• Application Note

ACQUITY UPLC HILIC Gradient Separation of Ascorbic Acid and Isoascorbic Acids

ACQUITY UPLC HILIC Gradient Separation of Ascorbic Acid and Isoascorbic Acids
  • Waters Corporation
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This is an Application Brief and does not contain a detailed Experimental section.

Abstract

This application brief highlights the gradient separation of ascorbic acid and isoascorbic acids using ACQUITY UPLC BEH Amide Columns.

Introduction

Experimental

Test Conditions

Columns:

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

Part Number:

186004801

Mobile Phase A:

50/50 MeCN/H2O with 10 mM CH3COONH4 and 0.02% CH3COOH, pH 5.0

Mobile Phase B:

90/10 MeCN/H2O with 10 mM CH3COONH4 and 0.02% CH3COOH, pH 5.0

Flow Rate:

0.2 mL/min

Injection Volume:

5.0 μL (PLNO)

Sample Concentration:

30 μg/mL each

Sample Diluent:

75/25 MeCN/MeOH with 0.2% HCOOH

Column Temperature:

25 °C

Weak Needle Wash:

95/5 MeCN/H2O

Detection:

UV @ 260nm

Sampling Rate:

20 points/sec

Filter Time Constant:

0.2

Instrument:

Waters ACQUITY UPLC with ACQUITY UPLC PDA Detector

Gradient

Results and Discussion

The compounds used in this syudy are:

  1. Isoascorbic acid
  2. Ascorbic acid

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