Hello everyone,
I want to extract metabolites from placental tissue for my study. We are using biphasic method and LC-MS platform. The tissue we have is stored in RNAlater. I was wondering if it can be used for the metabolomics or not!! Does RNAlater has any effect on the metabolites of the tissue??!!
Thanks a lot!
Dear Arpita
I did not try this, so I please consider my answer as an opinion, but I would not use this tissue for metabolomics. RNAlater contains EDTA to inhibit RNAse and other enzymes that use metals as co-factors, but not all enzymes. Metabolite conversion is a very fast process, so I would not trust the obtained profile. Also, RNAlater is a high-salt solution, so I would expect to have problems with this during LC-MS run itself too. In summary, most likely you can extract metabolites out of this tissue, but when you will analyze them, I am not sure how real and meaningful results you will get.
Hi Arpita,
I back and concurr with Albert Batushansky 's wisdom here too!
Imagine if RNAlater enters the tissues, then the metabolites from fluids also come out of the tissue into RNAlater fluid. And not systematically, but very random process.
Plus, RNAlater processing needs a > Over night 4 C percolation of RNALater into the tissues and then frozen into -20C/-80C. Means, during the 4 C storage of tissues with RNAlater a lot of thawing of tissues/ activation of all sorts of enzymes will take place. Say, all glucose being used up, or proteins degraded into amino acids, dipeptides by proteases, and RNAases released a lot of nucleotides/ nucleosides/ riboses/ other sugars back into the tissue/ solution. Thus, all metabolite data will be painting a false picture.
I am also not very convinced of using paraffin dissected tissues or FPPE stored samples or formalin fixed bulk tissues, as there would be losses into formaldehyde/ such solutions/ matrices. But studies where the phenotype is very strong such as cancer vs non-cancer you may still see a discriminating set of metabollites, say BCAA or lactate, but others worth not reporting! : )
So highly suggest not to use it for metabolomics studies.
Thanks,
Biswa
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