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- Posts: 2
- Joined: Wed Jan 04, 2012 12:00 am
Hello all,
I am trying to design a workflow to measure concentrations of photorespiratory intermediates in seawater (eg. glycolic, glyoxylic, malic, formic, oxalic, succinic, citric, isocitric, cis-aconitic acids). Environmental concentrations will probably be low nanomolar to even picomolar so SPE or other pre-concentration as well as exclusion of competing inorganic salts will be needed. 2,4-dinitrophenylhydrazine hydrochloride (DNPH) will be the derivitizing agent in the presence of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride as a coupling agent. The solvent will be buffered acetone with ion-pairing using tetrabutylammonium hydroxide and tetradecyltrimethylammonium bromide for the counter ions, pH adjusted to 4.5 with phosphoric acid. I want to measure the hydrazones with absorption around 380 to 400nm. I am looking to select appropriate columns for these hydrazone derivatives keeping sensitivity in mind. Unfortunately many of the available resources for this assay are severely outdated. I'm a rookie so I would really appreciate some suggestions or even an alternative assay. Thanks in advance!
Best,
John
I am trying to design a workflow to measure concentrations of photorespiratory intermediates in seawater (eg. glycolic, glyoxylic, malic, formic, oxalic, succinic, citric, isocitric, cis-aconitic acids). Environmental concentrations will probably be low nanomolar to even picomolar so SPE or other pre-concentration as well as exclusion of competing inorganic salts will be needed. 2,4-dinitrophenylhydrazine hydrochloride (DNPH) will be the derivitizing agent in the presence of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride as a coupling agent. The solvent will be buffered acetone with ion-pairing using tetrabutylammonium hydroxide and tetradecyltrimethylammonium bromide for the counter ions, pH adjusted to 4.5 with phosphoric acid. I want to measure the hydrazones with absorption around 380 to 400nm. I am looking to select appropriate columns for these hydrazone derivatives keeping sensitivity in mind. Unfortunately many of the available resources for this assay are severely outdated. I'm a rookie so I would really appreciate some suggestions or even an alternative assay. Thanks in advance!
Best,
John
