Metabolomic study of the red algae Chondrus crispus
Instrument: Precellys® 24 Dual with Cryolys cooling device to have a constant temperature of – 20 °C within the homogenization chamber using liquid nitrogen.
Sample: Red algae (Chondrus crispus)
Protocol:
- Precellys kits: 03961-1-008 (MK28-R).
- Sample: 3x3 replicates of MeJA elicited & control Chondrus crispus.
- Extraction solvent: 1 ml ice-cold [MeOH/H2O (8:2)] for 200mg fresh tissue (added after homogenization).
- Each lysing tube was added of 150 mg Fontainebleau sand.
- Run the Precellys 24 at 1x30s 6800rpm - 1x50s 6000rpm - 1x30s 6800rpm - 1x90s 6200rpm (with 5min break between each run).
- Extraction was carried out at +4°C during 1hour with constant agitation. Centrifugation 3000g, 15 min.
- Samples were injected in UHPLC/MS on Dionex U3000 RSLC coupled to a LTQ-Orbitrap Discovery (Thermo, Les Ullis). Data treatment with XCMS and Mev.
Results:
Our results clearly demonstrate the elicitation of the red algae Chondrus crispus by MeJA (results not shown).
Morever, the 9 different replicates display very reproducible metabolomic profiles (Figure 1), thanks to simultaneous grinding with Precellys 24. During this study, the Cryolys guaranteed a constant temperature of -20°C within the homogenization chamber and thus avoided an uncontrolled defrosting of the frozen samples.
Compare to manual grinding, simultaneous cryo-grinding avoided biological material degradation and allowed higher reproducibility, faster grinding and better metabolite extraction.
