I used a two-phase extraction protocol (essentially the Bligh-Dyer or Folch protocol) to fractionate C. elegans into aqueous metabolites, lipids, and protein. These worms have been fed bacteria containing heavy stable isotopes and then we performed a chase period. The goal is to determine the turnover of molecular fractions via Isotope Ratio Mass Spec IR-MS). However, this requires packing the dried product (dried via nitrogen drying) into a capsule.
I can do this no problem with the protein and aqueous metabolite fractions, but I am struggling to get the lipid fraction off of the glass tube. So far I've tried using various spatulas to scrape off the residue, but that hasn't worked. I suppose I could resuspend in chloroform and then try to lyophilize, but would prefer not to do that if possible. Does anyone have any suggestions?
When you have scrapped off as much of the residue as you can, then use dichloromethane as the solvent to wash the glass surface. It has a comparatively low boiling point (lower than chloroform), however, you will need to be sure that all the solvent has evaporated before attempting your IRMS analysis.
- Badges
- Science topic