• Application Note

Gradient Separation of Morphine and Metabolites on Atlantis HILIC Silica

Gradient Separation of Morphine and Metabolites on Atlantis HILIC Silica

  • Waters Corporation
Round white pill in hand; glipizide; diabetes drug

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

Abstract

This application brief demonstrates the gradient separation of morphine and its metabolites on Atlantis HILIC Silica column.

Introduction

The compounds used in this study are-

  1. 6-Acetylmorphine
  2. Morphine
  3. Morphine-3β-D-glucuronide
Structures of the compounds used in this study. Figure 1. Structures of the compounds used in this study.

Experimental

Test Conditions

Column:

Atlantis HILIC Silica, 2.1 x 50 mm, 3 μm

Part Number:

186002011

Mobile Phase A:

10 mM NH4COOH in H2O, 0.125% HCOOH in 50:50 ACN:H2O

Mobile Phase B:

10 mM NH4COOH in H2O, 0.125% HCOOH in 90:10 ACN:H2O

Flow Rate:

0.6 mL/min

Injection Volume:

5 μL

Sample Concentration:

25 μg/mL each

Sample Diluent:

75:25 ACN:MeOH with 0.2% HCOOH

Column Temperature:

30 °C

Detection:

UV @ 280 nm

Sampling Rate:

20 points/sec

Time Constant:

0.1

Instrument:

Waters ACQUITY UPLC with ACQUITY PDA

Gradient:

Time(min)

Profile

%A

%B

0.00

0.1

99.9

1.05

0.1

99.9

4.35

99.9

0.1

4.50

0.1

99.9

6.00

0.1

99.9

Results and Discussion

WA64070, August 2009

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