Soil Biology & Biochemistry 43 (2011) 1563e1568

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Soil Biology & Biochemistry

journal homepage: www.elsevier.com/locate/soilbio

Quantitative analysis of biochar in eld soil

Roger T. Koide*, Krittika Petprakob, Matthew Peoples
Department of Horticulture and Graduate Program in Ecology, The Pennsylvania State University, University Park, PA 16802, USA

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 21 October 2010
Received in revised form
4 April 2011
Accepted 7 April 2011
Available online 22 April 2011

Biochar is used with increasing frequency as a soil amendment because of its potentially benecial
effects on soil carbon sequestration, crop yield, nutrient leaching and greenhouse gas emissions. Simple
methods for the analysis of biochar in soil, however, are currently unavailable. Therefore, we have
adapted the loss on ignition method for this purpose. The technique requires knowledge of the
proportions of both biochar and biochar-free soil that are lost on ignition. One can use values determined
prior to the amendment of the soil with biochar, assuming that the values do not change after biochar is
incorporated in the soil. We tested these assumptions. Over the course of 15 months, the assumptions
proved to be valid under our test conditions. The technique accurately determined a wide range of
biochar concentrations in eld soil.
2011 Elsevier Ltd. All rights reserved.

Keywords:
Analysis
Biochar
Carbon sequestration
Loss on ignition
Soil organic matter
Quantication

1. Introduction
Thermochemical conversion (pyrolysis) is one route for the
production of liquid fuel from biomass (Boateng et al., 2006). One of
the byproducts from pyrolysis is biochar, which may be useful as
a soil amendment due to its unusual chemical and physical characteristics. Biochar contains high concentrations of carbon that can
be rather recalcitrant to decomposition, so it may stably sequester
carbon (Glaser et al., 2002). It can increase soil aeration (Laird,
2008) and reduce soil emissions of N2O, a greenhouse gas
(Spokas et al., 2009; Singh et al., 2010). It has increased crop yield
through various mechanisms including stimulation of benecial
soil microbes such as mycorrhizal fungi (Warnock et al., 2007),
increase of soil base saturation (Glaser et al., 2002; Major et al.,
2010a,b), increase in water holding capacity (Glaser et al., 2002;
Steiner et al., 2007), and retention of nutrients in the portion of
the soil column containing roots, thus improving nutrient use
efciency (Chan et al., 2007; Steiner et al., 2008).
There are multiple situations in which it might be important to
measure the amount of biochar in soil. Effective and fair carbon
sequestration compensation schemes may require documenting
both the temporal and spatial stability of biochar. Biochar is capable

* Corresponding author. Tel.: 1 814 863 0710; fax: 1 814 863.6139.

E-mail addresses: rxk13@psu.edu (R.T. Koide), kup146@psu.edu (K. Petprakob),
peoplma@gmail.com (M. Peoples).
0038-0717/$ e see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.soilbio.2011.04.006

of migrating in soils (Major et al., 2010a,b) and, while it is generally

assumed to be recalcitrant, a portion of most biochars, particularly
fresh biochars, is easily decomposable (Lehmann et al., 2009).
Therefore, repeated analyses of biochar in time and space become
necessary. Moreover, because biochar has the potential to improve
crop growth and reduce leaching and greenhouse gas emissions
(see above), its cost-effective use will require knowledge of the
relationship between biochar quantity and the magnitude of these
benets.
Despite the clear need for a simple and routine way to quantify
biochar in soils, the complex chemistry of biochar and the difculty
in distinguishing biochar from other forms of organic matter has
resulted in methods that are either extremely labor intensive or
require specialized instrumentation, both of which are impractical
for routine analysis by most scientists (Manning and Lopez-Capel,
2009). For example, biochar in soils has been quantied by handsorting (Zackrisson et al., 1996), in situ analysis via scanning calorimetry, NMR spectroscopy or infrared spectroscopy (Schmidt and
Noack, 2000; Manning and Lopez-Capel, 2009; Nguyen et al.,
2009), analysis of molecular markers (Manning and Lopez-Capel,
2009), or by preferential removal of inorganic and non-biochar
organic C by selective oxidation or acid treatment followed by
analysis of residual organic material (presumably equivalent to
biochar) by NMR, optical, or mass spectroscopy, or thermal
conductivity (Schmidt and Noack, 2000; Elmquist et al., 2004;
Manning and Lopez-Capel, 2009). None of these methods is
suited to routine analysis of biochar in soils by most agronomists or

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R.T. Koide et al. / Soil Biology & Biochemistry 43 (2011) 1563e1568

ecologists engaged in soil amendment studies. Our goal, therefore,

was to adapt the familiar loss on ignition method for use in the
routine analysis of biochar in soil.
2. Material and methods
2.1. Development of the method
Our method for analysis of biochar in soils is based on the loss
on ignition method for the analysis of soil organic matter (Heiri
et al., 2001). Loss on ignition is considered to be superior to wet
combustion for the estimation of soil organic matter because it
accounts for a greater fraction of the organic matter present in the
sample, but it can overestimate soil organic matter if there is
substantial carbonate present in the sample (Byers et al., 1978). A
temperature of 550  C is generally assumed to oxidize the
conventional (non-thermally altered) organic matter in most soils
(Ball, 1964; Heiri et al., 2001). In preliminary tests we determined
that 550  C also thermally oxidized the organic portion of biochar
produced from hardwood via conventional charcoaling (slow
pyrolysis) methods. We therefore subsequently used 550  C in all
experiments of this study.
Because both conventional soil organic matter and biochar are
oxidized in the same process, the weight that is lost on ignition
from a soil sample containing biochar can be given in the following
equation:

LI OM CHLI

(1)

where:
LI is the total weight of the sample that is lost on ignition,
OM is the weight of the native, non-biochar organic matter
resident in the soil (frequently referred to as thermally unaltered
organic matter, Schmidt and Noack, 2000) that is lost on ignition, and
CHLI is the weight of the biochar in the soil sample that is lost on
ignition.
The portion of the biochar in the soil sample exposed to 550  C
that is not lost on ignition is the mineral component of biochar or
ash. Our soil, described below, did not contain measurable
concentrations of carbonate.
In the absence of biochar, OM is operationally dened as that
portion of the soil that is lost on ignition (Heiri et al., 2001). It is
assumed to be the non-biochar organic matter resident in the soil.
Therefore, in a soil sample containing biochar:

OM qW  CH

(2)

where:
q is the proportion (by weight) of pure soil (without biochar)
that is lost on ignition,
W is the weight of the soil sample containing biochar, and
CH is the weight of the biochar in the sample (including that
which is lost on ignition plus the ash component of the biochar that
remains after combustion).
CHLI, the weight of biochar lost on ignition, can be given by the
following equation:

CHLI y CH

(3)

where:
y is the proportion (by weight) of biochar that is lost on ignition.
We assume this is the organic fraction of the biochar.
Substituting Eqs. (2) and (3) into Eq. (1), we obtain:

LI qW  CH y CH
Rearranging the equation to solve for CH, we obtain:

(4)

CH LI  q W=y  q

(5)

Thus, the weight of biochar in a soil sample can be calculated

from the weight that is lost on ignition from the soil sample containing biochar, the weight of the soil sample containing biochar,
the proportion of pure soil (not including the biochar) that is lost
on ignition (q), and the proportion of the biochar that is lost on
ignition (y).
2.2. Testing the assumptions of the method
The proposed method for biochar analysis assumes knowledge
of q, the proportion of the pure soil (without biochar) that is lost on
ignition, and y, the proportion of the pure biochar (without soil)
that is lost on ignition. We imagine that in most cases it will be
difcult to separate biochar from a soil sample, especially if the
biochar is ne, in order to routinely determine either q or y.
However, in cases in which biochar is experimentally added to soil,
samples of pure biochar can be analyzed to determine y and
samples of pure soil can be analyzed to determine q. In order for
that approach to be useful, however, one must assume that y and q
do not change as a consequence of residence of the biochar in the
soil. We tested these assumptions by adding biochar to soil in the
eld and, 15 months later, collecting eld samples and separating
the biochar from the remaining soil (see below) in order to determine whether q and y remained unaltered.
In June 2009, we rototilled to a depth of 25 cm a portion of a eld
formerly planted to corn at the Russell Larson Research and
Education Center. Its location is 40 420 45.6500 N, 77 570 26.2100 W.
This eld was not the source of the soil described in Section 2.3, but
it possesses the very same soil type. The soil is Hagerstown soil:
ne, mixed, semiactive, mesic Typic Hapludalfs (USDAeNRCS,
2010). The surface texture is a silt loam with subsurface textures
of a silty clay loam and silty clay. After rototilling, we laid out 4-1 m2
plots in each of three blocks. Two plots in each block were amended
with biochar while two plots were not and served as controls. Plots
were separated from each other by 2 m. The biochar plots received
approximately 5.7 kg (12.5 lbs) biochar, equivalent to approximately 56.8 tonne ha1. The biochar was produced from domestic
hardwood via conventional, slow pyrolysis and obtained from
Humphrey Charcoal (Brookville, PA, USA). In blocks 2 and 3 we used
mesh size #6 biochar; the pieces of biochar were smaller than
3.4 mm in the smallest dimension. In block 1 we used mesh size
#10 biochar; the pieces of biochar were smaller than 1.7 mm in the
smallest dimension. Other than size, the biochars were identical.
After spreading the biochar uniformly over the plots, we mixed it
into the soil with spades to a depth of approximately 25 cm, turning
the soil 3e4 times for thorough mixing. The area was then planted
to sweet corn (Delectable, Rupp Seeds Wauseon, OH, USA), and the
corn was harvested later that season. In May 2010, the same area
was planted to soybean (FS H535A90, Growmark, Bloomington, IL,
USA). In September 2010, when the biochar had been in the soil for
15 months, we collected soil samples to a depth of 15 cm from each
of the 12 plots using a standard 2.5 cm diameter soil sampler
(Oakeld Apparatus, Inc., Oakeld, WI, USA). Two cores were taken
from each control (no biochar) plot and four cores were taken from
each biochar plot.
We then separated out approximately 15 ml of biochar pieces
from each biochar plot soil sample. The separation was facilitated
by the fact that the biochar pieces were relatively large (see above).
We employed a 5 mm soil sieve to perform the initial separation of
biochar from soil. Most of the biochar in the sample remained on
the sieve. The biochar pieces and soil aggregates remaining on the
sieve were easily separated. The soil aggregates were retained as
part of the pure soil fraction. The smaller aggregates of soil passing

R.T. Koide et al. / Soil Biology & Biochemistry 43 (2011) 1563e1568

through the sieve were picked free of biochar using forceps. The soil
aggregates that passed through the sieve were added to the larger
aggregates remaining on the sieve and collectively comprised the
pure soil fraction. The smaller pieces of biochar were added to the
larger pieces retained by the sieve to comprise the biochar fraction.
The biochar samples separated from soil were cleaned of
adhering soil by placing them overnight in 50 ml polyethylene
centrifuge tubes lled with a 0.5% (by weight) aqueous solution of
sodium hexametaphosphate, (NaPO3)6. The tubes were then
shaken vigorously to dislodge the soil particles, and the biochar was
rinsed four times with distilled water. They were then allowed to
soak in distilled water for a few hours before being drained and
dried in an oven at 70  C for temporary storage.
We performed loss on ignition analyses of samples of control
plot soils, which never contained biochar (10 g), samples of pure
biochar separated from the biochar-amended soil plots (2.5 g),
samples of pure soil separated from the biochar-amended soil plots
(10 g), as well as three samples of biochar (2.5 g) that had never
been added to soil. Loss on ignition was determined as the difference in weight between the sample after drying at 105  C and after
heating in a mufe furnace at 550  C for 4 h. The values of q and y
were determined from these measurements according to the
explanations given beneath Eqs. (2) and (3), respectively. By
comparing biochar that had not been added to soil with biochar
that had been in the soil for 15 months, we tested the assumption
that y did not change. By comparing soil from the control plots with
soil separated from biochar-amended soil plots, we tested the
assumption that q did not change.
2.3. Testing the method
We tested our method by comparing actual and calculated
biochar contents of approximately 5 g samples of eld soil to which
we added varying amounts of biochar. There were two replicates
each of 1%, 2%, 5% and 10% biochar by weight. The biochar, supplied
by Humphrey Charcoal (Brookville, PA, USA), was the same as used
above (Section 2.2) but, in this case, the biochar was ground to
a ne powder with mortar and pestle before mixing with eld soil
in order to increase the homogeneity of the samples. The soil was
obtained from an agricultural eld of the Russell Larson Agricultural Research and Education Center of the College of Agricultural
Sciences, Penn State University, located in Rock Springs, PA, USA,
40 420 44.4300 N, 77 570 19.2900 W. This eld was not the source of the
soil described in Section 2.2, but it possesses the same Hagerstown
soil (Fine, mixed, semiactive, mesic Typic Hapludalfs) (USDAeNRCS,
2010). The soil was collected from the top 15 cm and air dried and
sieved to pass a 2 mm sieve prior to mixing with the biochar. In
addition to the four concentrations of biochar in soil (above),
duplicate samples of 100% soil (no biochar) and 100% biochar (no
soil) were also prepared for loss on ignition analysis in order to
determine the values of q and y. In all cases, weight loss on ignition
was calculated as the difference between the weight of samples
after drying at 105  C and their weight following 4 h at 550  C in
a mufe furnace (Barnstead Thermolyne 48000, Dubuque, IA, USA).
In the 5 cm (diameter) ceramic crucibles we employed for the loss
on ignition procedure, the 4 h duration of heating was sufcient to
completely oxidize 5 g sample of pure biochar to ash, and to
completely oxidize the biochar in the 5 g soil samples.

1565

the proportion of the pure soil lost on ignition (p 0.53) or y, the

proportion of biochar lost on ignition (p 0.88). There was no
signicant effect of the 15 month residence time of biochar in the
eld soil on q; according to the analysis of variance, the q of the soil
in the control plots (mean 0.045, se 0.002) was not signicantly
different (p 0.79) from that of the soil separated from the biocharamended soil plots (mean 0.046, se 0.002). Moreover, there was no
signicant effect of the 15 month residence time of biochar in the
eld soil on y; the y of the biochar taken from the biochar-amended
soil plots (mean 0.878, se 0.010) was not signicantly different
(p 0.42) from that of the biochar that had never been added to soil
(mean 0.861, se 0.009).
3.2. Testing the method
The average q of the duplicate pure soil samples was 0.0429 (se
0.0003). The average y of duplicate pure biochar samples was 0.861
(se 0.009). Using these values and Eq. (5), we found a linear relationship between actual and calculated biochar weights in the
prepared soilebiochar mixtures described above (Fig. 1). The
mixtures ranged from 0% biochar up to 10% biochar (by weight).
The slope was very close to one (0.960) and there was little scatter
in the data (r2 0.996). Therefore, the method proved to accurately
determine the actual weight of biochar in each sample over a wide
range of biochar concentrations in soil.
4. Discussion
There are currently no standard methods for the analysis of
biochar in soils. In most cases, biochar content of soils has been
determined by laborious or instrument-intensive techniques
(Zackrisson et al., 1996; Schmidt and Noack, 2000; Elmquist et al.,
2004; Manning and Lopez-Capel, 2009; Nguyen et al., 2009).
None of these is suitable for routine analysis in typical ecological or
agronomic studies due to the requirement for specialized apparatus
or the sheer labor involved in hand-sorting. Because of the
simplicity of the instrumentation involved (mufe furnace and
balance), our method of analyzing biochar in soil samples is ideally

3. Results
3.1. Testing the assumptions of the method
Despite using different size biochar in block 1 compared to
blocks 2 and 3, there were no signicant effects of block on either q,

Fig. 1. Plot of actual biochar weight in sample vs. calculated biochar weight in sample
based on use of Eq. (5), assuming q 0.0429 and y 0.861. The data points represent
duplicate samples of biochar mixed in eld soil at 0, 1, 2, 5 and 10% biochar by weight.

1566

R.T. Koide et al. / Soil Biology & Biochemistry 43 (2011) 1563e1568

suited to routine determinations. It is far simpler than other

methods that require more sophisticated instrumentation, the
results of which are difcult to interpret and do not yield biochar
weights directly (Schmidt and Noack, 2000; Manning and LopezCapel, 2009). Moreover, our method does not assume that
biochar and non-thermally altered soil organic matter are distinguishable either analytically or in terms of their recalcitrance to
oxidation. Instead, our adaptation of the loss on ignition methodology involves the thermal oxidation of all forms of organic
matter in the soil, including biochar.
The method does require knowledge of q, the proportion of
weight of pure soil that is lost on ignition and y, the proportion of
weight of biochar that is lost on ignition. Heterogeneity among soil
samples in organic matter content such as caused by random bits of
plant litter will result in variation in q from sample to sample, but
such variation cannot by itself alter the mean of the calculated
biochar content as long as a sufcient number of samples are
analyzed. While we have tested our method only on biochar
manufactured from hardwood via conventional, slow pyrolytic
charcoaling methods, it should also be applicable to other forms of
biochar that can be thermally oxidized using the conventional loss
on ignition conditions as long as y is determinable.
When biochar cannot be separated from soil samples in order to
determine q and y such as was done in this study, it may be
acceptable to use values for q and y determined prior to the addition of biochar to the soil. Even when biochar was incorporated into
a eld soil for a period of 15 months during which a typical cropping rotation occurred (corn and soybeans), q and y did not change
signicantly in the test soil using the test biochar. In other soils or
with other types of biochar, q and y could behave differently. The
assumptions of temporal stability of q and y would have to be
validated for each study.
Indeed, we recognize the potential for y to change following
the addition of biochar to soil. For example, y may be reduced if,
over time, ash comprises a greater proportion of the remaining
weight as the easily decomposable fraction of biochar (Lehmann
et al., 2009) decomposes or as the volatile fraction is lost (Brewer
et al., 2009). Moreover, as biochar ages in the soil, it may become
enriched in oxygen and depleted in carbon (Cheng et al., 2008;
Hammes and Schmidt, 2009; Nguyen et al., 2009), and this
could also possibly alter the value of y. All of these phenomena
could be inuenced by the quality of the biochar, which is inuenced by choice of feedstock and pyrolysis conditions (Brewer
et al., 2009).
The value of q could be altered by biochar addition to soil
because root growth can be stimulated by biochar in soil (Makoto
et al., 2010). Moreover, biochar can inuence microbial communities, and soil microbial communities may inuence the quality and
quantity of soil OM. For example, biochar may stimulate mycorrhizal fungi (Watrud et al., 1978; Saito, 1989; Ishii et al., 1999; Ezawa
et al., 2002; Yamato et al., 2006; Rondon et al., 2007; Warnock et al.,
2007), and mycorrhizal fungi produce a soil-aggregating material
called glomalin, a decomposition-resistant glycoprotein that
apparently comprises a large fraction of stable organic matter in
soils (Wright and Upadhyaya, 1996, 1998; Treseder and Allen, 2000;
Rillig et al., 2001). Mycorrhizal fungi also signicantly enhance soil
structure and erosion resistance due to the production of glomalin
and of vast lengths of hyphae (see Miller and Jastrow, 1992;
Gianinazzi and Schuepp, 1994; Boswell et al., 1998; Kabir and
Koide, 1999). Soil aggregation stabilizes organic matter against
rapid decomposition by reducing access by microorganisms (Elliott
and Coleman, 1988; Jastrow, 1996; Jastrow and Miller, 1998; Tisdall
and Oades, 1979; Tisdall et al., 1997; Jastrow et al., 2005). Soil
aggregation also reduces soil erosion, which further limits
decomposition of organic matter in the soil (Peterson et al., 1998).

If, by altering microbial activity, biochar inuences the quality of

thermally unaltered soil organic matter, its presence could, over
time, inuence the value of q.
While we did not observe signicant changes in q or y following
15 months of biochar in soil, because of the potential for change we
tested whether signicant under- or overestimations of either q or y
could result in signicant error in the calculation of biochar weight.
We approached this problem by calculating biochar weights rst by
assuming y 0.861 while assuming q was either 90% or 110% of the
assumed correct value of 0.0429 (q 0.0472 and q 0.0386,
respectively) and, secondly, by assuming q 0.0429 while
assuming y was either 90% or 110% of the assumed correct value of
0.861 (y 0.947 and y 0.775, respectively). In the rst instance,
the results are shown in Fig. 2. In the second, the results are shown
in Fig. 3.
Fig. 2 indicates that in our system a 10% error in the value of q
results in a nearly constant absolute error in biochar weight of
approximately 0.025 g across the range of tested weights of
biochar from 0 to nearly 0.5 g per sample (equivalent to concentrations of biochar from 0 to 10% of soil by weight). A systematic
absolute error may be acceptable, particularly for studies in which
comparisons among treatments are the major consideration.
Fig. 3 indicates that a 10% error in the value of y results in
a variable absolute error in the calculation of biochar weight. At the
lowest actual biochar weight (0.05 g, equivalent to 1% biochar
concentration) the absolute error is comparatively small (approximately 0.005 g). At the highest actual biochar weight (close to
0.5 g, amounting to 10% biochar concentration) the absolute error is
comparatively large (approximately 0.05 g). However, the percent
error was approximately the same across the entire range of tested
weights of biochar; a 10% error in y resulted in an approximately
10% error in the calculation of biochar weight, irrespective of biochar weight. Given our results, it does not seem likely that in the
experimental use of biochar in eld soils y would change as much
as 10%. Thus, the error in y for our test circumstances would appear
to be acceptable given the fact that no other analytical method is
capable of accurately determining biochar weight. However, our

Fig. 2. Plot of actual biochar weight in sample vs. three calculated values of biochar
weight in sample based on use of Eq. (5), assuming y 0.861 and three values of q
(0.0429 and 90% and 110% of 0.0429). The data points represent duplicate samples of
biochar mixed in eld soil at 0, 1, 2, 5 and 10% biochar by weight.

R.T. Koide et al. / Soil Biology & Biochemistry 43 (2011) 1563e1568

Fig. 3. Plot of actual biochar weight in sample vs. three calculated values of biochar
weight in sample based on use of Eq. (5), assuming q 0.0429 and three values of y
(0.861 and 90% and 110% of 0.861). The data points represent duplicate samples of
biochar mixed in eld soil at 0, 1, 2, 5 and 10% biochar by weight.

assumptions obviously require validation for each soil and biochar

type.
In some soils, 20e25 tonne biochar ha1 have been applied with
benecial effect (Major et al., 2010a,b; Jin et al., 2007). If we assume
that the biochar is placed in the top 15 cm of soil and that the soil
has a dry specic gravity of 1.2 g cm3, the concentration of biochar
applied at the rate of 25 tonne ha1 would be 1.4% of soil dry
weight, well within the range of this study (0e10%). We conclude
that this method, based on the loss on ignition methodology, is
capable of accurately determining biochar in eld soils at realistic
concentrations assuming that both y and q are known.

Acknowledgments
We thank Elizabeth Reid for help in laying out the plots of the
eld study and two anonymous reviewers for suggestions leading
to improvement of this manuscript. We acknowledge nancial
support from the Pennsylvania Soybean Promotion Board, the
Northeast Sun Grant, the USDA NIFA competitive grants program,
and the College of Agricultural Sciences and Department of Horticulture of the Pennsylvania State University.

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