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Tissue sample preparation for targeted metabolomics (aqueous extraction)

EP Elena Puris
ŠK Štěpán Kouřil
LN Lukáš Najdekr
SL Sanna Loppi
PK Paula Korhonen
KK Katja M Kanninen
TM Tarja Malm
JK Jari Koistinaho
DF David Friedecký
MG Mikko Gynther
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For aqueous extraction, frozen cortex and hippocampus samples (26.2 ± 4.26 mg) were transferred into bead beating tubes (Sample Tubes RB, QIAGEN) preloaded with one 5-mm steel bead (QIAGEN). Subsequently, a 50% (vol/vol) aqueous solution of pre-chilled methanol was added, with the adjusted volume based on the tissue weight (100 μL of solvent per 28 mg of tissue). After the samples had been frozen on dry ice, the lysis of the tissue and extraction of metabolites was performed using a bead beater (TissueLyser II, QIAGEN). The vibrating of the bead beater was 30 times/s for 40 s with 2 plus 2 cycles. The samples were centrifuged at 13,000 × g for 20 min at 4 °C. Aliquots of 100 μL of supernatant (28 mg of sample/100 μL of aliquot) were put into Eppendorf tubes and freeze-dried. The samples were stored at − 80 °C until the analysis.

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