppNotes Library (1888)
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- Frequently Asked Questions?? (3402)
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May 9, 2012
Tanya

AppNotes ANP/HILIC Methods / AppNotes Cogent Diamond Hydride / AppNotes Library / AppNotes Metabolites / ARTICLES, APPNOTES, FAQ

ATP & AMP Separated in Aqueous Normal Phase (ANP)

The chromatogram below shows an example of the separation possible with this Method for three Adenosine analytes using an Aqueous Normal Phase (ANP) Gradient.

Peaks:
1. Adenosine-3’,5’-Cyclic Monophosphate
2. Adenosine 5’-Monophosphate (AMP)
3. Adenosine 5’-Triphosphate (ATP)

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

Time (minutes)	%B
0	95
10	70

Post Time: 5 minutes
Injection vol.: 2μL
Flow rate: 0.3mL / minutes
Detection: UV @ 254nm
Sample Preparation: 0.3mg of each Nucleotide in 50% Acetonitrile / DI Water +12% Ammonia

Notes: Nucleotides are important Phosphate-containing compounds that are found in living cells and are associated with a broad array of metabolic and biological processes. They have significant roles in the synthesis of DNA and RNA, are involved in signal transduction pathways, function as coenzymes in biosynthetic pathways and serve as energy reservoirs in biological systems.

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