Methods for determining enzymes and metabolites of intermediary metabolism, from cytoplasmic and mitochondrial compartments, have  been discussed in the following publications: 

MH Chowdhury & L Agius. Epidermal growth factor counteracts the glycogenic effects of insulin in parenchymal hepatocytes. Biochem J. 247: 307-314, 1987 

L Agius, MH Chowdhury & KGMM Alberti. Regulation of ketogensis, gluconeogenesis and  the mitochondrial redox state by dexamethasone in hepatocyte monolayer culture. Biochem J. 239: 593-601, 1986 

L Agius, MH Chowdhury et al. Regulation of ketogenesis, gluconeogenesis, and glycogen synthesis by insulin and proinsulin in rat hepatocyte monolayer cultures. Diabetes. 35: 1286-1293, 1986

You want to perform the reaction without the enzyme that normally catalyzes it?

Is the enzyme reaction energetically unfavorable, but normally coupled to a second enzyme reaction that makes it more favorable energetically? If so, then the reaction may not happen by itself.

Are the substances stable to heating? Do you have to protect the reaction from oxidation by air?

Hi Adam,

This metabolite will be converted into oxidized form from its reduced state in reaction with hydrogen and NADPH. What are the materials and method should I follow for this reaction in-vitro? Are you capable to do this reaction in your lab? Can I have your E-mail for more fruitful discussion and potential collaboration for this project? please let me know.

Thank you.

E-mail: [email protected]

I have a lot of experience with enzyme-catalyzed reactions, including oxidoreductases, but not with the sort of thing you are trying to do. You should talk to someone in the Chemistry department of your university.

I will say, however, that your description of the reaction sounds incorrect. Reacting the metabolite with hydrogen and NADPH should be a reduction, rather than an oxidation.

Hi Adam,

This reaction is not a kind between the reactants to observe the products. We want the natural reaction in the mitochondria and thereby want to observe the enzymatic changes in the presence of the drug. 

We would be so grateful if you provide your private contact so that we could discuss our confidential idea. This is my email id: [email protected]

Looking forward to hear from you soon for a good collaboration if there is any opportunity. 

Best,

Shams

Hi Shams,

I would be happy to share ideas with you and help if I can. We can discuss your experiment using the private message channel on RG.

Adam.