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A COMPARISON OF FIELD METHODS OF

DETERMINING THE AVAILABLE
PHDSPHORUS 0F SOILS

THESIS FOR THE DEGREE OF M. S.

Clarence A. Engberg
1932

 

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COMPARISON OF FIELD TmTHCDS OF
DETERMINING THE AVAILABLE PHOSPHOEUS OF SOILS
By
CIARENCE A. EECEEEC
A THESIS
PRESENTED To TIE FACULTY
OF
MICHIGAN STATE COLLEGE
OF
AGRICULTURE AND APPLIED SCIENCE
IN
PARTIAL WIHMEIH OF THE REQUIREMENTS
FOR TIE

DEGREE 0F MASTER OF SCIENCE

East Lansing

1932

THESIS

nflT”"‘" “‘rn=*;~7fi
av;~._..Uz;L_‘.J.u....—i.i

The writer wishes to Express sincere gratitude to
Dr. C. H. Spurs-gr for his advice during tie experimental work and
for criticism of this thesis. Also to Dr. C. E. Miller, and
other members of tie Soils Department for their kind advice and

help.

INTRODUCTION

considerable progress has been made in recent years in
the development of methods for determining the amounts of the
soluble or the so-called available1 nutrients of plants contained
in field soils. Almcst all of the studies pertaining to the
testing of these methods were conducted in the field in connection
with long-time experimntal fertilizer trials. Although the results
of these researches are of practical value to agriculture, no
accurate methods have yet been devised for the determination of the
mounts of available plant nutrients in each separate field. field
plat experimentsIare still essential in order to establish the I
broad general principles of soil fertilisation. If the experimental
fields were located in a region of uniform soil type and the soil
treatments applied by the farmers were uniform in character it
might be possible to determine the specific fertilizer needs by
field experiments. In a region where the soil types vary within
short distances, however, the use of scattered experimental plats
gives no reliable guide for the application of connercial fertilisers
to the soil. Also, in 81W one region there is usually too much
variation in the soil treatment given to individual fam fields to
admit of an accurate determination of the fertiliser requirements
by means of experimental plate.

(1) The term available phosphorus as used in this thesis means
the mount of phosphorus determined by the tests used.

Tanners are interested, generally, in methods for
determining the proper treatment of soils for more profitable
cr0p production and simple, easily applied, chemical soil tests,
by means of which the deficiencies of plant nutrients could be
determined, would be valuable to them. In recent times soil
chemists have developed a number of rapid tests for determining
the amount of some of the soluble plant nutrients in soils, that
may be used either in the laboratory or in the field.

A considerable amount of work has been done in the development
of tests for the amount of available soil phosphorus. The determi-
nation of the amount of available soil phosphorus by means of some
of the more recent methods proposed by soil chemists depends on the
developnent of a blue color when the molybdenum of ammonium phospho-
molybdate is reduced by a suitable reducing reagent.

Several of these methods have been developed quite recently
and a comparison of some of them, using soil samples from the
same locations on unfertilized field plats and plats fertilized with

phosphatic fertilizers is the object of this thesis.

-1-
HISTORICAL REVIEW

Some of the first researches on the determination Of
phosphorus in solutions, were carried out by bio-chemists who
have attempted to determine the amount of phosphorus in plants,
blood and urine by colorimetric methods. as original method
for determining phosphorus in aqueous solutions was proposed by
Deniges (AI) in 1920. Deniges used a 10% ammonium molybdate
solution to which was added an equal volume of strong sulphuric
acid, and as a reducing agent 0.1 gram of pure tin dissolved
in 2 cc. of concentrated hydrochloric acid with the aid of a 14$
copper sulphate solution and then made up to a volume of 10 cc.
In 1981, l‘lorentin (6) altered the composition of the ammonium
molybdate solution used by Deniges so that it was composed of
10% amonium molybdate solution to which was added three volumes
of sulphuric acid. made from equal parts of strong acid and
water. This solution however must be stored in the dark in
order to prevent reduction by light.

Later, Atkins (l) revived and further investigated the
methods of Deniges (u) and Florentin(6) and applied his modified
method to the study of soil extracts. This method consisted of
making a colorimetric comparison of a standard phosphate solution
with an aliquot part Of the diluted soil extract, after the addition
of 2 cc. of reagent A followed by 5 drops of reagent 3 to 100 cc. of
each solution. Atkins' reagent A consisted of 100 cc. of a 10%

solution of amonium molybdate with 300 cc. of 50% sulphuric acid.

.2...
Reagent B is a stannous chloride solution prepared daily in the
some manner as reconmended by Deniges.

For the determination, 10 grams of air-dried soil is passed
through a 100 mesh sieve and shaken for 3 or u hours with so cc.
of conductivity water. 10 cc. of this liquid is centrifuged until
clear. and 5 cc. of the clear extract is then diluted to 100 cc.
with the conductivity water. the liquid is then treated with
reagents A and 3. The color of the unknown solution, which has been
developed by reagent 3, is compared with tint of a standard solution.
Atkins introduced the use of Bismarck brown in the case of slightly
colored solutions and in doing so eliminated the greenish tint present
in some extracts.

In 1926, Lonstein (ll) proposed a method for determining the
amount of available phosphorus in the soil which eliminates the
disturbing effects due to the presence of silica and iron compounds
in the extracts. The iron and silica are separated from the other
constituents present by igniting the dry residue from the acid
extract of the soil and extracting the ignited residue with sulphuric
acid; the silica is filtered off and the dissolved iron is removed
by the calcium acetate method.

Some of the workers in this field have attempted to use a
reducing agent other than tin or stannous chloride. Bell and Doisy (2)
have proposed twdroquinone as a suitable reducing agent for phospho-
molybdic acid. rho color produced by the reduction is intensified by
mking the solution alkaline and the greenish tint due to the presence

of quinone is removed by the addition of sodium sulphite. who two

.3-
solutions used by Bell and Daisy (2) were (a) a 2% solution of
twdroquinone in 0.1% sulphuric acid and (b) a 15% solution of
sodium sulphite added to four volumes of 10% sodium carbonate
solution. By this test it was possible to Obtain a distinct
blue color with 0.05 mg. of phosphorus in a volume of 100 cc.

In using the test equal volumes of molybdate and hydroquinone
solutions are added to the solution to be tested followed by
five volumes of the sulphite solution. The color is then matched
against the color of the standard solution.

In 1920. Fislne and Subbarrow (5) used 1:23’4 amino-naptll.
sulphonic acid instead of hydroguinone as a reducing agent. A
canparison Of the two methods seemed to show that the 182$
mnino-naptho sulphonic acid gave more accurate results in the
presence of much more inhibiting material than was allowable when
twdroquinone was used.

Partner and Judge (13) compared the methods of Fish and
Subbarrow with Denige's coerrulo-molybdate method as modified by
Atkins. they found that small amounts of phosphorus could be deter-
mined by the use of both methods, but they also found that the
Deniges method was five times as sensitive as that of ll'islne and
Subbarrow. In a study of salt influences on these methods it was

found that none of the cannon salts in the soil affected the color.

Parker and Judge recomended the use of the Deniges modified method

because of its greater sensitivity.
In the determination of available phosphorus, in soils. there

has been a great disagreement among workers in the method of extraction.

 

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In lngland and the European countries If citric acid has been used a.
a. standard extracting solution. In the United States a variety of
solutions have been used for the extraction, some of the workers
using 0.2 I. nitric acid and others using water extracts or weak
sulphuric acid. . The 0.2 I. nitric acid method was developed in this
country mainly by Peter and ivorittuh). stoddart (20) and l‘raps (7)
and this method, with the is citric acid method developed in Inglend
have been the most successful methods proposed.

Russell and Prescott (16) carried on some work using different
acids for the extraction of soil phosphoms. They found that the
strong acids such as hydrochloric, nitric. and sulphuric acid were less
potent than equivalent concentrations of the weaker acids such as citric
and oxalic acids. 0.1 II. Weehloric acid and nitric acid brought out
the least phosphorus, while 0.1 H. oxalic acid brought out the most
phosphorus. Ms m be explained on the basis of adsorption, the
acids that are adsorbed the most themselves dissolve more phosphorus.
no authors also found that when the time of extraction was reduced
to minim the interpretation of the results would be simplified,
because the composition of the liquid has suffered minim change by
interaction with the soil.

Ball and Plymen (9) studied the effects of different solvents
on soil phosphorus and case to the conclusion that weak solvents give
more trustworthy infomation as to the requirements of the soil for
fertilisers than do strong solvents. According to Hall and Plymsn (9)
a suitable solvent should fill three requirements: (a) the quantity

of phosphorus should show a wide variation in passing from a. good soil

'5'
to a. poor soil: (b) the quantity of phosphorus extracted should

be sufficient for exact determinations; (c) the action of the
solvents used should not be effected by the presence of variable
constituents in the soil, such as calcium carbonate and organic matter.

Hibbard (10) , in 1931. stated that citric and oxalic acids
should not be used as extractants as it is necessary to remove then before
the solution is tested by the molybdate method. Ilse, he states that
the best kinds of acid to use, in the estimation of the available
phosphorus in soils is one that is neither strong, highly ionized, nor
slightly buffered, since such an acid will not hold the pH or the P0”,
concentrations constant. To be comparable all extracts should have
almost the some pH and the mount of acid needed for any one soil
should be determined by experiment. . a 1:5 extract made by weak acid
solvents was found by 31be to represent much better than strong
acid extract the phosphorus supplying power of soils as indicated by
the growth of plants on them.

Shedd (17) proposed a short. rapid test in 1920 which made
use of 0.2 N. nitric acid. In his method ten grams of air-dried soil
is added to 25 cc. of 0.2 I. nitric acid and the whole shaken every
minute for five minutes. The solution is then filtered until clear
into a test tube; 1 or 2 cc. of 60$ monium nitrate solution is then
added, and 5 cc. of ordinary molybdate solution. The contents of
the tube are then heated to 60°0, shahen several time and allowed to
stand about 30 minutes at room temperature. The results are then
read as 'large', ”fair“, “moderate“, or “very moderate' in decreasing

order of the amount of ammonium phospho-molybdate obtained. Shsdd (21)

.5.
did not attempt to place the method on a quantitative basis at that
time.

Iruog (22) (23) has proposed a laboratory method, which has
been used quite extensively in lisconsin, for the determination of
available phosphorus in soils. 1 0.002 N. sulphuric acid solution,
buffered to a pH of 3 is used in the extraction. fruog states tlmt
this method correlates quite well with growth of plants under lisconsin
conditions.

In Germany two workers have introduced methods for estimating
the fertiliser requirement of soils by means of the growth of plants.
Neubauer's method (12) is based upon the. rapid absorption of soluble
plant nutrients by young rye seedlings. Bye seedlings are grown in
the soil for eighteen We and are then harvested and analysed for
potash and phosphorus. Neubauer determined the' limiting Value" of
the soil under German conditions, as 215 mg. of I20 and 6 mg P205.
Ihcrnton (13) studied the Neuhauer method and set the 'limiting value”,
for Indiana farming conditions, at 1‘ mg. of P20 and 10 mg. of [205.
In general Thornton found that the Neubauer method correlated with the
pot and field tests that were carried out by him.

Hitscherlich's method, which is described by Stewart (19) is
based on the theory that there is a quantitative relationship between
the yield and the concentration of mitrients producing it. i'his method
has met with success in Germany, but it is a slow and expensive method
for determining the fertiliser requirement of the soil.

lone of the methods, which have been reviewed are applicable

to use in the fields. Truog has collaborated with the Ieuotte Chemical

Company in producing a method which can be used in the field. Spurway (18)

-7-

has developed a method of determining the water soluble phosphorus

of the soil. This method can be used in either the field or laboratory.
Bray (3) has proposed a test for determining the relative amounts of

phosphorus in the soil. All of these field methods will be discussed

further under procedure and methods.

  

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Distribution of Fertility Plats

PLATE I . LOCATION OF PMS

-3-
PROCEDURE AND METHODS
Soil Samples

The soil samples used in this comparison were obtained from
various station experimental plate and cooperative experimental fields
in different parts of the state as shown by Plate 1. live samples
were taken from each plat at a depth.of plow layer and.thoroughly
mixed. The soils were passed over a leh sieve and the pebbles and
coarse materials taken.out. .Lll of the soils were then tested in the
laboratory by the use of the various methods under the same conditions.

By following this plan it was possible to investigate a.nnmber
of soil types from.different sections of the state. The crop yields
for the plate were obtained from the records of the fertility work of
the Soils Department.

Description of Methods

Spurway's water-soluble phosphorus method: In Spurwny's (18) test
for water-soluble phosphorus, the use of glassware is eliminated
except for the reagent bottles. It is possible to use tap or well
water, if free from.phosphorus and.areenic. The determinations are
made by means of a.ndcro-chemical phosphorus test based on the reaction
of Deniges. The molybdate reagent is made by dissolving five grams of
O. P. amonium molybdate in 50 cc. of distilled water. Il'his solution
of amonium molybdate is then poured into 50 cc. of pure, concentrated
nitric acid which.mnst be phosphorus free. Then dilute this mixture
with.lOO cc. of distilled water. A.blank test should.be made on
this reagent at regular intervals of time and if a‘blue color develops
it is necessary to add.3 to h cc. of nitric acid to the whole quantity

of reagent. Should the blue color persist it is best to discard all of

(\

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.9-
the reagent and make up a fresh supply. The tin pencil used for
reducing the molybdate in the test is a rod of pure tin about
three-sixteenths of an inch in diameter and three or four inches
long pointed at one end.

The color chart used was developed by Spurway (18) , by
testing phosphorus solutions of known strengths and comparing the
colors developed with the color charts of Bidgeway (15).

In performing the test a piece of waxed paper is folded
lengthwise, and opened to form a trough. Some of the soil is
I placed in the upper end of the trough chose to the hand and should
fill the paper to the edges and extend about three-quarters of an
inch lengthiee on the paper. A slight cavity is then made in the
rear of the soil: this cavity will form a reservoir to hold the water
used in the extraction. Iater is then dropped behind the soil smple
in the trough, slowly and carefully allowing the soil to tales up the
water drop by drop, the controlling the movement of water by tipping
the end of the paper slightly downward. When the clear extract appears
at the end of the trough a drop may be placed on a separate flat piece
of waxed paper. To this drop of extract is added an equal quantity of
molybdate reagent and the liquids mixed by stirring with the corner
or another waxed paper. The mixture is then stirred with the tip of the
tin pencil for ten seconds. If phosphorus is present in the extract,
a blue color develops. and the intensity of this color is a measure
of the quantity of phosphorus present in the extract. The maxim
color will develop in a short time after which the color will gradually

fade. According to the author the test can be used in several lines

of research pertaining to soil phosphorus.

-10-

Illinois phosphate test: Bray (3) has developed a.method which is
commonly called the Illinois phosphate test. The solution, with
‘which the test is carried out, is prepared by dissolving 100 grams

of highest purity ammonium.molybdate in 850 cc. of distilled water.
This solution, after being filtered and cooled, is added with.constant
stirring to a cold.mixture of 1700 cc. of concentrated hydrochloric
acid (36%) and 700 cc. of water. This is the stock solution. The
solution as used for the test is made up by diluting 120 cc. of the
stock solution to 1000 cc. with.distilled water. '1 tin rod, as
recommended by Spurway (18) is used in the test.

The test is made by shaking one part of soil with three
parts of the solution in a small vial or test tube. Just enough
shaking to mix the soil and solution is required. .After settling,
which takes about five minutes, the clear solution is stirred gently
with a tin rod until the maximum.intensity of color is reached. The
varying amounts of phosphorus are indicated by the various shades
and color intensities which increase with.the amount of phosphorus
present. The four amounts of phosphorus are listed as ”high",
Imedium”. "doubtful” and ”low" in available phosphorus. According
to the author these colors fade very little on standing. The color
is compared with.a color chart to determine the relative amount of
phosphorus in the soil.

Truog-LaMotte method: The TruogblaMotte phosphorus test requires three
reagents which are called it“. "B“, and “C”. The measuring spoon which
is included in the outfit is filled level, with the soil. This spoon-

ful of soil is placed in one of the extraction tubes. Distilled

-11..

water is added to the mark, which is about one inch from the bottom.
Then three drops of reagent “A“ are added, the tube immediately stop-
pered and shaken for one minute. The soil suspension is then filtered
into a comparison tube which has a 3 cc. mark. [After 3 cc. of filter-
ed extract is obtained, three drops of reagent ”B“ are added to the
extract and the tube immediately shaken to mix the contents. A

volume equal to a pinhead of reagent '0”, which is a powder, is now
added and the tube again shaken. ‘After one minute the maximum color
is developed and may be compared to the standard colors on the color
chart. It is possible to read the pounds per acre of available
phosphorus from this chart. It is recommended by Truog (22) that

for general farming conditions in northern states, silt loam, sandy
loam, and clay loam soils should contain at least 75 pounds of readily
“available” phosphorus per acre and sandy soils 50 pounds per acre.
Truog's laboratory method: Truog's (22) (23) laboratory method has
been used with success in Wisconsin. The sulphuric acid solution,
which.is umed for the extraction, is prepared by making a stock
solution of exactly 0.1K sulphuric acid by titrating against standard
alkali. The extracting solution is then prepared by diluting convenient
volumes of the 0.1N sulphuric acid solution to 0.002N and buffering to
a,pfi of 3 by adding 3 grams of ammonium.sulphate per liter. The
'amonium molybdate-sulphuric acid solution is prepared by dissolving
25 grams of ammonium.molybdate in 200 cc. of water heated to 60°C.,
and filtered. 280 cc. of phosphorus- and arsenic-free sudphuric acid

is diluted to 800 cc. with distilled water. After both solutions have

.12..

been cooled slowly add the ammonium molybdate solution to the
sulplmric acid solution. When the resulting solution has cooled
to room temperature it is diluted to 1000 cc.

Stannous chloride solution is prepared by dissolving 25
grams of stannous chloride (SnCl .23 0) in 1000 cc. of dilute (10%
by volume) hydrochloric acid solution. This solution is then stored
in a bottle with a side Opening at the bottom arranged with a glass
stopcock for delivering the solution in drops. The solution is
then protected from the air by floating a layer of mineral oil over
the surface.

It is necessary to prepare a standard phosphate solution
to use in comparison of the colors. 0.2195 grams of recrystallized
potassium dihydrogen-phosphate is dissolved and diluted to 1000 cc.
This solution will contain 50 ppm. of phosphorus and is too concen-
trated for use. A second stock solution is prepared by diluting
50 cc. of the original solution to 500 cc. This second stock solution
will then contain 5 ppn. and can be used for making the standard
colorimetric solution. To prepare this standard take 5 cc. of the
second solution, dilute to 95 cc. with distilled water and add 1} cc.
of the ammonium molybdate-sulphuric acid solution and mix. To this
mixture add 6 drops of stannous chloride solution to develop the color
and the solution is ready for use. It contains 0.25 ppm. of phosphorus.

In the extraction process two grams of soil and W cc. of
0.®2 N. sulphuric acid are placed in a bottle or Erlenmeyer flask of

suitable size and shaken for 30 minutes. The soil suspension is filtered

-13-
off using No. he Whatman filter paper.(1) The filtrate is discarded
until it comes through clear. For comparing the colors, intensities
of the standard, and unknown solutions, a Klett calorimeter is satis-
factory instead of the ordinary Nessler tubes as specified by Truog.
M8 cc. of the clear filtrate are placed in an.Erlenmeyer flask, then
2 cc. of the ammonium.molybdate-sulphuric acid solution are added and
the mixture shaken well. To this mixture three drops of the stannous
chloride solution are added and the whole shaken again. The standard
and the unknown solutions are then placed in the colorimetet tubes
which.are adjusted'until the colors matdh. The calculations are easily
made. If 50 is the reading of the unknown solution and 19.5 the
reading of the standard solution the amount of available phosphorus
will be 19° x 50 or 19.5 ppm. on air dried soil. In
50 reading of unknown
order to convert parts per million to pounds per acre plow layer it
is necessary to mmltiply by the factor 2 in the case of heavy soils,

2.5 for sandy soils, and.somewhat less than 2 for mucks.

 

(1)
Trucg recommended the use of S. S. 589 filter paper, but Whatman

Nb. #2 gives good results.

41+-
DISCUSSION OF RESULTS

There is a variation in the limiting values of plant
nutrients for normal crop growth in different soil areas under
different climatic conditions and these limiting values may be
established only after a detailed study of all the factors involved.
In this discussion, however, the limiting values recommended by
the respective originators of the several phmsphorus tests will be
‘used. The values as set by the several investigators are as follows:

Truog-LaMotte method: 75 pounds per acre of available
phosphorus for heavy soils and 50 pounds per acre for sandy soils.

Spurway's water-soluble phosphorus method: 0.5 pm. of
water-soluble phosphorus in the soil extract.

Illinois phosphate test: "Doubtful“.

Truog laboratory method: Approximately 20-25 ppm. of
available phosphorus in sandy soils and 35-h0 ppm. in heavier soils.

The Mankowski fertility series, the data from which are
shown in Table l and Figure 1 (1) is located on Roselawn sandy loam
soil, which is of low fertility in the virgin state. Both Spurway's
method and Truog's laboratory method showed that this soils was low
in available phosphorus. The Truog-LaMotte method and the Illinois
phosphate test showed no deficiency in the available phosphorus content
of this soil. An application of phosphatic fertilizer to this soil
produces a decrease rather than an increase in yield. In the case

of plat 202, which received 200 pounds of superphosphate per acre

 

(1)

The graphs represent comparative values and the form of the

curve a
is the main means of comparison.

-15-

both Spurway's method and Truog's laboratory method showed this

soil to contain more available phosphorus than the soil from plat
201, yet the quantity of phosphorus was the minimum quantity
required for suitable plant growth. The other methods also showed
an increase in the amount of available phosphorus. However, with this
addition of fertilizer plat 202 produced a smaller yield of potatoes
than did the plat which received no phosphatic fertilizer. When
MOO pounds of superphcsphate were added to the soil as was done on
plat 203 all of the methods, with the exception of the Illinois
phosphate test, indicated an increase in the quantity of available
phosphorus. The yield from plat 203 was greater than that from 202
but it was still lower than the yield from plat 201.

Spurway's method showed an increase in phosphorus in the
sample from plat 205 over that in the sample from plat 20h. Similar
results were obtained by the Truog laboratory method. The quantity
of phosphorus was less in plats 20M and 205 than in plat 203. The
results from.Spurway's and Truog's laboratory method correlated quite
well with the fertilizer applications but do not correlate with the
crop response to phosphatic fertilizers. The data from.the Truog-
Leflotte method and the Illinois phosphate test correlated the closest
with the crop response to applications of phosphatic fertilizer.

The Peeble fertility series, the results from which are
shown in Table 2 and Figure l, is located on Onaway sandy loam soil.
The Truog-LaMotte method, Truog's laboratory method, and Spurway '3
method showed that the soil was deficient in available phosphorus,
while the Illinois phosphate test indicated that the soil was well

supplied with phosphorus. In this series there was a good correlation

-16-

between the results from all the methods, with the exception of

the Illinois phosphate test, and the crop response to phosphate
applications. In the case of plats 208 and 20h, to which N00 and 600
pounds of superphosphate were added, respectively, the Truog labora-
tory method, Spurway's method, and the Truog-LaMotte method show no
increase in the amount of available phosphorus. These results correlate
with the fact that there was no noticeable increase in crop yield on
these plats. However, when 800 pounds of superphosphate were added
to plat 205 there was an increase in the quantity of available phos-
phorus in the soil and a noticeable increase in the crop yield.
Throughout this series there was quite a.marked correlation between
the results obtained by all of the methods, with the exception of

the Illinois phosphate test, and the crop response to fertilizer
application.

Table 3 and Figure 2 show the results from the Elsby
fertility series. In this series none of the methods showed a defic-
iency of available phosphorus and there was very little crop response
to fertilizer applications. The only plats showing a larger yield
than the check plat, which received no fertilizer, are plats 5 and 7,
which received 1000 pounds and 2000 pounds per acre of ”—16-8 ferti-
lizer, respectively. This increase may have been due to the potash
and nitrogen added to the soil in the fertilizer. All the methods,
with.the exception of the Illinois phosphate test, showed increases
in the quantities of available phosphorus with increasing applications

of fertilizer until 750 pounds of M—lE-S fertilizer had been added.

-17-
The Truog-Motto method might have shown a further increase but
this was the limit of the color chart. The Illinois phosphate
test indicated that the soil was “high“ in all plots. The Truog
laboratory method indicated an increase in available phosphorus
after 1500 pounds of 1t-16--8 fertilizer had been applied.

The results of the Bird fertility series, which are shown
in Table h and Figure 3, indicated a fair relation between the data
obtained from the Truog-Watts method and Spurway's method, and
the crap response to phosphatic fertilizers. The Illinois phosphate
test and Truog's method both showed that there was a deficiency of
available phosphorus in this soil. The results from the Truog-
LaMotte method and Truog's laboratory method showed a good relation
to the yield from plot 3, which received 500 pounds of L164
fertilizer. The yield from plot 3 was less than the yield from
plot 2, which received only 250 pounds per acre of 15-16-8 fertilizer
and both of the tests also showed a decrease in the quantity of
available phosphorus present. There seems to be no explanation for
this decrease in yield and quantity of available phosphorus, but it
does show the relation between the two methods and the crop yield.
Again there was a gradual increase in available phosphorus as shown
by the Truog-Motto method and Spurway's method as the fertilizer
applications were increased. The results from the Illinois phosphate
test indicated that the soil was “high“ in available phosphorus with
the exception of plat l, which was "medium” in available phosphorus.

The results from the Truog laboratory test were inconsistent through-

-15-
out as the results showed a decrease in the quantity of available
phosphorus as the fertilizer application was increased.

The Demerest fertility series, the results of which.are
shown in Table 5 and.Figure h, was located on Roselawn sandy loam
soil, a soil which is low in phosphorus. The results from all of
the methods with the exception of the Illinois phosphate test
showed this soil to be low in available phosphorus. A.decided
increase in.yield was obtained when 375 pounds of 2-16—6 fertilizer
wereaadded to this soil. In the case of plat 501 Truog's laboratory
method was the only method the results from which indicated a de-
ficiency of phosphorus. When 750 pounds of 2-16-6 fertilizer were
added there was an increase in yield over that of plat 501. _The
results from all the methods, with the exception of those from the
Illinois phosphate test which were "high” throughout, showed increasing
amounts of available phosphorus as the fertilizer application was
increased. The best relation in this respect was between the results
from.the Truog laboratory method and the fertilizer applications.

Table 6 and Figure 5 showed the results from the Hopfer
fertility series, which is located on Onaway sandy loam soil. With
the exception of the Illinois phosphate tests, the results from all
the methods showed the soil to be low in available phosphorus. However,
there was no increase in crop yields to fertilizer applications. In
fact, a decrease in yield occurred when fertilizer was added. All of
the results from.the different methods agreed with the fertilizer
applications, as the quantity of available phosphorus increased with

increasing fertilizer applications. When the fertilizer application

-19-

was increased from 500 pounds to 1000 pounds per acre the quantity
of available phosphorus was doubled as shown by the Truog-IeMotte
method, Spurway's method, and Truog's laboratory method.

The results from the Orr fertility series are shown in
Table 7 and Figure 5. There was sufficient phosphorus in the soil
according to the results from the Illinois phosphate test and the
Truog-LaMotte test. The results from Spurway's method and Truog's
laboratory method showed that there was a minimum amount of phos-
phorus available in plats 601, 602, and 60h. When 1000 pounds of
2—16-6 fertilizer were added both of the methods then showed that
there was sufficient available phosphorus present for plant growth.
The yield was also increased when 1000 pounds of 2-16-6 fertilizer
were added so it would seem that the phosphorus-fixing power of
this soil was high. If this is true Spurway's method and Truog's
laboratory method must indicate the power of phosphorus fixation
that is characteristic of many soils.

Table 8 and Figure 6 present the results from the Griswold
fertility series, which was located on Isabella sandy loam. The
results from the Illinois phosphate test indicate that there was a
deficiency of available phosphorus in this soil. The results from
the Tmog-LeMotte and Truog's laboratory method indicated that there
was a deficiency of phosphorus in the check plat, which received no
superphosphate. The results from Spurway's method indicated that
the available phosphorus content of the soil was low in plate 201,
202, 203, and 20h. There was an increase in yield until Goo pounds

of superphosphate was added. Then there was a decrease of 60 bushels

-20-
per acre in the yield from that of plat 203 which received 400
pounds of superphosphate. The soil samples taken from plat 20h
also showed a decrease in available phosphorus by the Truog
laboratory method. When 800 pounds of superphosphate were added
the yield increased #2 bushels per acre over that of plat 20h.
This might indicate that in some way some of the phosphorus was
fixed by the soil and thus decreased the yield. It seems safe
to state that there was a relation between the results from.Spurway's
method and the crOp response to phosphatic fertilizers.

In Table 9 and Figure 7 are shown the results from.the
College phosphorus plats. The results from Spurway's method and
Truog's laboratory method show that there was a deficiency of phos—
phorus in this soil and a slight increase in crap yield was obtained
with.phosphatic fertilizers. The results from the Truog-LaMotte
method showed that the minimum amount of phosphorus necessary for
crap growth was present in this soil. The Illinois phosphate test
results showed that the available phosphorus was "high" in this
soil. With the addition of increasing amounts of phosphorus to
the soil, varying from 0 pounds of superphosphate to 250 pounds
of 3-6h-10 there was a change in the quantity of available phosphorus
as indicated by the Truog-LaMotte method and Spurway's method. There
was a slight change indicated by Truog's laboratory method of deter-
mining available phosphorus. It would appear that this soil also
has a high fixing power for phosphorus applied to it. If this is true
the Illinois phosphate test does not seem.to be sufficieptly sensitive
to distinguish between phosphorus which is so fixed by the soil that
it is unavailable to plants and that phosphorus which is available to

the plant.

.21-
DISCUSSION OF RESULTS FROM SOIL SAMPLES TAKEN AT VARIOUS DEPTHS

The soil samples used in this comparison were obtained
from the fertility plate on the College experimental farm. The
samples were obtained at the various depths by digging holes, in'
the various plats, to a depth of 12 inches. The samples were then
taken from the walls of holes at the depths indicated in the tables.
Samples were taken on two dates. May 9 and August 1% of the same
owner. The crops had been harvested when the August 19‘ samples
were taken.

The results from the IF plat, which received no fertilizer,
are shown in Table 11 and Figure 8. Truog's laboratory method
showed only two samples of those taken May 9 as containing enough
phosphorus for good plant growth. All of the other methods showed
the samples from all of the depths to contain enough phosphorus for
good plant growth.

The results from the Truog-LaMotte method showed that the
August 1’4 samples were all low in available phosphours. All of the
methods showed that the samples taken August 11} at depthe of eight
to ion inches and ten to twelve inches were low in available phos-
phorus. There was a fair agreement in the quantities of phosphorus
found by the Truog-LaMotte method and Spurway's method.

_The results shown in Table 12 and Figure 9 are from the
23' plat which was fertilized the previous fall with )480 pounds per
acre of 20% superphosphate. The Truog laboratory method showed soil

from all of the depths low in available phosphorus, with the exception

of the lurface layer. The Illinois phosphate test showed all samples

.22-
"high! or “medium" in available phosphorus. ‘All of the samples
taken May 9, with the exception of those from the eight to ten
and ten to twelve inch depths contained enough available phosphorus
for plant growth according to the Truog-LaMotte test. Spurway‘s
method showed adequate phosphorus in the soil from the one to two
inch depths and the minimum quantity of phosphorus necessary for
plant growth in all of the remaining samples. There is a fair
relation between the results from all of the methods with the exception
of the Illinois phosphate test.

The results from the BF plat which was fertilized in the
spring with “80 pounds of 20% superphosphate are shown in Table 13
and Figure 10. The results from the Illinois phosphate test indicated
that there was sufficient phosphorus present for plant growth, in the
soil from all depths with the exception of that from the eight to ten
and ten to twelve inch depths. The surface soil according to Truog's
laboratory method and the Truog-leMotte method was well supplied with
available phosphorus on May 9, while Spurway's method showed that the
minimum amount of phosphorus necessary for plant growth was present.
$011 from the eight to ten and ten to twelve inch depths was veny
deficient in phosphorus according to the results from all of the methods.
All the methods showed an increase in the quantity of available phos-
phorus in the two lower depths from May 9 to August IN.

The IP plat was fertilized with 250 pounds of 3-8-10 fertilizer
per acre. The results from this plat are shown in Table l“ and Figure
11. The results from all of the methods, with the exception of the
Illinois phosphate test, showed a deficiency of phosphorus in the soil

of this plat. The Illinois phosphate tests showed that all of the

-23-

samples taken May 9 were "high" in phosphorus and those taken
August 134 were “medium" in phosphorus. The results from all of
the other methods also indicated a decrease in the quantity of
available phosphorus from May 9 to August 11+.

The results of the 2P plat, which received no fertilizer,
are shown in Table 15 and Figure 12. According to all of the
methods, with the exception of the Illinois phosphate test, the
samples taken both May 9 and August 1’4 were low in phosphorus as
only the minimum amount of phosphorus necessary for plant growth
was present. The Illinois phosphate test indicated all of the soils
as "high" or "medium" in available phosphorus.

The 3P plat, the results of which are shown in Table 16
and Figure 13, was fertilized with 250 pounds of 3-16-10 fertilizer.
The results from this plat are very similar to those received from
the 2P plat. All of the methods agreed or correlated in that the
quantity of available phosphorus in the surface layer decreased from
the time of the first sampling to the time of the second sampling.

The results from the up plat, which was fertilized with
250 pounds of 3-32-10 per acre, are shown in Table 17 and Figure
11‘. The results from all of the methods agreed fairly well although
the Illinois phosphate test showed all of the samples to contain
somewhat more phosphorus than did the other methods.

Table 18 and Figure 15 present'the results from the 5P
plat which was fertilized with 250 pounds of a 3-94-10 fertilizer.
Again all of the data agreed in showing that all of the samples
were relatively high in available phosphorus.

Throughout this series all of the methods, excepting the

-2l+-
Illinois phosphate test, agreed with each other in the quantities
of available phosphorus found. When a plat had been heavily
fertilized with a phosphatic fertilizer the results from all of the
methods indicated an increase in available phosphorus. However,
the Illinois phosphate test does not indicate a larger quantity and
so the results from it could not correlate closely with those from

the other methods.

-25- .
THE USE OF BUFFERED AND UNBUFFEPED EXTRAOTIITG SOLUTIONS
IN THE TRUOG LABORATORY METHOD
Truog's laboratory method (1’4) calls for the use of a

0.002 N. sulphuric acid solution, buffered to a pH of 3 with
ammonium sulphate for the extraction of available phosphorus from
soils. The writer attempted to determine if there was any dif-
ference in results when an unbuffered solution was used instead
of the buffered solution. No changes were made in the method
except that unbuffered 0.002 N. sulphuric acid solution was used
instead of a buffered 0.002 N. sulplnlric acid solution.

In Table 15 and Figures 8, 9, and 10 are shown the results
obtained from this work. Very little difference was found in the
amounts of phosphorus extracted by the two different solutions but

in the majority of cases the unbuffered solutions extracted more

phosphorus than the buffered solutions.

-26-
GENERAL DISCUSSION OF RESULTS

In this discussion the results will be taken up according
to the location of the plate in the different sections of the State
and with regard to the various soil types. The lower peninsula of
the State can be divided into the northern lower peninsula and.southern
lower peninsula. Otsego and.Antrim counties are included in the northern
lower peninsula group, while Kent, Montcalm, Ionia, Ingham, and
Kalamazoo counties are included in the southern lower peninsula. The
Mankowski, Demerest, Peeble, and Hopfer fertility series are in the
northern half of the lower peninsula. The Mankowski series showed a
relation between the TruogbLaMotte method and Spurway's water-soluble
phosphorus method. The crop yields were quite variable and the Truog-
LaMotte method did not correlate well with them. The Demerest fertility
series shows a very good response to fertilizer. The relation between
the Illinois phosphate test and the response to fertilizer is very poor
as the Illinois phosphate test shows a high phosphorus content throughp
out the series. When the Truog-LaMotte method and Spurway's method
show a soil deficient in available phosphates, in the majority of cases
that soil will respond to phosphatic fertilizers. The relation between
the Truog laboratory method and the response to fertilizer is good.
In the Peeble fertility series a fair relation is shown between the
Spurway, Truog-LaMotte method, and Truog's laboratory method and the
crop response to fertilizer. There is a poor relation between the
results of the Illinois phosphate test and the crop response to fertilizers.
The same results were found with the Hapfer fertility series.

In the southern.part of the lower peninsula are the Orr,

Elsby, Griswold, Bird, College plats, and the Kalamazoo fertility series.

-27-
In this section of the State the Illinois phosphate test again
shows few correlations. This test often shows the soil as “high”
in available phosphorus and yet there is a variable crop response
to the fertilizers. .Also, the test seems to lack sensitivity as
it fails to show the great differences in the amount of available
phosphorus, present in the soil due to fertilizer applications.
The Truog laboratory method works well in the southern part of
the lower peninsula. The results from.the method correlate with
the crop response of the plots to fertilizer treatments and this
method also shows differences in the amounts of available phos-
phorus in the soils studied. The Truog-LaMotte method and Spurway's
water—soluble method both.generally show a correlation with the
crop yields and fertilizer applications of the various fertility
plats. These methods show differences in the amounts of available
phosphorus for the plate. If the crop,yields are increased hy
phosphate fertilizers both of these methods generally show the
'untreated plat as being low in available phosphorus. From these
comparisons there is very little difference between the results
obtained by these two methods on soils in the various parts of the
lower peninsula.

In Table 19 is shown a comparison between the Truog-
LaMotte method of determining available phosphorus and Truog's
laboratory method of determining available phosphorus. The largest
number of soils are shown as containing 20-30 ppm. of available
phosphorus by the Truog laboratory method and the same soils contain

75 pounds of available phosphorus per acre by the Truog-LaMotte

-25-

method. .As the amount of available phosphorus in the soil increases
by the Truog laboratory method the amount of phosphorus increases by
the Truog-lei.iotte method.

In Table 20 is shown a comparison between the Illinois
phosphate test and the Truog laboratory method. The Truog laboratory
method shows N5 soils as containing 30-h0 ppm. of available phos-
phosus while the Illinois phosphate test shows the same soils as
'high”. Truog‘s laboratory method shows no soils as containing
10-20 ppm. of available phosphorus, while the Illinois phosphate
tests shows these same soils as ”medium”. 10-20 ppm. of available
phosphorus would indicate that the soil is low in available phos-
phorus.

In Table 21 is shown a comparison between Spurway's water-
soluble phosphorus method and Truog's laboratory method. The
largest number of soils are shown as containing 10-20 ppm. of avail-
able phosphorus in the soil by Truog's laboratory method while
Spurway's method shows that the water extracts of those same soils
contain 0.5 ppm. of phosphorus. .Also ”0 of the soils contain 20-30
ppm. of available phosphorus by the Truog laboratory method while
they are shown as containing 0.5 ppm. of phosphorus in the water
extract by Spurway's method. As the amount of water-soluble phos-
phorus increases by Spurway's method the amount of available phos-
phorus increases by Truog's laboratory method.

The results from the Illinois phosphate test did not
correlate with the yield data from the plate receiving fertilizer

treatments. Usually, if the soil was deficient in phosphorus the

-29-
Illinois phosphate test showed it as ”high” or "medium” in avail-
able phosphorus. Fraps (8) found that the Illinois phosphate
test was quite variable with different analysts and with the same
analyst at different times. Fraps also found that soils containing
30 ppm. of active phosphoric acid were classed as high by this method.
The active phosphoric acid in this case is the amount removed from
the soil by 0.2 N nitric acid.

The Truog-LeMotte method showed a fair cerrelation with
the crop results from the different soil plate. It showed similar
results in both the northern and southern part of the lower peninsula.

Similar results were obtained with.Spurway's water-soluble
phosphorus method. This method correlated quite well with.the Truog-
LaMotte method in.most of the cases studied. When one of these
methods showed a deficiency of phosphorus the other method would
show similar results. Spurway's water-soluble phosphorus method also
showed similar correlations when used in connection with soils from
the northern or southern part of the State.

The Truog laboratory method in.most cases correlated qhite
well with the results obtained from the different fertility series
throughout the State. It can be said that the Truog laboratory method
showed the variations in phosphorus content between the different
plats more closely than the other methods, but this difference can be
accounted for by the wider range of results obtainable with this
method. In the determination of the plant nutrient deficiencies of

soils by chemical tests it is perhaps not necessary to distinguish

-30-

between small differences. The results may vary considerably and
still indicate roughly the condition of the soil with respect to
the supply of plant nutrients. This method, however, did correlate
quite well with the Truog-LaMotte method and Spurway's water-

soluble phosphorus method.

-31-
'CONCLUSIONS
.A comparison of four methods, for the determination of

the available phosphorus of soils, was made on soil samples taken
from different station fertility plots.“ Three of the methods,
Spurway‘s water-soluble phosphorus method, the Illinois phosphate
test, and the Truog-Lahotte method are fundamentally field tests,
'while the fourth is Truog's laboratory method.

The results from the Illinois phosphate test showed a
poor correlation with the crop response of the soils to phosphatic
fertilizer treatments. :The test showed very little difference
between soils deficient in phosphorus and those containing a large
amount of phosphorus as judged from fertilizer applications and
crop yields. The Illinois phosphate test as used in this research
has shown a tendenqy to show phosphorus deficient plate as high in
phosphorus. The limiting value of phosphorus in this test in most
cases could be established as ”medium“ for this test. Very few
soils were found by this method.which showed "low" or doubtful".

The results from the Truog-LaMotte method showed a fair
correlation with the crop response of the soil to phosphatic
fertilizers.

The results from Truog's laboratory method showed a fair
correlation with the crop response of the soil to phosphatic
fertilizer. However, when large amounts of phosphorus were present
in the soil as indicated by the Truog-LaMotte method and fertilizer
applications, Truog's laboratory method did not indicate this large

amount. Only one standard is used in this method and that may

.32-

account for this variation due to the wide difference between the
depth of color developed by the standard and the unknown. Because
of this wide variation it is not possible to read as accurately
when large amounts of phosphorus are present.

The results obtained by Spurway's water-soluble phosphorus
method correlate fairly well with the results obtained by the use
of phosphatic fertilizers.

The limiting values as established by Truog-LaMotte
method, Spurway's water-soluble phosphorus method, and Truogis
laboratory method correlate quite well with all of the results
obtained in this study. The Truog-LaMotte method and Spurway's
water-soluble phosphorus method could both be recommended as field
methods of determining phosphorus deficient soils under Michigan
conditions. Truog's laboratory method could be recommended for
use in.the laboratory to determine the available phosphorus of
soils although.it is not as rapid as the Truog-LaMotte method and
Spurway's water-soluble phosphorus method.

The main difference between all of these methods is in the
method of extraction of the phosphorus from the soil, and it is thmught
a better means of extraction should be developed-a.method which would
show more clearly the true status of the available phosphorus in the

soil under different moisture contents and fertilizer treatments.

(1)

(2)

(3)

(u)

(5)

(6)

(7)

(8)

(9)

(10)

(11)

-33-
Literature Cited

Atkins, W. R. J. The rapid determination of available
phosphate in soil by the caemlo-molybdate reaction of
Deniges. Journal of Agricultural Science, )4:192-197,
19?. .

Bell, R. D. and Daisy, A. L. Rapid colorimetric methods for
the determination of phosphorus in urine and blood.
Journal of Biological Chemistry, nu:55-67, 1920.

Bray, R. H. A field test for available phosphorus in soils.
Illinois Agricultural Experiment Station Bulletin 337, 1929.

Deniges, D. An extremely sensitive reaction of phosphate
and arsenates and its application. Compt. Rind., 171:802-
son, 1920. (Abstracted in Chemical Abstract, 15: 218, 1921.)

Disks, 0. H. and Subbarrow, Y. The colorimetric determination
of phosphorus. Journal of Biological Chemistry, 66: 375-1400,

1925.

Florentin, D. The determination of phosphate in waters.
Ann. Chem. Anal. Chem. Appl.. 33295-296. (Abstracted in
Chemical Abstract, 16: 201, 1922.)

Drape. G. S. Active phosphoric acid and its relation to the
needs of the soil for phosphoric acid in pot experiments.
Texas Agricultural Experiment Station Bulletin 126: l.

Traps, G. S. How reliable are existing chemical methods for
determining soil deficiencies in ash constituents of plants.
Journal of Americal Society of Agronomy, 23: 337-352, 1931.

Hall, A. D. and Plymen, F. J. Determination of available
plant food in soils by use of weak acid soluents. Transactions
of Chemical Society, 81: 117-1uu, 1906.

Hibbard, P. L. Chemical methods for estimating the availability
of soil phosphate. Soil Science, 31: l+37—H67, 1931.

Lonstein, 1. Rapid, colorimetric determinations of phosphates
in soils and vegetation. Journal South Africa Institute, 10:

' “9-50, 19270

(12)

Neubauer, H. Determination of the potash phosphate requirements
of soils by the seedling method. Trans. 2nd Comm. Interm. Soc.
5911 Sci., a: 159-170. 1929. (Chem. Abst. 2h: M573. 1930)

(13)

(1h)

(15)

(15)

(17)

(18)
(19)
(20)
(m
(22)

(23)

-311...

Parker, F. W. and Fudge, J. 1'. Soil phosphorus studies:

I The colorimetric determination of organic and inorganic
phosphorus in soil extracts and the soil solution. Soil
Science, 21+: 109-118, 1927.

Peter, A. u. and Ave-ritt, s. n. Soils. Kentucky
Agricultural Experiment Station Bulletin 126, 1906.

Ridgeway, R. A. Color standards and nomenclature. Hoen and
Company, Baltimore, Maryland.

Russell, E. J. and Prescott, J. A. The reaction between
dilute acids and the phosphorus compounds of the soil.
Journal of Agricultural Science, 8: 65-110, 1916.

Shedd, O. M. A short test for easily soluble phosphates
in soils. Soil Science 11: 111-122, 1921.

Spurway. C. H. A test for water-soluble phosphorus.
Michigan Agricultural Experiment Station Technical Bulletin
101. 1929.

Stewart, R. The manurial requirements of soils. (Two
modern methods of estimation). The Scottish Journal of
Agriculture, 12: No. 3. 1929.

Stoddert, G. I. Soil acidity in its relation to lack: of
available phosphates. Journal Industrial and Engineering
Chemistry, 1: 69-71%, 1909.

Thornton. 8. 1. Experiences with the Neubauer method for
determining mineral nutrient deficiencies in soils. Journal
of American Society of Agrononw. 33: 195-208, 1931.

Truog, E1111 and Meyer, A. H. Improvements in the Deniges
colorimetric method for phosphorus and arsenic. Industrial
and Engineering Chemistry, 1: 136-139, 1929.

Truss, nail. The determination of the readily available
phosphorus of soils. Journal of American Society of Agronomy,

«can noes no 3M .

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