Extraction of blood Plasma/Serum/CSF - 100 μl

Abstract

A protocol for extraction of blood plasma/serum to create dry samples.

Reagents

  • 30 μl methoxyamine in pyridine (15 μg/μl) 30 μl heptane 30 μl MSTFA 1% TMCS
Extraction buffer
  • MeOH/H2O (90:10 v/v) Incl. IS (7 ng/μl)

Equipment

  • GC-vials speedvac ice bath shaking machine

Instructions

  1. Melt the frozen plasma/serum samples for 5-10 minutes, room temp (100 μl sample).Add 900 μl Extraction mix* on an ice bath.Cool the samples on an ice bath for 2h (cold room,4-5°C).Centrifuge for 10 min, 4°C, 14000 rpm.Transfer 950 μl supernatant to new container vials (1.5 ml).Use 200 μl supernatant and transfer to GC-vials.Evaporate the samples until dryness, using a speedvac.
GC/MS Derivisation
  1. Add 30 μl methoxyamine in pyridine (15 μg/μl).Shake for 10 min in a shaking machine.Put the samples in an oven, 70°C / 60 minutes.Let the reaction continue for 6 h (room temperature).Add 30 μl MSTFA 1% TMCS, vortex, reaction time 1 h (room temp).Add: 30 μl heptane (including methyl stearate 15 ng/μl), vortex. GC-TOF analysis

Citation:
Linus Malm. Extraction of blood Plasma/Serum/CSF - 100 μl. The Molecular Methods database. Thu, 09/03/2015 - 08:26. Acc.nr nvpot10.
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