BIOPHARMACY PRACTICAL

FOURTH STAGE
Lab 2\ preparation of calibration curve of salicylic acid with the application of
statistics
By ph. Mina Emaad
 Calibration curve
◦Calibration curve :is a curve prepared from a series of standard solution and used as a
reference curve to obtain the concentration of unknown sample of the same drug.
◦The curve should be fitted either by :
1. Eye fitting
2. Least square fitting (most reliable method)
 ◦ Application of calibration curve :
 1- Analysis of concentration
 2- Verifying the proper functioning of an analytical instrument or a sensor device such as an ion
selective electrode
The X axis is the concentration (independent variable).
The Y axis is the absorbance (dependent variable).
To measure the concentration of any substance, we have many methods , one of them is spectrophotometry
method for measurement of concentration of the sample by applying certain waves either :
A. Vesible light, which range from 400-700nm.

B. Ultraviolet light, which range from 200-400 nm .

Spectronic 20 is an instrument which read the absorption for colored solution and used for quantitative
analysis to find the concentration of the sample by regreation analysis.
◦
Experimental procedure
1. Prepare 250 ml of stock solution of sodium salicylate containing the equivalent of 200 mg S.A\100 ml .
2. From the stock solution of sodium salicylate use volumetric procedure to accurately prepare solutions
containing the equivalent 50,40,30,20,10 mg S.A\100ml
3. Take 1 ml of each solution and add 5 ml of color developing reagent , mix the solution .
4. Determine the absorbance against suitable blank at 530 nm.
5. Plot absorbance verses concentration of salicylic acid.
6. Apply theregrration line equation to calculate the slop and intercept.
Notes :
◦ A blank solution is a solution containing no analyte usually used to calibrate instruments such as
a colorimeter.

◦A stock solution is a concentrated solution that will be diluted to some lower concentration for actual use.
Stock solutions are used to save preparation time, conserve materials, reduce storage space, and improve
the accuracy with which working lower concentration solutions are prepared.