I'm using methanol, chloroform (1:3) method to extract lipids from serum for lipidomics study using NMR and mass spectroscopy. I need to remove debris from my solvent containing methanol and chloroform, but I am not sure which centrifugal filter membrane to choose for. There are nylon, cellulose acetate, PVDF and PTFE. Can someone please suggest the membrane?
Dear Sourav, extracting lipids from serum your debris (!) is most likely proteins. Simply by adding water to your extract, proteins (debris) will be at the interface of solvents and you can then easily obtain a debris-free organic extract. If you plan on continuing your work in lipid metabolomic, it might be best to do 2-step extraction, during 1st step don't remove solvent close to the buffy layer (leave a small volume next to the debris layer, and then add a 2nd volume of organic solvent to extract the remaining (Again leaving a small volume close to buffy layer). Introducing a 2nd extraction step will improve the extracting efficiency (at a cost of time) but then whatever losses you may have will occur in the less concentrated extract and therefore negligible. Hope this helps, Ana
Dear Ana, thank you very much for your suggestion! Let me try this 2-step extraction method. 1 more thing, is it OK to use plastic labwares (eppendorf tubes, pipette tips etc) while extracting lipids or do I need to use glasswares as suggested by some?
NOPE, avoid all ("cheap") plastic otherwise you're more likely to acquire LCMS of plasticizers and you do not want that. You can use material that is more resistant to organics bleaching such as PTFE material. You can find ("good") plastic material (eppendorf tubes, pipette tips) that does not bleach plasticizers. Try and convince some of the companies to give material for you to test in your lab and once you decide, stick with them for all purchases in your lab. Ana
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