Biopharmaceutics & Pharmacokinetic II (PHA312)

Lab Report

Date: 27/1/2022 Lab No: 1

Title
Spectrophotometric Analysis of Salicylic Acid.

Aim:
- To examine and confirm Beer's law using the sodium salicylate solution as an example.
- To draw the calibration standard curve of Sodium Salicylate.
- To find out the equation used to describe the straight line of the calibration curve.
- To determine the concentration of unknown sample from the standard calibration curve of
sodium salicylate.

Materials & Instruments used:

- Salicylic acid. - Distilled Water.
- Spectrophotometer. - Volumetric Flasks [25ml] Each.
- Graduated Pipettes. - Container for Disposal Samples.
- Cuvettes. - Dropper.

Procedure:
1 - Calculate the required volume for other concentrations.
2 - Measure the Abs for each solution & for unknown sample.
3 - Present Data on graph paper and find out the equation of the calibration curve for sodium
salicylate at 310 nm.
4 - Determine the concentration of unknown sample from the standard calibration curve of
sodium salicylate.
Results:

Absorbance Concentration
2.5 0.225
5 0.239
10 0.274
15 0.311
20 0.35
Unknown Sample A 0.240
Unknown Sample B 0.276

Calculations:
0.025% =
0.025 g 100 ml
Xg 1 ml
(0.025 x 10 ) / 100 = 250 ug/ml
6

.............................................................................

- For preparation of the solutions, we use [C1 V1 = C2 V2] equation:

(250) x V1 = (2.5 ug/ml) x (25 ml) = 0.25 ml
(250) x V1 = (5 ug/ml) x (25 ml) = 0.5 ml
(250) x V1 = (10 ug/ml) x (25 ml) = 1 ml
(250) x V1 = (15 ug/ml) x (25 ml) = 1.5 ml
(250) x V1 = (20 ug/ml) x (25 ml) = 2 ml
.............................................................................

- Concentration of unknown samples:

A:
y = 0.0072x + 0.2044
0.240 = 0.0072x + 0.2044
= 4.94 ug/ml

B:
y = 0.0072x + 0.2044
0.276 = 0.0072x + 0.2044
= 9.94 ug/ml

Conclusion:
- As the concentration increases, absorbance increases.