• Application Note

Beta Blockers in Rat Plasma

Beta Blockers in Rat Plasma
  • Waters Corporation
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Doctor with mask and gloves holding test tube with separated plasma

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

Abstract

This application brief demonstrates the analysis of beta blockers in rat plasma using XTerra Columns. 

Introduction

The compounds analyzed in this study are:

  1. Atenolol
  2. Pindolol 
  3. Metoprolol 
Compounds

Experimental

Conditions

Column:

XTerra MS C18 4.6 x 20 mm, 2.5 µm

Part number:

186001889

Mobile phase A:

0.1% TFA in H2O

Mobile phase B:

ACN

Flow rate:

3.0 mL/min

Injection volume:

20 µL

Sample concentration:

0.1 mg/mL of atenolol 0.05 mg/mL of pindolol 0.1 mg/mL of metoprolol

Temperature:

Ambient

Detection:

UV @ 220 nm

Instrument:

Alliance 2695, 2996 PDA

Gradient Table

Gradient Table

Protein Precipitation Procedure

Analytes*: 

  • Atenolol (10 mg/mL in MeOH) 
  • Pindolol (5 mg/mL in MeOH/H2O) 
  • Metoprolol (10 mg/mL in MeOH) 

*prepared in strong conc. to spike into plasma 

Spiked Plasma Sample: 

  • 50 µL of Atenolol 
  • 50 µL of Metoprolol 
  • 50 µL of Pindolol 
  • 4750 µL of Rat Plasma 
  • 100 µL of H3PO4 
  • 5000 µL total sample volume 

Protein Precipitation: 

  • Mutiple samples prepared from 5000 µL spiked plasma
    350 µL of Spiked Plasma sample 
    1000 µL of acetonitile 
    1350 µL total sample volume 
  • Samples were centrifuged at 3000 RPM for 30 minutes
  • Supematent transferred to culture tube and evaporated 
  • Sample reconstituted in 350 µL water and injected

Results and Discussion

Results

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WA20738.016, June 2002

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