• Application Note

Pharmaceutical Compounds in Plasma

Pharmaceutical Compounds in Plasma
  • Waters Corporation
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This is an Application Brief and does not contain a detailed Experimental section.

Abstract

This application brief demonstrates analysis of pharmaceutical compounds in plasma.

Introduction

The compounds used in this study are – 

  1. Caffeine
  2. Practolol
  3. N-acetyl procainamide
  4. Propranolol
  5. Methoxyverapamil
  6. Amitriptyline

Experimental

LC Conditions

Column:

Xterra MS C18, 2.1 x 20 mm IS, 3.5 μm, (p/n: 186001923)

Mobile phase A:

10 mM NH4HCO3, pH 10

Mobile phase B:

Methanol

Flow rate:

0.4 mL/min

Injection volume:

20 μL

Sample concentration:

5 μg/mL

Temperature:

Ambient

Detection:

MS

Instrument:

Waters 277 Sample Manager, Waters 1525 Binary HPLC Pump and Waters Micromass Quattro Ultima

Gradient

Gradient.

MS Conditions

Quattro Ultima

ES+ MRM cone (V):

5.0

Capillary:

3.5

Source temp. (°C):

150

Desolvation temp. (°C):

400

Cone gas flow (L/Hr):

50

Desolvation gas fLow(L/Hr):

550

LM resolution 1 & 2:

13.5

HM resolution 1 & 2:

13.5

Ion energy 1:

0.4

Ion energy 2:

0.8

Multiplier (V):

650

Sample Preparation

SPE:

Oasis HLB μElution Plate, (p/n:186001828BA) p/n:186001828BA

Condition:

200 μL Methanol

Equilibrate:

200 μL Water

Load:

150 μL Spiked rat plasma with 2% H3PO4, diluted with 1:1 Water

Wash:

200 μL 5% Methanol in Water

Elute:

50 μL Methanol

Dilute:

100 μL Water

Results and Discussion

Compounds (MRM Transitions, Cone voltage (V), CID (eV)

  1. Caffeine (195 > 138; 20; 20)
  2. Practolol (267 > 190.1; 30; 20)
  3. N-acetyl procainamide (278 > 205.1; 25;20)
  4. Propranolol (260 > 183; 25;18)
  5. Methoxyverapamil (485.2 > 165.1; 45; 30)
  6. Amitriptyline (278 > 233.1; 25; 17)

The top figure is the total ion current, followed by the extracted ion signals for each of the three analytes.

Featured Products

WA31787.19, June 2003

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