• Application Note

Ostro Sample Preparation Products: Recovery

Ostro Sample Preparation Products: Recovery

  • Waters Corporation

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

Abstract

As part of the evaluation of the Ostro plate, recovery for a broad range of analytes in human and rat plasma was evaluated to judge how well potential target compounds could be quantified.

Benefits

The Ostro plate demonstrates excellent recovery across a variety of conditions

Introduction

Recovery determination is important for quantitative methods in that it provides a measure of the actual amount of analyte obtained from the matrix using a particular sample preparation device or technique. As part of the evaluation of the Ostro plate, recovery for a broad range of analytes in human and rat plasma was evaluated to judge how well potential target com-pounds could be quantified.

Experimental

Samples

  • Human or rat plasma fortified at high, medium, and low concentrations
    • Low concentration range: 0.2-40 ng/mL
    • Medium concentration range: 2-400 ng/mL
    • High concentration range: 20-4,000 ng/mL
  • 3 loading volumes:
    • 50 μL plasma: 150 μL of 1% HCOOH in ACN
    • 100 μL plasma: 300 μL of 1% HCOOH in ACN
    • 200 μL plasma: 600 μL of 1% HCOOH in ACN

Analyte mixture

  • Fluticasone
  • Niflumic Acid
  • 9-OH Risperidone
  • Propranolol
  • Clonazepam
  • Ketoprofen
  • Imipramine
  • Naproxen
  • Valethamate
  • Salicylic Acid
  • Terfenadine
  • Caffeine

Protocol

  • Load plasma onto Ostro plate
  • Load extraction solvent
  • Aspirate 3X using electronic pipette set at same volume as extraction solvent
  • Pull through using vacuum set at 15” Hg for 5 minutes
  • Dry down using nitrogen gas
  • Reconstitute in same volume as plasma (50 μL, 100 μL, or 200 μL)
  • Inject 8 μL into 10 μL loop

Results and Discussion

Analyte Recovery 50 μL Plasma Loaded: Rat Plasma

Analyte Recovery 100 μL Plasma Loaded: Rat Plasma

Analyte Recovery 200 μL Plasma Loaded: Rat Plasma

Analyte Recovery 50 μL Plasma Loaded: Human Plasma*

Analyte Recovery 100 μL Plasma Loaded: Human Plasma

Analyte Recovery 200 μL Plasma Loaded: Human Plasma

* To improve recovery for small sample volumes, you may increase the ratio of organic solvent to plasma. Use of Waters Positive Pressure-96 Processor has also been shown to improve recovery.

Conclusion

The Ostro plate demonstrates excellent recovery across a variety of conditions. In this evaluation, low, medium, and high concentrations of a range of compounds were examined and were consistently recovered using the device. This robustness was shown in both rat and human plasma.

720003454, September 2010

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