Water Photometric Analysis

Analysis: AMMONIA – NITROGEN (NH4 – N) Standard method 418-B

A. Basic theory:
The Nesslerization reaction is specific reaction for the determination of dissolved ammonia ions. The
yellowish to brownish color can be measured with photometric equipment in a range of wave length
from 400 to 500 nm.
B. Reagents:
1. Rochelle salt solution (stabilizer):
50 g potassium sodium tartrate KNaC4H4O6.4H2O is dissolved in 100 ml. of dist. Water. To remove
ammonia which may be present in the salt boil of appr? 30 ml and make it up to 100 ml again when it is
cooled.
2. Nessler reagent:
Dissolve 100 g Mercury iodide and 70 g potassium iodide in appr. 200 ml of dist. water. This solution is
mixed slowly to a cool mixture of 160 g NaOH in 500 ml dist. water. Finally fill up to 1000 ml with dist.
water.
C. Apparatus:
Spectrophotometer:
Wave length: 405 nm
Light pass: 10 mm
Nessler tubes: 50 ml
Measuring pipettes: 1; 2; 5 ml
D. Disturbances:
Turbidity and color in water may cause too high results, also high amounts of calcium and magnesium
may precipitate showing a non specific turbidity. Residual chlorine has to be removed

E. Procedure:
1. Pretreatment
If necessary remove residual chlorine with sodium thiosulfat solution (3.5 g Na2S2O3-5H2O) in 1000
ml dist. water) 1ml of solution will remove 1 mg/L of Cl2 when using 500 ml sample. Turbidity has to
be removed by pre filtration of the sample. If distributing amounts of calcium and magnesium or
other metal ions are present these compounds must be precipitated at PH of 10-10.5 (with NaOH) and
filtered out.
١
 2- Formation of color
Use 50 ml of sample (or a suitable) portion filled up to 50 ml in a graduated Nessler tube. Add 0.05 to
0.1 ml (2-3 drops) Rochelle solution and mix well. Add 1ml of Nessler reagent and mix well.
Remark:
If the amounts of Ammonia can be expected high enough (higher than 2mg/L) in a slightly turbid sample it
is preferable to diluted the sample and to reduce by this also the turbidity.
3-Photometric measurement
After a time of 10 minutes the color has developed and can de measured in the spectrophotometer at
wavelength of 405 nm against a blank sample of dist. Water treated with Rochelle salt and Nessler
reagent.

F. calibration of the photometer.

1- NH4-N stock solution
Dissolve 3.819 g anhydrous NH4Cl dried at 100 0C for 2 hours in ammonia free dist. water and fill up
to 100 ml.
1ml of stock solution contains 1 gm/L NH4-N
2- Standard solution
Dilute 10 ml of (1) to 1000 ml of ammonia free dist. Water 1 ml of standard solution contains 0.01 mg
NH4-N
3- Calibration standards
Prepare standards of 0.1, 0.2, 0.3, 0.4, 0.5 mg/L NH4-N.

Volume of Volume of Amount of Concent.of Measured

Standard No
stand. sol. Dist .water NH4-N/ 50ml stand. Absorbance
1 0.5 ml 49.5ml 0.005 mg 0.1 mg/L
2 1.0ml 49.0 ml 0.010 mg 0.2 mg/L
3 1.5 ml 48.5 ml 0.015 mg 0.3 mg/L
4 2.0 ml 48.0 ml 0.020 mg 0.4 mg/L
5 2.5 ml 47.5 ml 0.025 mg 0.5 mg/L

Treat the standards with reagents as described in section E and measure the absorbance of the formed
color against a blank sample of dist. Water plus reagents.
Plot the result as calibration curve absorbance via concentration calculate the slop of the calibration curve
in the linear range for the use as factor to calibrate the photometer in concentration.

٢
Example:

Abs. in
abs. unit

Example1
Example2

Concenter in mg/L

Example 1: tana1 = 0.3 mg /L 0.075 abs .units =4.00 mg/L abs. units
Factor 1 = 4.00

Example 2: tana2 = 0.4 mg/L / 0.058 abs .units = 6.90 mg/L abs .units
Factor 2 = 6.90

٣
G .calculations
The calculation of the concentration with the factor of the calibration:
C=F. a. 50/v
C: concentration of NH4-N in mg/L
F: factor in mg /L.abs. units
A: absorbance measured in the sample
V: volume of sample used for the analyses (between 1 and 50ml)

Remarks
by using the factor calculated in section F the photometer can be programmed to give the results directly in
concentration in mg /L when 50ml of sample was used , if less than 50 ml was used this has to be considered
by multiplying the result with the dilution factor .

Example:

If 10ml of sample was diluted to 50 ml, the result given on the photometer display has to be multiplied with
5.

٤
 Analysis: NITRATE –NITROGEN (NO3-N)

A. Basic theory

Nitrate forms with potassium – sodium tartrate a yellowish complex that can be photometrically
measured at a wavelength of 420 nm

B. Reagents:

1- potassium sodium tartrate solution

60gm potassium sodium tartrate and 400 g sodium hydroxide are mixed in appr. 500 ml dist.
water when cooled this solution is filled up in a 1000ml volumetric flask to the mark. Keep the
solution in a plastic bottle.
2- sodium salicylate
0.5 g sodium salicylates are mixed to 100ml of dist water this is not stable and has to be
prepared fresh before the analysis.

3- Al2 (SO4)3- solution:

Dissolve 120g Al2(SO4)3 in 1000 ml dist water.

4- Soda solution:
Dissolve 100 mg Na2CO3 and 50gm NaOH into 300ml of dist. Water
.
5- Sulfuric acid conc.

C. Apparatus

Spectrophotometer

Wavelength 420.0nm
Light pass: 10 mm
Beakers : 50-150 nm
Measured pipettes: 1, 10, 20 ml

D. Interferences

Turbidity and color may give too high results. High amounts of organic substances disturb the analysis

٥
E. Procedure
1- Pretreatment:
Turbidity, color and organic substance can be reduced by a precipitation with Al2 (SO4)3 solution. Add
to 100ml of the sample 5 ml of Al2(SO4 )3 solution and neutralized afterwards by drop wise dosing the
soda solution to a PH of 6.5 -7 while intensively mixing then filter the sample for the analysis.

2- Formation of color

Pipits 20 ml of sample or a suitable portion diluted to 20 ml in a beaker and add 1ml of the sodium
salicylate solution. Evaporizes the sample slowly at 105 0C in a drying oven. The residue will be cooled
before adding 2ml of conc. H2SO4 after 10 min. add first 15 ml dist. Water then 15 ml of potassium
sodium tartrate solution and wait 10 min. until the color has fully developed.
3- Photometric Measurement:
The specific color can be measured at 420 nm against a blank sample of dist. Water which has been
treated like the sample.

f. calibration of the photometer:

1- NO3-stokcksolution
Dissolve 720.8 mg potassium nitrate and dilute to 1000 ml. 1 ml of stock solution contains 0.1mg
NO3-N=0.443 mg NO3
2- Standard solution:
Dilute 100 ml of stock solution to 1000 ml with dist. Water.
1 ml of standard solution contains 0.01 mg NO3-N = 0.0443 mg NO3

3-calibration standard:

Volume of Volume of Amount of Measured

Standard NO. Conc. of stand
stand. Sol. Dist.water NO3-N/20ml Absorbance
1 0.4ml 19.6 ml 0.004mg 0.200 mg/L
2 0.8 ml 19.2 ml 0.008 mg 0.400 mg/L
3 1.2 ml 18.8 ml 0.012 mg 0.600 mg/L
4 1.6 ml 18.4 ml 0.016 mg 0.800 mg/L
5 2.0 ml 18.0 ml 0.020 mg 1.000 mg/L
6 3.0 ml 17.0 ml 0.030 mg 1.500 mg/L
7 4.0 ml 16.0 ml 0.040 mg 2.000 mg/L
8 6.0 ml 14.0 ml 0.060 mg 3.000 mg/L
9 8.0 ml 12.0 ml 0.080 mg 4.000 mg/L
10 10.0 ml 10.0 ml 0.100 mg 5.000 mg/L

٦
Treat the prepared standard with reagents as described in section E and measure the absorbance of the
formed color against a blank sample of dist.water plus reagents plot the results as calibration curve
absorption via concentration. Calculate the factor of the calibration curve in the linear range for the use to
program the photometer to give the results in concentration readings.

Example:

Example

Abs.
In abs
units

Linear range

Conce. In
mg/L

G- calculation:

The calculation of the concentration is done with the factor of the calibration curve:
C= f.a.20/v
C = concentration of NO3-N in mg/L

F = factor in mg/L abs .units.

A =absorbance measured in the sample

V = volume of sample used for the analysis (between 1 and 50ml).

٧
Remarks
By using the factor calculated in section f the photometer can be programmed to read the results directly in
concentration in mg/1 when 20 ml of sample was used. If les than 20 ml was used this has to be considered
by multiplying the result on the photometer display with the dilution factor.
Example
If only 50 ml of sample was used and filled up with 15 ml of dist. water to required volume of 20 ml, the
result has to be multiplied by 4.

٨
 Analysis: PHOSPHATE (PO4-P) standard method 425 D

A- Basic Theory:
Dissolved ortho- phosphate (PO4) reacts under acid considered with ammonium molybidate to form a
hteropoly molypdo-phosoatic acid. In presence of vanadium a vandomolybdophosphatic complex with
yellow color is built. This color is specific for the phosphate concentration and can be measured with a
photometer in a range of wavelength between 400-490 nm.
In low concentration ranges usually measured at 400 nm
B- Reagents
1- vandate-molybdate solution
Solution A; Dissolve 25 g ammonium molybdate (NH4)6MO7O24H2O) into 400 ml of dist. water.
Solution B: Dissolve 1.25 g ammonium metavandate ( NH4VO3) in 250 ml of Dist. water by
heating it to boiling. Cool and add 330ml of conc. HCl.
Pour solution A to the cooled solution B and dilute to 1000 ml.

C-Apparatus:
Spectrophotometer:
Wavelength: 400nm
Light pass: 10ml
Nessler tubes; 50ml
Measuring pipette: 10ml

D-Interferences

The PH-value of the sample should be in the range between PH = 4 and PH = 10 turbidity and color will
affect the result.
Measuring cylinders: 50ml

E .procedures:
1- pretreatment:
Turbidity may be reduced by filtration and dilution of the sample. Color has to be removed if
necessary by shaking 100 ml of sample with a ppr. 200 mg of phosphor free activated carbon and
filtering.

2- Formation of color:
Measure 35ml of sample or a suitable smaller portion into a 50ml Nessler tube. Add 10ml of vandate-
molybdate reagent, fill up to the mark and mix well, the color will form within 10 minutes.
٩
 3- Photometric measurement:
The specific yellow color can be measured at 400nm against a blank sample of 40ml dist. water +
10 ml reagent.

f- Calibration of the photometer:

1- Phosphate stock solution:
Dissolve 4.39 g of hydrates potassium dihydrogen phosphate (K2H2PO4) in dist .water and dilute
to 1000 ml.
1 ml of stock solution contains 1.000 mg P /L
2- Standard solution:
Pipette 10 ml of stock solution into a 1000 ml volumetric flask and fill up to the mark with dist. water
3- Calibration standards:

standard Volume of volume of dist. amount of PO4- measured

Conc.of stand
no. stand. sol. water P/35 ml absorbance

1 0.7ml 35 ml 0.007 mg 0.2 mg/L

2 1.4 ml 34 ml 0.014 mg 0.4 mg/L
3 2.1 ml 33 ml 0.021 mg 0.6 mg/L
4 2.8 ml 32 ml 0.028 mg 0.8 mg/L
5 3.5 ml 31.5 ml 0.035 mg 1.0mg/L
6 7.0 ml 28 ml 0.070 mg 2.0 mg/L
7 10.5 ml 25 ml 0.105 mg 3.0 mg/L
8 14.0 ml 21 ml 0.140 mg 4.0 mg/L
9 17.5 ml 17 ml 0.175 mg 5.0 mg/L

Treat the prepared standards with 10 ml of reagent and fill up to the 50 ml mark as described in section E
and measure the absorbance of the formed color against a blank sample.
Plot the results as calibration curve absorbance via concentration. Calculate the factor of the calibration
curve in the linear range for the use to program the photometer to give the results in concentration
readings.

١٠
EXAMPLE:

Example

Abs.
In abs
units

Linear range

Conce. In
mg/L

Factor: tan = 2.0 mg/L / 0.500 abs. units

= 4.00 mg/L abs. units

١١
G – Calculations:
The calculation of the concentration from a measured absorbance may be done with the factor of the
calibration curve:
C = f. a. 35
V
C = concentration of PO4-P in mg/L
f= factor in mg/L abs. units
a = absorbance measured in the sample
V= volume of sample used for the analysis (Between 1 and 35 ml)

Remarks:
By using the factor calculated in section F – the photometer can be programmed to give the results
directly in concentration readings. When according to this method 35 ml of sample was used.
If less than 35 ml was used this has to be considered by multiplying the result on the photometer
display with the dilution factor
Example:
If only 5 ml of sample was used the result has to be multiplied by 7

١٢
 Analysis: SULFATE (SO4)-2
A-Basic theory:
Sulfate- ions may form in a slightly hydrochloric acid media (PH = 4.5) in presence of barium ions a
precipitation of Barium sulfate if the Barium sulfate turbidity is caused by uniform crystals turbidity
can be used as specific parameter for the determination of the sulfate concentration by the means of a
photometer .
B- Reagents
1. Conditioning reagent
Dilute 75g NaCl in 300ml of dist. water and 30ml HCl - case
Add 100ml ethyl alcohol (95 %) and 50 ml glycerol
2- BaCl2- crystals (20-30 mash)
C-Apparatus:
Spectrophotometer
Light- pass: 10mm
Wavelength: 420nm
Stopwatch (for calibration)
Erlenmeyer flasks: 250 ml
Measuring cylinder; 100ml
Measuring pipette: 5ml
D. disturbances
Turbidity and color in raw water may cause too high results.
E- procedure:
1- Pretreatment
If water is turbid filter before the analysis. Turbidity caused by colloidal suspended matters or
colors has to be compensated by using the original sample and 5ml of conditioning reagent as blank
sample.
2- formation of BaSO4 - turbidity :
Measure 100ml of water sample into a Erlenmeyer flasks (or a suitable portion filled up to 100ml).
Add exactly 5 ml conditioning reagent and mix intensively. Add appr. 0.2mg BaCl2 crystals and mix
for the homogenous formation of BaSO4 turbidity 1 minute.
3- photometric Measurement :
The turbidity is measured with the photometer at 420 nm in a 10mm sample cell against a blank
sample of dist. water which has been treating with conditioning reagent and BaCl2. If the water
sample is colored or turbid compensate the zero absorbance with 100ml of original water containing
only 5ml of conditioning reagent. Before filling the measuring cell with the sample intensively mix
the content of the Erlenmeyer. Wait for the reading exactly two minutes

١٣
Calibration of the photometer;

1- SO4-stock solution
0.02 H2SO4, 1 ml of stock solution contains 0.96mg SO4.

2- SO4 – standard solution

Dilute 104,1 ml of (1) to 1000ml with dist.water
1 ml of standard solution contains 0.1mg SO4.
3- Calibration standards
Prepare standards of 10 , 20 , 30 , 40 , 50 mg / L

standard Volume of volume of dist. amount of Consenter .of measured

no. stand. sol. water SO4/100 ml stand absorbance

1 10 ml 90 ml 1 mg 10 mg/L

2 20 ml 80 ml 2 mg 20 mg/L

3 30 ml 70 ml 3 mg 30 mg/L

4 40 ml 60 ml 4 mg 40 mg/L

5 50 ml 50 ml 5 mg 50 mg/L

Treat the standards as described in section E and measure the absorbance of the BaSO4 – turbidities
against a blank sample of dist. water plus reagent.
Plot the results of the measurement as calibration curves absorbance via concentration.
Calculate the slope of the calibration curve in the linear range for use as factor.

١٤
 Abs. unit

Concent.mg/L

Example 1: tan α1 = 30.0 mg/l /0.600 abs. units = 50 mg /L abs. units

Example 2: tan α2 = 40mg/l /0.300 abs. units = 133.33 mg /L abs. units

G. Calculations:

The calculation of the concentration is done with the factor C= f x a. 100

V
C= concentration of SO4 in mg/L f = factor in mg/L abs. units
a = absorbance measure in the sample
v = volume of the sample used for the analysis (between 1 and 100 ml)

Remarks
By using the factor calculated in section F the photometer can be programmed to give the results in
calculation of SO4 in mg/L
When 100 ml of sample was used .if less than 100 ml of sample was used this has to be considered by
multiplying the reading of the photometer display by the dilution factor

Example:
If only 5 ml of sample was used and filled up to the mark of the nessler tube the result has to be
multiplied by 20
١٥
 Analysis : SILICA (SiO2)

Molybdate reactive silica standard method 426 B

A-Basic theory:

Ammonium-molybdate reacts at PH –value of appr. 1.2 With silica and phosphate to produce heteropoly
acids. Oxalic acid will destroy the molybdophosphoric acid but not molybdosilica acid.
The intensity of yellow color is then proportional to the present silica concentration

B- Reagents:

1- Hydrochloric acid (diluted 1:1)

2- Ammonium molybdate reagents:

Dissolve ammonium molybdate 10 gm (NH4)6 Mo7O24.H2O) in dist. water with stinning and gently
warming and dilute to 100 ml .filter if necessary and adjust the PH at 7 - 8 with NaOH .store the
solution in a plastic bottle.

3- Oxalic acid solution

Dissolve 10 g oxalic acid (H2C2O4. 2H2O) in distilled water and dilute to 100 ml

C- Apparatus:

Spectrophotometer
Wavelength: 410 nm
Light- pass: 10 mm
Measuring tubes; 50 mm
Measuring pipette: 1.2 ml

D - Interferences:

Strongly colored or turbid samples have to be pretreated. High concentrations of phosphor may give to
high results.
The method is not determining the molybdate UN reactive silica

E –Procedures:

1- pretreatment:

Turbidity may be reduced by filtering the sample – color has to be treated with activated carbon
and filtration.
Low turbidities or colors can be easily reduced by dilution of the sample if the silica concentration is
high enough.

١٦
 2- formation of color:

Measure 50 ml of sample or a suitable smaller portion filled up to 50 ml into a nessler tube. Pipet in
the given orders and in rapid succession 1.0 ml HCl (1:1) , 2.0 ammonium molybdate reagent and
after mixing wait 5 minutes before add 2 ml of oxalic acid . measure color after a time of 2 minutes
but before 15 minutes.
3- photometric measurement:

The specific color can be measured at a wavelength of 410 nm against a blank sample of 50 ml dist.
water and reagents.
Within a time of 5 to 15 minutes the color will be stable.

F –calibration of the photometer:

1- stock silica solution

Dissolve 4.73 g sodium metasilicate Nona hydrate (Na2SiO3.9H2O) into dist.water and dilute to 1000 ml .

1 ml of stock solution contains 1 mg SiO2

2- standard solution
Dilute 10 ml of stock solution to 1000 ml dist .water
1 ml of standard solution contains 0.010 mg SiO2
Store stock and standard solutions in plastic bottles.

3- calibration standards
Prepare calibration standards of 1.0, 2.0, 3.0, 4.0, 5.0, 7.5, 10.0 mg/L
SiO2 in 50 ml Nessler tubes

Volume of volume of dist amount of concenter.of measured

standard no.
stand. sol. water SiO2/50 ml stand absorbance

1 5ml 45 ml 0.05 mg 1.0 mg/L

2 10 ml 40 ml 0.10 mg 2.0 mg/L
3 15 ml 35 ml 0.15 mg 3.0 mg/L
4 20 ml 30 ml 0.20 mg 4.0 mg/L
5 25 ml 25 ml 0.25 mg 5.0 mg/L
6 37.5 ml 12.5 ml 0.375 mg 7.5 mg/L
7 50 ml 0 0.50 mg 10.0 mg/L

Treat the prepared standards with the reagent as described in section E and measure the absorbance of
the formed color against a blank sample within a time between 5 and 15 minutes.
Plot the results as calibration curve absorbance via concentration and calculate the factor of the
calibration curve in the linear range.
Use this factor to program the photometer to give the results in concentration readings or to calculate
absorbance readings into concentrations.

١٧
 Abs. units

Concen. mg/L

Factor: tan α= 10 mg/L / 0.350 abs .units = 25.57 mg/L abs. units

G. Calculations:

The calculation of the silica concentration from a measured absorbance signal may be done with the
factor of the calibration curve:
C= f . a . 50
V
C= concentration of SiO2 in mg/L
f = factor in mg/L abs. units
a = absorbance measure in the sample
v = volume of the sample used for the analysis (between 1 and 50 ml)

١٨
 Analysis : IRON (total: Fe - II, Fe – III)
Standard method 310 A

A-Basic theory:

The iron II-ion forms with 3 molecules of phenanthroline at pH 3.2 to 3.3 chelate complex of range-red
color. The intensity of color is specific for the concentration of iron II. To measure total iron the iron –
III has to be reduced into the famous form. If only iron –II shall be measured the reduction reaction is
not necessary. The amount of iron III can be calculated by the concentration of total iron –minus iron-II

B- Reagents:

1- Hydrochloric acid conc. . . .not containing more than 0.00005% of iron

2- hydroxyl amine chloride solution

Dissolve 10g NH2 OH.HCl in 100 ml of dist. water

3- ammonium acetate buffer solution

Dissolve 250 g NH4C2H3O2 in 150 ml dist.water and add 700 ml of concentrated (glacial) acetic acid

4- sodium acetate buffer solution

Dissolve 200 g NaC2H3O2.3H2O in 800 ml of dist.water

5- phenantroline solution
Dissolve 100 mg 1.10- phenantroline monohydrate (C12H3N2.H2O) in 100 ml of dist.water by low
heating not over 80 OC. Add appr. 2 drops of HCL to the dist. water heating is unnecessary if 2 drop
of conc. HCl are added to the water. Heating is unnecessary if 2 drops of conc. HCl are added to the
water.

c- Apparatus:

Spectrophotometer
Wavelength: 510 nm
Light- pass: 10mm
Erlenmeyer flasks: 125 ml
Measuring cylinder; 50ml
Measuring pipette: 1, 2, 5,10ml

D –Procedures:

1- pretreatment:

The interference of strong oxidizing agents as cyanides, nitrites and polyphosphates will be
eliminated by the initial boiling and 1 ml of excess hydroxylamine solution. In the presence of
interference heavy metal ions a larger excess of phenantroline is required.

١٩
 2- formation of color:

Measure 50 ml of sample into an Erlenmeyer flask (or on suitable portion diluted to 50 ml). Add 2
ml of HCl and 1 ml of hydroxylamine and heat to boiling.

Containing boiling until the volume is reduced to appr. 20 ml

Cool and transfer the sample quantitively to a 50 ml nessler tube.
Add 10 ml of ammonium-acetate buffer, 2 ml phenantroline solution and fill up to the mark.
Mix thoroughly and wait for the development of color for 10 min
For the determination of ferrous iron the analysis can be made without adding the reducing agent.

3- Photometric measurement:

After 10 min the intensity of color can be measured with a spectrophotometer at 510.0 nm against a
blank sample of dist. water treated with the same reagents but without boiling.

E – Calibration of the photometer

1- iron- stock solution

Add slowly 20 ml of conc. H2SO4 to 50 ml of dist.water
Dissolve 1.404 g of ferrous ammonium sulfate (Fe (NH4)2 (SO4)2.6H2O) and add the drop wise 0.10
KMnO4 until a faint pink color persists .dilute to 1000 ml with dist.water
1 ml of stock solution contains 0.2 mg of Fe

2- iron standard solution

Pipette 5 ml of standard into 1000 ml volumetric flask and fill up to the mark with dist.water
1ml of standard solution contains 0.001 mg Fe

3- calibration standards
Prepare standards of 0.05, 0.10, 0.20, 0.30, 0.40, 0.50, 0.75, 1.0 mg/L Fe

Volume of volume of dist amount of Fe concenter.of measured

standard no.
stand. sol. water in 50 ml stand absorbance

1 2.5ml 47.5 ml 0.0025 mg 0.05 mg/L

2 5.0 ml 45.0 ml 0.0050 mg 0.10 mg/L
3 10.0 ml 40.0 ml 0.0100 mg 0.20 mg/L
4 15.0 ml 35.0 ml 0.0150 mg 0.30 mg/L
5 20.0 ml 30.0 ml 0.0200 mg 0.40 mg/L
6 25.0 ml 25.0 ml 0.0250 mg 0.50 mg/L
7 37.5 ml 12.5 ml 0.0375 mg 0.75 mg/L
8 50.0 ml 0.0ml 0.0500 mg 1.00 mg/L

Treat the prepared standards with the reagents as described in section D and measure the
absorbance of the formed color against a blank sample of dist .water plus reagents.
Plot the results as calibration curve absorbance via concentration. Calculate the slop of the
calibration curve in the linear range for the use as factor to calibrate the photometer in
concentration.
٢٠
 Abs. in
abs. units

Linear range of calibration

Conce.in
mg/L

Factor: tan α= 0.5 mg/ L / 0.09 abs .units = 5.55 mg/L abs. units

G. Calculations:

The calculation of the concentration is done with the factor of the calibration curve:
C= f x a. 50
V
C= concentration of iron in mg/L Fe
f = factor in mg/L abs. units
a = absorbance measure in the sample
v = volume of the sample used for the analysis (between 1 and 50 ml)

Remarks

By using the factor calculated in section E the photometer can be programmed to give the results directly in
concentration in mg/L, when 50 ml of sample was used. If less than 50 ml was used this has to be considered
by multiplying the result on the photometer display with the dilution factor

Example

If only 20 ml sample was used and filled up to 50 ml in the nessler tube, the result has to be multiplied by
2.5
٢١
 Analysis : CHLORINE (residual CL2)
Standard method 409

General introduction:

Chlorination of water serves primarily to destroy or des activate disease producing micro organisms.
A secondary benefit is the improvement of water quality by the reaction with ammonia, iron, manganese,
nitrite, sulfide and some organic substances. But adverse effects may be the intensifying of taste and color
and formation of chlorinated hydrocarbons.
Chlorine applied in its elemental from initially undergoes hydrolysis to form free available chlorine,
consisting of aqueous molecular chlorine, hypochlorous acid and hypochlorite ion. The portion of each free
chlorine form is pH- dependant. At the pH value of the most waters the hypochlorous acid and hypochlorite
form will dominate. Free chlorine reacts with ammonia and certain nitrogenous compounds to form
combined available chlorine.
Both free and combined chlorine may be present simultaneously.
For the determination of free and combined chlorine basically 2 determination methods are available:

Method 1: jodometric method (409A)

Titration of iodine liberated by chlorine from a potassium iodide solution with a standard
thiosulfate solution (0.01 n or 0.025 n Na2S2O3)

Method 2: colorimetric method

A) orthotolidine method (409D)
B) DPD method (409 F)

Selection of methods

The iodometric method is suitable for measuring chlorine concentrations greater than 1mg /1 in treated
water. Orthotolidine and DPD method are suitable for lower concentration from 0.05 mg/L to 4.0 mg/L.
DPD (N, N-diethyl-p-phenylenediamine) is superior to neutral orthotolidine as indicator.
The produced color is more stable. Fewer reagents are required and fully response of chloramines in
neutral solutions is obtained.

٢٢
 Analysis CHOLORINE (free Chlorine)
Stabilized neutral orthotolidine method (SNORT)

Reagents
1- neutral orthotolodine reagent
Add 5ml conc. HCl to 100 ml chlorine demand free dist. water into 10ml of this acid solution adds
20mg mercuric chloride (HgCl2). 30 mg disodiume ethylendiamine tetra acetate dehydrate and 1.5mg
orthotolidine dihydrochloride and dilute to 1000 ml with dist. water. Store the solution in a brawn glass
bottle in the dark. Avoid contact with rubber and direct sunlight. Do not use longer than 6 months.
Handel this reagent with extremes care.

2- Buffer – stabilizer reagent :

Dissolve 34.4 gm dipotassiume hydrogen phosphate (K2HPO4) 12.6 g potassium dihydrogne phosphate
(K2HPO4) and 8.0 g di (2ethylhexyle) sulfosuccinate in a solution of 500 ml dist.water and 200 ml
diethylene glycolmonobutyl ether. Dilute to 1000 ml.

Apparatus:

Spectrophotometer
Wave length: 625 nm
Light path: 10 mm
Procedure:
Use 0.5 ml neutral orthotolidine reagent and 0.5 ml stabilizer buffer reagent for 10ml of sample.
Place the reagent in a sample tube and add with gentle mixing 10 ml of sample.
Measure the color within 2 minutes against a blank sample of dist water and reagents.
To compensate natural color or turbidity add 5 ml arsenate (5.0g Na AsO2in 1000 ml dist. water) to 100ml
of sample and this as blank sample to the reagents.

Calibration of the photometer:

Calibrate the photometer with standards of known chlorine concentration in the range of 0.05 to 4.0
mg/L.
Make the calibration standards from a standardized hypochlorite solution by dilution with dist. water.

٢٣
 Analysis: CHOLORINE (free chlorine)
N, N-diethyl-p-phenylenediamine method (DPD)

Reagents:

1- DPD indicator solution

Dissolve 1 g N, N –diethl-p-pheniyleneoxalate or 1.5 g p-amine-N-N-diethylaniline sulfate in
chlorine free dist .water, containing 8 ml H2SO4 (1:3) and 200 mg disodium ethylene diamine tetra
acetate dehydrate. Fill up to 1.00 ml with dist.water and store in a brown glass bottle. Discard the
solution when discolored

2- phosphate buffer solution

Dissolve 24 g anhydrous disodium hydrogen sulfate (Na2HPO4) and 46 g anhydrous potassium
dihydrogen phosphate (KH2PO4) in dist. water
Mix with 100 ml dist. water in which 800 g disodium ethylenediamin tetra acetate dehydrates have
been dissolve. Dilute to 1.000 ml and add 20mg HgCl2 to prevent mold growth.

Apparatus:

Spectrophotometer
Wave length: 515 nm
Light path: 10 mm

Procedure

Place 0.5 ml of buffer reagent and 0.5 ml of DPD indicator solution in a test tube. Add 10 ml of sample and
mix. Measure the color immediately in the photometer against a blank sample of dist. water and reagents

Calibration of the photometer

Calibrate the photometer with standards diluted from a hypochlorite solution of known concentration of
free chlorine in the range of 0.05 to 4.0 mg/L

٢٤
 Analysis: ALUMINIUM (Al)
Standard method 302 B

A-Basic theory:

Dissolved aluminum will form in a buffered solution at pH 6 with eriochrome cyanine R a red to pink
complex. The intensity of color is influenced by aluminum concentration reaction time, pH value
temperature and alkalinity.

B- Reagents:

1- eriochrome cyanine R stock solution

Dissolve 100 mg eriochrome cyanine (or 150 mg eriochrome cyanine R) into 100 ml of dist water.
Adjust the pH of the solution to about 2.9 with acetic acid (1:1)

2- working dye solution

The stock solution has an excellent stability of at least one year
Dilute 10 ml of the stock solution to 100 ml with dist.water
This working dye solution may be kept stable for appr. 6 months

3- sulfuric acid 0.02 N

4- Ascorbic acid
Prepare daily fresh a solution of 0.1 g ascorbic acid diluted to 100 ml with dist.water

5-buffer reagent
Dissolve 136 g sodium acetate ((NaC2H3O2.3H2O) in dist water, add 40 ml of 1N acetic acid and dilute
to 1000 ml with dist. water

C – Apparatus:

Spectrophotometer
Wavelength: 535 nm
Light- pass: 10 mm
Nessler tubes: 50 ml
Measuring pipette: 1, 5,10ml

D – Interferences:

Color and turbidity affects the result.

Interfering iron and manganese will be eliminated by ascorbic acid.
Fluoride and polyphosphates increase errors with increasing amounts and with increasing
concentration of aluminum
Below 0.10 mg/l F and a concentration less than 0.5 mg/l all these errors are neglectable.

E – Procedure:

1- pretreatment

Color and turbidity of water will be compensated by a special blank sample. Iron and manganese is
eliminated by ascorbic acid solution. Fluoride and phosphate have to be destroyed when excessive
amounts are present. In low concentration as to be expected in the raw and treated water no errors
٢٥
 should be given by this. Place a 25ml sample or a suitable smaller portion diluted to 25ml into a test
tube or porcelain dish. Add a few drops of methyl orange indicator and titrate with 0.02N sulfuric
acid to a faint pink color. Record this reading for the further analysis.
2- Formation of color

Measure in each of tow Nessler tube 25ml of sample or a suitable smaller portion to 25ml as used for
the acid titration. To each of the sample add the same volume of 0.02N sulfuric acid found in the
titration and 1ml in excess. One of both samples will be used as blank to compensate color or
turbidity .to this blank add 1ml of EDTA- solution (0.01M) to complex the aluminum.
Then add to each sample in the given order 1ml ascorbic acid, 10ml buffer reagent and 5ml working
dye solution. Mix and fill immediately by fill up to the 50ml mark with dist. water.
3- Photometric measurement:

The color will be formed within 5 min. And will be stable for appr. 15min. measure the absorbance
of the color after 10min. at a wavelength of 535nm against the EDTA treated blank sample.

f- Calibration of photometer;
1- stoksolution :
Dissolve 8.792 g aluminum potassium sulfate (Al k (SO4)2.12H2O) in dist. water and dilute to
1000ml.
1ml of stoksolution contains 0.5mg Al.

2 – Standard solution:
Dilute 10ml of stock solution to 1000ml with dist.water.
1ml of standard solution contains 0.005mg Al.

3- Calibration standards:
Prepare calibration standards of 0.05, 0.10, 0.15, 0.20, 0.25, 0.30 mg/1 in 25 ml samples.

Volume of Volume of dist. Amount of Concenter of Absorbance

Standard No.
standard sol. water AL/25ml AL measured
1 0.25 ml 25ml 0.00125 mg 0.05 mg/1
2 0.50 ml 25m 0.0025 mg 0.10 mg/1
3 0.75 ml 25m 0.00375 mg 0.15 mg/1
4 1.00 ml 25m 0.0050 mg 0.20 mg/1
5 1.25 ml 25m 0.00625 mg 0.25 mg/1
6 1.50ml 25m 0.0075mg 0.30 mg/1

Treat the prepared calibration standards with reagent as described in section E. but use a sample of
dist. water plus reagent as blank sample. Fill all samples to 50ml and measure the color after a time of
10min. in the photometer. Plot the results as calibration curve absorbance via concentration and
calculate the calibration factor in the linear of that curve.

٢٦
 Analysis: MERCURY (Hg)
Standard method 315-B

A- Basic theory:
Mercury ions react with dithizone solution in chloroform to form an orange color. The various shades
of orange are measured in a spectrophotometer and unknown concentrations are estimated from a
factor given by a calibration curve.

B- Reagents
1- potassium permanganate solution
Dissolve 5 g KMnO4 in 100 ml of dist.water

2- potassium per sulfate solution

Dissolve 5 g K2S2O8 in 100 ml dist.water

3- hydroxylamine hydrochloride solution

Dissolve 50 g NH2OH.HCl in 100 ml dist.water

4- dithitone solution
Dissolve 6 mg in 1000 ml chloroform

5- potassium bromide solution

Dissolve 40g KBr in 100 ml of dist.water

6- phosphate-carbonate buffer solution

Dissolve 150 g Na2 HPO4.12H2O and 25g anhydrous K2CO3in 1000 ml dist.water. extract
with 10 ml portions of dithizone until the last portion remains blue.
C - Apparatus:
1- Spectrophotometer
Wavelength: 490 nm
Light- pass: 10 mm

2- Separatory funnels

D –disturbances:
Copper, gold, divalent palladium and silver reacts with dithizone in acid solution. Copper is separated
during the procedure by remaining in the organic phase while mercury is left in the aqueous phase.
The mercury dithizonate should be measured quickly, because it is photo sensitive.
E – Procedure:
1- pretreatment

a) preparation of mercury stock solution

Dissolve 135.4 g of HgCl2 in 700 ml dist.water, add 1.5 ml conc. HNO3 and make up to 1000
ml with dist.water
1 ml = 100 µg Hg
b) preparation of mercury standard solution
Dilute 10.0 ml stock solution to 1000 ml dist.water
1ml = 1 µg Hg
Prepare immediately before use

٢٧
 c) preparation of calibration curve pipette 0 ,2, 6, 8, 10 ml of standard solution into separate
beakers
To each beaker add 500 ml dist.water, 1ml KMnO4 solution and 10 ml conc. H2SO4. Stir
and bring to boil. If necessary, add a few drops KMnO4 until a pink color persists. Then
add continuously by 5 ml K2S2O8 solution on and allow cooling for 0.5 hr. Add one or more
drops of NH2OH. HCL solution to discharge the pink color.
When the sample is cool transfer each solution on to a separatory funnel. Add 25 ml
dithizone solution and shake the funnel vigorously transfer each organic layer to another
funnel and repeat this extraction at least three times
Make sure that the color in the last dithizone layer is an intensive blue as the original
dithizone solution. Wash the accumulated dithizone extraction in the funnel by shaking
with 50 ml of 0.25 N sulfuric acid. Transfer the washed dithizone extract to another funnel.
Add 50 ml 0.25 N H2SO4 and 10 ml KBr solution, shake vigorously to transfer the mercury
dithionate from the organic layer to the aqueous layer. Discard the lower dithionate layer
and wash the aqueous layer with a small volume of chloroform
Discard the chloroform. Transfer 20 ml phosphates carbonate buffer solution to each
separatory funnel and add 10 ml standard dithizone solution. Shake thoroughly and after
separation transfer the mercury dithizone to beakers
d) preparation of the samples
Take 500 ml of the water to be analyzed and 500 ml of dist.water for the blank. Complete
all the steps written under (c)
f -calibration:
Take the beakers from no. (c) And transfer the contents to a 10 mm sample cell and measure the
absorbance. On linear graph paper plot absorbance versus micrograms of mercury
Calculate the factor as follows:
Absorbance unit
f = ‫ـــــــــــــــــــــــــــــــــــــــــــ‬
Conc. Of mercury

Because there are 5 samples, take the average of all 5 factors and note this factor.

G –measurement:

Because you took the same volume for the standards as for the sample the concentration of mercury
is:
-
c= f. abs .unit

-
f = average of all 5 factors from calibration curve.

٢٨
 Analysis: CADMIUM (Cd )
Standard method 305-B

A - Basic theory:
Cadmium ion under suitable conditions reacts with dithizone to form a pink to red color, that can be
extracted with chloroform.
This extract can be photo metrically measured.

B - Reagents

1- sodium potassium tartrate solution

Dissolve 250 g Na K C4 O6.4H2O in dist water and fill up to 1000 ml

2- sodium hydroxide-potassium cyanide solutions

a) Solution 1: dissolve 400g NaOH and 10g KCN in 1000 ml dist.water (this solution is stable for
1 or 2 months) store in a polyethylene bottle.
b) Solution II: dissolve 400 g NaOH and 0.5g KCN in 1000 ml dist.water – store solution in
polyethylene bottle.

CAUTION: KCN is extremely poisonous. Never use mouth pipettes to deliver cyanide solutions.

3- hydroxylamine hydrochloride solution

Dissolve 20 g NH2OH.HCl in 1000 ml of dist.water

4- dithizone stock solution

Dissolve 100 mg dithizone in 1 L chloroform. Keep in brown bottle in a refrigerator until required
and use while still cold.

5- tartaric acid solution

Dissolve 20g H2C4H4O6 in 1000 ml dist.water keep in a refrigerator and use while still cold.

6- thymol blue indicator solution

Dissolve 0.4 g thymol solution –ephthalein sodium salt in 100 ml dist.water

C - Apparatus:

1- Spectrophotometer
Wavelength: 518 nm
Light- pass: 10 mm

2- separatory funnels

3- Glass ware
All needed glass ware should be cleaned with 1:1 HCl and rinsed with dist.water

٢٩
D –disturbances:

There are no disturbance under the concentration of there metal ion which are normally found in
surface water. Ordinary room lighting dose not affect the cadmium dithizonate color.

E – Procedure:

1- preparation of cadmium solution

a) stock cadmium solution
Weigh 100.0 mg pure cadmium and dissolve in 20 ml dist.water plus 5 ml conc. HCl.heat this
composition to dissolve the metal. Then fill up to 1000 ml with dist.water and store in a
polyethylene bottle
1.0 ml = 100 µg Cd

b) Standard cadmium solution

Pipette 10 ml of stock solution, add 10 ml conc. HCl and fill up to 1000 ml with dist.water
Prepare this standard short before using
1 ml =1 µg Cd

2- standard dithizone solution

Dilute 100 ml stock dithizone solution to 1000 ml chloroform
Keep in a brown bottle in a refrigerator but allow to warm to room temperature before use.

3- preparations for calibration curve

Pipette 0, 2, 4, 6, 8, 10 µg Cd into a series of separatory funnels. Add dist.water up to 25 ml
Add reagents in the following order, mixing after each addition:
1 ml sodium potassium tartrate solution
5 ml NaOH-KCN solution I
1 ml hydroxylamine hydrochloride solution
15 ml stock dithizone solution
Drain the chloroform layer into a second funnel containing 25 ml cold tartrate acid solution
Add 10 ml chloroform to the first funnel, shake for 1 min. and drain into the second funnel again
Shake the second funnel for 2 min. and discard the chloroform layer. Add 5 ml chloroform, shake
for 1 min. and discard the chloroform layer. Then add in the following order:
0.25 ml hydroxyl hydrochloride solution
15 ml dithizone standard solution
5 ml NaOH-KCN solution II

٣٠
 And shake immediately. Filter the chloroform layer through cotton into a sample cell of the
photometer, which must be dry.
Read the absorbance at 518 nm and plot a calibration curve
4- pretreatment for the samples
Take 200 ml of the water to be analyzed
Add 0.5 ml conc. HCl and evaporate to 20 ml. Add a few drops of thymol blue indicator solution and
then 6N NaOH until the indicator just turns yellow at pH app. 2.8
Make up to 25 ml with distilled water. Take a standard cadmium solution with 6.0 g Cd in 25 ml and
a blank.
Proceed as written in chapter E 3 above
The standard cadmium solution is used for controlling the calibration curve
Read the absorbance and compare with the calibration curve.

٣١
 Analysis: Fluoride ions SPADNS
Standard method 414-C

A - Basic theory:

The colorimetric method is based on the reaction between fluoride and zirconium dye-lake. The fluoride
reacts with the dye lake dissociating a portion of it into a colorless complex anion (Zr F6)-2 and the dye

B – Reagents:

1- stock fluoride solution

Dissolve 221.0 mg anhydrous sodium fluoride NaF in dist water and dilute to 1000 ml
1 ml = 0.1 mg F

2- standard fluoride solution

Dilute 100 ml stock fluoride solution to 1000 ml with dist.water
1 ml = 0.01 mg F

3- SPADNS – solution
Dissolve 958 mg SPADNS in dist.water and dilute to 500 ml
This solution is stable if protected from direct sunlight
4- zirconyl- acid reagent
Dissolve 133 mg zirconyl chloride octahydrate ZrOCl2.8H2O in 25 ml dist. water. Add 350 ml conc.
HCl and dilute to 500 ml with dist. water
5- Reference solution
Add 10 ml of SPADNS – solution to 100 ml dist.water. Dilute 7 ml conc. HCl to 10 ml and add to the
diluted SPADNS – solution. This reference solution is used for setting the zero at the
spectrophotometer and is stable
6 - Mix 500 ml of SPADNS – solution with 500 ml of zironyc-acid reagent. This acid zironyc SPADNS
reagent is stable for 2 years
7 - Sodium arsenate solution
Dissolve 5.0 mg NaA5O2 and dilute to 1000 ml with dist.water
Caution
NaA5O2 is extremely poisonous
Take care to avoid investigation

٣٢
C - Apparatus:
1- Spectrophotometer
Wavelength: 570 nm
Light- pass: 10 mm
D – Interferences
In the general, the colorimetric methods are susceptible to the same interfering substances, but to
different degrees (see table below)

Substances Conc. mg/L Type of error

Alkalinity (ppm) CaCO3 5000 -

Al3+ 0.1 -

Cl- 7000 +

Fe3+ 10 -

(NaPO3)6 1 +

(PO)3- 16 +

(SO4)2- 200 +

Chlorine interferes this and has to be removed by adding NaA5O2 solution

E – Procedure:
1- preparation for calibration curve:
Prepare standards in the range of 0-1.4 mg/L F- by diluting the standard fluoride solution

Conc. mg/L 0.1 0.2 0.3 0.4 0.5 0.8 1.0 1.2 1.4

Volume of
ml 0.5 1.0 1.5 2.0 2.5 4.0 5.0 6.0 7.0
standards

This table is valid for 50 ml volume

For example: for 0.3 mg/L F- you take 1.5 ml of standard solution and 48.5 ml dist.water
Set the photometric at zero with the reference solution and take the reading of the absorbance
immediately. Plot a curve and calculate a factor as the average of all 9 factors (f)
Note the temperature of the standards
2- pretreatment of the sample:
To remove the residual chlorine
Add 50 ml of NaA5O2 solution for 0.1 mg Cl2. Use a 50 ml sample
٣٣
 Res.Cl2(total) mg/L 0.5 0.6 0.7 0.8 0.9 1.0 1.1 1.2 1.3

volume of NaA5O2 ml 5 6 7 8 9 10 11 12 13

Adjust the temperature of the sample to the same value as the standards for the calibration curve.
To 50 ml add 10 ml of the acid-zyrconyl- SPADNS – solution
Mix well and take the reading of the display after setting the instrument to zero as written before.

F – Calculation:
Conc. F mg/L = f. Abs. unit
Only if the volume of standards is equal to the volume of the sample

٣٤
 Determination of Nitrite

Reagents:

1- Naphthylamine
0.1g of (N-(1- Naphthyl ethylene diamine dihydrochloride) (C12H14N2.2HCl)
Dissolve in 1.0 ml conc. HCl then dilute to 100 ml
2- sulfanilic acid
Dissolve 0.6 g of sulfanilic acid in 70 ml hot dist.water & cool. The solutions then add 20 ml conc.
HCl dilute to 100 ml with dist.water and mix thoroughly
3- Sod. acetate buffer solution
(2M) dissolve 16.4 g CH3COONa or (27.2 g CH3COONa.3H2O) in dist.water and dilute to 100 ml
with dist.water. Filter necessary
4- Sod. Nitrite " stock solution"
Dissolve 0.246 g anhydrous NaNO2 in nitrite free dist.water and dilute to 1 liter
1.0 ml = 0.05 mg N
Preserve by adding 1.0 ml chloroform

5- Standard sod. nitrite solution

Dilute 10.0 ml from stock solution to 1 liter
Add 1.0 ml chloroform
1.0 ml = 0.0005 mg N
Volume of sample 50.0 ml

Volume of Volume of dist.

Standard No. Conc. of st.ppm
standard sol. water
1 0.5ml 49.5 ml 0.005
2 1.0 ml 49.0 m 0.01
3 1.5 ml 48.5 m 0.015
4 2.0 ml 48.0 m 0.02
5 2.5 ml 47.5 m 0.025
6 3.0 ml 47.0 m 0.03

٣٥
Procedure:

1- 50.0 ml sample (clear sample) + 1.0 ml sulfanilic acid , mix thoroughly (pH~ 1.4)
2- Standing 3-10 min (5 min)
3- Add 1.0 ml N-(1- Naphthyl ethylene diamine dihydrochloride)
4- Add 1.0 ml sod .acetate buffer. mix well at this point (pH= 2.0 – 2.5)
5- Measure the reddish – purple color after 10-30 min (10 min)
At λ = 530 nm
λ→ 520 nm

٣٦
 Analysis: Nitrite – Nitrogen (NO2-N )
Standard method 420
A - Basic theory:
The nitrite concentration is determined through the formation of a reddish purple azo dye produced
at pH 2.0 to 2.5 by the coupling of diazotized sulfanilic acid with N-(1-naphthyl)-ethelene-diamine
dihydrochloride
This method is suitable for determination of nitrogen nitrite in the range of 1-25 µg/1 N-
Photometric measurement with a light path of 10 mm allows a determination in the range of
2-20 µg/L NO2- -N
B – Reagents:
1- sulfanilamide reagent
Dissolve 5 g sulfanilamide in mixture of 50 ml conc. HCl and about 300 ml dist.water. Dilute to 500
ml with dist.water
The solution is stable for many months
2- N-(1-naphthyl)-ethelene-diamine dihydrochloride solution
Dissolve 500 mg of this salt in 500 ml dist.water
Store in a dark bottle. Renew the solution monthly or immediately when it develops a brown
coloration
3- HCl-solution (1:3)
4- Stock nitrite solution
Dissolve 1.232 g NaOH in dist water and dilute to 1000 ml with dist.water
1ml = 0.25 mg NO2- -N
Preserve with 1 ml chloroform. Standardization of stock solution
Pipette 50 ml 0.05 N KMnO4, 5 ml conc. H2SO4 and 50 ml of stock nitrite solution into a glass
Stoppard bottle
Shake carefully and warm up to 70 oC- 80 oC. Discharge the permanganate color by adding
0.05 N sodium oxalate (3.35 g Na2C2O4) in 1000 ml dist.water in 10 ml portions
Titrate with KMnO4 (0.05 N) to the faint pink end point.
A =] (BxC) – (DxE) ] .7
F
A= mg/ml NO2-N in stock solution
B= ml (total) of 0.05 N KMnO4
C= 0.05
D= ml of Na2C2O4
E= 0.05
F= ml of stock NaNO2 solution taken for titration
Each 1ml of 0.05 N KMnO4 consumed by the nitrite corresponds to 0.35 mg NO2- -N
٣٧
 5- Intermediate nitrite solution
Take as volume for this solution
g = 12.5/A dilute this volume G (app.50ml) to 250 ml with dist. water
1ml = 50 µg NO2- -N
This solution is not stable it has to be prepared fresh for the measurement
6-nitrite standard solution
Dilute 10 ml of intermediate nitrite solution to 1000 ml with dist.water
1ml = 0.5 µg NO2- -N
C – Apparatus:
Spectrophotometer
Wavelength: 543 nm
Light- pass: 10 mm
D – Interferences
Suspended solids should be removed by filtration before color development. Chemical incompatibility
makes it unlikely, that nitrite, free available chlorine and nitrogen trichoride will coexist in water. See
method 409 F for determination of nitrogen trichloride.
E – Procedure:
To 50 ml sample add 1 ml sulfnilic acid solution
Allow the reagent to react for a period of more than 2 min. but not longer than 8 min
Add 1 ml 1-naphthyl-ethylenediamine solution and mix immediately
Measure the absorbance (or concentration with a factor) between 10 min and 2 hrs. After wards.
F –calibration of photometer

Prepare standards from a freshly prepare nitrite standard solution

The volume of the standards should be 50 ml with concentration of 1.0, 2.0, 4.0, 7.0, 10, 15, 20, 25 µg/l
Plot a curve and calculate the factor

٣٨
 Determination of phosphate

Reagents
1 – H2SO4:-
120 ml of Conc. H2SO4 + 750 ml .dist. water, cool the soln. and dil to 1L of dist water
2 – SnCl2.2H2O :- ( freshly prepared )
2.5 gm of SnCl2.2H2O is dissolved in 10 ml of conc. HCl, dil to 100 ml dist H2O, then filter
3 – Amm. Molybdate -strong acid soln : -
Dissolve 25 gm of (NH4)6Mo7O24.4 H2O in 175 ml dist. H2O/Add 310 ml .conc.– H2SO4 to 400 ml dist
H2O, Cool , add the Molybdate soln. and dil to 1 L H2O .

Standards:-
1 – St. Phosphate soln.:-
Dissolve 0.7164 gm of KH2PO4 which has been dried in an oven at 105 0C in dist. H2O and dil. The soln. to 1
L with dist. H2O.
Dil 100 ml of this soln. to 1 L dist. H2O
1 ml = 0.05 mg of PO4

Procedure:-
50 ml of sample free from color and turbidity + 1 drop of ph.ph. Indicator, if pink color appears, put H2SO4
Dropwise to discharge the color, and then add 2 ml of Amm. Molybdate soln. with stirring + 0.25 ml ( 5
drops ) of SnCl2 soln. measure the color at 690 nm after 10 min.

Another method

Stannous chloride reagent:-

Dissolve 2.5 gm of SnCl2.H2O in 100 ml glycerol. Heat in water bath and stir with glass rod to hasten
dissolution. This reagent is stable and requires neither preservatives nor special storage.

(100 ml sample + 4 ml Amm. Molybdate + 0.5 ml (10 drops) SnCl2 soln. wait 10 min. and measure)

٣٩
٤٠