Experiment No.

5 Spectrophotometric Determination of Phosphate in Detergent

Experiment No. 5
SPECTROPHOTOMETRIC DETERMINATION OF
PHOSPHATE IN DETERGENT

Objectives

1. To apply spectrophotometric technique in determining the amount of phosphate in detergent

Reagents

Conc. H2SO4 molybdate solution reducing solution distilled water

Antifoaming agent

Apparatus

50-mL round bottom flask condenser Hot plate iron stand

100-mL volumetric flask graduated cylinder 50-mL volumetric flask
10-mL volumetric flask 1mL pipette 5-mL pipette

Procedure

I. Hydrolysis of Detergent

Weigh 0.500 g sample of your detergent and transfer it to 100-mL round bottom flask. Add two
or more drops of antifoaming agent. Slowly add 10mL 2M H2SO4 into the flask. Add boiling chips into
the flask and attach a reflux condenser into it. Gently boil the mixture for 30 mins (start the timer when
the solution starts to boil). Make sure to keep the volume constant by adding distilled water if necessary.
Cool the mixture and quantitatively transfer it to 100 mL volumetric flask. Add one more drop of
antifoaming agent and dilute the mixture with distilled water up to the mark. Transfer 5.00 mL of the
hydrolyzed sample into a 100-mL volumetric flask and dilute it with distilled water up to the mark.

II. Color Development of hydrolyzed sample via Molybdenum Blue Method

Pipet 5.00 mL of this solution into a 10-mL volumetric flask, add 1.00 mL of molybdate solution
and 0.4 mL of reducing solution and dilute the mixture up to the mark. Stopper the volumetric flask and
mix the solution via repeated inversions. Remove the stopper and allow the solution to stand for 20
minutes. Get the Absorbance of the solution. Repeat this procedure to make triplicate results.

Experiment No. 5 Spectrophotometric Determination of Phosphate in Detergent

III. Determination of Phosphate Concentration in sample

1. Standard Calibration Method

Prepare 100.0 ppm standard phosphate solution by dissolving appropriate amount of dried
(1100C) KH2PO4 in 500 mL (0.05g KH2PO4) of 0.1% H2SO4 (0.25 mL conc. H2SO4 per 500 mL).
From this solution prepare five (5) 100.0 mL standards ranging from 1.00 ppm to 25.0 ppm
phosphate. From this solution transfer V of standard required in mL in a 100 mL Volumetric flask
and add 10.00 mL of molybdate solution and 4.0mL reducing agent. Mix and let it stand for 20 mins.
Get the Absorbance of the solution. Use these data to construct a calibration curve for standard
solution (A vs concentration) and use it to compute the amount of phosphate in your sample. Note
that in measuring the absorbance, you need to determine first the wavelength at maximum
absorption. Make three readings for every absorbance.

2. Standard Addition Method

Determine the amount of phosphate in all your samples using standard addition method.
Select one standard concentration for this method. From the prepared standard solutions choose
one concentration and use it to make 5 data points of standard addition according to the table
below:

Volume of Volume of Volume of Volume of

analyte Sample Molybdate Reducing Total Volume
standard detergent solution Agent
0.00 mL 5.0 mL 1.0 mL 0.4 mL 50.0 mL

2.0 0mL 5.0 mL 1.0 mL 0.4 mL 50.0 mL

4.00mL 5.0 mL 1.0 mL 0.4 mL 50.0 mL

6.00 mL 5.0 mL 1.0 mL 0.4 mL 50.0 mL

8.00 mL 5.0 mL 1.0 mL 0.4 mL 50.0 mL

10.0 mL 5.0 mL 1.0 mL 0.4 mL 50.0 mL

Construct the Standard addition curve by plotting (A vs Vstd). Derive the expression for the
x-intercept and from it determine the concentration of phosphates in your sample.