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For more detailed information please refer to the BUCHI Kjeldahl Guide.
www.buchi.com/kjeldahl/en/applications/literature/
In addition to this short guide you can find detailed information including our
application database at: www.buchi.com/kjeldahl/applications
And to download BUCHIs practical tablet guide or Kjeldahl calculator for iOS,
Android and Windows Phone 7: www.buchi.com/kjeldahl/apps
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Sample
preparation
Grinding
Sample tube size
Weighing table
Kjeldahl Tablets
Digestion:
Digestion
Digestion parameters
Neutralization/Alkalinization:
Distillation
Distillation:
Receiver:
Titration
Result
Titration:
Calculation
LIMS
KjelLink
The digestion mixture is alkalized with NaOH prior to distillation to free up the ammonia.
The ammonia is steam distilled
into an acidic receiver solution.
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Sample preparation
Sample tube selection and weight
Micro
Sample weight
< 0.2 g
Sample volume
23 mL
H2SO4
25 mL
Kjeldahl Tablets
1 (Micro)
300 mL
Sample weight
0.1 5 g
Sample volume
< 200 mL
H2SO4
530 mL
Kjeldahl Tablets
500 mL
Sample weight
>4g
Sample volume
< 400 mL
H2SO4
> 10 mL
Kjeldahl Tablets
Recommendation
Benefit
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Sample preparation
Weighing
The actual weight depends on
Sample
preparation
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Sample preparation
Usage of the weighing table
4
0.5
0.125
N [mg]
per glas
N [%]
0.5
0.01
0.03
0.05
0.10
0.40
3.6
2.0
0.04
0.10
0.20
0.40
1.60
14.3
2.5
0.05
0.13
0.25
0.50
2.00
7.0
0.14
0.35
0.70
1.40
5.60
10.0
0.20
0.50
1.00
2.00
8.00
50.0
1.00
2.50
100.0
2.00
0.05
0.1
0.5
2
2.9
3.6
10.0
14.3
3
3
1.8
5.0
7.1
1.4
7.1
14.3
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Sample preparation
Addition of chemicals and Kjeldahl Tablets
Sample
preparation
1.
2.
3.
4.
Safety aspects
Digestion time
Ecological aspects
Foam formation of the sample
For more demanding samples or samples that are high in fat or carbohydrate see
page 10.
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Sample preparation
Weighing table for solid and liquid samples
Solid samples
Sample: weight [g]
5
0.125
N [mg] N [%]
0.01
0.05
0.1
0.5
0.5
0.01
0.03
0.05
0.10
0.40
3.6
2.0
0.04
0.10
0.20
0.40
1.60
14.3
2.5
0.05
0.13
0.25
0.50
2.00
3.6
1.8
7.0
0.14
0.35
0.70
1.40
5.60
10.0
5.0
10.0
0.20
0.50
1.00
2.00
8.00
14.3
7.1
50.0
1.00
2.50
7.1
100.0
2.00
14.3
2.9
1.4
Procedure:
A: Select N% of sample
B: select titrant concentration
C: Choose weight in order that the titrant consumption can be expected
between 3 and 17 mL
Liquid samples
Sample
[mL]
Titrant
N [%]
N mg/L
Titrant [N]
0.10.6
0.10
0.060.4
0.10
10
100200
0.01
25
50100
0.01
50
2050
0.01
100
1020
0.01
250
510
0.01
400
<5
0.01
8
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Sample preparation
Kjeldahl Tablets overview
Article
Composition
Weight
Titanium
# 11057980
Benefit:
Recommendation:
Time saving
Optimal compromise between environmental and performance
priorities.
3.71 g
1.59 g
Benefit:
Recommendation:
Missouri
# 11057982
4.98 g K2SO4
0.02 g CuSO4 5 H2O
Benefit:
Recommendation:
ECO
# 11057983
3.998 g K2SO4
0.002 g CuSO4
Benefit:
Recommendation:
Eco-friendly
Most environmentally friendly catalyst, due to the very low copper content
Antifoam
# 11057984
0.97 g Na2SO4
0.03 g Silikon Schaumhemmer
Benefit:
Recommendation:
Copper Micro
# 11057985
1.5 g K2SO4
0.15 g CuSO4 5 H2O
Benefit:
Recommendation:
5g
4g
1g
1.65 g
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Sample preparation
Amount of sulfuric acid
The amount of H2SO4 is given by:
1. Conversion of K2SO4 to KHSO4 (K2SO4 is a component of Kjeldahl Tablets)
ca. 23 mL
2. Consumption by organic matter
Organic matter
H2SO4/ g
[mL]
Example:
Salami
Fat
9.7
27.3%
Protein
4.9
20.6%
Carbohydrates
4.0
0.0%
4. Remaining volume
= amount of used Kjeldahl Tablet (e.g. 10 g Kjeldahl tablets = 10 mL H2SO4)
H2SO4 volume =
conversion + (total consumption by org. matter) + evaporation + remaining volume
3 mL + (3.97 + 1.51 + 0.00) mL + 1 mL + 10 mL = 18.48 mL ~18 mL
10
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The digestion
Hints
Inlet for ambient air
Optimal conditions are achieved when the condensation zone remains 5 cm below the constriction of the sample tube.
For foaming samples one Kjeldahl Tablet Antifoam or stearic acid can be used.
To reduce the digestion time H2O2 can be added.
When samples crystalize despite optimal H2SO4/catalyst ratio the suction power of the
Scrubber should be reduced.
The Scrubber must only be placed on the left of the digester.
Minimal use of chemicals due to micro-Kjeldahl (only SpeedDigester).
For liquid samples boiling rods prevent boiling delays.
Boiling rods can in contrast to boiling stones, also be used for the following automated analysis
via the KjelSampler.
Slowly increasing the digestion temperature reduces foam formation of problematic samples.
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Digestion
Digestion parameter: SpeedDigester (IR)
TKN using 500 mL sample tube
Digestion
Temp. [C]
T nominal
T sample
1)
Step
Preheating
380
Digestion
520
Time [min]
Cooling
1)
Temp. [C]
T nominal
T sample
45
10
50
25
60
50
80
100
135
250
185
400
30
Preheating
480
8.5
Digestion
480
550
490
8.5
9.5
8.5
Cooling
10
10
65
30
Temp. [C]
Time [min]
Preheating
480
8.5
Digestion
480
500
480
8.5
9.5
8.5
Cooling
10
5
45
15
Time [min]
12
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Digestion
Digestion parameter: KjelDigester (only 300 mL tubes)
Standard Kjeldahl digestion using Kjeldahl Tablets
To avoid foaming and over splashing of the
sample, it is heated up slowly (3 steps)
Temp. [C]
T nominal
Step
Temp. [C]
Preheating
300
Digestion
340
15
420
105
Cooling
Time [min]
35
T sample
Time [min]
Time [min]
Step
Temp. [C]
Preheating
330
Digestion
420
60
Cooling
25
T nominal
T sample
Time [min]
13
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Digestion
Highest sample throughput
Reduce heat-up / cool-down periods
Block: Tablets
IR: Tablets
65 min
120
1. Sample preparation
per batch: 40 min
batch
#1
batch
#2
batch
#3
batch
#4
batch
#5
batch
#6
Mixer B-400
batch #2
batch #3
batch #6
batch #4
cooling
batch #5
cooling
batch #4
cooling
batch #3
cooling
KjelDigester K-449
3. Distillation / Titration
per batch: 100 min
batch #2
cooling
batch #1
cooling
2. Digestion
per batch: 100 min
batch #5
(unattended)
KjelMaster K-375,
KjelSampler K-376 / K-377
t [min]
Start 8:00 am
Finish 5:00 pm
14
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Distillation
Overview
General overview
1. Dilution of the acidic digestion mixture
2. Alkalinization to convert NH4+ in NH3
3. Steam distillation to drive out the NH3
4. NH3 collection in acidic receiver
H2O addition
NaOH addition
H2O steam addition
acid (H3BO3 or H2SO4)
Distillation
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Distillation
Parameter for boric acid titration
Step
Why
How much
Rule-of-the-thumb
Step 1: Dilution
H2O dist.
4 mL per mL
used H2SO4
Step 2: Alkalinization
NaOH 32%
Conversion of NH4+ in
NH3 (gaseous)
1590 mL
4.5 mL per mL
used H2SO4
4070 mL
180300 s
Distillation time:
180 s with KjelMaster
240 s with others
Step 5: Collection
NH3
For the following analysis potentiometric as well as colorimetric titration can be chosen.
16
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Distillation
Parameter for back titration
Step
Why
How much
Rule-of-the-thumb
2590 mL
4 mL per mL
used H2SO4
1590 mL
4.5 mL per mL
used H2SO4
1020 mL
180300 s
Distillation time:
180 s with KjelMaster
240 s with others
Step 1: Dilution
H2O dist.
Step 2: Alkalinization
NaOH 32%
Conversion of NH4+ in
NH3 (gaseous)
17
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Distillation
Hints
After digestion the samples must cool-down to 50100 C before they can be further processed.
Optimal NaOH concentration is 32%
For very low N-contents 2% H3BO3 with KCI (3 g/L) should be chosen as receiving solution,
to achieve lower detection limits.
With KjelSampler samples can be processed overnight.
With the IntelliDist function of the KjelMaster preheating of the instrument can be avoided.
Distillation and titration can be synchronized by means of the Online-Titration.
Waste from the sample tube and the receiver can be collected separately with the
KjelMaster.
18
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Titration
Overview
Boric acid titration
Potentiometric titration
Colorimetric titration
1. Standard
2. Online
1. Standard
a) endpoint titration
Start volume before titration can be used for all titration variants.
Back titration
Potentiometric Titration
1. Standard
Advantage
a) endpoint titration
Disadvantage
Titration
Endpoint
pH of boric acid must be adjusted to 4.65
Startpoint
More boric acid required (compared to end
point titration), to detect the startpoint
Start volume
Huge time saving, higher sample throughput
Only for well-known titration volume
19
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Titration
Endpoint and Startpoint titration (potentiometric)
Endpoint titration
Parameter
What
Setting
Rule-of-the-thumb
1. Algorithm
Dosage steps
for titration
Optimal
2. Start volume
Titrant
HCI / H2SO4
3. Titration
Titrant
4. Endpoint
pH-value
pH 4.65
What
Setting
Rule-of-the-thumb
4. Startpoint
pH is
detected
pH-value before
NaOH addition
What
Setting
Rule-of-the-thumb
1. Algorithm
Dosage steps
for titration
Optimal
2. Start volume
Titrant NaOH
3. Titration
Titrant
4. Endpoint
Titrant NaOH
pH-value
Back titration
pH 7.00
20
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Titration
Endpoint titration (Colorimetric)
Endpoint titration
Parameter
What
Setting
Rule-of-the-thumb
1. Algorithm
2. Start volume
Titrant
HCI / H2SO4
3. Titration
Titrant
4. Endpoint
Color change
21
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Titration
Hints
Endpoint Titration Boric Acid
B(OH)3 + 2 H2O B(OH)4 + H3O+ (pKa = 9.24)
B(OH)3 + 2 H2O
Sher Indicator
Endpoint pH 4.65
B(OH)4 + H3O+
22
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Result calculation
Hints
The KjelMaster K-375 performs the calculation automatically.
The PC software KjelLink facilitates error-free sample data editing. In addition, the results can be transferred to a PC for printing or if necessary for recalculation.
To determine the optimal method parameters, the BUCHI Kjeldahl-calculator can be used as a
helpful tool.
[V(1) - V(Bl)] F c f M(N)
w(N) =
m 1000
% N = w(N) 100%
% P = w(N) PF 100%
Example (Titrant is 0.25 M H2SO4,f=1.000):
weight fraction of N
consumption of titrant, sample, [mL]
average consumption of titrant, blank, [mL]
molar reaction factor (1 = HCI, 2 = H2SO4)
concentration of titrant [mol /L]
factor of titrant
molecular weight of N (14,007 [g/mol])
sample weight [g]
conversion factor (mL in L)
Protein factor
% of weight of N
% of weight of protein
Result
w(N):
V(1):
V(Bl):
F:
c:
f:
M(N):
m:
1000:
PF:
% N:
% P:
23
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