The method should be fast, specific and accurate and it should detect the concentrations under the 0,5 mg of H2O2 per milliliter in cell culture media. And I would like to measure it with UV VIS spectrometer. Is it necessary to use a enzyme based method?
Kind regards, Natasa
Mrs/Miss Hojnik,
Please find below two approaches based on OsVIII and PtII complexes:
1. Spectrophotometric determination of hydrogen peroxide
with osmium(VIII) and m-carboxyphenylfluorone, M. Hoshino, S. Kamino, M. Doi, S. Takada, S. Mitani, R. Yanagihara, M. Asano, T. Yamaguchi, Y. Fujita, Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 117 (2014) 814–816
2. Mechanism of tetrachloroplatinate(II) oxidation by hydrogen peroxide in hydrochloric acid solution, P. Murray,K. Koch, R. van Eldik, Dalton Trans., 2014, 43, 6308–6314
Hi Natasa,
There is a very simple spectrophotometric method using xylenol orange and Fe2+. You can find the original procedure in this paper:
Dringen, R., Kussmaul, L., Hamprecht, B., 1998. Detoxification of exogenous hydrogen peroxide and organic hydroperoxides by cultured astroglial cells assessed by microtiter plate assay. Brain Res. Prot. 2, 223–228.
I used it several times in different aqueous solutions and it worked wonderfully. The beauty of this protocol is that you can use 96-well microplates and process large number of samples simultaneously. Also, the complex is colored, so it can be quantified at 570 nm.
If you need further details, don't hesitate to send me a message.
Best,
Sandra
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