Science of the Total Environment 658 (2019) 1106–1116

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Science of the Total Environment

j ou rn al ho mep age: w w w . e l s e v i e r . co m / l o c a t e / s c i t o t e n v

Quantifying biochar content in a field soil with varying organic matter

content using a two-temperature loss on ignition method
Seyyed Ali Akbar Nakhli, Sriya Panta, Joseph D. Brown, Jing Tian, Paul T. Imhoff ⁎
Department of Civil and Environmental Engineering, University of Delaware, Newark, DE 19716, USA

H I G H L IG H T S
G R A P H I C A L A B S T R A C T

• Two-temperature loss on ignition

(LOI) method developed to quantify
biochar in soil.
• Spatial variation of soil organic matter
(SOM) content is accounted for in the
method.
• Method applied to biochar-amended
roadway soil and error was b3%.
• Method is applicable for soils with low
to moderate SOM content.
• Method is applicable for biochars
pyro- lyzed at high temperatures with
low ash content.

a r t i c l e i n f o
a b s t r a c t
Article history:
Received 27 July 2018 While the use of biochar as a soil amendment for agronomic and environmental management is gaining popular-
Received in revised form 7 November 2018 ity, quantification of biochar in soil is still challenging. The objective of this work was to develop a fast, simple
Accepted 11 December 2018 and inexpensive method to quantify biochar content in field soil with varying organic matter content – the
Available online 14 December 2018 two- temperature loss on ignition (LOI) method. In this approach, biochar mass fraction in a biochar-
amended soil is computed by measuring the dry mass of biochar/soil mixture after heating sequentially at
Editor: Baoliang Chen two temperatures: low temperature (LT), and high temperature (HT). This method requires the LOI profile for
pure soil and pure bio- char that are representative of soil and biochar in the field. Although the soil LOI
Keywords: profile may vary due to spatial variation in soil organic matter (SOM) content, the method only requires that
Biochar
the relative soil LOI at LT with re- spect to LOI at HT is uniform because of similarity in SOM chemical
Quantification
composition. In this method, LT and HT are selected such that the maximum difference in LOI exists at these
Loss on ignition
Soil organic matter
temperatures between pure soil and biochar. The method was tested by quantifying the biochar content in
Spatial variation roadway filter strips with and without a wood bio- char pyrolyzed at high temperature (550 °C). The estimates
of biochar content from the method matched inde- pendent measurements for soils with low (−0.23 ± 0.09 CI
%, CI = 95% confidence interval, versus actual 0%) and high (3.9 ± 0.3 CI% versus actual 4.0 ± 1.1 CI%) biochar
mass fraction. The method is applicable when SOM con- tent is low to moderate (e.g. b15%) and mostly
composed of labile organic compounds, and when biochars are pyrolyzed at moderate to high temperatures
(i.e. N400 °C) and composed of relatively low ash content (e.g. b30%).
© 2018 Elsevier B.V. All rights reserved.

1. Introduction

* Corresponding author. Biochar is a pyrogenic carbonaceous material (PCM) produced from

E-mail address: imhoff@udel.edu (P.T. Imhoff). thermochemical conversion of biomass in an oxygen-limited

https://doi.org/10.1016/j.scitotenv.2018.12.174
0048-9697/© 2018 Elsevier B.V. All rights
reserved.
 S.A.A. Nakhli et al. / Science of the Total Environment 658 (2019) 1106–
environment (pyrolysis) purposefully for use as a soil amendment for
agronomic or environmental purposes (Lehmann and Joseph, 2015).
Important defining features of biochar compared to the parent
material are depleted levels of hydrogen (H) and oxygen (O) and
elevated level of organic carbon (Corg) that is dominated by aromatic
structures (International Biochar Initiative, 2012; Keiluweit et al.,
2010). These materials offer high resistance to biotic and abiotic
degradation pro- cesses, thereby ensuring that any beneficial effects
of biochar can en- dure for decades to millennia (Enders et al., 2012;
Zimmerman, 2010). Slow turnover rates and high carbon (C) content
of biochar along with its potential beneficial impacts on soil
physicochemical properties and biota have prompted interest in its
land application. Biochar may be- come a key component in integrated
agricultural/environmental strate- gies to build soil, enhance water
quality, and increase agricultural productivity, while sequestering C
and thus mitigating global climate change (Harvey et al., 2012; Imhoff
and Nakhli, 2017; Lehmann et al., 2011; Lehmann and Joseph, 2015;
Zhang et al., 2012).
Understanding the dynamics of and the interactions within
biochar-
soil systems requires knowledge of biochar quantity in soil (Koide et
al., 2011; Lehmann and Joseph, 2015). Even if mixed with soil
homoge- neously, biochar content may become temporarily and
spatially variable under field conditions. Although biochar generally is
considered to be recalcitrant, it may be lost in soil over time due to
decomposition and leaching of the incompletely charred fraction and
semivolatile organic compounds that coat biochar surfaces (Dong et
al., 2017; Yi et al., 2015; Zimmerman, 2010). Biochar also can migrate
vertically and hori- zontally in soil by surface runoff, wind, water
infiltration, and tillage (Dong et al., 2017; Obia et al., 2017). In
addition, imperfect and incom- plete mixing initially and during land
management practices such as tillage and sowing in subsequent years
may lead to heterogeneous dis- tributions (Olmo et al., 2016). These
factors suggest the necessity of rou- tine analyses of biochar content,
which requires a fast, inexpensive, and accurate procedure for biochar
quantification in soil.
Quantification of biochar is often accomplished by distinguishing
biochar from two other forms of C - inorganic carbon (C inorg) and non-
biochar Corg (Poot et al., 2009; Raya-Moreno et al., 2017). However,
the complex chemistry and diverse composition of biochar and soil C
pools makes it difficult to quantify biochar in natural environments
(Keiluweit et al., 2010; Koide et al., 2011). Several techniques have
been used to quantify, isolate, and characterize biochar in soil. These
methods include in-situ visual categorization based on soil darkening
using diffusive reflectance spectroscopy (Olmo et al., 2016),
measuring the 13C isotope signature and total organic carbon content
(Obia et al., 2017), oxidizing the soil using chemothermal or wet
oxidation methods (Brä ndli et al., 2009), and loss on ignition (LOI)
(Dong et al., 2017; Koide et al., 2011). Here and below, LOI refers to
heating a sample of a material at a specified temperature in air,
allowing components to volatilize and/ or combust, until sample mass
ceases to change. Raya-Moreno et al. (2017) compared several
techniques including LOI, dry-combustion, strong and mild acid
potassium dichromate oxidation, acid hydrolysis, and peroxide
oxidation to assess biochar C in soil and concluded LOI, dry-
combustion, and mild acid oxidation were the best proxies for bio-
char C quantification.
In general, the techniques for quantification of PCM, of which bio-
char is one example, can be grouped into five major classes: physical
methods such as flotation and density separation, chemical tech-
niques like dichromate and ultra-violet oxidation, thermal methods
like LOI and chemo-thermal oxidation, spectroscopic techniques
such as nuclear magnetic resonance and mid-infrared spectroscopy,
and molecular marker methods such as benzene polycarboxylic acid
and pyrolysis gas chromatography – mass spectrometry (Brä ndli
et al., 2009; Hammes et al., 2007; Lehmann and Joseph, 2015; Poot
et al., 2009). However, most of these methods are expensive, labor
intensive, and require specialized instrumentation (Koide et al.,
2011; Olmo et al., 2016). No method is ideal (Lehmann and Joseph,
When loss on ignition is applied to soil, the mass loss has been
pro- posed as a reliable, fast, accurate and inexpensive method for
estimating soil organic matter (SOM) (Ghabbour et al., 2014; Heiri
et al., 2001; Salehi et al., 2011). The LOI technique was first adopted
for quantifying biochar content in field soil by Koide et al. (2011). In
their approach, Koide et al. (2011) measured LOI for pure soil and
biochar after heating at 550 °C for 4 h, prior to biochar-amendment.
LOI measurements of biochar-amended soil were then used to
estimate the biochar content based on distinct LOI values for pure
soil and biochar. In applying the LOI method, Koide et al. (2011)
assumed that SOM did not vary spatially and remained constant with
time, even after biochar amendment. While this situation may be
valid for agricultural soils that are fre- quently mixed by plowing,
the organic matter in soil typically varies spatially and usually
decreases with depth (Turer and Maynard, 2003; Zhan et al., 2013).
Koide et al. (2011) also assumed that the LOI value for biochar
remained stable over time and demonstrated this was the case for
the 15-month duration of their study. However, they acknowl- edged
that this assumption might not be valid for all biochars, since bio- char
aging might change its chemistry and subsequently the mass of
biochar that is combusted with LOI measurements (Koide et al.,
2011). In this research, a new method based upon LOI
measurements at two temperatures is evaluated for determining
biochar content in biochar-amended soil. Because measurements are
based upon LOI data at two temperatures, the method does not
require spatial or tem- poral uniformity of SOM. The theory behind
this two-temperature LOI method is presented first. The accuracy of
the method is then evaluated by applying it to samples extracted from
a field site where side-by-side filter strips, one with and one without
biochar, were constructed. Fi- nally, the applicability of the method
for different soils and biochars is
discussed.
2. Theory
The methodology is developed based on assumed LOI profiles for
soil and biochar illustrated in Fig. 1. For soil samples collected in close
prox- imity, it is likely that SOM chemical composition is similar
regardless of variation in total SOM content. This assumption was
made for the three samples in Fig. 1. In this case, the relative soil mass
loss with respect to the mass loss at a selected reference high
temperature (HT), where most organic matter is removed, should
approach an approximately uniform trend for all soil samples (Fig.
1B). Moreover, the thermally labile frac- tion of organic matter in soil
is typically much greater than that found in biochar produced at
moderate to high heat treatment temperatures (HTTs) (i.e. N400 °C).
Thus, the relative LOI profiles are distinctly differ- ent for soil and
biochar as shown in Fig. 1B. In the proposed method, the difference in
the relative mass loss with respect to a particular HT be- tween soil
and biochar is most pronounced at a particular low temper- ature
(LT). Temperatures that exceed those used to ash biochar are not
considered here, since the methodology uses differences in organic
car- bon between soil and biochar, rather than differences in mineral
composition.
In this methodology, four dimensionless LOIs are defined:
LT MDT −MLT
LOI b b
;:
¼ b MDT
b
HT M −MHT
DT
LOI
¼ b b b
;
MDT
b
ð1Þ
s s
LOI LT ¼ M DT − MLT ;
M
s DT
s
HT M DT −MHT
LOIs ¼ s s
M sDT
where LOIα is the LOI for material β when heated at temperature α, Mα
 S.A.A. Nakhli et al. / Science of the Total Environment 658β (2019) 1106– β
2015). is mass of material β after heating at temperature α, and β = b for pure
 S.A.A. Nakhli et al. / Science of the Total Environment 658 (2019) 1106–

100
6
A relative soil mass loss at LT with respect to the mass loss at HT
(RLOIs) should be constant
80 5
Biochar
with Respect to DT (%)

LOIsLT

Soil Mass Loss with

Biochar Mass Loss

RLOI¼ ≈ constant ð Þ5s

60 4 LOIsHT

Respect to DT (%)
3 With this assumption, to estimate the biochar content of a
biochar/ soil mixture, M DT, M LT,and M HT are measured. Using these
three mea-
LT HT
m m m
40
surements along with measured or estimated LOIb , LOIb , and RLOIs
2 ob- tained from pure materials, Eqs. (2)–(5) are solved for the
Soil four
unknowns MDT, MDT, LOILT and LOIHT (see Section S2 Supporting Informa-
20 b s s s
1
tion). The biochar mass fraction in a sample (bf) is then expressed as
0 0 DT
RLOIs MDT −MHT þ MLT −MDT
DT LT HT M m m m m ð6Þ
b ¼m
B
100 ¼ MDT RLOI · LOIHT −LOILT
f
with Respect to Mass Loss at HT (%)

MDT s
m b b

80
The approach outlined above is similar to that of Koide et al.
Relative Mass Loss

(2011) where LOI measurements at a single temperature were used

60
40
20
0
DT LT
Temperature (°C)
Fig. 1. Schematic of (A) loss on ignition profile and (B) relative loss of ignition profile
with respect to loss on ignition at high temperature for hypothetical soils and biochar,
where the biochar has relatively low ash content. DT, LT, and HT are drying temperature
and selected low and high temperature, respectively. Line color represents the natural
color of biochar (back) and soil (brown), where darker brown represents higher soil
organic matter. (For interpretation of the references to color in this figure legend, the
reader is referred to the web version of this article.)
biochar and β = s for pure soil. Note that these LOIs are defined with
re- spect to the sample mass at the drying temperature (DT), which is
the temperature at which all moisture is assumed to be driven off.
If a given dry biochar/soil mixture (β = m) is heated sequentially
at two temperatures (LT and HT) and maintained for sufficient time at
each temperature to achieve constant mass, the masses of the heated
samples can be expressed using the LOIs defined above for pure
biochar and soil:
MDT DT DT
m ¼ Mb þ M s
to deter- mine the biochar content of a biochar/soil mixture.
However, in this new approach the assumption of constant LOI for
biochar-free soil re- quired by Koide et al. (2011) is unnecessary.
Instead, LOI measurements at two temperatures are used and it is only
necessary to assume that RLOIs is uniform within all biochar/soil
mixtures. The validity of this two-temperature LOI method for
quantifying biochar content was tested at the field site studied in this
work.
HT 3. Materials and methods
3.1. Field site description
The project site is located south of the Chesapeake and Delaware
Canal in Delaware (USA) along the northbound lane of Rt. 896 at the
Bethel Church Road on-ramp (Lat: 39° 31′ 48.19″ N, Long: 75° 44′
9.65″ W). Two filter strips each 6.1 × 1.8 × 0.3 m (length × width
× depth) abutting the roadway were modified in November 2015 to
as- sess the impact of biochar amendment on stormwater runoff. The
filter strips are shown in Fig. 2 with associated trench drains,
instrumentation shed, and piping for periodic collection of runoff
samples and monitor- ing the filter strips performance in runoff
reduction. One filter strip was tilled to 30 cm depth, while the other
was tilled and amended with Soil Reef™ biochar (The Biochar
Company, Berwyn, PA) to achieve 4% (w/ w) in the top 30 cm soil.
This biochar was produced from Southern Yel- low Pine by pyrolysis
at 550 °C for 10 min. Biochar was used as is from the manufacturer
for application to the field site. The two plots were separated from
each other by 1–2 m. Stormwater runoff from a 93 m 2 impervious
roadway area was evenly distributed into the two filter strips during
rainfall events. A detailed description of the preparation of both filter
strips is discussed in Section S1 Supporting Information. The biochar
content of the tilled + biochar filter strip was estimated to be 4.0 ±
1.1 CI% (w/w), where the ±1.1 CI% defines an interval hav-
ð2Þ ing a confidence level of approximately 95%.

MLT ¼ MDT 1−LOILT þ MDT 1−LOILT ð3Þ 3.2. Soil and biochar sampling
m b b s s

To assess the spatial variation of soil LOI profiles and to determine

m b b s s
RLOIs, four soil samples collected at different locations in the field and
M ¼ M 1−LOI
HT
þ M 1−LOI DT HT DT HT
at different times were used: two intact core samples (samples 1 and
where M and MDT in Eqs. (2)–(4) are for the biochar and soil, respec-
DT
2 in Table 1) from the top 30 cm soil of the tilled filter strip collected
b s
tively, in the biochar/soil mixture and not separate samples of the pure
materials as in Eq. (1). LOILT and LOIHT are assumed invariant
S.A.A. Nakhli with of
et al. / Science bio-
the Total Environment 658 (2019) 1106–
in September 2017, a well-mixed tilled soil sample collected immedi-
ately after tillage in November 2015 (sample 3), and a soil sample
b b
char content of the biochar/soil mixture and are measured indepen- from excavated soil that was stored in a stockpile during construction
dently with pure biochar. Because spatial variation is an inherent (Fig. 2) and collected in June 2017 (sample 4). Core samples were col-
characteristic of SOM in soil, LOILT and LOIHT will presumably vary lected using an AMS compact slide hammer (Model 400.96, AMS, Inc.,
s s
among different soil samples. If, however, the chemical composition of American Falls, ID) with a 2.86 × 58.9 cm plated soil recovery probe
SOM is assumed uniform throughout all biochar/soil mixtures, the (Model 424.07, AMS, Inc.) and mixed thoroughly before use. Two
 S.A.A. Nakhli et al. / Science of the Total Environment 658 (2019) 1106–
Fig. 2. Field site layout showing locations of tilled filter strip, tilled and biochar-amended filter strip, top soil stockpile, intact soil cores (eight in each plot), and weathering pots
that contained biochar exposed to environmental conditions. Trench drains adjacent to roadway were covered with a geomembrane for stormwater experiments.
additional soil samples from the topsoil stockpile collected in
February 2017 and 2018 (samples 5 and 6) were used to evaluate the
accuracy of the two-temperature LOI method in estimating biochar
content. Plant matter and root hair were removed from soil samples,
after which samples were pulverized and sieved through a 2-mm
sieve. The sieved soils were oven dried at 105 °C for 24 h and stored
in sealed dark containers until use.
To assess the effect of biochar aging on its LOI profile and obtain
the
LOILT and LOIHT constants, biochar and biochar + soil samples were
b b
placed in clay pots at the field site and sampled through 25 weeks. As
will be shown below, 25 weeks was a sufficient period to observe
initial changes and then stabilization of the LOI profile. Before using
biochar, the biochar was rinsed at a 1:50 biochar/deionized water
(w/w) ratio until the electrical conductivity (EC) of the rinsate
decreased below 100 μs/cm. Rinsing removes minerals and fractions
of biochar that have not been completely charred, and semivolatile
organic compounds coating biochar surfaces during pyrolysis
(Lehmann and Joseph, 2015; Yi et al., 2015). The EC of 100 μs/cm was
selected based upon prelimi- nary laboratory experiments, where it
was found to be a good indicator of when most readily leachable
solutes were removed. After rinsing, the biochar was oven dried at
105 °C for 24 h and stored in a sealed dark plastic container until use.
Six clay pots (opening diameter = 12 cm, bottom diameter = 8
cm, internal depth = 10 cm) were filled with rinsed biochar, and
six clay pots (opening diameter = 21 cm, bottom diameter = 15 cm,
internal
Table 1
Sample descriptions, samples IDs, collection dates, and initial air-dried masses used in LOI tests.
Sample type Description (no. of samples)
Field soil (no biochar) Mixed tilled soil core, 0–30 cm (2)
Mixed tilled soil (1)
Stockpile (3)
Biochar aging pot Biochar, week 0, 6, 8, 10, 25 (5)
Soil + 4% biochar by mass, week 0, 6, 8, 10, 25 (5)
Intact core from tilled regions Tilled, 0–50.5 cm (8)
Tilled + biochar, 0–50.5 cm (8)
depth = 18 cm) were filled with mixed tilled soil (sample 3) amended
with 4% (w/w) rinsed biochar. The pots were placed in the field (see
Fig. 2) in November 2016 by excavating soil and inserting the clay
pots at the ground surface, where they were exposed to
environmental conditions similar to those of the biochar + tilled filter
strip. At weeks 0, 6, 8, 10, and 25, one pot of biochar and one pot of
biochar-amended soil were removed. Biochar was removed from the
center of each pot for each sampling period, and the sample masses
removed are reported
in Table 1. For biochar-amended soil, forceps were used to carefully ex-
tract biochar particles, which by necessity consisted of larger particles
that could be distinguished from the soil. This biochar was rinsed
with deionized water to remove visible soil particles.
To assess the reasonableness of the two-temperature LOI
procedure for quantifying biochar content and to assess any vertical
variation in SOM and biochar distribution, eight intact field cores
were collected from the tilled and tilled + biochar filter strips in
March 2017 (Table 1). Cores were collected at locations shown in Fig.
2 following procedures described above with one difference – cores
were driven to 50.5 cm depth to collect full 30 cm cores. Because of
compaction dur- ing coring, a 30-cm long core always resulted in b30-
cm of material. Based on measurements of sample length during
coring, soil compac- tion was determined to be uniform along the core
length. Each intact core was cut into six equal sections (each
representing 8.4 cm depth), and plant matter and root hair removed
from the core sections. The lower core sections had negligible
amounts of visible root hair. Because
Sample ID Collection date Sample mass (g)
Sample 1 and 2 Sep-2017 13–17
Sample 3 Nov-2015 13–17
Sample 4–6 Jun-2017, Feb-2017, Feb-2018 13–17
– Nov-2016 to May-2017 Week 0: 3–4, week 6: 5–6, week 8: 5–6,
week 10: 11–13, week 25: 12–15
– Mar-2017 14–16
– Mar-2017 14–16
 S.A.A. Nakhli et al. / Science of the Total Environment 658 (2019) 1106–

variation of biochar content with depth was of primary interest, sam-

3.5. Statistical analysis
ples at each depth increment in each filter strip were combined to
achieve sufficiently large sample masses for analysis. This provided
In the two-temperature LOI method, the unknown biochar mass
six samples each for the tilled and tilled + biochar filter strips.
fraction in a sample (bf) is computed from mixture sample masses
heated to DT, LT, and HT along with independently measured LOILT, LOI-
b b
HT
, and RLOIs using Eq. (6). Uncertainty in bf arises from uncertainty in
3.3. Soil and biochar characterization
the sample measurements (M mDT, M m LT
,Mm HT
) and in estimates of the
LOI parameters for biochar and soil (LOILT, LOIHT, RLOI ). The uncertainty
b b s
Rinsed biochar and the mixed tilled soil sample (sample 3) were in bf was estimated using propagation of uncertainty (Taylor and Kuyatt,
used to characterize soil and biochar physicochemical properties. Parti- 1994), where the uncertainty in sample measurements (M DT, M LT,
m m
cle size distributions were measured following ASTM D422 – 63 with M mHT) was estimated from the accuracy of mass measurements, and
(soil) or without (biochar) a dispersing agent. Biochar skeletal and en- un- certainty in LOI parameters (LOILT, LOIHT, RLOI ) were estimated
from b b s
velop densities were characterized by mercury intrusion porosimetry 1–4 and biochar aging pots were performed in triplicate. Duplicate LOI
analysis (Micrometrics Analytical Services, Norcross, GA, USA). Soil tests were used for soil samples 5 and 6 and intact core samples from
par- ticle density was measured by water pycnometer according to both filter strips.
ASTM D854 – 14. The Brunauer–Emmett–Teller (BET) surface area was
deter- mined by N2 adsorption at 77 K using Micromeritics ASAP 2020
analyzer after degassing the samples under vacuum at 95 °C for 960
min. The el- emental (CNHS) analysis was performed using CNHS
Elementar Cube. Carbonate content was measured following ASTM
D4373 – 14. Biochar fixed C, volatile matter and ash content were
determined following ASTM D1762 – 84. Biochar O content was
calculate by subtracting the sum of CNHS elements and ash content
from total mass according to ASTM D3176 – 15. By definition, O
content calculated according to the ASTM standard does not include O
in the mineral matter or in the ash. All analysis were performed in
duplicate. The thermogravimetric (TG) thermogram for the rinsed
biochar was obtained in air (flow rate = 10 mL/min) using a
differential scanning calorimeter with capabilities for simultaneous
TG-DSC analysis (SDTQ600; TA Instruments, New Cas- tle, DE) at a
temperature ramp of 20 °C/min.

3.4. Loss on ignition test

In the LOI method, several factors such as ignition temperature,

ex- posure time, sample mass and position of the sample in the
furnace may affect the LOI results (Ghabbour et al., 2014; Heiri et al.,
2001). Gen- erally, higher temperature, longer time and larger sample
mass leads to more precise and less sample-dependent LOI
measurements for diverse soil samples (Ghabbour et al., 2014;
Hoogsteen et al., 2015). Accord- ingly, in this research LOI
measurements were performed using a rela- tively long ignition time
(~12 h) with large sample masses (Table 1) for all samples. In
addition, replicate samples were placed in different positions in the
furnace, center or rear, to minimize any possible bias in LOI
measurement due to sample position.
A muffle furnace (Thermos Scientific, Model no. F48015-60) with
heating range 100–1200 °C, accuracy ± 3 °C, and uniformity ± 3.6
°C was used to conduct all LOI measurements. The manufacturer's
guide- lines were followed to calibrate the oven. A thermocouple
(Omega, se- rial no. P12A526K4FA) was used to check oven
temperatures (110 and 200 °C) throughout the testing period, and
measured temperatures were always within ±2 °C of the oven set
temperature.
For each LOI profile test, a known mass of sample (Table 1) was
se- quentially heated to 110 (=DT), 200, 300, 350, 400, 450, 500, and
550
°C. The duration of heating was 24.0 ± 0.6 h at 110 °C, and 12.0 ±
0.8 h at other temperatures. The heating time of 24 h was selected be-
cause it is the typical duration used to remove moisture from soil
sam- ples. Samples were heated for 12 h at higher temperatures
because mass loss was found to be negligible for heating between 12
and 24 h in preliminary tests. Between each temperature, samples
were removed and allowed to cool in a desiccator before measuring
sample mass. LOI profile measurements were made for different soil,
biochar, and mix- ture samples described above which are also listed
along with the sam- ple masses in Table 1. LOI tests for soil samples
 S.A.A. Nakhli et al. / Science of the Total Environment 658 (2019) 1106–
data reported below for aged biochar and soil (see also Section S3
Supporting Information). Tukey's Honestly Significant Difference
test (Tukey's HSD) was used to compare the means of biochar LOI
obtained from biochar aging experiments at each temperature
(Kao and Green, 2008).

4. Results

4.1. Soil and biochar physicochemical properties

Physicochemical properties of mixed field soil collected after

tillage in November 2015 (sample 3 in Table 1) and rinsed biochar
are pre- sented in Table 2. The soil is a sandy loam with smaller
particles than biochar: soil d50 = 0.21 mm, while biochar d50 =
0.70 mm. Biochar is less dense than soil and has significantly
higher specific surface area.
Elemental analysis showed that biochar is rich in C with Corg
content of 76.7 ± 0.7 SE% (SE = one standard error of the mean)
and H/Corg and O/Corg molar ratios of 0.13 and 0.10, assuming all
Cinorg exists as carbon- ate and calculated O content does not
include O in the mineral matter and ash content. A proximate
analysis revealed that 81.0 ± 0.4 SE% and 19.0 ± 0.2 SE% of
biochar ash free content is fixed C and volatile matter,
respectively. Soil Corg (2.63%) is significantly less than biochar Corg
and therefore it is expected to have significantly less mass loss
after burning all organic matter at high temperature. The soil
H/Corg molar ratio was 1.8. Clay and carbonate content of soil and
biochar are sufficiently small (b3 and 1%, respectively) to neglect
the loss of struc- tural water from clay minerals and Cinorg that
might occur at 550 °C after the initial burning of organic matter
(Heiri et al., 2001). Therefore, LOIs at 550 °C were considered
measures of the organic matter content of soil and biochar, which
was also assumed by Koide et al. (2011).

4.2. Aged biochar loss on ignition profile

LOI measurements for biochar retrieved from clay pots with

100% biochar over 25 weeks are shown in Fig. 3 for four
temperatures: 200 (A), 300 (B), 400 (C), and 550 °C (D). All
reported mass losses are with respect to dry mass at 110 °C.
There was a slight decrease in the mass loss at 200 °C and a slight
increase in the mass loss at 300 and 550 °C between weeks 0 and
6. The mass loss at 400 °C also increased slightly between week 6
and 8, but otherwise, no clear change in LOI with time was
observed after 8 weeks. Tukey's HSD indicated that at α
= 0.05 there was a significant difference in mass loss with heating
at 400 °C between weeks 0–8 and at 550 °C between weeks 0
and 6 (Kao and Green, 2008). There were no significant
differences in the mass loss at 200 and 300 °C from weeks 0 to 25.
Thus, over a 25-week aging period in the field, the biochar LOI
profile did not change signifi- cantly after 8 weeks.
Biochar particles were also retrieved from clay pots of biochar-
amended soil and similar LOI profiles measured. Mass losses for
those biochar particles at 200 °C was always more (b9%) and at all
other tem- peratures were less (b7%) than those collected from
samples with bio- char alone, respectively. Scanning electron
microscopy (JSM-7400F, high-resolution scanning electron
microscope) was used to examine both sets of biochar particles
and showed small soil particles attached
 S.A.A. Nakhli et al. / Science of the Total Environment 658 (2019) 1106–

Table 2
Physicochemical properties of field soil (sample 3 in Table 1) and rinsed biochar. Values are means (standard errors of the mean) of duplicate measurements.

Type Particle size (% mass) Particle density N2 BET surface Component (% mass)a
(g/cm3) area
(m2/g)
Fine Sand Silt Clay Skeletal Envelope C H N S Ob Fixed Volatile Ash CO3−2
gravel C matter (calcite
equivalent)

Biochar 12.21 65.0 20.13 2.6 1.05 0.52 355 76.8 0.830 1.87 0.25 9.7 72.5 16.98 10.56 0.34
(0.16) (0.2) (0.15) (0.0) (0.12) (0.03) (6) (0.7) (0.014) (0.02) (0.00) (0.7) (0.4) (0.19) (0.18) (0.00)
Soil 0.0 71.3 26.3 2.3 2.59 – 1.68 2.67 0.39 1.04 0.12 – – – – 0.83
(0.0) (0.4) (0.4) (0.0) (0.00) (0.02) (0.00) (0.02) (0.04) (0.02) (0.02)

a
Expressed on a total dry mass basis.
b
Determined from %O = 100 − %(C + H + N + S + Ash) and does not include oxygen in the mineral matter or in the ash.

to biochar retrieved from biochar-amended soil that were not present

data from aged biochar are shown in Fig. 4A. Based on these LOI
in biochar-alone samples. A similar observation was reported by
data, SOM measured as mass loss on ignition at 550 °C is not uni-
Sorrenti et al. (2016): minerals and soil particles adhered to and/or
form among samples and ranged from 3.3 to 4.8%. For all soil sam-
were physi- cally trapped in biochar particles recovered from a
ples, though, regardless of SOM content a similar trend of LOI was
nectarine orchard after four years of environmental exposure. Since
observed: while mass loss increased continuously with increasing
these soil particles could not be removed by rinsing, LOI profiles for
temperature, the extent of increase in mass loss between succes-
biochar removed from biochar-amended soil were not used in the
sive temperatures correlated with SOM content (measured as
analyses below.
mass loss on ignition at 550 °C). This implies that soil mass loss at
Since biochar aging was minimal after 8 weeks and sample masses
any temperature normalized with respect to the mass loss at high
were largest for data collected for weeks 10 and 25 (see Table 1), data
temperature, 400 or 550 °C for each soil sample, might result in a
from weeks 10 and 25 were used to estimate LOI for aged biochar.
single curve.
LOIbT at T = 200, 300, 400, and 550 °C with respect to 110 °C were
Data in Fig. 4A were normalized with respect to mass loss at ei-
(42.97 ± 0.17 SE) × 10−3, (593.3 ± 0.3 SE) × 10−3, (879.6 ± 0.4
ther 400 or 500 °C, and the average relative mass loss with respect
SE)
to the high temperature (either 400 or 500 °C) are plotted in
× 10−3, and (894.4 ± 0.6 SE) × 10−3, respectively. These LOITb were
Fig. 4B. Error bars on the sample means (four samples) plotted in
used in all calculations below. The residual biochar mass
Fig. 4B represent ± SE and are so small as not to be visible. Thus, in-
remaining when heated at 550 °C for 12 h, 10.56 ± 0.06 SE%, was
dividual soil LOI shown in Fig. 4A scaled reasonably well with respect
consistent with
to mass loss at either 400 or 500 °C. Using data from each of the four
10.56 ± 0.18 SE% ash content of biochar when heated at 750 °C for 6 h
soil samples, average RLOIs at 200 and 300 °C with respect to LOI at
(Table 2) and indicates that minimal mass loss will occur by heating
400 °C was (24.5 ± 0.5 SE) × 10−2 and (69.6 ± 0.6 SE) × 10−2, re-
the biochar at temperatures between 550 and 750 °C.
spectively. The average RLOIs at 200, 300 and 400 °C with respect to
LOI at 550 °C was (21.3 ± 0.4 SE) × 10−2, (60.6 ± 0.7 SE) × 10−2
4.3. Soil loss on ignition profile
and (87.0 ± 0.4 SE) × 10−2, respectively.
The LOI profiles of the four soil samples collected at different
field site locations and times (samples 1–4 in Table 1) along with

4.7 60
a A a aB
a a
Mass Loss at 200 °C with

Mass Loss at 300 °C with

4.6 59
Respect to 110 °C (%)

Respect to 110 °C (%)

a
4.5 58

a
4.4 a 57

a a
4.3 56

0 0
0 6 8 1025 90 0 6 8 1025
88.2
a C a D
a a a
Mass Loss at 400 °C with

88 a
89.5 a
Mass Loss at 550 °C
to 110 °C (
Respect to 110 °C (%)

87.8
89
th Respectwi%)

87.6 c
b
88.5
87.4 b

0 0
0 6 8 10 25 0 6 8 1025
Biochar Weathering Time (Weeks) Biochar Weathering Time (Weeks)

Fig. 3. Loss on ignition data of aged biochar. Mass loss at (A) 200 °C, (B) 300 °C, (C) 400 °C, and (D) 550 °C with respect to 110 °C are shown for biochar aged for 0–25 weeks. Data
represent mean and error bars depict standard errors of the means (n = 3). For each mass loss plot at a given temperature, letters denote differences using Tukey's HSD test (α = 0.05).
 S.A.A. Nakhli et al. / Science of the Total Environment 658 (2019) 1106–

A
100 Biochar 6 40
Soil LT=200, HT=400 °C LT=200, HT=550 °C LT=400, HT=550 °C
Sample 1
Biochar Mass Loss with

with Respect to 110 °C (%)

80 Sample 2

Biochar Mass Fraction (%)

Relative Standard Error of
Sample 3 30
Respect to 110 °C (%)

Sample 4 4

Soil Mass Loss

60

20
40
2

20 10

0 0
0 100 200 300 400 500 B 0
0 3 6 9 12 15
with Respect to Mass Loss at HT (%)

100 Biochar
HT= 400 °C Sample Mass (g)
HT= 550 °C
80 Soil
Relative Mass Loss

HT= 400 °C Fig. 5. Estimated relative standard error (standard error/mean) of biochar mass fraction
HT= 550 °C using the two-temperature LOI method for LT = 400 °C/HT = 550 °C, LT = 200 °C/HT
= 550 °C, and LT = 200 °C/HT = 400 °C. The biochar content of the sample is assumed
60 4% (w/w).

40 decreased from 6.0% to 2.5% as the sample mass increased from 3 to

15 g. Sample masses ranged from 13 to 17 g (Table 1) for intact cores
20
from tilled and tilled + biochar filter strips collected in March 2017.
Thus, the relative error in estimated bf for those data is expected to be
~2.5%, and the estimated standard error to be ~0.1%. All calculations
0 below were conducted using LT = 200 °C and HT = 400 °C.
0 100 200 300 400 500 The accuracy of the two-temperature LOI method was evaluated
Temperature (°C) by measuring biochar mass fraction in two independent soil samples
amended with known biochar contents. Soil sample 5 and 6 (Table 1)
Fig. 4. (A) Loss on ignition profiles of two well-mixed intact soil cores (sample 1 and with SOM content of 4.8 and 3.2%, respectively, were amended with
2 collected Sep-2017), tilled and well-mixed soil (sample 3 collected at Nov-2015), rinsed biochar at the rate of 0, 2, 4, 6, and 8% (w/w). The measured
soil from stockpile (sample 4 collected at June-2017), and aged biochar (average of
bio- char content versus actual biochar content is shown in Fig. 6.
week 10 and 25). (B) Relative loss on ignition profile of average of four soil samples
with respect to loss on ignition at HT = 400 and 550 °C. Data represent (A) the mean
Although there is some scatter in the data at high biochar contents of
of three soil samples and six biochar samples and (B) the mean of twelve soil samples 6 and 8%, data are close to the 1:1 line for both tested soils. For
and six biochar samples; error bars represent ± one standard error of the mean are samples 5 and 6 without biochar, the measured biochar content was –
small and not visible for most data. 0.2 ± 0.3 SE% and – 0.09 ± 0.16 SE%, respectively. At 8% (w/w)
actual biochar mass fraction, the calculated biochar content for
sample 5 and 6 was bf=8.0
the best temperature pair with the smallest relative standard error was
4.4. Optimum temperature selection and method accuracy LT/HT
= 200 °C/400 °C. For this temperature pair the relative standard error
For the two-temperature LOI method, a low and a high heating
tem- perature are selected, with the primary requirement being that
the dif- ference in the relative mass loss with respect to a particular HT
between soil and biochar is most pronounced at a particular LT. Based
on soil and biochar LOI profiles (Fig. 4A and B), three possible
temperature pairs seemed reasonable. The two pairs LT/HT = 200
°C/400 °C and LT/HT
= 200 °C/550 °C were selected for analyses below because biochar
mass loss was significantly higher than soil mass loss between LT and
HT (Fig. 4B). The third pair LT/HT = 400 °C/550 °C was selected
because biochar mass loss was negligible compared to soil mass loss
for this tem- perature pair.
A propagation of uncertainty analysis was used to calculate the
esti- mated standard error of bf and the estimated relative standard
error (standard error/mean) as a function of sample mass. The results
are pre- sented in Fig. 5 for three selected temperature pairs,
assuming actual bf
= 4%. In all calculations LOIsT at T = 200, 400 and 550 °C were assumed
0.9, 3.5 and 4% (average of soil samples 1–4), respectively. For all
tem- perature pairs, the relative standard error decreased with
increasing sample mass from 3 to 15 g. For any given sample mass,
LT/HT = 400
°C/550 °C resulted in the highest and LT/HT = 200 °C/400 °C the
lowest uncertainty. Based on the propagation of uncertainty analysis,
 S.A.A. Nakhli et al. / Science of the Total Environment 658 (2019) 1106–
± 0.3 SE% and bf=7.6 ± 0.2 SE%, respectively.

1:1 Line
8 Sample 5: SOM = 4.8%

Measured Biochar Mass Fraction (%)

Sample 6: SOM = 3.2%

0 2 4 6 8
Actual Biochar Mass Fraction (%)

Fig. 6. Measured biochar content using two temperature LOI method (LT/HT = 200
°C/400
°C) versus actual biochar content for two stockpile soils with different SOM
(sample 5 collected at Feb-2017 and sample 6 collected at Feb 2018). Data
represent the mean of duplicate measurements; error bars represent ± one
standard error of the mean.
 S.A.A. Nakhli et al. / Science of the Total Environment 658 (2019) 1106–

4.5. Estimates of biochar mass fraction at field site

a minimum – 3.6% at 42.1–50.5 cm depth. The average estimated bio-
char content for depth of tillage (30 cm) was 4.8 ± 0.5 CI%. Thus, the
LOI profiles of tilled and tilled + biochar cores are shown in Fig. 7
Koide et al. (2011) method underestimated bf in the tilled filter strip
and relative LOI of tilled cores are illustrated in Fig. S1B. While the
(−1.59 ± 0.12 CI% versus 0%) and overestimated bf in the tilled + bio-
mass loss for the top three cores (0–25.2 cm depth) in biochar-
ochar filter strip (4.8 ± 0.5 CI% versus 4.0 ± 1.1 CI%).
amended filter strip was significantly higher than biochar-free filter
strip at temperatures N200 °C, no significant difference were
5. Discussion
observed in LOI profiles for the bottom three cores (25.2–50.5 cm
depth) between both filter strips. This result indicates the presence of
5.1. Method accuracy in estimating biochar mass fraction
biochar from 0 to
25.2 cm in the tilled + biochar filter strip. SOM content in the tilled
The two-temperature LOI method underestimated the average
filter strip (biochar-free) decreased with depth and ranged from 5.9%
bio- char content in the tilled region slightly: – 0.23 ± 0.09 CI% versus
at 0–8.4 cm depth to 1.7% at 42.1–50.5 cm depth. The SOM contents
in biochar-free samples span a wider range compared to the soil actual 0%. The estimated biochar content for 0–8.4 cm (− 0.78 ± 0.10
samples used to determine RLOIs (3.3–4.8%). SE%) de- viated more than the other soil layers from the expected 0%.
The mea- sured RLOIs for this layer was 11% higher than the
Results from the two-temperature LOI method are shown in Fig. 8.
estimated value used for the biochar content calculation, resulting in
Horizontal error bars represent ± one estimated SE and were
an underestimation of biochar content. The mass loss at low
deter- mined from the uncertainty analysis described above with bf
temperature is associated with loss of more labile SOM (numerator of
assumed to be equal to the estimated biochar content for all samples,
RLOIs in Eq. (5)), while mass loss at high temperature is associated
while ver- tical bars indicate the sampling range. For the tilled filter
with mass loss of more stable SOM (denominator of RLOIs in Eq. (5))
strip without biochar, bf ranged from – 0.8–0.1% with no
(Lopez-Capel et al., 2005; Ranalli et al., 2001; Siewert, 2001; Strezov
consistent trend with depth. The largest deviation from expected
et al., 2004). Therefore, the higher RLOIs in the top layer indicates that
zero biochar content was observed in the sample at 0–8.4 cm depth in
the ratio of the labile fraction to the stable fraction of SOM in this
which estimated biochar mass fraction was – 0.78 ± 0.10 SE%. The
layer was higher than in the deeper layers, presumably due to the
average biochar content for the top 50.5 cm was bf = − 0.23 ± 0.09
accumulation of plant litter, root hair, and particulate Corg from
CI% (α = 0.05). Thus, the two-temperature LOI method resulted in
stormwater runoff (Kö gel-Knabner, 2017).
an accurate estimate of the ac- tual biochar content (bf = 0.0%) in the
The average estimated biochar mass fraction in tilled + biochar
biochar-free filter strip.
filter strip was 3.9 ± 0.3 CI%, which was about 2.5% less than the
For the tilled + biochar filter strip, bf ranged from 0.4–5.6% for the
average of the incorporated biochar in the top 30 cm soil (4.0 ± 1.1 CI
top 30 cm, with no significant estimated biochar mass fraction at
%). While this small difference is certainly within measurement error,
25.2–30 cm depth (0.38 ± 0.11 SE%). This result implies that the it might also occur because of the tendency of the LOI method to
actual depth of mixing achieved by tillage of soil and biochar was slightly under- estimate biochar mass fraction, as was observed in the
~25 cm rather than the desired 30 cm. The average biochar content
tilled region without biochar (especially for topsoil), and/or because
for the top 30 cm was bf = 3.9 ± 0.3 CI% (α = 0.05) which is within the time of sam- pling was ~17 months after the addition of biochar.
the range es- timated for the site when it was constructed 15 Over this 17-month period, biochar mass loss might have occurred by
months before these field measurements: bf = 4.0 ± 1.1 CI%. leaching and decom- position of the labile fraction of biochar
Therefore, the two- temperature LOI method estimated biochar (Zimmerman, 2010), down- ward migration of biochar particles, or
mass fraction in the tilled
biochar erosion by surface runoff (Obia et al., 2016). Assessing biochar
+ biochar filter strip that is consistent with the estimated mass
biogeochemical stability re- vealed that leaching and decomposition
fraction when the site was constructed.
may account for b1% mass loss of the biochar in this study (see also
For comparison the method proposed by Koide et al. (2011) was
Section S4 Supporting Information). Downward migration is ruled out
ap- plied to estimate biochar content, using LOI values at a single
since negligible biochar was mea- sured underneath of the depth of
tempera- ture of 550 °C. LOI results for mixed tilled soil (sample 3)
tillage by the two-temperature LOI method. However, in addition to
and biochar at week 0 were used to estimate biochar content in intact
the tendency of the method to under- estimate biochar content at the
core samples from tilled and tilled + biochar filter strips. The
top layer, erosion of biochar particles might be a reason for the
estimated biochar con- tent in the tilled filter strip (biochar-free)
smaller estimated biochar mass fraction in the top layer (4.46 ± 0.10
ranged from 1.4% at 0–8.4 cm depth to – 3.7% at 42.1–50.5 cm, with an
SE%) than that in the second layer (5.63 ±
average of – 1.59 ± 0.12 CI% for the 0–50.5 cm soil. For the tilled +
0.10 SE%), as shown in Fig. 8B.
biochar filter strip, computed biochar content ranged from a maximum
6.6 ± 0.5 SE% at 8.4–16.8 cm depth to

12 12
Mass Loss with Respect to 110 °C (%)

0 - 8.4 cm A 0 - 8.4 cm B
8.4 - 16.8 cm 8.4 - 16.8 cm
10 16.8 - 25.2 cm 10 16.8 - 25.2 cm
25.2 - 33.7 cm 25.2 - 33.7 cm
33.7 - 42.1 cm 33.7 - 42.1 cm
8 42.1 - 50.5 cm
8 42.1 - 50.5 cm

6 6

4 4

2 2

0
0 100 0
200 300 400 500 0 100 200 300 400 500
Temperature (°C) Temperature (°C)
 S.A.A. Nakhli et
Fig. 7. Loss on ignition profile of (A) tilled soil cores (Mar-2017) andal.(B)
/ Science of the
tilled and Total Environment
amended 658 (2019)
with 4% biochar 1106–
(w/w) soil cores (March-2017) for samples collected at six depths.
Data represent the mean of two samples collected from a composite of eight cores. Error bars represent ± one standard error of the mean and most are small and not visible.
 S.A.A. Nakhli et al. / Science of the Total Environment 658 (2019) 1106–
Estimated Biochar Mass Fraction (%)
-1 0 1 2 3
0 0
10
20
Depth (cm)
30
40
50
Fig. 8. Estimated biochar mass fractions for (A) tilled and (B) tilled + biochar cores collected in March 2017 from the field site. Data represent the mean of two samples collected from a
composite of eight cores. Horizontal error bars represent ± one estimated standard error and were determined from the uncertainty analysis. Vertical bars indicate the sampling range.
5.2. Applicability of the two-temperature loss on ignition method
The proposed two-temperature LOI method for quantifying
biochar mass fraction in biochar-amended soils requires 1)
knowledge of the pure soil and pure biochar LOI profiles, which
represent soil and biochar under field conditions; 2) a distinctly
different LOI profile for soil and biochar between low and high
temperatures; and 3) a uniform RLOIs, which requires a uniform SOM
chemical composition. In this section the characteristics of soil and
biochar necessary to satisfy these require- ments are discussed.
5.2.1. Requirement 1: representative soil sample and stable biochar
The LOI profiles for pure soil and pure biochar found in field soil at
a wide range of temperatures are needed to assess the method
applicabil- ity for any given biochar/soil mixture. However, in most
cases removing
and separating soil and biochar particles from field samples of their mix-
ture is not feasible because biochar particles may be initially fine or
might disintegrate under field conditions (Spokas et al., 2014), soil
par- ticles and minerals adhere to biochar particles (Sorrenti et al.,
2016), and the presence of indigenous PCMs complicates the isolation
of bio- char particles (Lehmann and Joseph, 2015). When the pure
biochar and soil samples do not represent the biochar and soil
components in a biochar/soil mixture, error in computed biochar
mass fraction arises due to the difference between LOI parameters for
the pure soil and bio- char samples and those in the mixture (see
Section S5 Supporting Infor- mation for more detailed discussion).
If soil samples are not available prior to biochar addition, pure soil
samples can be taken from neighboring soils with similar vegetation,
texture, and climatic conditions, or from a control treatment that
exists in most research-based field studies and is representative of
soil in the biochar-soil mixture (Jobbá gy and Jackson, 2000; Quideau
et al., 2001). In either case, the priming or suppressing effect of
biochar on SOM mineralization should be minimal (Keith et al., 2011),
which may result in a difference between the LOI profile for the soil
component in a biochar/soil mixture and pure soil.
A critical characteristic of the biochar is that its LOI profile must
be stable over the duration of any field measurements. The
biogeochemical stability of the biochar LOI profile can be assessed by
elemental, proxi- mate, and thermogravimetric analyses and by
comparing the fixed C, volatile matter, O/Corg, H/Corg, and the
recalcitrance index (R50) values with criteria suggested by Enders et
al. (2012) and Harvey et al. (2012), respectively (see Section S4
Supporting Information). Generally, increasing HTT reduces volatile
matter content and H/Corg and O/Corg molar ratios, and increases fixed
C content (Enders et al., 2012; Keiluweit et al., 2010; Ronsse et al.,
2013). This trend is due to the tran- sition from slightly charred
biomass to char/charcoal, then to soot-like, and finally to graphitic
biochars with increasing HTT (Keiluweit et al.,
Estimated Biochar Mass Fraction (%)
4 5 6 -1 0 1 2 3 4 5 6
A B
10
20
30
Depth of Tillage Depth of Tillage and Biochar
40
50
2010). Therefore, biochars produced at higher HTTs (i.e. N400 °C) are
generally more environmental recalcitrant and have more stable LOI
than those produced at lower temperatures. Once a biochar is
selected that is expected to have a stable LOI profile, it is
recommended to rinse the biochar thoroughly, as was done in this
study, before LOI anal- ysis. Rinsing might remove the labile fraction
of biochar and make it more similar to biochar under field conditions
(Dong et al., 2017; Yi et al., 2015). For the most accurate
measurements, aging this rinsed bio- char in pots in the field before
LOI measurements is recommended.
5.2.2. Requirement 2: soil and biochar LOI parameters that differ
appropriately
In the two-temperature LOI method, LT and HT are selected such
that the maximum difference in LOI exists at these temperatures
between pure soil and biochar. That is, RLOI s ¼
s LOI =LOI must be
LT HT
s
significantly
different from the ratio LOILT =LOIHT. Thermal stability of soil and biochar
b b
is fundamentally a function of their chemical composition: reactions
at lower temperature are ascribed to the decomposition of more
thermally labile C (aliphatics, less aromatic rings), while higher
temperature reac- tions are attributed to thermal breakdown of more
thermally stable C (more aromatic rings) (Keiluweit et al., 2010;
Lehmann and Joseph, 2015; Lopez-Capel et al., 2005; Siewert, 2001).
Soils generally contain more thermally labile and less thermally stable
organic matter (humic substrates, black C) than biochars, due to
aromatization of biomass dur- ing pyrolysis. For example, according to
the Van Krevelen diagram, the soil H/Corg molar ratio from this study is
within the range of that for con- densed hydrocarbons and lignin (Kim
et al., 2003), while biochar H/Corg and O/Corg molar ratios are similar to
those for soot-like/graphitic chars (Hammes et al., 2006; Harvey et al.,
2012). Therefore, biochars are ex- pected to lose a smaller fraction of
their total organic matter when heated at low temperature than soils
(e.g. Fig. 1B).
Because high charring temperatures cause a pronounced degree of
aromatization in biochar (Keiluweit et al., 2010; Lehmann and Joseph,
2015), biochars produced at high HTT (i.e. N400 °C) are expected to
ex- hibit mass loss at low and high LOI temperatures that are
significantly different than most soils. This guideline is violated,
though, if indigenous black C and/or humic substrates make up a high
proportion of the soil organic carbon (SOC). In this case, the soil and
biochar LOI at LT and HT may be similar. Thus, to achieve significant
differences in LOI at LT and HT, biochars produced at high HTT
should be used with soils that have low amounts of indigenous black C
and/or humic substrates.
Finally, the two-temperature LOI method will result in inaccurate
re- sults if the LOI for soil is large while the LOI for biochar is small in a
given biochar/soil mixture. Biochars with high ash content when
sampled in soils with high SOM may result in inaccurate
measurements. To illus- trate this effect, the propagation of
 S.A.A. Nakhli et al. / Science of the Total Environment 658 (2019) 1106–
uncertainty analysis was used to es- timate the increase in relative
SE of biochar mass fraction if the biochar
 S.A.A. Nakhli et al. / Science of the Total Environment 658 (2019) 1106–
ash and SOM contents were simultaneously increased by a factor of
three or five for the biochar/soil mixture in this study. The relative SE
of biochar mass fraction in a 15 g soil sample increased from 2.5% to
9.2% by tripling the biochar ash and soil SOM (to 31.7 and 12.2%, respec-
tively). If biochar and soil SOM are quintupled (to 52.8 and 20.3%, re-
spectively), the relative SE of biochar mass fraction increases to
22.1%. Although ash content also increases with increasing HHT, it
varies more with feedstock than pyrolysis conditions (Enders et al.,
2012; Ronsse et al., 2013). Biochars produced from animal manure
(bull, poul- try litter, and digested dairy) and waste (food, papers)
contain higher ash content than wood-based biochars (Enders et al.,
2012). Therefore, the two-temperature LOI method may render
inaccurate results when is applied to estimate manure- and waste-
based biochars with high ash content (e.g. N30%) in soils with high
SOM content (e.g. N15%).
5.2.3. Requirement 3: uniform soil organic matter chemical composition
Although the two-temperature LOI method does not require
uniform
SOM content, it does require relative uniform RLOIs ¼ LOILT =LOIHT . For
example, a 10% change in RLOI s s
sfrom the assumed mean value for the
bio-
char/soil mixture in this work results in a constant absolute error of 0.5%
in bf, based upon the propagation of uncertainty analysis (see Section
S5 Supporting Information). Thus, uniformity in SOM chemical
composition is important for method applicability, at least as it affects
LOI at selected LT and HT.
The SOM is a function of several factors such as climate, land
cover, soil texture, and soil management practices (Jobbá gy and
Jackson, 2000; Quideau et al., 2001). Therefore, SOM chemical
composition might be similar at least for some fractions of
organic matter (i.e., labile or humic) in soils with the same
abovementioned factors, es- pecially for soils from the same horizon.
This will lead to similar RLOIs at different spatial locations within the
soil. The effect of spatial variations in RLOIs on estimated biochar
content can be assessed if a sufficient number of biochar-free soil
samples are analyzed for LOI at LT and HT. The variation of RLOIs for
the soil in this study is discussed in detail in Section S3 Supporting
Information.
5.3. Guidelines for selecting low and high temperatures
Biochar thermal stability is a function of charring temperature and in-
creases with increasing HTT (Harvey et al., 2012; Sun et al., 2014). By
comparing thermogravimetric thermograms of biochars reported in
the literature, a relatively similar trend for all biochars is observed
regardless of feedstock and pyrolysis temperature: biochars are
thermally stable below the ignition temperature, lose mass above the
ignition temperature over a particular temperature range (~200–250 °C)
that diminishes as the burnout temperature is approached, and above the
burnout temperature no further mass loss occurs (Fan et al., 2017;
Harvey et al., 2012; Sun et al., 2014; Zhou et al., 2014). The ignition
and burnout temperatures are higher for biochars produced at higher
temperatures. Thus, mass loss for biochars occurs at temperatures
that increase with HTT. This trend was observed for the biochar used
in this study in both the LOI (Fig. 4A) and thermogravimetric data (Fig.
S2). The ignition and burnout tempera- tures obtained from the LOI
method can be selected as LT and HT, respec-
tively, as long as RLOIs ¼ LOIsLT =LOIsHT is significantly different from the
ratio of LOILT =LOIHT . However, extra caution must be taken when
HT
b b
N 500 °C in soils with a high clay or C inorg content, since significant soil
mass loss might occur over the same temperature range that biochar
loses mass (Heiri et al., 2001; Wang et al., 2011).
6. Conclusions
A methodology was developed to determine the mass fraction of
biochar mixed within soil of unknown and varying organic matter
balance), the proposed method is ideally suited for routine quantifica-
tion of biochar in soil. The validity of the proposed methodology was
tested in adjacent filter strips at a field site with known biochar
content: bf = 0% in biochar-fee soil, and bf = 4.0 ± 1.1 CI% in biochar-
amended soil. Using the two-temperature LOI method, average
biochar content in the biochar-free soil was bf = −0.23 ± 0.09 CI%,
and in the biochar-amended soil was bf = 3.9 ± 0.3 CI%. Thus, the
method pro- duced estimates of biochar content that agreed with
independent measurements.
While the two-temperature LOI method shows much promise,
there are limitations. The method requires the LOI profile of the pure
soil and pure biochar that represent soil and biochar in the mixture.
The biochar should be stable and have minimal effect on SOM
mineralization. The method can be utilized for quantifying biochars
produced at high tem- peratures (i.e. N400 °C) with more confidence,
since these biochars are more environmentally recalcitrant. Another
requirement is the distinct difference in relative mass loss of the
biochar and soil at two tempera-
tures. While this requirement is often met because SOM is generally
more labile than biochar and biochar typically has higher organic con-
tent than soil, the method accuracy must be checked when soil has
high organic matter content (e.g. N15%) or significant quantities of
in- digenous black C and humic substances. The method should also
be checked when biochar is made from a feedstock with high ash
content (e.g. N30%), such as manure and food waste, and when
produced at low temperatures (i.e. b400 °C). The method also
requires relatively uniform SOM chemical composition. This
requirement may be satisfied for soils under the same climatic and
land management conditions, es- pecially if the depth of interest is in
the same soil horizon. Finally, the biochar ignition and burnout
temperatures where the mass loss starts and diminishes in pure
biochar LOI profile, respectively, may be selected as low and high
temperatures as long as other requirements of the two- temperature
LOI method are met.
Acknowledgments
The research was funded by the National Fish and Wildlife
Founda- tion, funded by the US Environmental Protection Agency
through the Chesapeake Bay Program, the Delaware Department of
Transportation, and the NAS Transportation Research Board through
award NCHRP-182.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.scitotenv.2018.12.174.
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