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)
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.
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.
Hierarchical clustering of all detected ions from of control and MeJA elicited Chondrus crispus samples.
Samples are in columns; each line represents a detected ion and its variations, from low amount (green) to higher amount (red). Here, only the 91 first ions are represented (2448 detected).