May 07, 2025

Public workspaceLyophilized plant extraction for LC-MS Analysis

DOI
https://dx.doi.org/10.17504/protocols.io.e6nvwe8zwvmk/v1
  • Kateřina Kučerová1
  • 1IOCB
  • Pluskal Lab
Kateřina Kučerová
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DOI: https://dx.doi.org/10.17504/protocols.io.e6nvwe8zwvmk/v1
Protocol Citation: Kateřina Kučerová 2025. Lyophilized plant extraction for LC-MS Analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.e6nvwe8zwvmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
Created: April 21, 2025
Last Modified: May 07, 2025
Protocol Integer ID: 131014
Keywords: lyophilized plant extraction for lc, lyophilized plant extraction, extracting metabolite, lyophilized plant material, plant material for lc, extraction, ms analysis, transfer to lc, ms vial
Abstract
This protocol describes a rapid and efficient method for extracting metabolites from lyophilized plant material for LC-MS analysis. Samples are homogenized, extracted with ethanol–water, dried, and reconstituted in water–acetonitrile before final centrifugation and transfer to LC-MS vials. The method is optimized for small-scale use and ensures compatibility with LC-MS workflows.
Attachments
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Workflow.png
86KB
Materials
Solvents
  • EtOH-H2O (75:25)
  • H2O-CH3CN (50:50)
Experimental Supplies
  • 2.0 ml Eppendorf tubes
  • Stainless Steel grinding beads
  • Dry ice
Labels
  • For weighted material
  • For the extracted material
  • For LC-MS vials
Instruments
  • Analytical balance
  • TissueLyser
  • Pipette (100–1000 μL) + tips
  • Centrifuge
  • Ultrasonic bath
  • Vortex
  • LC-MS system
Troubleshooting
Weight 25 ± 2.5 mg of lyophilized plant material into a 2.0 Eppendorf tube with round bottom
20m
Add a metal bead to each tube
30s
Homogenize in TissueLyser 30 sec, 25 rpm
30s
Add 2 000 μL of EtOH-H2O (75:25) and vortex to mix
10m
Incubate 3 min at 40 °C
3m
Homogenise in TissueLyser 60 sec, 25 rpm
1m
Centrifuge 5 min, 14 000 rpm
5m
Transfer 500 μL of supernatant into a new 2.0 Eppendorf tube
20m
Dry in SpeedVac
3h
Resuspend in 2 000 μL of H2O-CH3CN (50:50)
10m
Sonicate 15 sec and vortex 30 sec
10m
Centrifuge 5 min, 14 000 rpm
5m
Transfer supernatant into LC-MS vials
20m