• Application Note

Gradient Separation of Morphine and Metabolites on XBridge HILIC

Gradient Separation of Morphine and Metabolites on XBridge 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 Xbridge HILIC Columns.

Introduction

The compounds used in this study are:

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

Experimental

Method Conditions

Column:

XBridge HILIC, 2.1 x 50 mm, 3.5 μm

Part number:

186004432

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 ng/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

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WA64079, August 2009

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