Instrumental Analytical Methods Lab

Lab Report 2013

Anastazija Ristovska
Experiment 5
Photometric Determination of Phosphates

The purpose of this experiment was to form a complex phosphomolybden acid using a
phosphate ion PO43- and ammonium molybdate (NH4)6Mo7O24, and then reduce the complex
phosphomolybden acid with a reducing agent with the ultimate goal of preparing a series of
solutions with different concentrations the phosphate concentration of which can be determined
spectrophotometrically since the phosphate has been bound in a complex, and to then use these
series of standard solutions to prepare a standard calibration curve i.e. a linear fit curve.
Eventually the equation of the linear fit curve is used to find the unknown phosphate
concentrations of other solutions that has the phosphate bound in the same complex and under
the same chemical conditions.
Phosphates cannot be measured spectrophotometrically without introducing them in a
complex because they do not absorb in the UV-vis spectrum. Therefore we reacted them with
ammonium molybdate to form a phosphomolybden acid complex which can be easily detected
in the UV-vis range.
We used a blank sample to calibrate the instrument. The blank was prepared in a 25-ml
volumetric flask by adding 0,5 ml Na2S2O5(aq), 1 ml ammonium molybdate, 1 ml methanol, and 1
ml methyl alcohol, filling the flask with distilled water to the 25-ml volumetric mark.
We allowed the solutions to react for 30min in order for the complex-formation reaction
to take place and to be considered that it has reached completion. The kinetics of this reaction
are such that a 30-min time period was allowed to lapse before almost all phosphate ions could
be considered as bound in a complex.
The phosphomolybden acid complex has its absorption peak maximum at 600 nm and
therefore we use this wavelength for our measurements. This wavelength of 600 nm, however,
seems to have been rounded from a number such as 622 nm, as a random example (saying that
622 is approximately 600 nm); it is preferable the exact absorption peak maximum wavelength is
used instead of this rounded number, which means that we should do all measurements at other
wavelengths also.

[PO43-] (g/ml) Abs (600nm)

3,27
0,072
6
0,128
9
0,198
12
0,265
15
0,321

0,35

Absorbance (600 nm)

0,3
0,25
0,2
0,15
0,1
0,05
0
0

y = 0,0216x + 0,0016 R = 0,9987

unknown concentration solutions

sample
Abs (600nm)
1
0,245
2
0,226
3
0,238

8
[PO43-]

10

12

14

16

(g/ml)

[PO43-] (g/ml)
11.27
10,39
10,94

The linear fit equation is y = 0,0216x + 0,0016 with R2=0,9987 meaning that the model
explains for 99,87% of the variance in the system.
Calculations
[PO43-]=(Abs-0,0016)/0,0216=x g/ml
Sample 1: [PO43-]=(0,245-0,0016)/0,0216=11,27g/ml
Sample 2: [PO43-]=(0,226-0,0016)/0,0216=10,39g/ml
Sample 3: [PO43-]=(0,238-0,0016)/0,0216=10,94g/ml
Methol is just an old name for methanol CH3OH.