• Application Note

Separation of Nucleotide Phosphates on ACQUITY UPLC BEH Amide

Separation of Nucleotide Phosphates on ACQUITY UPLC BEH Amide
  • Waters Corporation
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DNA polymerase I

This is an Application Brief and does not contain a detailed Experimental section.

Abstract

This application brief describes the separation of nucleotide phosphates on ACQUITY UPLC BEH Amide column.

Introduction

The nucleotide phosphates used in this study are-

  1. Adenosine monophosphate (AMP)
  2. Uridine monophosphate (UMP)
  3. Adenosine diphosphate (ADP)
  4. Uridine diphosphate (UDP)
  5. Adenosine triphosphate (ATP)
  6. Uridine triphosphate (UTP)
Structures of the compounds used in this study.
Figure 1. Structures of the compounds used in this study.

Experimental

Test conditions

Column:

ACQUITY UPLC BEH Amide, 2.1 x 100 mm, 1.7 μm

Part Number:

186004801

Isocratic Mobile Phase:

70/30 ACN/H2O with 27 mM KH2PO4, pH 4.5

Flow Rate:

0.5 mL/min

Injection Volume:

5 μL (PLNO)

Sample Concentration:

shown on chromatogram

Sample Diluent:

80/20 ACN/H2O

Column Temperature:

25 °C

Weak Needle Wash:

95/5 ACN/H2O

Instrument:

Waters ACQUITY UPLC with ACQUITY PDA

Detection:

UV 260 nm

Sampling Rate:

20 Hz

Time Constant:

0.1 s

Results and Discussion

Figure 2. Sample Concentration
1- AMP (50 μg/mL), 2- UMP (50 μg/mL), 3- ADP (100 μg/mL), 4- UDP (100 μg/mL), 5- ATP (100 μg/mL), 6- UTP (100 μg/mL)

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