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Name of the Experiment: Determination of total nitrogen present in soil sample by modified
Kjeldahl method.
Principle:
The Kjedahl procedure generally employed for determination of total N involved
two steps:
Reaction In digestion:
The temperature of the digestion is very critical. If the temperature is
too low (below 360 o C), the release is slow or incomplete, and if too high (over 410o C),
some loss of NH3 from the mixture results. The reactions are given below:
0
− 410 C 0
→( NH 4 ) 2 SO4
Soil-N + H 2 SO4 360
The amount of N thus converted to ammonium sulfate may be determined by distilling it with
alkali and collecting the distilled ammonia in a standard acid (2% boric acid) and collected
ammonium is titrated against N 10 H 2 SO4 followed by finding out the strength of the acid.
The following reactions occur:
2
NH 3 + H 3 BO3
→ NH 4 .H 2 BO3 .
→ ( NH 4 ) 2 SO4 + H 3 BO3
NH 4 H 2 BO3 + H 2 SO4
Material:
(B) Reagent :
1. H 2 SO 4 , concentrated, 36N
2. Digestion mixture: In a mortar 100gm of anhydrous Na2SO4, 6.5 gm
of anhydrous CuSO4
3. 40% NaOH
4. Mixed indicator solution
5. 2% Boric acid mixed indicator solution
6. H 2 SO 4 of 0.1 N
Procedure:
1. 2gm of finely powdered soil was taken in a Kjedahl flask and 10 ml of distilled
water was added to it and left for sometimes.
2. Then 20ml of conc. H 2 SO 4 was added.
3. The flask was gently heated over a low flame in a digestion chamber for about 10
minute and then after off the burner followed by cooled.
4. After that 5 gm of digestion mixture was added, on the burner and temperature
was raised. The heat was confined within the portion of the flask that contained
liquid.
5. The liquid became colorless and another 1 hour heat was continued.
6. The digest was cooled and when cold, it was diluted with distilled water and
made up to volume in a 100 ml volumetric flask.
7. Then 10 ml of digest was distilled with 10 ml of 40% NaOH solution using
Kjeldahl’s distillation apparatus.
3
8. About 10-15 ml of distillate (NH3 gas) was collected in a 125 ml conical flask
containing 10 ml of 2% boric acid with mixed indicator.
9. The conical flask was removed and the delivery outlet of the distillation apparatus
was washed with distilled water.
10. The distillate was then titrated against the standard H 2 SO 4 until the solution
color became pink.
11. A blank experiment was also done simultaneously using the entire chemical
except soil.
Data:
Initial Final
burette burette Difference
No. of
reading reading (ml)
Observati Mean (ml)
(ml) (ml)
on
Sample 1. 9.6 12.6 3.0 3.0
Blank 1 15.2 15.5 0.3 0.3
Calculation:
Result:
Principle:
The total quantity of P in most soils is relatively small ranges from 0.03 to 03%.
Most soil P determination has two distinct phases:
First, the preparation of solution containing the soil P, and
Second, the quantitative determination of the P in this solution.
The choice of colorimetric method depends on the concentration of P in solution and
particular acid system involved in the analytical procedure.
→ H 3 P( Mo3O10 ) 4 + 12 H 2 O
H 3 PO4 + 12 H 2 MoO4
The complex, before reduction, gives a yellow hue to their water solution.
With high P concentration, a yellow precipitate is formed. In solution of low enough
concentration to be suitable for determination by reduction to form the blue color, the yellow
color is so faint that it is not noticed. At about 100 times this concentration, the yellow color
is suitable for spectrophotometer as for molybdosilicic acid color. The vanadomolybdo
phosphoric yellow color is greatly intensified by the vanadium component. The extent nature
of the yellow chromogen of the vanadomolybdophosphoric system is not known, but the
color is attributed to substitution of oxyvanadium and oxymolydenum radicals for the O of
PO4to give a heteropoly compound that is chromogic. The vanadomolybdophosphoric yellow
color method has the advantages of extreme simplicity, lower sensitivity, stability of color,
freedom from interferences with a wide range of ionic species in concentration up to 1000
ppm, and adaptability to HNO3, HCl, and H2SO4 or HClO4 system.
Apparatus:
1. Pipettes
2. volumetric flask
3. Spectrophotometer (400 to 490 nm)
Reagent:
2. Standard phosphorus solution of 0, 1, 2,5,10 ppm was made from primary stock
solution and secondary solution.
3. The following special combined HNO3- Van date molybdate reagent :
Solution-A and solution-B was freshly mixed before work. 400ml of solution-A and 600ml of
solution-B was mixed in the ratio of 1:1.5.
Procedure:
Data:
Calculation:
Principle:
Ba2++SO42-
→ BaSO4 ↓
Materials:
(b) Apparatus:
1. Measuring flask
2. Electric balance
3. Beaker
4. Volumetric flask
5. Pipette
6. Spectrophotometer, etc.
(c) Reagent:
Procedure:
5. Then 10 ml of standard solution from 0, 5, 10, 15 ppm was taken into four 2.5 ml
volumetric flask. Therefore, 3 ml of BaCl2 –Twen80 solution was added into every
volumetric flask was made to volume up to the mark.
6. The reading was then taken for the standard solution and the standard curve was
made where absorption were taken against the concentration in ppm.
7. Then the reading for the sample solution was taken.
8. Thus reading was put in the standard curve and sulfur content was determined from
the standard curve for sulfur.
Data:
Calculation:
Result:
Principle:
Total potassium in soils commonly ranges from 0.5 to 2.5%. Total potassium
in soil is determined by a wet digestion of the sample with nitric and perchloric acid followed
by flame photometry. It is a device that soaks out the sample through pipes. Up to burning on
the flame K+ gives characteristics color. In this zone other colors also produce if it contains
other element. A monochromatic filters adjoining to the flame select only the chromate of K+
and allows it to pass through it. Each penetrating wave makes a pulse when they strike
portable cell. This pulse no. is recorded by the machine and is displayed as digital signal. The
amount of pulse is directly related to the K+ content of the sample.
Material:
Procedure:
1. The extract was 10 times diluted by adding distilled water with 5ml digest and made
up to 50 ml volume in 50 ml volumetric flask.
2. The reading for potassium in a flame photometer from the diluted extract and standard
solution.
3. A standard potassium curve was prepared by using known solution of potassium.
4. Total potassium was determined from the standard potassium curve.
Data:
Calculation:
Result:
Principle:
Material:
1. Conical flask
2. Filter paper (whatman no.40)
3. Distillation apparatus
4. Micro burette
5. Measuring cylinder
6. pipette
7. Volumetric flask
(c) Reagent:
4. H2SO4 , 0.01 N
5. NaOH, 10%
6. 2% Boric acid
7. Mixed indicator solution
Procedure:
Data:
Calculation:
1000ml 1N H 2 SO4 ≡ 18 gm NH 4 + ≡ 14 gm N
14 × 0.3
∴0.3 ml 1N H 2 SO4 = gm N
1000
14 × 0.3 × 0.01
∴0.3 ml 0.00973 N H 2 SO4 ≡ gm N
1000
= 0.000042 gm N
∴ ( % ) of N in soil = 0.0042 gm
= 0.0042 ×1000 mg
= 4.2mg N
4 .2
So, CEC in meq / 100 gm soil = = 0.3 meq
14 100 gm soil
Result:
The cation exchange capacity of the soil sample is 0.3 meq/100gm soil.
13
Principle:
Materials:
Standard solutions of sodium of 0, 5, 10, 15, 20, 25, 30 ppm. Standard solution
was made by using Na2SO4 salt. At first 500ppm in 1000ml was made as primary stock
solution by taking 1.543 gm Na2SO4 in 1000 ml volumetric flask. Then secondary stock
solution was made from primary stock solution. And finally 5, 10, 15, 20, 25, 30 ppm was
made from by secondary stock solution by serial dilution.
Procedure:
1. The reading for Na was taken from flame photometer for both the extract and standard
solution. Besides blank reading was taken.
2. A standard curve of sodium prepared by using known concentration of sodium.
3. The amount of sodium was determined from the standard curve.
Data:
ppm Intensity
0 0
5 3.2
10 5.9
15 8.2
20 10
Sample 3.7
Calculation:
= 0.0023 meq Na
100 gm soil
Result:
meq Na
The amount of NH4OAc extractable sodium in sample is 0.0023 100 gm soil .
15
Principle:
Materials:
Standard solutions of sodium of 0, 5, 10, 15, 20, 25, 30 ppm. Standard solution
was made by using K2SO4 salt. At first 500ppm in 1000ml was made as primary stock
solution by taking 1.543 gm K2SO4 in 1000 ml volumetric flask. Then secondary stock
solution was made from primary stock solution. And finally 5, 10, 15, 20, 25, 30 ppm was
made from by secondary stock solution by serial dilution.
Procedure:
1. The reading for K was taken from flame photometer for both the extract and standard
solution. Besides blank reading was taken.
2. A standard curve of sodium prepared by using known concentration of K.
3. The amount of K was determined from the standard curve.
Data:
ppm Intensity
0 0
5 3
10 5.9
15 8.4
20 10
Sample 6.5
Reading for blank: 0 (intensity)
Reading for sample:
Intensity ppm from graph
6.5 11.25
16
Calculation:
= 11.25×100
10
= 112.5 µg Kg¯1
= 11.25 meqk/100 gm
39.1
= 0.287 meqk/100gm soil
Result:
Principle:
Materials:
1. Micro pipette
2. Small porcelains
3. volumetric flask
4. Pipette
5. Measuring cylinder
(c) Reagent:
1. NaOH 2.5N, 10gm of NaOH was taken in a 100 ml volumetric flask and
was made volume up to the mark with distilled water.
2. Standard EDTA solution, 0.02M. The molecular weight of EDTA is
372.254. Therefore, 1.8613 gm of EDTA was taken in 250ml volumetric
flask; dissolved and was made volume up to the mark. The solution was
0.02 M EDTA in 250ml.
3. Standard calcium solution , .02M
4. Murexide (ammonium turtarate ) indicator
Procedure:
1. 5ml of extract was taken in a small porcelain cup and 5 ml of 2.5 N NaOH was added
.
2. 0.02gm of murexide indicator was added which give the solution light pink color.
3. It was titrated against EDTA solution. At the end point color changed from pink to
light blue.
18
Data:
Calculation:
= 0.02×0.4×1×100 ×100
2.5×5
= 6.4%
19
Principle:
The lime requirement is the amount of lime that must be applied to acid
change soil pH from its present value to any desired value. The materials commonly used for
liming soils are Ca and /or Mg oxide, hydroxide, carbonates and silicates. The accompanying
anion must lower H+ activity and Al3+ in the soil solution. Liming reaction begin with the
neutralization of H+ in the soil solution by either OH-or HCO3- originating from the liming
materials . For example CaCO3behave as follows:
The rate of the reaction is directly related to the rate at the OH- ions are
removed from solution by H+ ions to made H2O.
Soil pH is the most important criteria determining the lime requirement of soil.
Other properties Such as texture, structure, percentage of organic matter amount and type of
clay affect lime requirement, CaCO3 is the most common agricultural lime but its incubation
in the lab is too much time consuming. To obtain relatively quick determination in laboratory
we use Ca (OH)2 for incubation experiment and convert the amount in CaCO 3. Buffering
capacity of the soil is the main considerable factors here. The more the buffering capacity, the
more the lime required to bring desired change.
Material:
1. Beaker ,50ml
2. pH –meter
3. Stirring rod
4. Electric balance
5. Para film
(c) Reagent:
1. Ca(OH)2
2. Standard buffer solution of pH 4 and 7.
3. Distilled water
Procedure:
20
Data:
Calculation:
We have raised the pH from 4.0 to6.8. From graph, to increase the soil pH
from 4.0 to 6.8, 0.12 gm Ca (OH)2 was needed per 20 gm soil.
Now, to get the calcium carbonate equivalent for Ca (OH)2 , CaCO3 and Ca(OH)2 should be
equal in term of acid neutralizing ability.
Ca (OH)2+2HCl=CaCl2+2H2O
CaCO3 +2 HCl = CaCl2+CO2 + H2O
21
Result:
To raise the pH from 4.0 to 6.8 of the supplied acid soil sample, the required
amount of Ca (OH)2 is 16.21 tons of calcium carbonate per hectors.
22
Name of the Experiment: Determination of total organic carbon in soil by Tyurin’s method.
Principle:
3 O2 + 3C → 3CO2
The amount of oxygen consumed during the oxidation of organic carbon is calculated
from the difference between the amount of dichromate taken and the amount remaining after
oxidation, this is determined by titration with a standard solution of Mohr’s salt.
Materials:
1. Beaker
2. Measuring flask
3. Burette, pipette, funnel
4. Hot plate
5. Conical flask
(c) Reagent:
Procedure:
5. The flask was then removed from the plate and the inner and outer surfaces of the
funnel and the neck and walls of the flask were carefully washed with about 10 ml of
distilled water.
6. The color of the liquid after the oxidation should be orange –yellow. A greenish color
of the liquid indicates insufficient oxidizing agent and the analysis must be repeated
to take small amounts of soil.
7. The unutilized K2Cr2O7 solution in the oxidation was titrated against 0.1 N Mohr’s
salt solution using phenylanthranilic acid as the indicator.
8. At the end point of the titration, the color was changed from brown to green.
9. A blank experiment was run at the same time using all the reagents except soil.
Data:
S ×V 1 × 10
So, S 2 = 1 1= = 0.547 N
V2 18.7
Calculation:
( B − T ) × 0.39 × f
% Organic C =
W
So , % Organic C =
(18.7 − 15.0) × 0.39 × 0.547
2
3.7 × 0.39 × 0.547
=
2
0.789
=
2
= 0.394 %
Result:
Principle:
The excess acid left after the oxidation may be determined volumetrically with standard
ferrous sulfate solution and the quantity of substances oxidized is calculated from the amount
of chromic acid reduced.
Materials:
1. Beaker
2. Measuring flask
3. Burette, pipette, funnel
4. Conical flask
(c) Reagent:
1. 1 N K2Cr2O7 solution
2. Conc. H 2 SO4 , 96%
3. Phosphoric acid
4. NaF
5. Ferrous sulfate solution
Procedure:
Data:
S ×V 1 × 10
So, S 2 = 1 1= = 0.547 N
V2 18.7
Calculation:
( B − T ) × 0.39 × f
% Organic C =
W
So , % Organic C =
(18.7 − 15.0) × 0.39 × 0.547
2
3.7 × 0.39 × 0.547
=
2
0.789
=
2
= 0.394 %
Result:
Principle:
The carbonate can be determined by decomposing them with and the amount
of CO2 evolved may be determined volumetrically. The CO3- may be reacted upon by a
known amount of standard solution and the acid required to decompose the CO3- may be
determined volumetrically.
Material:
1. Beaker ,
2. Stirring rod
3. Electric balance
4. Burette,
5. pipette,
6. funnel
(c) Reagent:
1. 1 N NaOH solution
2. 1 N HCl
3. Bromothymol blue indicator solution
Procedure:
Calculation:
=
(10.6 − 9.0) × 0.05 × 1 × 100 × 100
10 × 10
1.6 × 0.05 × 1 × 100 × 100
=
10 × 10
= 8%
Result:
Principle:
This reaction will proceed smoothly over a wide range of Mn concentration. The little
interference encountered can usually be readily eliminated. Permanganate is stable in the
presence of excess period ate. The intensity of the permanganate color developed will remain
unchanged in the absence of reducing agents and is readily reproduced. Organic matter and
chloride is the principal interfering agent that must be eliminated in analysis of soil. So,
reducing substances must be absent during the period ate oxidation of manganese and these
are destroyed with nitric or sulfuric acid and H2O2, before development of permanganate.
Chloride can be tolerated to some extent particularly if phosphoric acid is used; if perchloric
acid is used to evaporate the solution there will be little interference from chloride.
Apparatus:
1. Conical flask
2. Filter paper (whatman no.40)
3. Beaker
4. pipette
5. Hot plate
6. Spectrophotometer
7. Asbestos
Reagent:
1. H2SO4
2. H2O2, 30%
3. H3PO4, 85%
4. KIO4
5. NH4OAc
6. Standard Mn-solution
Procedure:
Calculation:
Result: