Analyse Spectrophotométrique d'un Mélange
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Résolution d'un mélange à deux composants par équations simultanées
Objectif
Déterminer les concentrations individuelles de deux colorants dans une solution mixte en utilisant des mesures d'absorbance à deux longueurs d'onde et des équations simultanées.
Contexte
When a solution contains two absorbing species with overlapping spectra, a single wavelength measurement cannot determine both concentrations. However, by measuring absorbance at two wavelengths (ideally the λmax of each component) and applying the additive property of absorbance (Beer-Lambert), a system of two simultaneous equations can be solved. This technique is the basis of multicomponent analysis in spectrophotometry.
Avertissements de sécurité
- Indicator dyes stain clothing
- Handle cuvettes by frosted sides only to avoid fingerprints
- Clean spills immediately
EPI requis
Matériaux
-
Methyl orange solution (0.01%) (100 mL)Component A
-
Bromothymol blue solution (0.01%) (100 mL)Component B
-
Unknown mixture of both dyes (50 mL)Prepared by instructor
-
Distilled water (200 mL)
Équipement
Procédure
Scan pure methyl orange from 350–700 nm. Record λmax (approximately 464 nm in acidic pH).
Scan pure bromothymol blue from 350–700 nm. Record λmax (approximately 615 nm at neutral pH).
Prepare 3 standard concentrations of each pure dye. Measure absorbance of each at both λmax values (464 and 615 nm).
Calculate the molar absorptivity (ε) for each dye at each wavelength from the standard curves: ε(A,464), ε(A,615), ε(B,464), ε(B,615).
Measure the absorbance of the unknown mixture at both wavelengths.
Set up the simultaneous equations: A(464) = ε(A,464)·C(A) + ε(B,464)·C(B), A(615) = ε(A,615)·C(A) + ε(B,615)·C(B). Solve for C(A) and C(B).
Verify your results by comparing the calculated total absorbance at several wavelengths with the measured mixture spectrum.
Résultats attendus
The simultaneous equations yield positive concentrations for both dyes. The reconstructed spectrum (sum of individual contributions) should closely match the measured mixture spectrum, validating the Beer-Lambert additivity principle.
Nettoyage
Pour dilute dye solutions down the drain with running water. Rinse all cuvettes and volumetric flasks thoroughly.
Détails
- Catégorie
- Colorimetry
- Difficulté
- Advanced (University)
- Durée
- 75 min
- Coût estimé
- $12,00
- Étapes
- 7
- Matériaux
- 4