1CHEM 121 LABORATORY EXPERIMENT DATE: NAME:

COLORIMETRIC ANALYSIS OF PHOSPHATE

OBJECTIVES

1. To determine the percent concentration of phosphorus in a commercial dishwasher

detergent.
2. To practice laboratory techniques and calculations.

INTRODUCTION

Phosphates have commonly been used in detergents. Presently, such use is limited or even
totally banned because of environmental concerns. Phosphates in detergents are in the form
of tripolyphosphate ion P3O105-. Tripolyphosphate ions act as a water-softening agent. Water
hardness is due to the presence of metal ions, e.g., Ca2+ and Mg2+, which form insoluble
precipitates with soap often seen around a bathtub. Tripolyphosphate ions form water-
soluble complexes with Ca2+ and Mg2+ [e.g., CaP3O103-(aq)] thus preventing the formation of
the precipitate.

However, phosphates are water pollutants. In rivers and lakes they cause eutrophication.
They serve as fertilizers for algae causing abnormally rapid growth. Following the growth,
huge masses of algae die and the oxygen dissolved in water is rapidly used during the decay
of the algae. This depletes the normal amounts of oxygen in the water and the oxygen-
dependent aquatic life, such as fish, disappears. This oxygen-depletion process is called
eutrophication. In addition to detergents, commercial fertilizers are also the source of
eutrophication of water.

In this laboratory, you will determine the percent by mass of phosphorus in a commercial
dishwasher detergent. You will use many familiar laboratory techniques and calculations.
You need to show the ability to work independently and you should spend some time planning
the experiment. You also should be able to develop and to carry out the necessary
calculations.

Phosphorus in the detergent is present in the form of phosphate. Phosphate will be detected
by a colorimetric method. Phosphate is not a colored compound by itself. However,
phosphate and ammonium molybdate form a blue phosphomolybdate complex. The
structure of this complex is not known with certainty. The intensity of the blue color is
directly proportional to the amount (concentration) of the phosphate in a sample. A
computer-interfaced MicroLAB spectrophotometer will be used to measure the concentrations
of our colored (blue) solutions. Spectrophotometer measures the amount of absorption of
the red light at 635 nm. The amount of absorbed light is directly proportional to the intensity
of the blue color which, in turn, is proportional to the concentration of phosphate (Beer’s law).
MicroLab software automatically generates a standard curve which is used to calculate the
molar concentration of phosphate in the detergent samples. The instructor will demonstrate
the use of the equipment.

The following is an example of the calculations in this experiment:

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 PROBLEM: Following the procedure described in this laboratory, the 1.00 g
sample of the detergent powder was dissolved in 100.0 mL of water. This stock
solution of the detergent was diluted 1:20. The molar concentration of the diluted
solution was measured 0.00300 M. Calculate the mass percent concentration of
phosphorus in the dry detergent powder. Show the calculations. Use correct
number of significant figures.

SOLUTION:
Since the detergent solution was diluted 1:20, the actual concentration (undiluted)
was:
20 x 0.00300 M = 0.0600 M phosphate

The volume of the stock detergent solution was 100.0 mL. We need to calculate
the amount of moles of the phosphate in this solution:
100.0 mL x 0.0600 mol = 0.00600 mol phosphate
1000 mL

There is one mole of elemental phosphorus (P) per one mole of phosphate.
Therefore, in our sample there is 0.00600 mole of P. We need to express that
amount of P in grams:
0.00600 mol P x 30.97 g P = 0.186 g P
1 mol P

Our sample of detergent weighs 1.00 g. Therefore, the percent (m/m)

concentration of P is:
0.186 g P/1.00 g sample x 100% = 18.6 % P (phosphorus)

The mass percent of phosphorus is 18.6 % in the detergent.

PROCEDURE

A. Preparation of standard curve.

The standard curve will include KH2PO4 solutions with four different concentrations: 0.00100
M, 0.00200 M, 0.00300 M, and 0.00400 M. These solutions should be prepared from 0.0100 M
KH2PO4 stock solution as follow:

a) 0.00100 M KH2PO4 - mix 2.00 mL of 0.0100 M KH2PO4 stock solution and 18.0 mL of
distilled H2O;
b) 0.00200 M KH2PO4 - mix 4.00 mL of 0.0100 M KH2PO4 stock solution and 16.0 mL of
distilled H2O;
c) 0.00300 M KH2PO4 - mix 6.00 mL of 0.0100 M KH2PO4 stock solution and 14.0 mL of
distilled H2O;
d) 0.00400 M KH2PO4 - mix 8.00 mL of 0.0100 M KH2PO4 stock solution and 12.0 mL of
distilled H2O;

Very carefully measure the amounts. Remember to mix solutions thoroughly by

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sealing them with Parafilm and inverting upside down several times. Label the test
tubes with standard solutions.

B. Preparation of the dishwasher detergent sample.

Dissolve 1.00 g of the detergent powder in about 40 mL of 0.5 M H2SO4 and GENTLY boil in an
Erlenmeyer flask for 30 min. Keep adding distilled H2O as needed to keep the volume fairly
constant. This process hydrolyzes tripolyphosphate ion breaking it into individual phosphate,
H2PO4- and HPO42-, ions. Cool the mixture and then transfer it into a 100-ml graduated
cylinder. Fill it up with distilled water to 100.0-ml mark and mix. This is a stock solution
of the detergent. For the colorimeter analysis use a 1:10 (2.00 mL + 18.00 mL H2O)
and 1:20 (2.00 mL + 38.00 mL H2O) dilution of this stock solution. Mix the diluted
solution well.

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C. Preparation of samples for absorption measurements with the
spectrophotometer.

All samples, blank, both the standard KH2PO4 solutions, and the diluted (1:10 and 1:20)
detergent solution, should be processed the same way:

1) using a 1.00 mL pipette put 1.00 mL of each sample into a test tube; for blank put
1.00 mL of distilled water
2) add 10.0 mL of HYDROXYLAMINE SOLUTION and 5.0 mL of AMMONIUM MOLYBDATE
SOLUTION. Mix well (cover the tube with Parafilm and turn it upside down);
3) after 2-4 min., add 1.00 mL of 6.50 NaOH. Mix well again;
4) allow to stand for about 10 min. and measure absorbance with the computer-
interfaced colorimeter set on red light (635 nm).

You will have the total of 9 test tubes: one test tube with the blank, 4 test tubes with standard
phosphate solutions (0.00100, 0.00200, 0.00300, 0.00400 M), and 4 test tubes with detergent
solutions (2 detergents A & B, each 1:10 and 1:20 dilution).

D. Measurements with the computer-interfaced MicroLAB spectrophotometer.

The instructor will demonstrate how to use the equipment. Briefly, first the blank” is placed
into the vial and the vial is inserted into ccolorimeter. EVERY TIME, CLEAN THE VIAL ON
OUTSIDE WITH PAPER TOWEL BEFORE YOU PUT IT INTO COLORIMETER. ANY
MOISTURE ON OUTSIDE THE VIAL WOULD RUIN COLORIMETER! In the 1st tab
“Calibration”, press “Read blank” button and the instrument reads the blank and sets
transmittance for all wavelength at 100 %. Then switch to “Absorbance [-logT]”on the task
bar and press the 2nd tab “Read.” Now, sequentially measure absorbance of the 4 standards
at 635 nm wavelength (click on the 635 nm red bar). First, add a standard to the vial and
insert the vial into spectrophotometer and then press “Add” button. In the pop-up window
type the ID of sample and the molar concentration, and press “OK” button. After reading
all 4 standards, switch to 3rd tab “Curve” and select “linear.” The computer will automatically
generate a standard curve. Switch to 4th tab “Read” and measure the absorbance of the 4
detergent samples. Finally, by clicking on the “X” for each unknown sample on the graph
read and record the exact molar concentration of each detergent sample.
Print the graph and include with your lab report.

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CHEM 121 LABORATORY REPORT DATE: NAME:

COLORIMETRIC ANALYSIS OF PHOSPHATE

Record the experimental data here:

Dishwasher A or B Sample dilution Molar conc. (M)

read from the
curve

A 1:10

A 1:20

B 1:10

B 1:20

M Calculate the mass percent concentration of phosphorus in the dry detergent powder. If
the concentrations of both detergent dilutions are on the curve, do a separate calculation for
each. If one of the concentrations falls outside the curve, disregard it. SHOW AND DESCRIBE
CLEARLY ALL THE STEPS! Use correct number of significant figures.

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