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Molecular sieve filtration: A method for separate measurement of intra- and extramitochondrial adenine nucleotides

1974; Elsevier BV; Volume: 62; Issue: 2 Linguagem: Inglês

10.1016/0003-2697(74)90181-x

1096-0309

Fritz Brawand, Paul Walter,

Metabolomics and Mass Spectrometry Studies

A simple technique of molecular sieve filtration is described for the separation of mitochondria (10–15 mg of protein) from the incubation medium. This method can be used for the determination of metabolites in the pellet and filtrate of the same incubation. Very little damage occurred to the mitochondria during filtration and the metabolic state was preserved. Measurements with 14C-labeled sucrose showed that only about 0.25 μl/mg protein of extramitochondrial fluid remained with the mitochondrial pellet on the filter. Results for adenine nucleotides, as well as for citrate and malate are presented.