Poster Note 42250
Carbon / Nitrogen Ratio in Soils
and Plants using The FLASH 2000
Organic Elemental Analyzer
Carbon / Nitrogen Ratio in Soils and Plants using The FLASH 2000
Liliana Krotz and Guido Giazzi
Thermo Fisher Scientific, Milano, Italy
Liliana Krotz and Guido Giazzi
Thermo Fisher Scientific, Milano, Italy
Overview
Overview
Purpose: Nitrogen
Nitrogen and
and carbon
carbon determination
determination by
by Organic
Organic Elemental
Elemental Analysis
Analysis (OEA).
(OEA).
Purpose:
Methods: Soils
Soils and
and plants
plants were
were analyzed
analyzed through
through an
an elemental
elemental analyzer
analyzer with
with
Methods:
automatic autosampler.
autosampler. The
The C/N
C/N ratio
ratio is
is calculated
calculated automatically
automatically by
by the
the dedicated
dedicated
automatic
Thermo
Scientific
Eager
Xperience
Data
Handling
Software.
Thermo Scientific Eager Xperience Data Handling Software.
Results: Data
Data collected
collected of
of nitrogen
nitrogen and
and carbon
carbon percentages
percentages and
and the
the relative
relative C/N
C/N ratio
ratio
Results:
from different
different soils
soils and
and plants
plants are
are showed
showed to
to assess
assess the
the performance
performance of
of the
the FLASH
FLASH
from
2000 Analyzer.
Analyzer.
2000
Introduction
Introduction
The carbon/nitrogen
carbon/nitrogen ratio
ratio of
of the
the organic
organic material
material added
added to
to the
the soil
soil affects
affects the
the rate
rate of
of
The
decomposition of
of organic
organic matter
matter and
and itit results
results in
in the
the release
release (mineralization)
(mineralization) or
or
decomposition
immobilization of
of soil
soil nitrogen.
nitrogen. IfIf the
the added
added organic
organic material
material contains
contains more
more nitrogen
nitrogen in
in
immobilization
proportion to
to the
the carbon,
carbon, then
then nitrogen
nitrogen is
is released
released in
in the
the soil
soil from
from the
the decomposing
decomposing
proportion
organic material,
material, while
while ifif the
the organic
organic material
material has
has aa less
less amount
amount of
of nitrogen
nitrogen in
in relation
relation
organic
to the
the carbon
carbon then
then the
the microorganisms
microorganisms will
will utilize
utilize the
the soil
soil nitrogen
nitrogen for
for further
further
to
decomposition and
and the
the soil
soil nitrogen
nitrogen will
will be
be immobilized
immobilized and
and will
will not
not be
be available.
available.
decomposition
In few
few words,
words, cellular
cellular carbon
carbon and
and nitrogen
nitrogen metabolism
metabolism must
must be
be tightly
tightly coordinated
coordinated to
to
In
sustain optimal
optimal growth
growth and
and development
development for
for plants.
plants. Furthermore,
Furthermore, C/N
C/N balance
balance is
is also
also
sustain
.
critical
for
the
ecosystem
response
to
elevated
atmospheric
CO
critical for the ecosystem response to elevated atmospheric CO22.
For these
these reasons
reasons the
the use
use of
of an
an accurate
accurate instrumental
instrumental analytical
analytical technique
technique is
is required.
required.
For
As the
the demand
demand for
for improved
improved sample
sample throughput,
throughput, reduction
reduction of
of operational
operational costs
costs and
and
As
minimization of
of human
human errors
errors is
is increasing,
increasing, itit is
is very
very important
important apply
apply aa simple
simple and
and
minimization
automated technique
technique which
which allows
allows fast
fast analysis
analysis with
with excellent
excellent reproducibility.
reproducibility.
automated
The Thermo
Thermo Scientific™
Scientific™ FLASH™
FLASH™ 2000
2000 Organic
Organic Elemental
ElementalAnalyzer,
Analyzer, based
based on
on the
the
The
dynamic flash
flash combustion
combustion of
of the
the sample,
sample, meets
meets effortlessly
effortlessly the
the wide
wide array
array of
of laboratory
laboratory
dynamic
requirements such
such as
as accuracy,
accuracy, day
day to
to day
day reproducibility
reproducibility and
and high
high sample
sample throughput.
throughput.
requirements
FI
FI
FIGURE 1.
1. Thermo
Thermo Scientific
Scientific FLASH
FLASH 2000
2000 NC
NC Soils
Soils Analyzer.
Analyzer.
FIGURE
Methods
Methods
The FLASH
FLASH 2000
2000 Elemental
Elemental Analyzer
Analyzer operates
operates according
according to
to the
the dynamic
dynamic flash
flash
The
combustion of
of the
the sample.
sample. Samples
Samples are
are weighted
weighted in
in tin
tin capsules
capsules and
and introduced
introduced into
into
combustion
the combustion
combustion reactor
reactor via
via the
the Thermo
Thermo Scientific™
Scientific™ MAS
MAS 200R
200R Autosampler
Autosampler together
together
the
with oxygen.
oxygen.
with
After combustion,
combustion, the
the produced
produced gases
gases are
are carried
carried by
by aa helium
helium flow
flow to
to aa second
second reactor
reactor
After
filled with
with copper,
copper, then
then swept
swept through
through aa H
H22O
O trap,
trap, aa GC
GC column
column and
and finally
finally detected
detected by
by
filled
thermal conductivity
conductivity detector
detector (TCD).
(TCD). (Figure
(Figure 2).
2). AAcomplete
complete report
report is
is generated
generated by
by
aa thermal
the Thermo
Thermo Scientific
Scientific Eager
Eager Xperience
Xperience Data
Data Handling
Handling Software
Software and
and displayed
displayed at
at the
the
the
end of
of the
the analysis.
analysis. Table
Table 11 shows
shows the
the analytical
analytical conditions
conditions used
used for
for analysis.
analysis. Figure
Figure 33
end
shows aa typical
typical NC
NC chromatogram.
chromatogram.
shows
Analytical Conditions
Conditions
Analytical
Left Reactor
Reactor Temperature
Temperature 950°C
950°C
Left
(combustion)
(combustion)
Right Reactor
Reactor Temperature
Temperature 840°C
840°C (reduction)
(reduction)
Right
Oven Temperature
Temperature 50°C
50°C
Oven
Sample Delay
Delay 12
12 sec
sec
Sample
Helium Carrier
Carrier Flow
Flow 140
140 ml/min
ml/min
Helium
Helium Reference
Reference Flow
Flow 100
100 ml/min
ml/min
Helium
Oxygen Flow
Flow 250
250 ml/min
ml/min
Oxygen
Oxygen Injection
Injection Time
Time 10
10 sec
sec
Oxygen
FIG
FIG
R
R
To
To
an
ana
Re
Re
Total Run
Run Time
Time 360
360 sec
sec
Total
TABLE 1.
1. Analytical
Analytical Conditions.
Conditions.
TABLE
Th
Th
ca
cal
Re
Re
an
and
Ta
ca
S
th
T
FIGURE
Thermo
Scientific
FLASH
2000
NCconfiguration.
configuration.
FIGURE
2.2.
Thermo
Scientific
FLASH
2000
NC
D
r
t
t
FIGURE
Typical
nitrogen
calibration
curve.
FIGURE
4.4.
Typical
nitrogen
calibration
curve.
r
3
n)
FIGURE
Typical
NC
chromatogram.
FIGURE
3.3.
Typical
NC
chromatogram.
Results
Results
demonstrate
the
performance
the
system
NC
Soil
configuration,
different
ToTo
demonstrate
the
performance
ofof
the
system
in in
NC
Soil
configuration,
different
analytical
tests
were
performed
and
showed
this
paper
using
five
instruments.
analytical
tests
were
performed
and
showed
in in
this
paper
using
five
instruments.
Repeatability
and
Reproducibility
were
confirmed
follow:
Repeatability
and
Reproducibility
were
confirmed
asas
follow:

Repeatability:
Thermo
Scientific
Soil
Reference
Material
was
analyzedin in
 Repeatability: Thermo Scientific Soil Reference Material
was
analyzed
triplicate
using
the
same
instrument
evaluate
the
accuracy
and
precision
triplicate
using
the
same
instrument
toto
evaluate
the
accuracy
and
precision
the
results.
The
average
the
data
obtained
was
compared
with
the
ofof
the
results.
The
average
ofof
the
data
obtained
was
compared
with
the
certified
values
and
the
acceptable
range,
following
the
technical
certified
values
and
the
acceptable
range,
following
the
technical
specification
the
system.
This
demonstrates
the
complete
conversion
specification
ofof
the
system.
This
demonstrates
the
complete
conversion
ofof
the
elements
without
any
memory
effect
changing
the
matrix.
the
elements
without
any
memory
effect
changing
the
matrix.
Reproducibility:
data
obtained
different
instruments
using
the
same
  Reproducibility:
data
obtained
ofof
different
instruments
using
the
same
reactors
and
same
gases
cylinders
were
compared
evaluate
the
reactors
and
same
gases
cylinders
were
compared
toto
evaluate
the
reproducibility
between
the
systems.
reproducibility
between
the
systems.
The
calibration
was
performed
with
4-5
mg
aspartic
acid
standard
using
factor
The
calibration
was
performed
with
4-5
mg
aspartic
acid
standard
using
KK
factor
asas
calibration
method.
Then,
three
analysis
50-100
mg
Thermo
Scientific
Soil
calibration
method.
Then,
three
analysis
ofof
50-100
mg
Thermo
Scientific
Soil
Reference
Material
were
performed
unknown.
Figure
and
show
typical
nitrogen
Reference
Material
were
performed
asas
unknown.
Figure
44
and
55
show
typical
nitrogen
and
carbon
calibration
curves.
and
carbon
calibration
curves.
FIGURE
Typical
carbon
calibration
curve.
FIGURE
5.5.
Typical
carbon
calibration
curve.
Table
shows
the
correlation
between
the
certified
NC
percentages
the
Soil
Table
2 2shows
the
correlation
between
the
certified
NC
percentages
ofof
the
Soil
Reference
Material
analyzed
unknown,
the
accepted
range
according
the
Reference
Material
analyzed
asas
unknown,
the
accepted
range
according
toto
the
technical
specification
the
system
and
the
experimental
data
obtained
triplicate
technical
specification
ofof
the
system
and
the
experimental
data
obtained
in in
triplicate
each
instrument.
data
are
acceptable
and
comparable,
confirming
the
forfor
each
instrument.
AllAll
data
are
acceptable
and
comparable,
confirming
the
repeatability
and
reproducibility
expected.
repeatability
and
reproducibility
expected.
Technical
Technical
Specification
Specification
FLASH
2000
Elemental
Analyzers
FLASH
2000
Elemental
Analyzers
Elem. Theor. Range
Elem. Theor. Range
%
N
C
%
(±)
(±)
T
M
1
%
%
1
2
RSD%
RSD%
%
%
2
RSD%
RSD%
3
%
%
4
RSD%
RSD%
%
%
4
RSD%
RSD%
5
%
%
RSD
RSD
%
%
0.187
0.187
0.187
0.186
0.02 0.187
0.187
0.186
0.02
0.187
C
1.688
1.687
1.699
1.697
1.691
1.688
1.687
1.699
1.697
1.691
1.708
1.680
1.701
1.694
1.683
1.697
0.02 1.708
0.679 1.680
0.304 1.701
0.212 1.694
0.302 1.683
0.595
1.708 0.679
1.690 0.304
1.706 0.212
1.704 0.302
1.703 0.595
1.697
0.02
1.708
1.690
1.706
1.704
1.703
0
0
0
0.186
0.186
0.186
0.186
0.185
0.186
0.820 0.186
0.185
0.188 0.820
0.186
0.188
0.186
5
N
0
0.186
0.186
0.186
0.186
0.185
0.186
0.311 0.186
0.185
0.185 0.311
0.186
0.185
0.186
3
TABLE2.2.
NC
data
Thermo
Scientific
Soil
Reference
Material.
TABLE
NC
data
ofof
Thermo
Scientific
Soil
Reference
Material.
2 Carbon / Nitrogen Ratio in Soils and Plants using The FLASH 2000 Organic Elemental Analyzer
0
0
Table 3 shows the ratio C/N for each NC determination in the five instruments
calculated automatically by the Thermo Scientific™ Eager Xperience Data Handling
Software, the values obtained are comparable indicating the excellent reproducibility of
the FLASH 2000 Elemental Analyzer.
Reference Material
Information
FLASH 2000 Elemental Analyzers – C/N ratio
1
2
3
4
5
23.41
23.68
23.69
23.38
23.39
23.51
23.36
23.40
23.40
23.38
23.43
23.13
23.47
23.29
23.60
TABLE 3. C/N ratio of Thermo Scientific Soil Reference Material.
FIGURE
Typical
calibration
curve. were analyzed to evaluate the
Different5.Soils
and carbon
Plants Reference
Materials
repeatability in a large range of concentrations. Table 4 shows the certified values and
the relative uncertainty, while Table 5 shows the experimental data obtained indicating
the excellent
Table
2 shows comparison.
the correlation between the certified NC percentages of the Soil
Elem.
N
2
0.133
RSD%
0.27
%
Loamy Soil
0.187
0.187
0.187
0.27
0.186
0.185
0.185
Sandy Soil
C
N%
1
%
0.02
Birch
1.697 Leaves
0.02
C%
Uncertainty (±)
0.023
1.61
0.09
Medium Organic
Content Soil
0.27
0.02
3.19
0.07
Loamy Soil
0.27
0.02
2.75
0.12
Chalky Soil
0.35
0.02
5.39
0.09
Sandy Soil
0.07
0.01
0.83
0.05
Birch Leaves
2.12
0.06
48.09
0.51
Orchard Leaves
2.28
0.04
50.40
0.40
Alfalfa
3.01
0.20
N.A.
-
FLASH 2000 Elemental Analyzers
Uncertainty (±)
Uncertainty (±)
C%
(±)
%
Medium
Organic
Content Soil
0.186
Uncertainty (±)
0.133
Specification
Low
Theor.Organic
Range
Content Soil
Chalky Soil
N%
Low Organic
Content Soil
TABLE 4. Certified Nitrogen and Carbon data of Soil and Plants Reference
Materials.
Reference Material analyzed as unknown, the accepted range according to the
technical specification of the system and the experimental data obtained in triplicate
for each instrument. All data are acceptable and comparable, confirming the
repeatability and reproducibility expected.
Reference Material
Information
Technical
Specification
Specification
1.688
1.708
1.708
0
0.35
0.07
2.12
0.679
RSD%
0.02
1.687
1.680
1.690
3
0.023
%
0.020.186
0.311
0.186
0.186
0.02
0.01
1.699
1.701
0.3040.06
1.706
1.61
RSD%
0
%
3.19
4
0.09 5
RSD%
0.186
2.75
0.185
0.188
5.39
0.820
0.83
1.697
1.694
0.212 48.09
1.704
%
0.07
0.186
0.12
0.186
0.186
0.09
0
0.05
1.691
0.302
1.683
0.51
1.703
Orchard Leaves
2.28
0.04
50.40
0.40
Alfalfa
3.01
0.20
N.A.
-
TABLE 2. NC data of Thermo Scientific Soil Reference Material.
RSD
%
0.595
TABLE 4. Certified Nitrogen and Carbon data of Soil and Plants Reference
Materials.
SD
%
0
95
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