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Table 1 A detailed overview of the four lipid extraction protocols used in the present study

From: A comprehensive comparison of four methods for extracting lipids from Arabidopsis tissues

Time for the extraction of 25 samples

Burgos et al. [27]

Hummel et al. [24]

Shiva et al. [23]

Welti et al. [18]

Total time

4 h

5 h

4 h + 24 h extraction time

12 h

1 h

Homogenize with 1 ml of CHCl3/MeOH/water (1:2.5:1)

Homogenize with 1 ml of MeOH:methyl-tert-butyl-ether (1:3)

Homogenize with 400 µl of 2-propanol with 0.01% BHT

Homogenize with 1 ml of 2-propanol with 0.01% BHT

1 h

Shake for 30 min at 4 °C

Spin down for 15 min at 4 °C at 13,200 rpm

Incubate for 10 min in a shaker at 4 °C

Incubate for 10 min in an ultrasonication bath at room temperature

Add 500 µl of water: MeOH (3:1)

Vortex and centrifuge at 13,200 rpm for 15 min

Heat the samples at 75 °C while shaking at 1400 rpm for 15 min

Cool to room temperature

Add 1.2 ml of CHCl3/MeOH/water (30/41.5/3.5, v/v/v)

Add 0.5 ml CHCl3 and 0.2 ml water

Heat at 75 °C while shaking at 1400 rpm for 15 min

1 h

 

Remove the upper organic phase containing lipids

 

Shake for 1 h at 1400 rpm at room temperature

4 h

   

Centrifuge at 1300 rpm for 15 min

Re-extract with 0.3 ml of CHCl3/MeOH (2:1) with 0.01% BHT, four times

2 h

   

Wash the combined extracts once with 0.4 ml of 1 M KCl followed with 0.7 ml of water

24 h

  

Shake for 24 h at 300 rpm and 25 °C

 

2 h

Dry down the organic phase in a SpeedVac

Dry down the organic phase in a SpeedVac

Dry down the solvent in a SpeedVac

Evaporate the solvents by SpeedVac

  1. BHT butylated-hydroxy-toluene