• Application Note

Gradient Separation of Nucleic Acid Bases on ACQUITY UPLC BEH HILIC

Gradient Separation of Nucleic Acid Bases on ACQUITY UPLC BEH HILIC
  • Waters Corporation
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Helix DNA molecule with modified genes

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

Abstract

This application brief demonstrates gradient separation of nucleic acid bases.

Introduction

Compounds that are used in this study includes:

  1. 5-Fluorouracil

2. Uracil

3. 5-Fluorocytosine

4. Cytosine

Structure of the compounds analysed
Figure 1: Structure of the compounds analysed.

Experimental

Test Conditions

Column:

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

Part Number:

186003461

Mobile Phase A:

20 mM CH3COONH4, 0.05% CH3COOH in 50:40:10 ACN:MeOH:H2O

Mobile Phase B:

4 mM CH3COONH4, 0.01% CH3COOH in 95:3:2 ACN:MeOH:H2O

Flow Rate:

0.790 mL/min

Injection Volume:

0.8 μL

Sample Concentration:

25 μg/mL

Sample Diluent:

75:25 ACN:MeOH with 0.2% HCOOH

Temperature:

30 °C

Detection:

UV @ 254 nm

Sampling Rate:

20 pts/sec

Time Constant:

0.1

Instrument:

Waters ACQUITY UPLC with ACQUITY TUV

Gradient

Time(min)

Profile

%A

%B

0.0

0.1

99.9

0.37

0.1

99.9

1.71

99.9

0.1

1.74

0.1

99.9

1.98

0.1

99.9

Results and Discussion

Chromatogram
Figure 2: Chromatogram of 1. 5-Fluorouracil 2. Uracil 3. 5- Fluorocytosine 4. Cytosine

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