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 |
1Â h | Â | Remove the upper organic phase containing lipids | Â | Shake for 1Â h at 1400Â rpm at room temperature |
4Â h | Â | Â | Â | Centrifuge at 1300Â rpm for 15Â min Re-extract with 0.3Â ml of CHCl3/MeOH (2:1) with 0.01% BHT, four times |
2Â h | Â | Â | Â | Wash the combined extracts once with 0.4Â ml of 1Â M KCl followed with 0.7Â ml of water |
24 h |  |  | Shake for 24 h at 300 rpm and 25 °C |  |
2Â 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 |