ROSALIE HILL

BIO 110- E. COLUNGA

LAB REPORT: THE SPECTROPHOTOMETER

February 24, 2011

PURPOSE:

Students will explain the difference between quantitative and qualitative analysis,
describe the relationship between absorbance and transmittance, use an absorbance curve
to determine the maximum absorbance and be able to use a standard curve to determine
the concentration of a solute in a solution.

RESULTS (Bromophenol Blue)

1.

Wavelength Absorbance
425 0.105
450 0.126
475 0.160
500 0.248
525 0.452
550 0.766
575 1.280
600 1.080
625 0.182

a. 575 (wavelength of maximum absorbance for bromophenol blue)

b. Yellow
2. Standard Curve for Bromophenol Blue

Tube Number Concentration Absorbance Units

1 0 0
2 0.001875 mg/ml 0.176
3 0.00375 mg/ml 0.488
4 0.005625 mg/ml 0.580
5 0.0075 mg/ml 0.592
6 0.009375 mg/ml 0.860
7 0.01125 mg/ml 0.930
8 0.013125 mg/ml 1.170
9 0.015 mg/ml 1.300

3. Unknown Letter: C
Absorbance Value 0.975

Conclusions

1. A portion of the light will be absorbed by the solute particles and some will be
transferred to the tube. What’s absorbed will be equal or close to the
concentration of the solution. Molecules are absorbing more light, the more
concentrated the solution is.
2. The standard curve explains a range to be able to determine the unknown.
3. Green is the color being transmitted and absorbed. The same color light that is
seen is the same color that is transmitted.
4. To calibrate the spectrophotometer.
5. The blank would be prepared with 5ml of water and the dye.
6. The final concentration of the dye is 0.05 mg/ml
7. 0.68 mg/ml.

SEE SEPARATE SHEET FOR STANDARD CURVE