• Application Note

Gradient Separation of Morphine and Metabolites on ACQUITY UPLC BEH HILIC

Gradient Separation of Morphine and Metabolites on ACQUITY UPLC BEH HILIC
  • Waters Corporation
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This is an Application Brief and does not contain a detailed Experimental section.

Abstract

This application brief demonstrates the gradient separation of morphine and metabolites on ACQUITY UPLC BEH HILIC Columns.

Introduction

The compounds used in this study are:

  1. 6-Acetylmorphine
  2. Morphine
  3. Morphine-3β-D-glucuronide

Experimental

Test Conditions

Column:

ACQUITY UPLC BEH HILIC, 2.1 x 50 mm, 1.7 μm

Part Number:

186003460

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:

1.235 mL/min

Injection Volume:

5 μL

Sample Concentration:

25 ng/mL each

Sample Diluent:

75:25 ACN:MeOH

Column Temperature:

30 °C

Sample Temperature:

15 °C

Weak Needle Wash:

ACN/H2O 95/5

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

0.51

0.1

99.9

2.11

99.9

0.1

2.19

0.1

99.9

2.91

0.1

99.9

Results and Discussion

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WA60132, August 2009

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