: Uv/Vis Spectroscopy: Acid-Base Equilibrium of Methyl Red

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Physical chemistry for the life science: Laboratory ( Chem 239) Physical chemistry with lab B52 Experiment 5: UV/VIS spectroscopy: acid-base equilibrium of methyl red Objective: Determine the acid base equilibrium of methyl red indicator by using UV\UIS spectroscopy. Introduction: m-Methyl red (m-MR), 3-[4-(dimethyl-amino)phenylazo] benzoic acid, is one of the three isomers of methyl red that is one of a large number of indicators based upon aminoazobenzene, which is still a useful volumetric indicator. This indicator has four possible basic centers to receive added protons with increasing of the acidity in aqueous solutions. Similarly m-MR has the same four basiccenters with only a change in the position of one group. In this work, we examine spectrophotometrically the different m-MR forms and structures in aqueous solutions at different acidity values. Procedure: 1. Stock solution of MR of unknown solution will be provided. 2. Using 1 ml micropipette add 1.0 MR in each four 10 ml volumetric flasks. 3. Prepare following MR solution in four flasks: flask 1: add 5 mL pH 4 buffer (graduated cylinder) and fill to the mark with water flask 2: add 5 mL pH 5 buffer and fill to the mark with water flask 3: add 5 mL pH 7 buffer and fill to the mark with water flask 4: add 1 mL 0.01M NaOH and fill to the mark with water. 4. Record the spectrum for these four solutions in range 400-700 nm. 5. Plot all four spectra on one graph. Results: Calculations: T(λ) = A(λ) =logIoI A = ε.l.C Conclusion: UV-visible spectroscopy is a valid, simple and cost effective method for determining the concentration of absorbing species if applied to pure compounds, and used with the appropriate standard curve. A standard curve relating absorbance to concentration can be developed for any compound, and used to determine the concentration of
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