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Chem 155 Unit 10 Page 182 of 316Limitations to Beer’s Law:1. Fundamental Deviations:1.1. Charge-charge, or simple electrostaticinteractions between atoms/molecules canchange the energy levels and symmetries ofmolecules, and therefore change ε(λ). Theseappear at high absorber concentration.1.2. Refractive Index – ε depends on refractiveindex (n) – so, in cases where comparisonsbetween absorbance in solutions of greatlydiffering n must be made, then you can re-writeBeer’s Law as: A = ε⋅n/(n 2 +2) 2 ⋅b⋅c.Why is charge-charge so different from Van derWaals?2. Chemical Deviations: Van der Waals bonds,hydrogen bonds, or other non-covalent bonds or“chemical interactions” can change the energylevels and symmetries of molecules, and thereforechange ε(λ). These appear at high concentration.3. Instrumental Deviations: Several instrumentalfactors are known to cause non-Beer’s lawbehavior:3.1. The monochromator effective bandwidth islarge compared to the spectral bandwidthsanalyzed.3.2. Stray light is getting into the detector.3.3. The light levels are too low for the detector toaccurately represent.Page 182 of 316
Chem 155 Unit 10 Page 183 of 316Limitations to Beer’s Law cont.:Too large of a monochromator effective bandwidthmeans that polychromatic radiation will be incident on yourdetector at any given supposedly monochromatic setting.This is depicted below. Analyzing for the moment just two ofthe wavelengths of light that strike the detector, we cancalculate the apparent aggregate absorbance assuming thatthe individual absorbances obey Beer’s law:Way overblown…Page 183 of 316
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