Lesson Objective

Explain the quantitative relationship between the light absorbance of a solution and the concentration of the absorbing chemical species.

How can we use light to non-destructively determine the exact concentration of a colored solution?
Why must a spectrophotometer be calibrated at the analytical wavelength of maximum absorbance (lambda max)?

Beer-Lambert Law
Absorbance
Transmittance
Molar Absorptivity (epsilon)
Path Length (b)
Cuvette
Calibration Curve
Analytical Wavelength (lambda max)

Learning Objective 3.13.A; Suggested Skill 2.E (Question and Method - Identify source of error).

Covers the Beer-Lambert Law (A = epsilon b c). Absorbance is directly proportional to concentration. Students must understand how to construct a standard calibration curve and analyze how standard laboratory errors (e.g., leaving fingerprints on a cuvette, adding extra water droplets) skew the final calculated concentration.

Support: Walk through a virtual simulator of a colorimeter, showing visually that a dark solution blocks more light (higher absorbance) than a pale, dilute solution.

Extension: Give students an experimental scenario where a student fails to wipe the outside of a cuvette before measuring. Have them trace how this extra light scattering impacts the recorded Absorbance reading and subsequent concentration calculation.

A student uses a spectrophotometer to find the concentration of an unknown sample of CuSO4(aq). They build a calibration curve by plotting Absorbance versus Concentration, obtaining a straight line equation: A = 42.5c.

  1. If the unknown sample yields an absorbance value of 0.340, calculate its concentration.

  2. If the student accidentally left water droplets inside the cuvette before filling it with the unknown solution, would the calculated concentration be falsely high, falsely low, or unaffected? Justify your answer.