SOIL, 10, 275–279, 2024
https://doi.org/10.5194/soil-10-275-2024
© Author(s) 2024. This work is distributed under
the Creative Commons Attribution 4.0 License.
SOIL
The six rights of how and when to test
for soil C saturation
Johan Six, Sebastian Doetterl, Moritz Laub, Claude R. Müller, and Marijn Van de Broek
Department of Environmental Systems Science, ETH Zurich, Zurich, 8092, Switzerland
Correspondence:
Johan Six (jsix@ethz.ch)
Received: 28 September 2023 – Discussion started: 7 November 2023
Revised: 12 February 2024 – Accepted: 28 February 2024 – Published: 15 April 2024
Abstract.
The concept of soil organic carbon (SOC) saturation emerged a bit more than 2 decades ago as our
mechanistic understanding of SOC stabilization increased. Recently, the further testing of the concept across a
wide range of soil types and environments has led some people to challenge the fundamentals of soil C saturation.
Here, we argue that, to test this concept, one should pay attention to six fundamental principles or “rights” (R’s):
the right measures, the right units, the right dispersive energy and application, the right soil type, the right clay
type, and the right saturation level. Once we take care of those six rights across studies, we find a maximum of
C stabilized by minerals and estimate based on current data available that this maximum stabilization is around
82
±
4 g C kg
−
1
silt
+
clay for 2 : 1-clay-dominated soils while most likely being only around 46
±
4 g C kg
−
1
silt
+
clay for 1 : 1-clay-dominated soils. These estimates can be further improved using more data, especially
for different clay types across varying environmental conditions. However, the bigger challenge is a matter of
which C sequestration strategies to implement and how to implement them in order to effectively reach this
82
/
46 g C kg
−
1
silt
+
clay in soils across the globe.
In recent years, several studies (e.g., Begill et al., 2023;
Salonen et al., 2023) have questioned the concept of soil car-
bon (C) saturation, i.e., organic C stabilized by soil minerals
(Hassink, 1997; Six et al., 2002). Here, we want to draw at-
tention to six fundamentals that we think one should be cog-
nizant about when testing and questioning soil C saturation.
1
The right measures
Soil C saturation is, by definition, a non-responsiveness
of soil C content upon an increase in C inputs and thus
should ideally be assessed by plotting stabilized soil C (e.g.,
mineral-associated organic C (MAOC)) versus C input (Six
et al., 2002). However, most datasets used to test saturation
contain a large range of soil textures, clay types, and cli-
mates, where C input from vegetation to soil (especially to
specific soil C fractions) is usually not known; thus, the re-
sulting plots of soil C versus C input are often not sensible
and thus inconclusive (see West and Six, 2007; Stewart et al.,
2007; Feng et al., 2014). Hence, the most practical way to
test saturation is plotting MAOC vs. silt
+
clay content as a
proxy for the reactive mineral phase in soil. Specifically for
fractions of soil C, in Stewart et al. (2007), it was correctly
argued that saturation can also be tested by plotting a frac-
tion of soil C, such as MAOC, versus total SOC. However,
this does not account for the above-mentioned confounding
factors when going across systems or biomes. Therefore, the
most elegant, accurate, and thus preferable way to test for
saturation is by plotting MAOC versus silt
+
clay content in
the right units.
2
The right units
As for any analysis, using the correct units is of great im-
portance. For C saturation, the best data analysis that can
be done is relating MAOC (
y
axis), expressed per unit to-
tal soil (i.e., g C kg
−
1
total soil or mg C g
−
1
total soil; the
emphasis is on per total soil, which is
<
2 mm fine soil) to
silt
+
clay content (in units of g silt
+
clay kg
−
1
soil or %)
(
x
axis). Then, to correctly quantify the maximum MAOC in
Published by Copernicus Publications on behalf of the European Geosciences Union.
276
J. Six et al.: The six rights of how and when to test for soil C saturation
the right unit, one should conduct a boundary analysis (or a
quantile regression; Koenker, 2005) where the slope of the
linear boundary curve is the maximum MAOC (expressed in
g MAOC per unit of silt
+
clay – i.e., the portion of the soil
<
50 or 63 µm; we remark here that it is not total soil but
silt
+
clay that is the right unit for maximum MAOC) (see
Feng et al., 2013). Depending on the exact units used on the
y
axis and
x
axis, a conversion might have to be done to put the
maximum MAOC in the preferred right unit (i.e., g C kg
−
1
silt
+
clay or mg C g
−
1
silt
+
clay or % C of the silt
+
clay).
It is also pertinent to have the curve (1) not predict beyond
the range of available silt
+
clay measurements; (2) be linear
because, from a mechanistic perspective, it does not make
sense that the capacity to stabilize C by individual silt and
clay particles is dependent on the total amount of silt
+
clay
in the soil; and (3) go through the origin because if there
are zero minerals (i.e., silt
+
clay) then there is, by defini-
tion, also zero MAOC – hence, a non-zero intercept is the-
oretically not possible. Nevertheless, the difference between
boundary lines without and with an
y
intercept could indicate
the degree of contamination of MAOC with POC and thus
which samples should possibly be omitted from the anal-
ysis because of too high levels of contamination. Here, we
want to give an example of how the units matter: Georgiou
et al. (2022) state that their data-driven maximum MAOC es-
timate of 86 g C kg
−
1
mineral (
=
silt
+
clay) is a notable up-
date to the model-predicted 45–50 g C kg
−
1
soil reported by
Cotrufo et al. (2019). However, the value of 86 has the unit
of g C kg
−
1
silt
+
clay, whereas the estimate of 45–50 has the
unit of g C kg
−
1
(total) soil; thus, these are not comparable.
3
The right dispersive energy and application
As for any soil C fractionation method, the separation of
MAOC is operationally defined; thus, methodological short-
comings should always be considered when interpreting the
data. For the testing of MAOC saturation, it is obviously per-
tinent to separate an as-pure-as-possible mineral fraction that
is not contaminated by, for example, particulate organic car-
bon (POC). The separation of the mineral fraction is done
by size and/or density separation after dispersing the soil
into primary particles. Crucial here is that enough disper-
sive energy (through shaking, ultrasonic dispersion, or any
other dispersive method) is applied to break up all soil aggre-
gates to release all sand-sized particles without applying too
much dispersive energy that would break up POC and con-
taminate the mineral fraction. In that sense, Amelung and
Zech (1999) did a very thorough assessment of the influ-
ence of ultrasonic dispersion on POM redistribution to the
silt
+
clay fraction. They concluded that 3 kJ for 10 g soil
is optimal and that
≥
5 kJ disrupts POC and contaminates
MAOC with POC. As an example, Begill et al. (2023) re-
port that they used 100 J mL
−
1
for 10 g soil in 150 mL wa-
ter. Hence, they disrupted 10 g soil with 15 kJ, 3 times the
maximum level of energy recommended by Amelung and
Zech (1999), thus most likely disrupting POC and contam-
inating MAOC. One could do a density flotation to remove
contaminating POC to get a more pure MAOC, but it is our
experience that one cannot float all of the POC without also
starting to float some MAOC because there is a continuum of
densities from POC to MAOC. Thus, even though the ideal
dispersive energy is soil dependent, we recommend limiting
the dispersive energy to the level recommended by Amelung
and Zech (1999) and, most importantly, to always perform a
visual inspection with a microscope of the POC fraction to
ensure the break-up of all soil aggregates (not concretions;
this is specifically important in tropical soils that have very
stable concretions (pseudo-sand) that cannot be broken up
without POC contamination of MAOC) and of the MAOC to
ensure a limited contamination (because no contamination is
impossible) of the MAOC with POC (especially in high-POC
soils). The principles of Amelung and Zech (1999) still hold
up to this day despite the fact that more recent works (e.g.,
Poeplau and Don, 2014; Graf-Rosenfellner et al., 2018) did
emphasize that, even if one stays below the recommended
maximum energy input, results between the applications of
the same total amount of energy input but at varying intensi-
ties and lengths can still significantly affect the overall result
of a soil carbon fractionation analysis.
4
The right soil type
When testing the limits of mineral C stabilization, con-
founding influences of other stabilization factors should be
avoided. For example, soil types characterized by anoxic
conditions such as Histosols, Gleysols, Stagnosols, Um-
brisols, and Cryosols should not be included in an analysis
of saturation of MAOC, even if they are currently drained or
melted. In such soils, MAOC has, at least in part, accumu-
lated due to oxygen limitation and not through mineral sta-
bilization. We are not stating that mineral stabilization does
not occur or is mechanistically different in such soils, but
the anoxic conditions confound the effect of stabilization by
interaction with the minerals. Furthermore, these soil types
often have high POC levels and thus an increased chance of
contamination of MAOC with POC during dispersion. Also,
Anthrosols and Technosols are not suitable to assess a “nat-
ural” limit because of all the anthropogenic processes that
are or were at play. Andosols should probably be treated in
their own category due to their specific soil properties (i.e.,
high levels of allophane and pyrophosphate-extractable alu-
minum) (see Beare et al., 2014, for a discussion on the soil C
stabilization mechanisms in Andosols). For the same reason,
Alisols could also be considered in their own category (or
together with Andosols) for a saturation limit analysis. For
Calcisols and Gypsisols, one obviously has to remove all in-
organic carbon before determining MAOC, but this can lead
to a high uncertainty in the estimate of MAOC; hence, cau-
SOIL, 10, 275–279, 2024
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J. Six et al.: The six rights of how and when to test for soil C saturation
277
tion should be applied. Of particular importance is that sam-
ples with geogenic C are not included (see Kalks et al., 2021,
for the prevalence of geogenic carbon in soils). Lastly, buried
soil layers, as found in Colluvisols, should probably not be
included due to their current or past conditions that can lead
to artificially high levels of MAOC. One could start thinking
that the saturation concept is then limited to few soils, but the
other 21 (out of 32) WRB soil groups do dominate across the
globe, and the soil types indicated to not be included in the
analysis are limited in range and are often associated with
specific positions within the landscape (except Cryosols, but
those are often not managed and are thus not suitable targets
to increase soil C content); even Gleysols cover only
∼
5 %
of the globe.
5
The right clay type
As has been done in most studies estimating a maximum of
MAOC (Hassink, 1997; Six et al., 2002; Feng et al., 2013;
Matus, 2021; Georgiou et al., 2022), the separation of soils
with different phyllosilicate clay types is necessary because
of their differences in reactivity and thus in their potential to
stabilize MAOC (as we indicated specifically for Andosols,
which are dominated by very reactive clays, in the previous
section). In that sense, it would be very interesting to have
estimates of the saturation level per clay type because then
we could define a saturation level for each soil based on the
amount of specific clays present in the soil. However, the
currently available data do not allow for that; thus, a sepa-
ration between the broad categories of 1 : 1 versus 2 : 1 clay
types is all that can be done. Similarly, what is still miss-
ing at larger scales is a better understanding of the role of
other, non-silicate clay-sized minerals for MAOC stabiliza-
tion, such as varying metal phases which do play a dominant
role in the mineralogy of many highly weathered soils. How-
ever, there are recent efforts to map clay type distributions
across the globe (see Ito and Wagai, 2017), and, hopefully
soon, the resolution of these maps will be fine enough for C
saturation analyses.
6
The right saturation level
Since many soils are not receiving high-enough C inputs to
come close to soil C saturation, particularly subsoils, it is
important to be cognizant of what level of MAOC can be
reached for a certain silt
+
clay content. A search through
the literature led to three publications that estimated the
maximum MAOC for 2 : 1-mineral-dominated soils with the
“right measures” and “right units”, as outlined above: Feng
et al. (2013) estimated 84
±
4 g C kg
−
1
silt
+
clay, Georgiou
et al. (2022) estimated 86
±
9 g C kg
−
1
silt
+
clay, and Ma-
tus (2021) estimated 81 g C kg
−
1
silt
+
clay (note: Matus
(2021) estimated it for all mineralogies and did have a small
intercept, but samples at the boundary line were 2 : 1 dom-
inated). Although there is overlap in data between the dif-
ferent studies, the similarity is striking, and they average
to a value of 84
±
3 g C kg
−
1
silt
+
clay. Very important to
note here is that Beare et al. (2014) also estimated a maxi-
mum MAOC for allophanic (153 g C kg
−
1
silt
+
clay) versus
non-allophanic soils (116 g C kg
−
1
silt
+
clay), but the non-
allophanic soils include Gleysols and some vitric Andosols.
Thus, it is not surprising that they have a higher estimate.
Nevertheless, their very high estimate for the allophanic soils
confirms that they should be treated in their own category.
For 1 : 1-mineral-dominated soils, we found two very sim-
ilar estimates (in part because of an overlap in data points
between the two studies): 43
±
4 g C kg
−
1
silt
+
clay (Feng
et al., 2013) and 48
±
6 g C kg
−
1
silt
+
clay (Georgiou et al.,
2022), averaging to 46
±
4 g C kg
−
1
silt
+
clay.
Here, we also want to give two examples of how consid-
ering the right saturation level is important. First, Salonen et
al. (2023) indicated that their soils sequestered more MAOC
than is suggested by saturation estimates and that the rela-
tionship between total soil OC and MAOC remained linear
without a flex point. A look at their data shows that they are
measuring a maximum MAOC of 60 g C kg
−
1
total soil for a
2 : 1-mineral-dominated soil with 80 % silt
+
clay (of which
68 % is actually clay). If we consider the average estimate
for maximum MAOC in 2 : 1-mineral-dominated soils of
84 g C kg
−
1
silt
+
clay and the 80 % silt
+
clay in their soil,
then we estimate saturation to be at 67 (
=
84
·
80 %) g C kg
−
1
total soil of MAOC. Hence, it is no surprise that Salonen et
al. (2023) did not see saturation of the MAOC because their
soils were below the limit. Second, Schweizer et al. (2021)
argued, based on the observation of a higher MAOC (ex-
pressed in the right unit of mg C g
−
1
fraction) in soils with
low clay content and an observed patchy and piled-up struc-
ture of SOM binding to clays, that clay content is not neces-
sarily a limiting factor for MAOC storage. However, all their
reported MAOC contents are below the 84 mg C g
−
1
fraction
(see Fig. 1e in their publication) and thus perfectly align with
our estimate of maximum MAOC.
Lastly, to show that getting the rights right is important, we
used the dataset (available at https://doi.org/10.5281/zenodo.
7966076) of Begill et al. (2023), who suggested that there
is no maximum MAOC content. Given that their dataset is
based solely on German soils, the right clay mineralogy is
2 : 1, with a maximum MAOC of 84
±
3 g C kg
−
1
silt
+
clay.
Given this, 159 out of their 189 soils are below the MAOC
saturation limit; only 30 (
<
16 % of all samples) are above
the saturation limit. Of those 30 soils, 13 are Gleysols, 4 are
Stagnosols, 3 are Anthrosols, 1 is a miscellaneous soil, and
1 soil is a Regosol with an extremely high silt
+
clay con-
tent (97 %) while not being a well-developed soil; hence, it
cannot be excluded that this soil contains geogenic carbon
stemming from the (likely) sedimentary rock parent mate-
rial. Thus, eight soils remain above the limit after consider-
ing the right soil type, which is 4 % of the soils in the dataset.
Furthermore, seven out of those soils are below 91 g C kg
−
1
https://doi.org/10.5194/soil-10-275-2024
SOIL, 10, 275–279, 2024
278
J. Six et al.: The six rights of how and when to test for soil C saturation
Figure 1.
Boundary analysis of mineral-associated organic C
(MAOC) versus silt and clay content of the fine soil based on the Be-
gill et al. (2023) dataset without Gleysols, Stagnosols, Anthrosols,
and “miscellaneous” soils (indicated by gray crosses). The bound-
ary line (in blue) is based on the 90th percentile (purple dots) with
a 95 % confidence interval (blue shade). Orange dots are all other
soils in the dataset, including Cambisols, Luvisols, Phaeozems,
Regosols, and Vertisols. Note that a much weaker trend between
MAOC and the silt and clay content is visible for the subgroup
of soils with gray crosses, indicating the presence of non-texture-
dependent factors (e.g., anoxic conditions) determining the mea-
sured MAOC content in those soils.
silt
+
clay, which is within 1 standard deviation of the value
of Georgiou et al. (2022), i.e., exactly what you would expect
in a normal distribution of samples and/or errors. However,
more interesting is that when all Gleysols, Stagnosols, An-
throsols, and miscellaneous soils are taken out of the dataset
and a boundary analysis is done, we find a perfect bound-
ary with an estimated maximum MAOC of 78
±
6 g C kg
−
1
silt
+
clay (see Fig. 1), which is a very similar estimate to
others for 2 : 1-dominated soils, thus supporting the use of
the rights to estimate rightly the maximum mineral stabiliza-
tion. Including this additional estimate in the three examples
above leads to an average for 2 : 1-mineral-dominated soils
of 82
±
4 g C kg
−
1
silt
+
clay or, thus, roughly 8 % C in the
mineral fraction.
7
Conclusions
In conclusion, we can confidently state that (1) there is strong
empiric evidence for a maximum of C stabilization by soil
minerals, and (2) without more mineralogy data, the esti-
mate of 82 g C kg
−
1
silt
+
clay for 2 : 1-clay-dominated soils
is rather solid, while the value of 46 g C kg
−
1
silt
+
clay for
1 : 1-clay-dominated soils has a larger uncertainty. Further-
more, from a practical standpoint, we conclude the follow-
ing:
1. Most “managed” soils have a substantial C saturation
deficit.
2. We should focus our efforts to sequester C (both as
MAOC and POC) in soils with a high silt
+
clay con-
tent that are far below the maximum MAOC.
3. In sandy soils, we should focus on how to get POC
stored because the MAOC will be saturated fairly
quickly (unless they have anoxic conditions).
4. When estimating potential rates and amounts of seques-
tering soil C with models, saturation dynamics should
be considered (see Stewart et al., 2008; Georgiou et al.,
2022) and made spatially explicit based on environmen-
tal conditions.
Data availability.
We only used data from Begill et al. (2023) that
are available at https://doi.org/10.5281/zenodo.7966076 (Poeplau,
2023).
Author contributions.
Conceptualization was done by JS. For-
mal analysis was done by CRM and JS. All the authors conducted
the investigation. JS wrote the initial draft. All the authors reviewed
and edited several drafts, including the final paper.
Competing interests.
At least one of the (co-)authors is a mem-
ber of the editorial board of
SOIL
. The peer-review process was
guided by an independent editor, and the authors also have no other
competing interests to declare.
Disclaimer.
Publisher’s note: Copernicus Publications remains
neutral with regard to jurisdictional claims made in the text, pub-
lished maps, institutional affiliations, or any other geographical rep-
resentation in this paper. While Copernicus Publications makes ev-
ery effort to include appropriate place names, the final responsibility
lies with the authors.
Financial support.
Funding to Claude R. Müller and Mar-
ijn Van de Broek was provided by a Swiss National Science
Foundation grant (SNSF; 2020; PZ00P2_193617/1). Funding to
Moritz Laub was provided by a Swiss National Science Foundation
grant (SNSF; 2017; 200021_172940).
Review statement.
This paper was edited by Bas van Wesemael
and Jeanette Whitaker and reviewed by two anonymous referees.
SOIL, 10, 275–279, 2024
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J. Six et al.: The six rights of how and when to test for soil C saturation
279
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SOIL, 10, 275–279, 2024