Assignment: Beer’s Law

1. Using a spectrophotometer to measure the concentration of a sample, the following data were
obtained: absorbance (A) of the standard solution was 0.55; A of the sample was 0.56 and
concentration of the sample used was 16.25mcg. The concentration of the standard was _______

2. The molar absorptivity constant of a particular chemical is 1.5/M·cm. What is the concentration of a
solution made from this chemical that has an absorbance of 0.72 with a cell path length of 1.1cm?

3. Procedure- Assay of Sulfisoxazole tablets

Preparation of Calibration Curve
 Dissolve 99.96 mg sulfisoxazole reference standard in 100 mL of 1N HCl.
 Transfer 25 mL sulfisoxazole solution to another container and add 75 mL of 1N HCl.
This serves as the “stock solution”.
 In four separate volumetric flasks, prepare the following solutions.
 VF1 – 11 mL stock solution + 20mL 3% vanillin + qs ad 100mL 1N HCl
 VF2 – 15 mL stock solution + 20mL 3% vanillin + qs ad 100mL 1N HCl
 VF3 – 20 mL stock solution + 20mL 3% vanillin + qs ad 100mL 1N HCl
 Get the absorbance at 390 nm.

Assay of Sulfisoxazole Tablets

 Weight 10 tablets of sulfisoxazole (500mg) = 6,523mg.
 Powderize and get a sample powder (100mg) = 99.05mg.
 Dissolve sulfisoxazole in 100 mL 1N HCl.
 Transfer 25 mL sulfisoxazole solution to another container and add 75 mL of 1N HCl.
This serves as the “tablet stock solution”.
 In a volumetric flask (VF4), transfer 15mL of the tablet stock solution, add 20mL of 3%
vanillin and sufficient amount of 1N HCl to make 100 mL.
 Get the absorbance at 390 nm.

Experimental Data
Weight of USP Reference Standard Sulfisoxazole = 99.96 mg
Weight of 10 Sulfisoxazole tablets = 6,523 mg
Weight of sample tablet = 99.05 mg
Solution # Concentration (ug/mL) Absorbance @ 390nm
1 26.13 0.232
2 37.76 0.315
3 48.17 0.589
4 ? 0.374

USP specification : Sulfisoxazole tablets contain NLT 98.0 and NMT 105.0 percent of the labelled claim.

Compute for:
a. Cu and U respectively
b. Amt./tab
c. % LA
d. Make a disposition.