How To Prepare 40 MM H2O2 In Phosphate Buffer (50 MM, PH 7.4)?

QuestionAnswers 14Similar questionsResearch that mentions Hydrogen PeroxideQuestionAsked 3 January 2017 Fiona Chung

• Swinburne University of Technology

How to prepare 40 mM H2O2 in phosphate buffer (50 mM, pH 7.4)?For hydrogen peroxide scavenging assays. I have calculated the vol needed to prepare 40 mmol/L H2O2 from 30% H2O2; the vol is 0.41 mL. Do I just add 0.41 mL of 30% H2O2 to 50mM, pH 7.4 phosphate buffer? Thank you in advance. Hydrogen PeroxideBuffer PreparationShare Facebook Twitter LinkedIn Reddit

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Stefanie MeyerAlfred-Wegener-Institut Helmholtz-Zentrum für Polar- und MeeresforschungHi Fiona,it depends on how much volume you want to prepare: You have 30% H2O2 (=9,79 mol/l=9790 mmol/l) and you want to have 40 mmol/l. The dilution factor is 244,75 (is concentration devided by shall concentration). If you want to prepare 1 liter, you devide 1 liter (=1000 ml) by the dilution factor (=244,75) and you got 4,086 ml. So, you take 4,086 ml of your 30% H2O2 for 1 liter of 40 mmol/l solution and fill it up with the buffer.Cite

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All Answers (14)

Stefanie MeyerAlfred-Wegener-Institut Helmholtz-Zentrum für Polar- und MeeresforschungHi,my calculation ist that You need 4,53 ml of Your 30% H2O2 to get one liter of 40 mM.Was that the question? Write me, if You need the whole calculation way.SteffiCite1 Recommendation Ahmed R. G.Beni-Suef UniversityI AGREED WITH DR. Stefanie Meyer Cite Daniel Carneiro MoreiraUniversity of BrasíliaIt is always a good idea to check the actual H2O2 concentration using a spectrophotometer. If your stock solution has been stored for a long time H2O2 may have decomposed. Moreover, free transition metals if present in your buffer may react with H2O2, decreasing its concentration.To do so, read your H2O2 solution against its solvent (phosphate buffer in this case) in a quartz cuvette at 240 nm. Using the molar attenuation coefficient for H2O2 at 240 nm 0.0394 mM-1 cm-1 (Aebi, 1984), you can calculate the H2O2 concentration in the cuvette. For instance, a 10 mM solution is expected to absorb 0.394 at 240 nm.Cite Srikanth Kondeizene life sciences pvt ltdif you need 1 litre then it is 1.36ml of 30% H2O2 for 40mM H2O2 in 50mM PHOSPHATE BUFFERCite Fiona ChungSwinburne University of TechnologyMr. Daniel, would you mind explain in detail on how to check the actual H2O2 concentration. How should I calculate the H2O2 concentration in the cuvette and do I need to prepare a blank and standard for checking the concentration? Cite Ummay Mahfuza ShaplaUniversity of Wisconsin–Madisonhow can i prepare 50 mM phosphate buffer of 7.4 pH using NaH2PO4. 2H20 and Na2HPO4 for H2O2 scavenging assay?Cite Stefanie MeyerAlfred-Wegener-Institut Helmholtz-Zentrum für Polar- und MeeresforschungC=50 mMM(NaH2PO4*2H2O) = 119,98 g/mol = 119,98 mg/mmolfor 1 liter:m = C * V * M = 50 mmol/l * 1 l * 119,98 mg/mmol = 5999 mg/l of NaH2PO4*2H2OM(Na2HPO4) = 141,96 g/mol = 141,96 mg/mmolfor 1 liter:m = C * V * M = 50 mmol/l * 1 l * 141,96 mg/mmol = 7098 mg/l of Na2HPO4Solve NaH2PO4*2H2O and Na2HPO4 in 900 ml water, adjust pH to 7.4 and add to 1 liter.Cite2 Recommendations Betty PC KhongRenci Community HospitalHi may I ask how do I dilute 3% H2O2 to get 50mM of H2O2?Cite Stefanie MeyerAlfred-Wegener-Institut Helmholtz-Zentrum für Polar- und MeeresforschungHello Betty,3% H2O2 is a concentration of 978,8 mM, so you have to dilute Your H2O2 with a factor of 19,6. If You want a have 1 ml of 50 mM, You have to take 51 µl of 3% H2O2 and 949 µl of water.Good luck!Cite Kesshmita JothiHi Betty, I am doing a H202 antioxidant assay using quartz cuvettes. How do I prepare 2.9 ml of hydrogen peroxide (10mM) using standard phosphate buffer (pH 7.4). Can i pls have the full calculations as well.Thanks. Cite2 Recommendations Zohra KebiliUniversity of GhardaiaStefanie Meyer please, how to adjust the pH? with HCl or with another solution?!!Cite Zohra KebiliUniversity of GhardaiaHello all,Please, I want to know why we can't use the calculation formula pH = pKa + log (molar concentration of acid / molar concentration of base)Thanks for helpCite Stefanie MeyerAlfred-Wegener-Institut Helmholtz-Zentrum für Polar- und MeeresforschungHi Zohra,thanks for the link, now I know what you mean. You're right, you can use the formular pH=pKa+log(conc acid/conc base). But this is only theory, in practise you will see that it is not exact.And for the question of the pH adjustment you first need to know in which direction you have to adjust: if the pH is above the pH you want, you have to take an acid; if the pH is under the pH you want, you have to take a base. And then you choose an acid or base with molecules you already have, means: with KH2PO4/K2HPO4 you have phosphate molecules, so you choose H3PO4 as acid and you have potassium molecules, so you take KOH for base.I hope this helps, otherwise please ask again.Good luck, SteffiCite1 Recommendation Stefanie MeyerAlfred-Wegener-Institut Helmholtz-Zentrum für Polar- und MeeresforschungHi Fiona,it depends on how much volume you want to prepare: You have 30% H2O2 (=9,79 mol/l=9790 mmol/l) and you want to have 40 mmol/l. The dilution factor is 244,75 (is concentration devided by shall concentration). If you want to prepare 1 liter, you devide 1 liter (=1000 ml) by the dilution factor (=244,75) and you got 4,086 ml. So, you take 4,086 ml of your 30% H2O2 for 1 liter of 40 mmol/l solution and fill it up with the buffer.Cite

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