• 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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For forensic toxicology use only.

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. 10-Hydroxymorphine
  2. Morphine-3β-D-glucuronide
  3. Morphine-6β-D-glucuronide
  4. Morphine
  5. Morphine N-oxide
  6. 6-Acetylmorphine

Experimental

Test Conditions

Column:

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

Part Number:

186003461

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:

0.5 mL/min

Injection Volume:

15 μL

Sample Concentration:

50 ng/mL each

Sample Diluent:

75:25 ACN:MeOH

Column Temperature:

30 °C

Instrument:

Waters ACQUITY UPLC with TQD

MRM:

Morphine 286 > 201

Morphine-3β-D-glucuronide 462 > 286

Morphine-6β-D-glucuronide 462 > 286

Morphine N-oxide 302 > 162

6-acetylmorphine 328 > 165

10-hydroxymorphine 302 > 58

Dwell Time:

25 ms

ISD:

10 ms

ICD:

10 ms

Gradient

Time (min)

Profile

%A

%B

0.00

0.1

99.9

5.00

99.9

0.1

7.00

99.9

0.1

7.10

0.1

99.9

10.00

0.1

99.9

Results and Discussion

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WA60130, August 2009

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