Mice brain preparation for HPLC

April 12, 2021, 07:43:07 AM

Hi

I am investigating Tryptophan metabolites in mice brain and colons. Do you have a method to recommend for this? I am leaning towards using perchloric acid. Your advice is much appreciated.

Jeannie

Re: Mice brain preparation for HPLC

Reply #1 – April 15, 2021, 11:03:22 PM

Hi! Your question has been tweeted by the MetSoc EMN (https://twitter.com/EMN_MetSoc/status/1382607928599965696)and received the following answers:

CAROLINA GONZALEZ: we use an extraction method for multiplataform analyses. We add MeOH:H2O 50% (1mg tissue:10uL solvent) for Mouse face Brain homogenization using a TissueLyser. Then, we mix 100uL of brain homogenate with 320uL MeOH + 80uL MTBE (for lipids) DOI: 10.1021/acs.jproteome.8b00816.

WENYUN LU: For tissue samples our preferred extraction method is first grind into powder then add 40:40:20 methanol:acetonitrile:h2o+0.5% formic acid then neutralize with 15% NH4 HCO3 followed by two rounds of centrifugation.
https://doi.org/10.1089/ars.2017.7014

Feel free to follow up with further Qs here or there!