Microchemical Journal 111 (2013) 4752

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Microchemical Journal
journal homepage: www.elsevier.com/locate/microc

Evaluation of some wet digestions methods for reliable determination

of total phosphorus in Australian soils
Benjamin Webb, Samuel B. Adeloju
NanoScience and Sensor Technology Research Group, School of Applied Sciences and Engineering, Monash University, Gippsland Campus, Churchill, Victoria 3842, Australia

a r t i c l e

i n f o

Article history:
Received 30 June 2012
Received in revised form 27 January 2013
Accepted 1 February 2013
Available online 9 February 2013
Keywords:
Phosphorus
Total P concentration
Soil
Wet digestion
Acid digestion

a b s t r a c t
Four common wet digestion methods have been evaluated for reliable determination of total phosphorus (P) concentrations in soil samples. Wet digestion of soil samples with nitric acid gave the highest recovery of total P concentrations with a percentage recovery of 90.10.9% (n=3). A lower percentage recovery of 87.01.4% (n=3)
was achieved by wet digestion of soil samples with sulphuric acid. The use of acid mixture or acidalkaline mixture
for wet digestion of soil samples gave phosphorus recoveries of 82.41.9% (n=3) and 85.42.1% (n=3) with
nitric acidsulphuric acid mixture and nitric acidpotassium persulphate mixture, respectively. Substantial improvement in phosphorus recoveries with wet digestion of soil samples with sulphuric acid was achieved by further treatment of digested soil samples with sulphuric acid, resulting in a recovery of 92.81.0% (n=3), which
was higher than possible with other acid and mixtures. The wet digestion of soil samples with sulphuric acid
was also the only method which met reactivity and safety considerations. The successful utilisation of wet digestion with sulphuric acid for reliable determination of total P concentrations in a range of soil samples from some
Australian dairy and beef rearing pastoral land is reported.
2013 Elsevier B.V. All rights reserved.

1. Introduction
Phosphorus (P) is an essential nutrient in natural ecosystems and
it is also critically important to agricultural systems [16]. It is required for many agricultural processes including photosynthesis, nitrogen xation, owering, seeding and fruit maturation [36]. Soils
in most parts of the world are low in P and decient for optimal agricultural production [1,48]. This deciency is often compensated for
by applying various sources of P to soil, and thereby increasing the
P status of the soil and its agricultural yield.
Unfortunately, this mode of addition of P to soils has also had severe
and detrimental effects on several waterways around the world, leading
to an increased rate of eutrophication [912] and common presence of
algal blooms which have been associated with various problems in
sh, livestock and human health [4,1214]. The presence of phosphorus
(as phosphate) and nitrogen (as nitrate) has also been implicated in
many cases with several reported outbreaks of blue-green algae
which can have devastating effects on waterways.
A very good indication of the soil P pools and management practices that may contribute to P enrichment of runoff and waterways
can be obtained by conducting soil P measurements [5,6,12,1517].
These measurements provide a basis for matching P inputs and agricultural crop demands [1,5,6,18]. Soil P measurements have been
used to assess both environmental and agronomic impacts of P concentration extractable from soil [6,1821].
Corresponding author. Tel.: +61 3 9902 6450; fax: +61 3 9902 6738.
E-mail address: Sam.Adeloju@monash.edu (S.B. Adeloju).
0026-265X/$ see front matter 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.microc.2013.02.001

Total P concentration in soil is an important parameter which accounts for all forms of P within the soil. This parameter is often used
to determine soil P status for phosphorus based fertiliser application
and for estimation of P exports or agricultural yield [5,6,22]. It also provides a useful indication of the overall and potential nutrient supply of
P, and has been used in relationship comparisons with other soil measurements. However, total P measurement is limited in that it does
not differentiate between plant available P and non-available sources,
such as organically bound from insoluble mineral P [23]. Nevertheless,
it is still a very signicant parameter for both environmental and agronomic considerations. The reliable determination of total P concentrations in soil is however not easy. It requires adequate and effective
decomposition of the soil matrix to ensure complete release of P into solution prior to analytical measurement.
A number of digestion methods have been proposed for the determination of total P concentrations in soil. In one method, Tan [24]
used a uoro-boric acid digestion which employed a specialised bomb
digestion vessels and hydrouoric acid. The original version of this
method [25] also required specialised platinum crucibles and utensils
for sodium hydroxide or sodium carbonate fusion [24]. Olsen and
Sommers [26] also proposed a sodium carbonate fusion method, and
this again required specialised platinum equipment. Another suggestion from Olsen and Sommers [26] involved wet digestion of soil with
a perchloric acid. This method was not only complex, but had a range
of safety issues and required a specialised perchloric acid fumehood.
For these reasons, this digestion method is rarely used, except in
specialised laboratories with adequate perchloric acid fumehood. Due
to safety concern, some of the available soil method handbooks have

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B. Webb, S.B. Adeloju / Microchemical Journal 111 (2013) 4752

deliberately not included methods that employ perchloric acid [23].

However, it is important to note that safety consideration is not only
limited to the use of perchloric acid, but also to the reactivity of some
acids and/or mixtures to soil samples.
In another study, Dick and Tabatabai [27] reported on the use of a
sodium hypobromite/sodium hydroxide solution for total P determination in soil samples. The mixture was heated to dryness on a sand
bath at 260 C, followed by addition of formic acid and sulphuric
acid. Bowman [28] also used ordered additions of sulphuric acid, hydrogen peroxide and hydrouoric acid to decompose soil samples for
total P determination. Both of these methods require considerable
manipulation of the sample and are labour intensive compared to
other methods [29].
The examples cited above highlight the current state of play with
the determination of total P concentrations in soil. This is obviously
not ideal for such a signicant parameter which is highly important
in assessing the inputs of P into farmland and waterways. In general,
most of the currently available methods require specialised laboratory conditions or equipment, dangerous chemicals and intensive labour. To address this issue and ensure ease of attainment of reliable
total P concentrations, there is a need for the development of a simpler soil digestion method that can be readily employed within standard laboratory conditions without safety concern, complex and
laborious processes.
This paper reports on a thorough evaluation and assessment of
four wet digestion methods carried out to enable identication and/
or development of a simple and direct approach for pre-treatment
of soil samples for reliable determination of total P concentrations.
The wet digestion methods considered are those that are readily
used for determination of other substances and that require the use
of simple laboratory equipment, glassware and reagents [30,31]. The
adequacy, efciency and choice of the digestion methods were
assessed on the basis of recovery efciencies for total P concentration
in soil samples, the ease of use, reproducibility and safety consideration. The application of the chosen method to the reliable determination of total P concentration in a wide range of soil samples from
two intensive agricultural areas was also considered.
2. Materials and methods
2.1. Reagents and standard solutions
All acids and reagents used were of analytical reagent grade solutions. All solutions were prepared and diluted with distilled deionised
water (18 M cm, Millipore, MA, USA).
Nitric acid (0.01 M) used for adjusting the pH was prepared by diluting concentrated nitric acid with distilled deionised water. Stock phosphate solution (1000 mg P/L) was prepared by dissolving 4.393 g of
potassium di-hydrogen orthophosphate in 1 L of distilled deionised
water. A standard phosphate solution (100 mg P/L) was prepared by diluting an aliquot of this solution further with distilled deionised water.
2.2. Instrumentation
All phosphate analyses were carried out by using an adapted method (SmartChem 140 Method 420-3651) modied for use with soil samples on the Westco SmartChem 140 automated wet chemistry discrete
analyser (Westco Scientic Instruments, Inc., Brookeld, CT, USA). This
method utilises an antimonyphospho-molybdate complex formed
through the reaction of ammonium molybdate, antimony potassium
tartate and dilute phosphorus solutions in an acid medium. Ascorbic
acid is added to reduce the complex to produce a blue coloured complex
measured at 880 nm. The resulting absorbance increased proportionally to P concentration in solution. The normal sample and reaction
diluant in the method was deionised water, but was changed in this
study to 0.01 M nitric acid to match the acidity of diluted sample digest.

2.3. Glassware
All glassware and other containers were washed, soaked in a 2 M
nitric acid for at least 7 days, rinsed three times with deionised water,
soaked in deionised water and nally soaked in 0.1 M hydrochloric
acid (HCl) until ready for use.
2.4. Heating sources
Standard laboratory hotplates with aluminium surfaces were used for
heating samples. The maximum temperature setting was used for each of
the methods or as safety permitted. The temperatures of the extracts
were recorded and their signicance for sample digestion is discussed
later.
2.5. Sample collection and preparation
Soil samples were collected from an irrigation bay (width 30 m,
length 300 m) at the Macalister Research Farm (3800S 14654E), a
dairy farm situated in the Macalister Irrigation District of south-east Victoria (Australia). The soil was a natric grey Sodosol [32] and carried pastures that contained perennial ryegrass (Lolium perenne), white clover
(Trifolium repens) and assorted invasive species including dock (Rumex
spp.) and distichum (Paspalum paspaloides) [11,33].
200 samples of 20 mm cores and 30 samples of 100 mm cores were
collected from the sampling sites using a grid pattern. The soil cores were
bulked for each depth to provide a composite sample for the sampling location. After collection, the soil was stored (4 C) in polyurethane bag
and transported to the laboratory. The bulked soil cores were air dried
(40 C), ground and passed through a 2 mm sieve. Samples were then
stored in polyethylene containers at ca. 20 C prior to analysis.
Other soil samples were collected from selected agricultural sites
within south-east Victoria, from 2 areas known as Maffra and Warragul.
These areas were selected as geographically close agricultural areas (approximately 120 km range) with varying agricultural management practices, particularly irrigation application. A total of 14 sites were sampled,
seven from each of the two areas. Soils were classied using an Australian soil classication system. The 7 sites in Warragul were of three different soil classications, as indicated later in the results. The 7 sites in
Maffra were also of 3 different soil classications, as indicated later in
the results.
2.6. Soil moisture content
The moisture content was determined by heating the soil samples
in a drying oven at 105 C, cooled in desiccators and weighed repeatedly until a constant mass was obtained. The total P concentrations
for each sample replicate were corrected for soil moisture content.
2.7. Digestion methods
Four wet digestion methods were investigated for reliable determination of total P concentrations in soil samples. These methods
were adapted from those reported previously by Adeloju et al.
[30,31] for trace metal analyses. Digestion of each sample was carried
out in triplicate. The specic soil digestion procedures for each of the
wet digestion methods are described as follows:
Method A: Nitric acid (HNO3) digestion: 0.3 g of soil sample was accurately weighed into a 100 mL Erlenmeyer ask. The ask containing
sample was then transferred to a fumehood, where 10 mL of HNO3
(70%) was added and a glass funnel was inserted into the neck of
the ask. The mixture was heated on a hotplate to approximately
125 C where nitrogen oxide fumes were evolved and the volume of
the mixture was reduced to approximately 2 mL. The ask was then

B. Webb, S.B. Adeloju / Microchemical Journal 111 (2013) 4752

allowed to cool for at least 3 min. This process was repeated twice
with another addition of 10 mL HNO3 each time. With the nal nitric
acid addition, heating was continued at 125 C until no further
nitrogen oxide fumes were given off. The funnel was then rinsed
with a small volume of deionised water into the ask and the contents were allowed to cool to room temperature. The contents
were transferred to a 25 mL volumetric ask and made up to the
mark with deionised water. Samples were allowed to settle overnight,
and an aliquot was taken by pipetting for analysis or storage. Samples
were then stored at 20 C in acid washed polyethylene containers
until ready for processing.
Method B: Sulphuric acid (H2SO4) digestion: 0.3 g of soil sample was
accurately weighed into a 100 mL Erlenmeyer ask. The ask containing sample was then transferred to a fumehood, where 10 mL
of H2SO4 (98%) was added and a glass funnel was inserted into the
neck of the ask. The mixture was heated on a hotplate to approximately 160 C until sulphite fumes were evolved and the volume of
the mixture was reduced to approximately 2 mL. The ask was then
allowed to cool for at least 3 min. This process was repeated 4 more
times with 1 mL H2SO4 addition each time. With the nal addition,
heating was continued at 160 C until no further sulphite mist was
given off. The funnel was then rinsed with a small volume of
deionised water into the ask and the contents were allowed to
cool to room temperature. The contents were transferred to a
25 mL volumetric ask and, the digestion ask was rinsed carefully
by using deionised water, and the content of the volumetric ask
was mixed to avoid an acidwater inversion layer. The content of
the ask was allowed to cool before being made up to the mark
with deionised water. Samples were allowed to settle overnight,
and an aliquot was taken by pipetting for analysis or storage. Samples were then stored at 20 C in acid washed polyethylene containers until ready for processing.
Method C: Mixture of nitric and sulphuric acids (HNO3H2SO4)
digestion: 0.3 g of soil sample was accurately weighed into a
100 mL Erlenmeyer ask. The ask containing sample was then
transferred to a fumehood, where 10 mL of HNO3 (70%) and 1 mL
H2SO4 (98%) were added and a glass funnel was inserted into the
neck of the ask. The mixture was heated on a hotplate to approximately 130 C where nitrogen oxide fumes were evolved and the
volume of the mixture was reduced to approximately 2 mL. The
ask was then allowed to cool for at least 3 min. This process was repeated 3 times with 10 mL HNO3 addition each time. With the nal
addition, heating was continued at 130 C until no further nitrogen
oxide fumes were given off. The funnel was then rinsed with a
small volume of deionised water into the ask and the contents
were allowed to cool to room temperature. The contents of the Erlenmeyer ask were transferred into a 25 mL volumetric ask and
made up to the mark with deionised water. Samples were allowed
to settle overnight, and an aliquot was taken by pipetting for analysis or storage. Samples were then stored at 20 C in acid washed
polyethylene containers until ready for processing.
Method D: Mixture of nitric acid and potassium persulphate (HNO3
K2S2O8) digestion: 0.3 g of soil sample was accurately weighed into
a 100 mL Erlenmeyer ask. The ask containing sample was then
transferred to a fumehood, where 10 mL of concentrated HNO3
(70%) and 4 mL K2S2O8 (10% m/V) were added and a glass funnel
was inserted into the neck of the ask. The mixture was heated on
a hotplate to approximately 120 C where nitrogen oxide fumes
were evolved and the volume of the mixture was reduced to approximately 2 mL. The ask was then allowed to cool for at least

49

3 min. This process was repeated with 10 mL HNO3 addition

each time. With the nal addition, heating was continued at
120 C until no further nitrogen oxide fumes were given off. The
funnel was then rinsed with a small volume of deionised water
into the ask and the contents were allowed to cool to room temperature. The contents of the Erlenmeyer ask were transferred
into a 25 mL volumetric ask and made up to the mark with
deionised water. Samples were allowed to settle overnight, and
an aliquot was taken by pipetting for analysis or storage. Samples
were stored at 20 C in acid washed polyethylene containers
until ready for processing.
2.8. Recovery spikes
For each digestion method, a series of standard spikes were employed
to assess recovery efciency. The spike additions employed were: 0
(blank), 500, 1000 and 1500 mg P/kg. The spiked amount was added to
individual soil samples prior to the addition of the digestion acid or
mixture. Each sample spikes were also repeated in triplicate.
2.9. Sample preparation and calibration
A 2 mL aliquot of each sample digest was diluted in a 25 mL volumetric ask with 0.01 M HNO3 solution prior to analysis. The resultant solution was then analysed for P concentration by using the
SmartChem 140 discrete analyser. The results of the replicate samples
were then combined and analysed statistically.
Calibration was carried out using a six point calibration with 0.0,
0.5, 1.0, 2.0, 3.1 and 5.0 mg P/L. The SmartChem 140 discrete analyser
automated system prepared a linear calibration plot through the dilution of a 100 mg P/L stock solution. The R 2 values for the calibration
plots ranged from 0.9841 to 0.9922.
2.10. Method/reagent blanks
Blank digestions of reagents were prepared for each of the 4 wet
digestion methods. The same digestion procedure outlined for each
digestion method was followed for each blank without the addition
of soil sample. Final concentrations were corrected for the blank solution P concentrations for each method.
The average blank contribution of the digestion solutions from
each method were: method A (0.19 0.04%), method B (0.20
0.04%), method C (0.15 0.02%) and method D (0.20 0.05%). Evidently, there were no signicant differences between the blank corrections for the four wet digestion methods.
2.11. Working environment
All sample preparation and analyses were conducted in controlled
laboratory conditions at 22.0 0.5 C. All digestions were undertaken
in standard laboratory fume hoods.
3. Results and discussion
3.1. Recovery study
As a rst step in assessing the effectiveness of the four chosen wet
digestion methods for reliable determination of total P concentrations
in soil, their recovery efciencies were compared and evaluated. As
the two broad type of soil samples collected from the sampling sites
in this study are clay loam and sandy loam, we chose to use a
clay loam for the comparison because the soil samples collected
from 13 of the 14 sampling sites were clay loam of different colour
variations. It was not expected that the colour variation will alter

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B. Webb, S.B. Adeloju / Microchemical Journal 111 (2013) 4752

the ability to acid digest the clay loam samples. The chosen digestion
method will be subsequently applied to the sandy loam sample.
Soil samples digested with only concentrated HNO3 (method A)
gave the highest percentage recovery for total P concentration, averaging 90.1 0.9%. On the other hand, soil samples digested with only concentrated sulphuric acid (method B) gave a slightly lower recovery of
87.0 1.4%. Contrary to expectation, the use of a mixture of concentrated nitric and sulphuric acids (method C) and a mixture of concentrated
nitric acid and potassium persulphate (method D) gave even lower recoveries of 82.4 1.9% and 85.4 2.1%, respectively. In particular, the
lowest recovery efciency achieved with method C was somewhat surprising. The expectation was that the increased strength of the two
strong acid mixtures would enable better decomposition of the soil matrix and, at least, produce similar or better percentage recovery of total P
concentration compared to that obtained with the use of only concentrated nitric acid.
To clarify the above unusual trend of the recovery efciencies
obtained for the four wet digestion methods, further investigation was
conducted with unspiked soil samples. Interestingly, when these four
digestion methods were applied to the determination of total P concentrations in unspiked soil samples, the data in Table 1 revealed that the
phosphate concentrations found increased in the following order:
Method B >

Method C > Method A > Method D:

This trend suggests that the use or inclusion of H2SO4 may be benecial for more effective release of organically bound P in soil. By taking
the total P concentration obtained for the soil sample with method B as
optimum (100%), the estimated recoveries of P in the unspiked soil
samples for the three other methods were 93.1% (method A), 95.9%
(method C) and 76.1% (method D). Based on this assumption, a notable
but signicant observation is that the application of methods B and C to
spiked soil samples (with 87.0 1.4% and 82.4 1.9% recoveries for
methods B and C, respectively) results in a net decrease of 1314% in
each case. As the presence of H2SO4 is a common factor in both digestion
methods, this observation seems to suggest that the lower recoveries
obtained in the spiked soil samples may be due to an impact of this
acid on the inorganic P spike. A possible link that can be deduced from
this observation is that the much higher digestion temperature
achieved with H2SO4 only (method B) or as a mixture (method C)
may result in losses of the inorganic P spikes during soil digestion. As
can be clearly seen in Table 1, the digestion methods which include

Table 1
Recovery efciency of wet digestion methods for total P concentrations in a grey clay
loam Dermosol.
Sample
description
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
a
b
c
d

A unspikeda
A spike 1a
A spike 2a
A spike 3a
B unspikedb
B spike 1b
B spike 2b
B spike 3b
C unspikedc
C spike 1c
C spike 2c
C spike 3c
D unspikedd
D spike 1d
D spike 2d
D spike 3d

Max digestion
temp. C

P spike conc.
(mg P/kg)

Total P
(mg P/kg)

Recovery
efciency (%)

125

0
500
1000
1500
0
500
1000
1500
0
500
1000
1500
0
500
1000
1500

1777 56
2229 113
2683 133
3111 59
1908 86
2346 62
2788 68
3189 55
1829 62
2249 76
2656 147
3034 89
1452 23
1893 36
2296 68
2720 44

90.5
90.7
89.0

87.5
88.0
85.4

84.0
82.7
80.3

88.1
84.4
84.5

160

130

120

Concentrated HNO3 (3 10 mL) additions only (n = 3).

Concentrated H2SO4 (1 10 mL and 4 1 mL) additions only (n = 3).
Concentrated HNO3 (410 mL) and concentrated H2SO4 (11 mL) additions (n=3).
Concentrated HNO3 (310 mL) and 10% w/v K2S2O8 (14 mL) additions (n=3).

H2SO4 (methods B and C) achieved higher solution temperatures, up

to 40 C higher than with other two digestion methods. Evidently, this
is why method B gave the highest phosphate concentrations in the
unspiked soil samples. Interestingly, the achieved solution temperature
with the four digestion methods increased in the following order:
Method B > Method C > Method A > Method D
which correlated well with the trend observed for the total P concentrations in the unspiked soil samples.
Notwithstanding the observed differences in the recovery efciencies, the total P concentrations found in the unspiked soil samples
within 020 mm depth by the four different digestion methods were
comparatively close to those previously obtained at the same farm in
2001 (1528 mg P/kg) and 2004 (1432 mg P/kg) [10,11]. The higher
concentrations found with methods A, B and C may be attributed to increasing P inputs into the sampled paddock between 2004 and 2007.
3.2. Choice of digestion method
Apart from the recovery efciencies achieved with the four wet digestion methods for total P concentrations in spiked and unspiked soil
samples, proper selection of a digestion method must also consider relevant and/or associated safety issues. One of the important considerations
in proposing a digestion method that can be readily employed in most
laboratories for the determination of total P concentration in soils is the
degree and extent of reactivity caused by the digestion medium or mixture during the digestion process. Highly reactivity digestion process can
result in unexpected losses of samples and, in extreme cases, can result in
accidental acid splash, corrosive reactions or other undesirable consequences. It is, therefore, highly essential that the chosen digestion method for soil samples is moderately reactive, while still ensuring complete
recovery of total P concentrations in soils.
The three wet digestions methods (methods A, C and D) which
employed HNO3, either alone or as a mixture, were found to be very reactive, causing a sudden release of pressure when heated to the maximum temperatures. A pressure build-up below the ne particulate of
the soil samples occur during digestion and led to the sudden pressure
release. This can pose a serious safety concern as the digest can escape
in some cases from the heated ask. This was particularly a problem
with method A which used only HNO3 for soil digestion. In comparison,
the pressure release and associated popping effect was somewhat subdued in method C and this appears to be due to the inclusion of H2SO4 in
the mixture at the initial stage. However, more popping effect was observed with the required subsequent additions of HNO3 during digestion with this method. As a precaution, irrespective of the HNO3 based
digestion method, care must be exercised and the heating temperature
may need to be reduced to minimise potential hazard, but this will lead
to a considerable extension of the digestion time. In contrast, no pressure build-up or popping effect was observed for sample digestion
with method B, as the boiling temperature of H2SO4 was much higher.
On the basis of recovery efciency achieved for total P concentrations in unspiked soil samples and safety considerations, method B
was chosen as the best of the four wet digestion methods for further investigation. Even with spiked soil samples, the recoveries obtained by
this method was only slightly lower than those obtained with method
A and were higher than those of methods C and D. Possible improvement in the recovery efciency achieved with method B for determination of total P concentrations was further investigated by examining the
effect of additional treatment with H2SO4.
3.3. Extended H2SO4 digestion (method B +)
The possibility of improving the recovery efciency achieved for
spiked soil sample with method B was investigated by subsequent treatment of the initial sample digest with several 1 mL H2SO4 additions.

B. Webb, S.B. Adeloju / Microchemical Journal 111 (2013) 4752

Table 2
Inuence of increasing addition of H2SO4 on recovery efciency of total P in a grey
sandy loam Dermosol.
Sample
description
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
a

B5
B5
B5
B5
B6
B6
B6
B6
B7
B7
B7
B7

unspiked
spike 1
spike 2
spike 3
unspiked
spike 1
spike 2
spike 3
unspiked
spike 1
spike 2
spike 3

Additionsa
(1 mL H2SO4)

Spike conc.
(mg P/kg)

Total P
(mg P/kg)

Recovery
efciency (%)

5
5
5
5
6
6
6
6
7
7
7
7

0
500
1000
1500
0
500
1000
1500
0
500
1000
1500

1083 15
1539 32
2000 37
2449 12
1092 18
1549 38
2018 16
2468 20
1123 13
1585 46
2062 38
2505 19

91.3
91.8
91.1

91.4
92.6
91.7

92.3
93.9
92.1

Please note: 0100 mm depth samples (n = 3) were used.

96

Recovery Efficiency (%)

Method B, as described in materials and methods, involved digestion of

soil sample in 10 mL concentrated H2SO4 followed by four 1 mL additions. Following exactly the same procedure, the effect of increasing
the 1 mL H2SO4 additions beyond four on the recovery efciency for
total P concentration from spiked samples was of particular interest.
The new methods (B5, B6, and B7 listed in Table 2 with 5, 6 and 7 additions of 1 mL H2SO4, respectively) were carried out on soil samples collected from 0 to 100 mm depth instead of the 020 mm depth used for
comparison of the four wet digestion methods. For this reason, it was obvious that the total P concentrations obtained for these unspiked soil
samples were lower than those reported in Table 1.
The data in Table 2 show that the total P concentrations in the
unspiked soil samples increased slightly with each 1 mL H2SO4 addition
and treatment, indicating small improvement in the release of organically bound P. Also the percentage recoveries obtained for the spiked
soil samples improved slightly with each acid addition. Evidently, the
average spike recovery obtained increased to 91.4 0.4% for method
B5, to 91.9 0.6% for method B 6, and further to 92.8 1.0% for method
B7. Fig. 1 shows the effect of increasing addition of H2SO4 on the average
recoveries of total P concentration in the spiked soil samples. These results demonstrate that a net improvement in recovery efciency of 4
6% was achieved by further treatment of sample digest with 13 additions of 1 mL H2SO4. Furthermore, it is worth noting that the average
spike recoveries obtained with methods B5, B 6, and B7 were all higher
than that of method A. Fig. 1 also shows that the recovery of P with
this approach increased considerably between the fourth and fth
1 mL additions and then levelled out with the sixth and seventh additions. This observation clearly suggests that further acid additions will
not lead to more improvement in recovery and was therefore not
considered.
These results also suggest that the lower P recoveries obtained
earlier for the spiked soil samples collected from 0 to 20 mm depth
may be due to other soil composition. The 020 mm soil is likely to
contain higher amounts of organic material than those from 0 to
100 mm depth. This could mean that the soil samples collected
from 0 to 20 mm depth were more difcult to digest. Interestingly,
recovery study of spike 0100 mm depth soil samples by the original
method B digestion method gave an average recovery comparable to
the original results obtained with the 020 mm depth samples
(87.0% 1.4% for 020 mm depth versus 87.2% 0.9% for 0
100 mm depth). However, it is recommended, based on the results
obtained in this section that method B should be employed with at
least ve 1 mL H2SO4 additions. Table 3 shows that this approach
was successfully employed for the determination of total P concentration in soil samples collected from two intensive agricultural areas.
Evidently, the total P concentrations in these soil samples are
inuenced by factors such as the soil characteristics, quantity, frequency and length of P fertiliser application.

51

94
92
90
88
86
84
3

Number of 1 mL H2SO4 Additions

Fig. 1. Average spike recovery (%) versus increasing number of 1 mL H2SO4 additions
for wet digestion of grey sandy loam Dermosol with method B. n = 3.

4. Conclusions
The careful evaluation of the four wet digestion methods for reliable
determination of total phosphorus (P) concentrations in soil samples
has demonstrated that the criteria for selecting an appropriate method
must include safety consideration, in addition to the achievable recovery efciency. Wet digestion with H2SO4 was found to be less reactive,
most effective and safe for total P determination in soil. Further treatment of pre-digested soil samples with H2SO4 led to substantial improvement in phosphorus recovery, achieving a percentage recovery
of 92.8 1.0%. This digestion method was successfully utilised for the
reliable determination of total phosphorus concentrations in various
soil samples from dairy and beef rearing pastoral areas.

Acknowledgements
One of the authors (Ben Webb) wishes to acknowledge the provision of an Australian Postgraduate Research Scholarship through
Monash Research Graduate School (MRGS) and other support provided by the School of Applied Sciences and Engineering. The authors
also acknowledge the research funding provided for this project by
Dairy Australia.

Table 3
Total phosphorus concentrations in soil samples collected from intensive agricultural
areas in Warragul and Maffra (S-E Victoria, Australia).
Sample
site

Total P
(mg P/kg)

Soil
characteristica

Period post fert.

appln. (day)

Last fert. appln.

(mg P/ha)

W1
W2
W3
W4
W5
W6
W7
M1
M2
M3
M4
M5
M6
M7

1838
1941
3009
2680
1779
2062
1103
1936
2381
1447
1470
1775
2000
1743

GCLD
GCLD
RCLF
RCLF
RCLF
GCLD
GSLD
GRCL
GRCL
GRCL
GRCL
GYCL
GCLS
GCLS

365
365
365
365
45b
20
730+
365
45
1000+
730+
60
130
130

24.8
24.8
24.8
24.8
3.7b
14.8
n/a
11.8
14.5
n/a
n/a
13.2
7.7
7.7

n/a = not available.

a
Soil description; GCLD=grey clay loam Dermosol, RCLF=red clay loam Ferrosol,
GSLD=grey sandy loam Dermosol, GRCLS=grey/red clay loam Sodosol, GRCLS=grey/
yellow clay loam Sodosol, GCLS=grey clay loam Sodosol.
b
Site W5 has only received a single fertiliser application (3.7 kg P/ha) in three years
prior to sampling.

52

B. Webb, S.B. Adeloju / Microchemical Journal 111 (2013) 4752

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