Journal of Soil Science and Plant Nutrition

https://doi.org/10.1007/s42729-020-00259-1

ORIGINAL PAPER

Phosphorus Extraction with Soil Test Methods Affected by Soil P

Sorption Capacity
Gilmar Luiz Mumbach 1 & Luciano Colpo Gatiboni 2 & Daniel João Dall’Orsoletta 3 & Djalma Eugênio Schmitt 4 &
Patrícia Pretto Pessotto 1 & Clovisson Menotti Boeira de Oliveira 5

Received: 17 January 2020 / Accepted: 30 April 2020

# Sociedad Chilena de la Ciencia del Suelo 2020

Abstract
The aim of this study was to evaluate the phosphorus (P) extraction ability of the methods Mehlich 1 (M-1), Mehlich 3 (M-3),
anion exchange resin in strips (AER-ST), and anion exchange resin in spheres (AER-SP), as a function of clay content, P rates,
and incubation time. Experimental units were built with 20%, 40%, 60%, and 80% of clay and five different phosphorus rates as
follows: 0, 100, 200, 400, and 600 mg P kg−1, with subsequent incubation for 0, 30, and 90 days. After that, the samples were
subjected to the analysis of extractable P using the M-1, M-3, AER-ST, and AER-SP extractors. All P extraction methods
evaluated were sensitive to clay content and to the amount of added P. The input of P increased the recovery of this nutrient
by all extractors, but this was less effective when the clay content of the soil was increased. In general, the ability of phosphorus
extraction from the soil followed the order: M-1 > AER-SP > M-3 > AER-ST. The longer contact time of P with the soil, lower is
the extracted P content, regardless of the extraction method, this being more evident in clayey soils. All extraction methods tested
are sensitive to clay content, incubation time, and to the P concentration applied.

Keywords Buffer capacity . Mehlich 1 and Mehlich 3 . Anion exchange resin

1 Introduction 2019), resulting in low nutrient efficiency and a high depen-

The predominant soils in Brazil present a high degree of
weathering and a high P adsorption ability (Bortoluzzi et al.
2015; Roy et al. 2017; Withers et al. 2018). The P added to the
soil might be adsorbed with a high degree of affinity between
the element and the soil particle (Roy et al. 2017; Cheng et al.
* Gilmar Luiz Mumbach
gilmarmumbach@hotmail.com
1
Department of Soil Science and Natural Resources, Soil Science Post
Graduation Program, Santa Catarina State University, 2090 Luiz de
Camões Avenue, Lages, SC 88520-000, Brazil
2
Department of Crop and Soil Sciences, North Carolina State
University, Raleigh, NC, USA
3
National Council for Scientific and Technological Development
(155424/2018-1), 2090 Luiz de Camões Avenue, Lages, SC, Brazil
4
Department of Agriculture, Biodiversity and Forest, Federal
University of Santa Catarina, Curitibanos, SC, Brazil
5
Department of Soil Science and Natural Resources, Soil Science Post
Graduation Program, 2090 Luiz de Camões Avenue, Lages, SC,
Brazil
dence on the input of fertilizers (CQFS – RS/SC 2016;
Menezes-Blackburn et al. 2017).
The interpretation of the availability of P in soils in the
South of Brazil is performed through the Mehlich 1 (M-1)
and Mehlich 3 (M-3) extractors (CQFS – RS/SC 2016). The
M-1 is easily used and present a good calibration for the pre-
dominant soils in the region, but it overestimates the P content
extractable in soils with the application of natural phosphates
and has a low extraction ability in clayey soils (Freitas et al.
2013), resulting in different values according to the clay con-
tent of the soil. The clay fraction and its mineralogy have a
great influence on the retention of P to the solid phase of the
soil (Bortoluzzi et al. 2015; Vásconez and Pinochet 2018) due
to the high reactivity of the constituents of this fraction, such
as iron and aluminum (hidr)oxides (Camargo et al. 2013; Fink
et al. 2016).
As an alternative to the M-1 method, the calibration of the
M-3 extractor was developed for the South region of Brazil,
which is less affected by the source of fertilizer used (Oliveira
et al. 2015), besides being a multielemental extractor, in which
other elements such as potassium (K), calcium (Ca), magne-
sium (Mg), sulfur (S), and some micronutrients might be

analyzed, in addition to P (Mehlich 1984). There are also P
extractors that act by ion exchange, such as the anion ex-
change resin (AER), which might be performed with strips
(AER-ST) or spheres (AER-SP), being methods recommend-
ed in other Brazilian states. However, few works have evalu-
ated the influence of clay content on the extractive ability of
these methods (Mumbach et al. 2018).
The time of contact of the phosphate fertilizers with the soil
affects the amount of this mineral that remains available to the
plants. In soils with a history of phosphate fertilization, the
efficiency of the applied fertilizers tends to be higher, com-
pared with natural and non-fertilized soils (Barrow and
Debnath 2014; Hopkins et al. 2018; Withers et al. 2018). In
these cases, the amount applied might be gradually reduced,
based mainly on the amount of exported grains of the crop of
interest (Barrow 2015; CQFS – RS/SC 2016). The increase in
phosphate fertilization efficiency in soils with a history of
fertilization occurs due to the decrease of the P binding energy
to the adsorption sites (Oliveira et al. 2014; Roy et al. 2017;
Withers et al. 2018), as well as to the increase in the negative
electric potential of the soil (Barrow and Debnath 2014;
Barrow 2015).
The clay content, the amount of P applied, as well as the
time elapsed since the application of P might interfere with the
amount this element extracted by M-1, M-3, AER-ST, and
AER-SP. However, there are still no studies evaluating the
sensitivity of these methods in soils with the same mineralogy
differing only in the clay content. In this sense, the main hy-
potheses of the study are as follows: (i) all tested P extraction
methods are sensitive to the clay content of the soil; (ii) the
amount added and the time of contact with the soil affect the
available amount of P, regardless of the extraction method.
Therefore, the objective of this study was to compare the
capacity of P extraction from the soil by the methods
Mehlich 1, Mehlich 3, and anion exchange resin in strips
and spheres, as a function of the P concentration, incubation
time, and soil clay content.
2 Materials and Methods
The experiment was conducted at the Agroveterinary Sciences
Center of the Santa Catarina State University (CAV-UDESC),
localized in Santa Catarina state, in Southern Brazil. The study
was carried out with a soil from the subsurface layer (Bw
horizon – 140/220 cm) from a Ferralsols (WRB 2014), with
800 g kg−1 of clay, collected in native field area, located at the
municipality of Campos Novos – SC (27°22′27”S, 51°04′
27”W). The mineralogy of the soil clay fraction used is com-
posed predominantly of kaolinite, goethite, hematite, and
clays in a 2:1 ratio with hydroxy-Al interlayers, with reduced
amount of gibbsite. Still, the oxidic forms found show high
crystallinity (Almeida et al. 2003).
J Soil Sci Plant Nutr
After collection, the soil was dried in an oven with a con-
stant temperature of 60 °C for 72 h and after sieved in 2.0-mm
mesh. Then, the soil was mixed with different amounts of
sieved sand in 1.0-mm mesh and retained in a 0.5-mm mesh,
previously washed with HCl 0.1 mol L-1 and distilled water.
For the composition of the experimental units, mixtures of
sand and soil were made in the proportions of 75% sand +
25% soil; 50% sand + 50% soil; 25% sand + 75% soil; and 0%
sand + 100% soil, representing soils with 20, 40, 60, and 80%
clay, respectively.
After the creation of the soils, they were incubated for
30 days at 80% of field capacity for stabilization of their
physical-chemical properties and re-incubated for another
30 days with acidity correction (Filler lime with 98.11%
PRNT) to raise the pH H2O to 6.0. The main characteristics
of the soil samples, established after mixing it with sand, are
presented in Table 1, and more details about the original soil
might be obtained in Oliveira et al. (2014).
After the incubation period, the samples were dried in an
oven with a constant temperature of 60 °C for 72 h and after
sieved on a 2-mm mesh. Then, the soils with different clay
contents were packed in 500-g plastic pots, with each pot
forming an experimental unit. The experimental design used
was completely randomized, in a 4 × 5 × 3 factorial scheme,
with four soils with different clay contents, to which five rates
of P were added as follows: 0, 100, 200, 400, and 600 mg P
kg−1 of soil, using a solution of KH2PO4 PA as source. After
adding P, the experimental units were subjected to three incu-
bation periods (0, 30, and 90 days after incubation (DAI)),
being the date “0” referred to the collection carried out on
the same day of the application of the treatments. Four repe-
titions were used for each treatment tested. During the incu-
bation period, the samples were kept with soil moisture close
to 80% of field capacity.
After each incubation period, the experimental units were
dried in an oven with forced aeration at 65 °C, ground and
sieved on a 2.0-mm mesh sieve. Samples of the experimental
units were submitted to extraction of P by four extraction
methods: (1) Mehlich 1 (M-1) (0.05 mol L−1 of HCl and
0.0125 mol L−1 of H2SO4) (Mehlich 1978); (2) Mehlich 3
(M-3) (0.2 mol L −1 of CH 3 COOH, 0.25 mol L −1 of
NH4NO3, 0.015 mol L−1 of NH4F, 0.013 mol L−1 of HNO3
and 0.001 mol L−1 of EDTA) (Mehlich 1984); (3) anion ex-
change resin in strips (AER-ST) (Tedesco et al. 1995); and (4)
anion exchange resin in spheres (AER-SP) saturated with
NaHCO3 (Miola 1995). After the extraction, the extractable
P was determined by colorimetry at a wavelength of 882 nm
(Murphy and Riley 1962).
The applied P recovery rate (% of P recovered) represents
the percentage amount of P recovered by each extraction and
was calculated for the four extraction methods according to
the P rates and clay levels. The P recovery rate was calculated
by Eq. 1:
J Soil Sci Plant Nutr

Table 1 Chemical and particle size attributes of the experimental units created from mixing the soil with different amounts of sand

Soil Clay Silt Sand TOC(1) pH H2O Ca(2) Mg(2) Al(2) H + Al K(3) P(3) Pmax*(4)
g kg−1 1:1 cmolc dm−3
mg kg−1

200 200 54 746 Nd 5.4 0.1 0.1 0.2 1.7 8.0 1.7 353.8
400 400 53 547 Nd 5.3 0.8 0.5 0.1 2.0 7.0 1.9 1000.6
600 600 58 342 2.0 5.8 2.5 1.4 0.0 2.0 9.0 1.8 1630.1
800 800 57 143 3.0 5.7 3.5 2.1 0.0 2.2 10.0 1.6 2111.5

*Data published in Oliveira et al. (2014). Nd: not analytically detected. (1) TOC: total organic carbon (Walkley-Black method). (2) Extracted with the KC1
1 mol L−1 . (3) Extracted through Mehlich 1. (4) Pmax: maximum phosphorus sorption capacity by Langmuir isotherms. Soil “800” represents the original
soil sample, with no added sand

The four methods of P extraction presented high correla-

ðPdetermined – P original in the soilÞ
% of P recovered ¼  100 ð1Þ tions between them (p < 0.05) (data not presented). The P
P added
extracted by the different extraction methods correlated nega-
where “P determined” represents the amount of P extracted in tively with the clay content and positively with the P rates. On
the treatment that received a certain P rate and “P original in the other hand, the P extracted by the different extraction
the soil” represents the non-fertilized condition; the “P added” methods showed no correlation with the incubation time (data
is represented by the amount of P applied in each sampling not presented).
unit. These three variables are considered in the calculation,
each expressed in mg kg−1, respectively.
The data were initially submitted to a normality test,
3.2 P Extraction According to the Time of Incubation
being transformed by cubic root. Later, they were sub-
and the Soil Clay Content
mitted to the analysis of variance (ANOVA) and, when
The amount of P extracted by the methods varied according to
significant, they were compared by the Tukey’s HSD
the incubation time and clay content (Table 3). In general, the
test of mean and linear regression, for the qualitative
largest amounts of P were extracted by the Mehlich 1 (M-1)
and quantitative variables, respectively. For all tests per-
method, followed by the AER in spheres (AER-SP), Mehlich
formed, the minimum probability error was set at 5% of
3 (M-3), and AER in strips (AER-ST).
significance (α = 0.05). The original data were used to
The incubation time affected the amount of P recovered by
interpret the results. The statistical analysis was per-
the extractors. After 90 days of incubation, considering the
formed using the Sisvar 5.6 (Ferreira 2014).
soil with only 20% clay, there was a reduction of 9.2, 11.0,
and 4.4% in the P extracted by the methods M-1, M-3, and
AER-SP, respectively. The AER-ST method presented an op-
posite behavior, with an increase of 16.4% in the comparison
3 Results
between soil samples collected on the day of incubation and
90 days after it. However, the 80% clay soil presented a re-
3.1 Overview of Results
duction of 48.5, 63.4, 59.7, and 28.8% in the P extracted
90 days after the incubation by the methods M-1, M-3,
The reduction of the soil clay content has altered some
AER-ST, and AER-SP, respectively. In general, considering
soil attributes, especially the reduction of the adsorption
the average rates applied and the clay content, after 90 days,
capacity of P (Table 1), as already highlighted by
the average reduction in extraction capacity was of 26.8%,
Oliveira et al. (2014). There were significant effects
36.9%, 18.9%, and 22.0% for the M-1, M-3, AER-ST, and
for all causes of variation tested on the availability of
AER-SP methods, respectively. The results suggest that all
P in the soil, except for the interaction between the P
extractors are influenced by time, recovering less P in longer
extracted by the different extraction methods, clay con-
incubation periods, especially with high amounts of clay
tent and incubation time (Table 2). Based on this, the
content.
two triple interactions that presented significant behavior
All the extractors evaluated were sensitive to soil
were unfolded. The triple interaction between clay, P
texture, recovering less P in soils with higher amounts
rate and time was disregarded, since it did not meet
of clay. In general, when the soil increased from 20 to
the main objective of this work, i.e., to assess the in-
80% clay, considering the averages of the applied rates
fluence of factors on P extraction by the tested methods.
and of the incubation times, the average phosphorus
 J Soil Sci Plant Nutr

Table 2 Analysis of Variance

(test F of ANOVA) for the P Variable/interactions DF(1) SS(2) MS(3) Fc(4) P value(5)
extracted depending of the
variables analyzed in the study Methods 3 50.4 16.8 230.1 < 0.0001
Clay 3 301.6 100.5 1375.3 < 0.0001
P rates 4 1234.8 308.7 4223.8 < 0.0001
Time 2 8.4 4.2 57.6 < 0.0001
Methods*Clay 9 18.4 2.0 28.0 < 0.0001
Methods* P rates 12 15.0 1.2 17.1 < 0.0001
Methods*Time 6 1.8 0.3 4.2 < 0.0001
Clay* P rates 12 137.5 11.4 156.8 < 0.0001
Clay*Time 6 2.7 0.4 6.1 < 0.0001
P rates*Time 8 3.0 0.4 5.1 < 0.0001
Methods*Clay* P rates 36 17.4 0.5 6.6 < 0.0001
Methods*Clay*Time 18 3.6 0.2 2.7 < 0.0001
Methods* P rates*Time 24 1.6 0.1 0.9 0.597
Clay* P rates*Time 24 4.5 0.2 2.6 < 0.0001
Methods*Clay* P rates*Time 72 8.0 0.1 1.5 0.005
Error 720 52.6 0.1 – –
(1) (2) (3) (4) (5)
Degrees of freedom. Sum of squares. Mean square. Calculated F value. Probabilistic value of
statistical significance

extraction capacity decreased 78.5%, 93.6%, 94.1%, and
78.6% for the M-1, M-3, AER-ST, and AER-SP
methods, respectively. Our results suggest that the re-
covery of P by the extractors reduces sharply with the
increase of clay, with the AER-ST extractor being the
most sensitive to this condition.
3.3 Extraction of P According to the Rates and the
Clay Content of the Soil
The extraction methods proved to be sensitive to P rates,
extracting higher amounts of this nutrient with the increase
of the rate applied (Fig. 1). There was a higher sensitivity to

Table 3 Amounts of P extracted by the Mehlich 1, Mehlich 3, anion exchange resin in strips (AER-ST), and anion exchange resin in spheres (AER-SP)
methods, according to the clay content and incubation times, considering the average rates of the nutrient applied

Time Extraction methods Extracted P, mg kg−1

Clay (%)

Days 20 40 60 80 Regression for clay content

0 Mehlich 1 69.6 aA 31.1 aA 27.2 aA 20.2 aA y = 3.0 + (1306.3**/x) R2: 0.97

Mehlich 3 55.4 abA 22.5 bA 12.2 bA 5.2 cA y = 119.7**e(−0.04**x) R2: 0.99
AER–ST 63.8 bcB 26.9 bA 14.7 bA 6.2 cA y = 65.6–0.62**x R2: 0.98
AER-SP 52.4 cA 41.0 aA 31.2 aA 14.6 bA y = 136.4**e(−0.038**x) R2: 0.99
30 Mehlich 1 67.5 aA 27.5 aAB 21.5 aAB 12.8 aB y = 110.5*e(−0,029*x) R2: 0.89
Mehlich 3 46.9 bAB 18.8 bA 8.9 cB 2.8 cB y = 114.8**e(−0,045**x) R2: 0.99
AER–ST 53.9 bB 29.7 bA 12.2 cbA 2.6 cB y = 122.9*e(−0,034*x) R2: 0.96
AER–SP 61.2 bA 34.8 aA 13.0 bB 9.5 bB y = 136.7*e(−0,043*x) R2: 0.94
90 Mehlich 1 63.2 aA 24.6 aB 19.4 aB 10.4 aB y = 107.9*e(−0,031*x) R2: 0.90
Mehlich 3 49.3 bB 16.7 bB 6.0 cC 1.9 bB y = 143.3**e(−0,053**x) R2: 1.0
AER–ST 76.3 aA 31.1 aA 7.2 cB 2.5 bB y = 62.0–0,7**x R2: 0.97
AER-SP 50.1 bA 34.1 aB 19.4 bB 10.4 aAB y = 237.0*e(−0,055**x) R2: 0.98

Equal lowercase letters between methods within each time condition and clay content do not differ by the Tukey’s HSD test (p < 0.05). Equal capital
letters for the same method between times in each clay content condition do not differ by the Tukey’s HSD test (0.05). *Significant at p < 0.05. **
Significant at p < 0.01
J Soil Sci Plant Nutr

P M-1 = 0.314**Rate - 0.004**Clay x Rate R²: 0.840 P M-3 = 0.244**Rate - 0.003**Clay x Rate R²: 0.893

180 180

150 0 150

)
)

-1
-1
P extracted (mg kg

P extracted (mg kg
30 0
60 50
120 120
90 100
120 150
90 150 90 200
180 100
60 60 120
600 600
500 500

-1
-1
30 30

g
400 400

gk

gk
,m

,m
300 300
0 0

fP
of
70 200 70 200

o
tes

tes
60 60
50 100 Ra a 50 100 b

Ra
40 40
Clay, 30 0 Clay, 30 0
% 20 % 20

P AER-ST = 0.344**Rate - 0.004**Clay x Rate R²: 0.869 P AER-SP = 0.294**Rate - 0.003**Clay x Rate R²: 0.904

180 180

150
)

150
-1

0
P extracted (mg kg

0
kg )

30
-1

120 30 120 60
P extracted (mg

60
90
90
90 90 120
120
150
150
180
60 180 60
600 600
500 500
-1

30 30

-1
g

400 400
gk

g
gk
,m

300 300
0 0

,m
fP

200 70 200

fP
70
o

60
tes

60

o
50 100 c 50 100 d
tes
Ra

40 40
Clay, 30 0 Clay, 30 0 Ra
% 20 % 20

Fig. 1 P extracted by the methods Mehlich 1 (P M-1) (a), Mehlich 3 (P M-3) (b), anion exchange resin in strips (AER-ST) (c), and anion exchange resin
in spheres (AER- SP) (d) as a function of the interaction between the applied P rates and the clay content of the soil, ** Significant at p < 0.01

the increased rate by the AER-ST extractor, followed by M-1, recovered P occurred for all extraction methods and for all ap-
AER-SP, and M-3, with average increments of 0.34, 0.31, plied P rates. Excess was observed for the AER-SP extraction
0.29, and 0.24 mg kg-1 of P for each mg kg−1 of the nutrient method, in which significant differences occur only between the
applied, respectively. However, with the increase in clay con- two soils with the greatest clay amplitude. Generally, considering
tent, there is a reduction in the recovered P content, regardless the average P rates applied and only the soils with 20% and 80%
of the method considered, overlapping the effect caused by the of clay, the amount of P recovered varied from 20.8 to 3.8% for
increasing rates of this nutrient. M-1, 14.9 to 0.6% for M-3, 15.6 to 0.7% for AER-ST, and 17.6
The percentage of the recovered P was sensitive to the clay to 2.9% in AER-SP, respectively.
content and to the concentration of added P, for all extraction Unlike what was observed for clay, the increase in the rate
methods (Fig. 2). The percentage of P recovery by the methods of applied P resulted in an increase in the percentage of recov-
was relatively consistent throughout the time, because of that, ered P, for all soils. This increase in recovered P was observed
Fig. 2 shows only the results after 30 days of incubation. With for all extraction methods. Within each clay class, the recov-
the increase in the clay content of the soil there was a significant ered P was statistically higher with the highest rate applied,
reduction in the amount of P recovered. This reduction in compared with the two lowest rates. Generally, with the
 J Soil Sci Plant Nutr

45 45

Aa
36 36
P Recovered (%)

27 27
Ab
Ab Ac

18 18
Ac
Ac Abc
ABa
ABab Bb
Ba
Ac
9 Bab 9 Bab
Bc Bbc Ca Bab
Bab Ca BCa
Bb Ba BCa
Ba Ab
a ABa Ba Ca Ca b
Ba Ba
0 0
100 200 400 600 100 200 400 600

45 45
20% of clay
40% of clay Aa

36 36 60% of clay
80% of clay
P Recovered (%)

Aa
Aa
27 27
Aa
Bab
18 18 Ba
Ba

Ab Ab

9 9 Ac Ca
Bab Ca
Ac ABb Ca
BCab ABa Ba Ca Ca
ABa
ABc ABb
Ba
Bb Bab
Ba Ca Da c Bb Ba d
0 0
100 200 400 600 100 200 400 600
P rates (mg kg-1) P rates (mg kg-1)
Fig. 2 Percentage of phosphorus recovery extracted by the methods clay content and lowercase within phosphorus rates, indicate statistical
Mehlich 1 (a), Mehlich 3 (b), anion exchange resin in strips (c), and differences between treatments using the Tukey’s HSD test (p < 0.05).
anion exchange resin in spheres (d), in relation to P rates and soil clay The dark bars on the columns represent the standard error of the mean
contents, after 30 days of incubation. Different letters, uppercase within

application of the lowest P rate (100 mg kg-1), the amount of reduction in the extracted P, being more expressive in
recovered P varied from 13.5 to 2.8% for M-1, from 7.0 to the first 30 days after P application. In general, the
0.1% for M-3, from 5.9 to 0.3% for AER-ST, and from 7.0 to reaction of the added P with the soil particles is fast,
2.2% for AER-SP, for soils with 20 and 80% clay, respective- with an initial increase due to the dissolution of fertil-
ly. However, with the application of the highest P rate izers (Motaghian et al. 2020), leading to a posterior
(600 mg kg-1), the amount of recovered P was considerably significant reduction in the extracted fraction and, con-
higher, ranging from 33.9 to 5.2% for Mehlich 1, from 18.5 to sequently, to the accumulation in non-labile forms
1.1% for Mehlich 3, from 27.5 to 0.9% for AER-ST, and from (Santos et al. 2016; Roy et al. 2017). After this rapid
31.6 to 3.7% for AER-SP, for soils with 20 and 80% clay, adsorption phase, the process continues, however, more
respectively. slowly, mainly by the penetration of this nutrient into
the mineral imperfections (Barrow and Debnath 2018).
In this context, according to Barrow (2015), the process
4 Discussion of P adsorption into the soil might be described in two
phases. The first stage is characterized by the binder
4.1 Availability of P: Effect of Time of Contact exchange, where the most P avid sites are exposed,
taking place almost immediately after adding the nutri-
The influence of time of incubation in the availability of ent. The second phase of the adsorption process occurs
P, for all the considered extractors, showed a gradual slowly, being marked by the diffusion of P into the
J Soil Sci Plant Nutr
imperfections of the adsorbing minerals and by the in-
crease of the energy of affinity of the nutrient with the
adsorption sites.
The response of the time of contact of the fertilizer with the
soil might vary, being influenced by factors such as the soil
characteristics (Zehetner and Miller 2006; Vásconez and
Pinochet 2018) and even the extractor solution (Broggi et al.
2010). Assessing changes in P availability during 360 days
after fertilization, Vásconez and Pinochet (2018) observed
that the reduction in extractable forms occurs more signifi-
cantly in the first 60 days of incubation. Furthermore, no cor-
relation was notes with the clay content, indicating the impor-
tance of the mineralogy of the clay fraction on the availability
of P. Santos et al. (2016), evaluating the effect of incubation
time, P rates, and different soil types on the P recovery capac-
ity, observed a very heterogeneous behavior of the “incuba-
tion time” factor, with a small reduction rate, although not
systematically. Also in this study, it is important to note that
the greatest influence of time was observed in the first 30 days
after incubation, a fact also observed in the present study,
which might be attributed to the first phase of the P adsorption
process, which is responsible for the highest percentage of
retention of this nutrient (Barrow 2015).
It is also worth noting that the rate of reduction of P avail-
ability over time may be dependent on the P adsorption po-
tential of soils, as well represented in Table 3. Soils with
higher clay content or with a predominance of minerals that
have greater potential retention of P results in lower amounts
of extractable P. Zehetner and Miller (2006), comparing two
soils in the Andes region of Ecuador, one with an altitude
above 2700 m and with a predominance of amorphous min-
erals and another at a lower altitude and with a predominance
of halloysite, observed that, after 30 and 180 days of applica-
tion of P rates, only 31 and 19% of the nutrient were recovered
in the higher altitude soil and around 75% in the soil with a
predominance of more crystallized minerals. In addition to
mineralogy, authors also attribute the highest P adsorption in
soil at an altitude above 2700 m to lower pH values, which
represent a greater potential for positive electrical charges.
4.2 Availability of P: Effect of the Clay Content
The increase in clay content causes a reduction in the amount
of P extracted in the soil. This phenomenon occurs due to the
higher total Fe and Al content in more clayey soils and, con-
sequently, of Fe and Al hydroxides, a factor that increases the
adsorption capacity and reduces the amount of P extractable
(Camargo et al. 2013; Oliveira et al. 2014; Bortoluzzi et al.
2015; Santos et al. 2016). Based on this, the adsorption capac-
ity of P by a soil is not explained only by its clay content, since
the predominant mineralogy has also a major influence
(Zehetner and Miller 2006; Bortoluzzi et al. 2015).
According to Wuenscher et al. (2015), the quantity and
crystallinity of iron oxides affect the behavior of P extraction
methods, in association with other factors such as pH and soil
texture. Studies show that clay alone cannot predict satisfac-
torily the availability of P in a group of contrasting soils; other
parameters, such as Pmax and remaining P, are much more
closely related (Broggi et al. 2010). In this study, in order to
evaluate the isolated participation of the clay content, only one
type of soil was used, which, mixed with sand, allowed the
creation of soils with different granulometric compositions,
but with equivalent mineralogy.
The dilution of the clay content, by the application of sand,
also resulted in dilution of the organic matter content of the
soil (Table 1). Organic matter can influence the dynamics of P
adsorption in soil in different ways, either by contributing to
the predominance of more amorphous minerals and with
greater reactivity with soil (Zehetner and Miller 2006) or by
reducing adsorption through the release of humic compounds
that block adsorption sites (Almeida et al. 2003).
The increase in soil clay content reduced the extraction
power of P in all methods in the following order: M-1 > M-
3 > AER-SP > AER-ST (Fig. 1). The effects of particle size
composition on the extraction capacity are acknowledged in
the literature (Bortolon and Gianello 2008; Freitas et al. 2013;
Mumbach et al. 2018). It is known that the M-1 extractor loses
extraction ability in soils with high clay content (Bortolon
et al. 2011), largely due to the exhaustion of the constituent
acids of the extracting solution (Bortolon et al. 2011), taking
this fact into account in manuals that recommend fertilization.
The availability of P in the Brazilian states of Rio Grande do
Sul (RS) and Santa Catarina (SC), as it happens in other re-
gions, is classified in clay classes, and the values of the critical
levels are lower for the more clayey soils (CQFS - RS/SC
2016), due to the abovementioned phenomenon. According
to the existing literature, other methods such as M-3 and ion-
exchange methods would not exhibit this same behavior or,
when they did occur, they would be affected to a lesser extent
(Oliveira et al. 2015).
The extraction behavior between the methods changes ac-
cording to the amount of clay in the soil. Considering the acid
dissolution extractors, the amount of P extracted by M-1 was
higher than M-3. However, the content of P was lower with
the higher clay content and with the lower P rates, conditions
where the adsorption potential of P is maximized (Oliveira
et al. 2014; Barrow 2015). In tropical and subtropical soils,
it is known that the largest extraction of P by M-3 compared
with M-1 only occurs in soils with up to 40% clay (Bortolon
and Gianello 2008; Mumbach et al. 2018). In more clayey
soils, the behavior is reversed (Bortolon and Gianello 2008;
Mumbach et al. 2018).
The extraction process with AER is considered to be of
good accuracy, with a better correlation with the amounts
absorbed by the plants compared the acid dissolution methods
(van Raij et al. 2009), no chemical alteration of the soil

constituents, and less influence of the clay fraction (Bortolon
et al. 2011; Steiner et al. 2012). In the present study, AER
methods only extracted more P, compared with the acid dis-
solution methods, in soils with 40% clay. The reduction in the
amounts extracted with the increase of clay, especially com-
pared with M-1, indicates that the granulometric composition
also affects the ion exchange methods (Mumbach et al. 2018).
Therefore, for better accuracy, the clay content should be tak-
en into account when assessing the availability of P for all the
evaluated methods. Higher amounts of P were extracted by
AER-SP than with the AER-ST; observed mainly for soils
with 80% clay, this might be attributed to the larger specific
surface of the spheres compared with the strips, delaying the
resin saturation with P.
4.3 Availability of P: Effect of the Amount of Applied
Fertilizer
The amount of P extracted increased linearly with the amount
of P applied to the soil. With the increase of the applied rate,
the most P avid sites become saturated and there is a gradual
reduction of the binding energy between the components
(Oliveira et al. 2014; Barrow and Debnath 2018; Khan et al.
2019). This lower affinity between P and the soil particles
might lead, even if in a less expressive way, to a more equiv-
alent behavior between the different extractors of the nutrient.
The increase in the P recovered as a function of the appli-
cation of P might be attributed to the blocking of parts of the
adsorption sites (Oliveira et al. 2014; Roy et al. 2017). As
already pointed out, in soils with low P content, the fertilizer
added to the soil and not absorbed by the plants is quickly
adsorbed, mainly by the sites more avid for P (Oliveira et al.
2014). However, in soils with a history of phosphate fertiliza-
tion, the adsorption of the subsequently added P starts to occur
in sites with lower binding energy and, consequently, a higher
amount tends to remain in forms of higher lability (Roy et al.
2017; Withers et al. 2018; Khan et al. 2019). In addition to the
saturation of the most avid sites, the P previously added to the
soil increases the negative electrical potential of the soil, since
it acts as a potential-determining ion, impairing the adsorption
of the nutrient added later (Barrow and Debnath 2018). Thus,
soils with high P availability present higher fertilization effi-
ciency, due to the higher desorption ability of the nutrient
(Barrow and Debnath 2014; Barrow 2015). However, the in-
crease in the percentage of P recovered with the increase in
applied rates, even though it is observed in all soils, it has its
magnitude varying according to the adsorption capacity. Soils
with higher P adsorption capacity result in lower percentages
of P accumulated in labile forms, regardless of the applied rate
(Zehetner and Miller 2006).
Field studies show that soils with a history of fertilization
tend to have a higher efficiency for applied fertilizers (Barrow
and Debnath 2014; Roy et al. 2017). This legacy of P present
J Soil Sci Plant Nutr
in the soil, even though largely unavailable to the annual cycle
plants, enables the reduction of the amount of fertilizer ap-
plied, based mainly on the amounts exported by crops
(Barrow and Debnath 2014; Barrow and Debnath 2018;
Withers et al. 2018). The gradual reduction of the P addition
limited to the amount exported by the crops, in soils with built
fertility, is a way to reduce the dependence on a non-
renewable input and reduce costs, especially in the Brazilian
weathered soils (Withers et al. 2018).
In soils of constructed fertility, the rational application of P,
based exclusively on the demand of the agricultural species,
can reduce nutrient losses, minimizing problems such as water
eutrophication (Zhang et al. 2017). The continuous and exces-
sive P addition, and the consequent reduction of its affinity to
soil particles (Barrow 2015; Khan et al. 2019), can lead to
vertical displacement of the nutrient, especially in soils with
less adsorption capacity (Guardini et al. 2012). In addition,
under a no-tillage system, the superficial application of P
causes an elevation of the nutrient in this layer, increasing
the risk of losses due to runoff (Schick et al. 2017).
5 Conclusions
The tested extraction methods correlate positively, indicating
an equivalent potential for use in soil characterization labora-
tories, provided they have calibration for the region of interest.
The methods Mehlich 1, Mehlich 3, anion exchange resin in
strips, and anion exchange resin in spheres are sensitive to the
clay content of the soil. This change in the extraction capacity
with the increase in clay content needs to be considered by the
fertilizer recommendation systems, when based on any of
these methods. The availability of P decreases with the incu-
bation time, especially in clayey soils. Soils receiving more
phosphate fertilizers and with less clay content have a higher
percentage of P extracted. Thus, soils with a history of fertil-
ization may present greater efficiency in the use of the applied
nutrient, which implies the reduction of phosphorus rates to
meet the plants’ needs.
Compliance with Ethical Standards
Conflict of Interest The authors declare that they have no conflict of
interest.
References
Barrow NJ (2015) Soil phosphate chemistry and the P-sparing effect of
previous phosphate applications. Plant Soil 397:401–409
Barrow NJ, Debnath A (2014) Effect of phosphate status on the sorption
and desorption properties of some soils of northern India. Plant Soil
378:383–395
J Soil Sci Plant Nutr
Barrow NJ, Debnath A (2018) Three residual benefits of applying phos-
phate fertilizer. Soil Sci Soc Am J 82:1–9
Bortolon L, Gianello C (2008) Interpretação de resultados analíticos de
fósforo pelos extratores Mehlich-1 e Mehlich-3 em solos do Rio
Grande do Sul. Rev Bras Cienc Solo 32:2751–2756
Bortolon L, Gianello C, Welter S, Almeida RGO, Giasson E (2011)
Simultaneous extraction of phosphorus, potassium, calcium and
magnesium from soils and potassium recommendations for crops
in southern Brazil. Pedosphere 21:365–372
Bortoluzzi EC, Pérez CAS, Ardisson JD, Tiecher T, Caner L (2015)
Occurrence of iron and aluminum sesquioxides and their implica-
tions for the P sorption in subtropical soils. Appl Clay Sci 104:196–
204
Broggi F, Oliveira AC, Freire FJ, Freire MBG, Nascimento CWA (2010)
Adsorption and chemical extraction of phosphorus as a function of
soil incubation time. Rev Bras Eng Agr Amb 14:32–38
Camargo LA, Marques Júnior J, Pereira GT, Bahia ASRS (2013) Clay
mineralogy and magnetic susceptibility of Oxisols in geomorphic
surfaces. Sci Agric 71:244–256
Cheng Z, Chen Y, Gale WJ, Zhang F (2019) Inorganic phosphorus dis-
tribution in soil aggregates under different cropping patterns in
Northwest China. J Soil Sci Plant Nutr 19:157–165
Comissão de Química e Fertilidade do Solo – CQFS RS/SC (2016)
Manual de calagem e adubação para os Estados do Rio Grande do
Sul e Santa Catarina, 1st edn. SBCS/NR, Porto Alegre 376 p
Ferreira DF (2014) Sisvar: a guide for its bootstrap procedures in multiple
comparisons. Cienc Agrotecnol 38:109–112
Fink JR, Inda AV, Bavaresco J, Barrón V, Torrent J, Bayer C (2016)
Adsorption and desorption of phosphorus in subtropical soils as
affected by management system and mineralogy. Soil Tillage Res
155:62–68
Freitas IF, Novais RF, Villani EMA, Novais SV (2013) Phosphorus ex-
tracted by ion exchange resins and Mehlich-1 from Oxisols
(latosols) treated with different phosphorus rates and sources for
varied soil-source contact periods. Rev Bras Cienc Solo 37:667–677
Guardini R, Comin JJ, Schmitt DJ, Tiecher T, Bender MA, Rheinheimer
D, Mezzari CP, Oliveira BS, Gatiboni LC, Brunetto G (2012)
Accumulation of phosphorus fractions in typic Hapludalf soil after
long-term application of pig slurry and deep pig litter in a no-tillage
system. Nutr Cycl Agroecosyst 93:215–225
Hopkins BG, Fernelius KJ, Hansen NC, Eggett D (2018) AVAIL phos-
phorus fertilizer enhancer: meta-analysis of 503 field evaluations.
Agron J 110:389–398
Khan A, Lu G, Zhang H, Wang R, Lv F, Xu J, Yang X, Zhang S (2019)
Land use changes impact distribution of phosphorus in deep soil
profile. J Soil Sci Plant Nutr 19:565–573
Mehlich A (1978) New extractant for soil test evaluation of phosphorus,
potassium, magnesium, calcium, sodium, manganese and zinc.
Commun Soil Sci Plant Anal 9:477–492
Mehlich A (1984) Mehlich 3 soil test extractant: a modification of
Mehlich 2 extractant. Commun Soil Sci Plant Anal 15:1409–1416
Menezes-Blackburn D, Giles C, Darch T, George TS, Blackwell M,
Stuller M, Shand C, Lumsdon D, Cooper P, Wendler R, Brown L,
Almeida DS, Wearing C, Zhang H, Haygarth PM (2017)
Opportunities for mobilizing recalcitrant phosphorus from agricul-
tural soils: a review. Plant Soil 427:5–16
Miola GR (1995) Extração de P, K, Ca e Mg do solo por diferentes
métodos e avaliação da disponibilidade de fósforo para as plantas.
140p. Dissertação (Mestrado) - Universidade Federal do Rio Grande
do Sul, Porto Alegre
Motaghian H, Hosseinpur A, Safian M (2020) The effects of sugarcane-
derived biochar on phosphorus release characteristics in a calcareous
soil. J Soil Sci Plant Nutr 20:66–74
Mumbach GL, Oliveira DA, Warmling MI, Gatiboni LC (2018)
Quantificação de fósforo por Mehlich 1, Mehlich 3 e Resina
Trocadora de Ânions em solos com diferentes teores de argila.
Ceres 65:546–554
Murphy J, Riley JP (1962) A modified single solution method for the
determination of phosphate in natural waters. Anal Chim Acta 27:
31–36
Oliveira CMB, Gatiboni LC, Miquelluti DJ, Smyth TJ, Almeida JA
(2014) Capacidade máxima de adsorção de fósforo e constante de
energia em Latossolo Bruno em razão de diferentes ajustes do
modelo Langmuir. Rev Bras Cienc Solo 38:1805–1815
Oliveira CMB, Gatiboni LC, Ernani PR, Boitt G, Brunetto G (2015)
Capacidade de predição da disponibilidade de fósforo em solo
com aplicação de fosfato solúvel e natural. Científica 43:413–419
Roy ED, Willing E, Richards PD, Martinelli LA, Vazquez FF, Pegorini L,
Spera SA, Porber S (2017) Soil phosphorus sorption capacity after
three decades of intensive fertilization in Mato Grosso, Brazil. Agric
Ecosyst Environ 249:206–214
Santos HC, Oliveira FHT, Souza AP, Salcedo IH, Silva VDM (2016)
Phosphorus availability as a function of its time of contact with
different soils. Rev Bras Eng Agr Amb 20:996–1001
Schick J, Bertol I, Barbosa FT, Miquelluti DJ, Cogo NP (2017) Water
erosion in a long-term soil management experiment with a humic
Cambisol. Rev Bras Cienc Solo 41:1–13
Steiner F, Lana MC, Zoz T, Frandoloso JF, Fey R (2012) Extraction
methods and availability of phosphorus for soybean in soils from
Paraná state, Brazil. Semina Cienc Agrar 33:1005–1014
Tedesco MJ, Gianello C, Bissani CA, Bohnen H, Volkweiss SJ (1995)
Análise de solo, plantas e outros materiais. Universidade Federal do
Rio Grande do Sul, Porto Alegre 174 p
van Raij B, Cantarella H, Quaggio JA, Prochnow LI (2009) Ion exchange
resin for assessing phosphorus availability in soils. Better Crops 93:
23–25
Vásconez G, Pinochet D (2018) Residual value of the phosphate added to
ecuadorian and chilean soils with different phosphorus retention
capacity. J Soil Sci Plant Nutr 18:60–72
Withers PJA, Rodrigues M, Soltangheisi A, Carvalho TS, Guilherme
LRG, Benites VM, Gatiboni LC, Sousa DMG, Nunes RS,
Rosolem CA, Andreote FD, Oliveira Junior A, Coutinho ELM,
Pavinato PS (2018) Transitions to sustainable management of phos-
phorus in Brazilian agriculture. Sci Rep 8:1–13
World Reference Base for Soil Resources - WRB: A framework for
international classification, correlation and communication (2014)
Food and Agriculture Organization of the United Nations.
IUSS/ISRIC/FAO, Rome (World Soil Resources Reports, 106)
Wuenscher R, Unterfrauner H, Peticzjka R, Zehetner F (2015) A com-
parison of 14 soil phosphorus extraction methods applied to 50
agricultural soils from Central Europe. Plant Soil Environ 61:86–96
Zehetner F, Miller WP (2006) Soil variations along a climatic gradient in
an Andean agro-ecosystem. Geoderma 137:126–134
Zhang W, Jin X, Liu D, Lang C, Shan B (2017) Temporal and spatial
variation of nitrogen and phosphorus and eutrophication assessment
for a typical arid river — Fuyang River in northern China. J Environ
Sci 55:41–48
Publisher’s note Springer Nature remains neutral with regard to jurisdic-
tional claims in published maps and institutional affiliations.