V A N E R E M E T A L . : JOURNAL OF A O A C INTERNATIONAL V O L . 81, No.
1,1998 25
AGRICULTURAL MATERIALS
Determination of Water in Forages and Animal Feeds by Karl
Fischer Titration
TERRI VAN EREM and NANCY THIEX 1
South Dakota State University, Department of Chemistry and Biochemistry, Box 2170, Brookings, SD 57007-1217
JORG POHMER
Allied Signal, Inc., 101 Columbia Rd, PO Box 1053, Morristown, NJ 07962-1053
W. MICHAEL POFFENBARGER
Hoechst Celanese Corp., U.S. Highway 43 North, PO Box 64, Bucks, AL 36512
VICTOR SMITH and ELENA PATEL
Brinkmann Instruments, Inc., PO Box 1019, Westbury, NY 11590-0207
Oven methods for determining moisture (volatiles)
in forages and other animal feeds are empirical.
The moisture concentration obtained depends
upon the time and temperature the sample was
dried and is influenced by the presence of other
volatiles than water. A validated reference method
to measure water in forages and animal feeds
could be used to evaluate the appropriateness of
oven methods for various types of animal feeds
and forages. Karl Fischer titration is a well-estab-
lished method for determining water. However,
thorough extraction of water from forages and
feeds is a challenge because they often contain
cellular structures that release water slowly. Water
was successfully extracted into methanol-forma-
mide (50 + 50) by high-speed homogenization and
then titrated directly at 50 C with a one-component
Karl Fischer reagent based on imidazole. The
method is described in detail, results of day-to-day
repeatability and laboratory-to-laboratory repro-
ducibility are reported, and preliminary compari-
son data between oven methods are provided.
here is a great deal of discussion and disagree-
T ment in the analytical community as to which
oven method is the best indicator of the moisture
contents of forages and animal feeds. Because of the
empirical nature of oven methods, moisture concentra-
tions obtained with a given method depend on the time
and temperature the sample was dried and are influ-
Received January 23, 1997. Accepted by RN August 11, 1997.
Author to whom correspondence should be addressed.
Published as South Dakota Agricultural Experiment Station Jour-
nal Series 2971.
This paper reports results of research only. Mention of a trade-
mark, proprietary product, or vendor does not constitute a guarantee
or warranty of the product and does not imply approval to the
exclusion of other products or vendors that may also be suitable.
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enced by the presence of volatiles other than water. An
analytical tool is needed to evaluate the appropriate-
ness of various oven methods for different types of
animal feeds and forages. A method specific for water
could be considered a reference method. The goal of
this study was to establish an accurate, reproducible,
yet practical method for determining water in dry,
ground forages and animal forages and feeds. The
method should have reasonable throughput so that it
could be used routinely in feed and forage laboratories.
A second objective was to make a preliminary compari-
son of the method to various oven methods for forages
and animal feeds.
Karl Fischer titration is a well-established general
method for determining water (1,2). However, a tech-
nique to efficiently extract water from these types of
materials and to subsequently analyze the extract by
Karl Fischer titration was not available. We tried un-
successfully to extract samples with methanol in a Spex
Model 8000 Mixer/Mill with 3-1.27 cm diameter steel
balls. Methods using this approach for different materi-
als were reported (3,4). We experienced problems with
leaky seals leading to loss of solvent and dilution error.
Error also was introduced during transfer of solvent
into the titration flask, especially when humidity was
high. Throughput was very low because of the laborious
procedure. This method was not desirable for routine
(non research) applications. We considered another
approach, which involves heating samples in N,N-di-
methylformamide (DMF) at 90°C for 60 min (5). We
found this approach undesirable because of potential
error introduced by atmospheric moisture because
DMF is hygroscopic, and because it involves direct
handling of DMF.
A number of advantages are associated with high-
speed homogenization of forage and animal feeds sam-
ples directly in the Karl Fischer titration vessel in
methanol-formamide (50 + 50) combined with subse-
26 VAN EREM E T AL.: JOURNAL OF AOAC INTERNATIONAL VOL. 81, No. 1, 1998
quent titration at elevated temperatures. The tech-
nique had been reported before (6). It provides quanti-
tative extraction of water from cellular structure in a
short time. Other components coextracted with water
(e.g., oils and carbohydrates) do not interfere because
the Karl Fischer method is based on a specific chemical
reaction consuming water.
Proposed Karl Fischer Method
Scope
The proposed method is applicable for determina-
tion of water in dry, ground animal feeds and forages.
Reagents
(a) Karl Fischer reagent.—One component, based on
imidazole, with titer of ca 5 mg H 2 0 / m L reagent,
Hydranal Composite 5 (Riedel-de Haen 34805, St.
Louis, MO), or equivalent.
(b) Methanol.—Anhydrous, for moisture determina-
tions, water content not to exceed 0.05% (EM Science,
Gibbstown, NJ).
(c) Formamide.—ACS reagent grade (Fisher Scien-
tific, Fair Lawn, NJ).
(d) Solvent.—50% methanol and 50% formamide.
(e) Sodium tartrate dihydrate.—Primary standard
(water content, 15.66 + 0.05%), Hydranal standard
sodium tartrate-2-hydrate (Riedel-de Haen 34803, St.
Louis, MO), or equivalent.
Apparatus
(a) Karl Fischer titration-homogenization system.—
Metrohm 720 KFS Titrino (Brinkman Instruments, Inc.,
Westbury, NY), includes titration vessel LP with water
jacket (50-150 mL), snap-in buret unit (10 mL),
703 titration stand with pump, cable with timer
(Titrino-Polytron), or equivalent.
(b) Circulating water bath.—Capable of maintaining
50° ± 1°C (Fisher Scientific, Pittsburg, PA).
(c) Homogenizer.—Brinkmann Polytron homoge-
nizer with PTA 20TSM foam-reducing generator with
saw teeth and knives. Assembled with generator ex-
tending into the titration vessel and adjusted to be ca
1 in. from the bottom of the titration vessel. Set to
provide a homogenization speed of 24 000 RPM.
(d) Balance.—Analytical, electronic, sensitive to
0.1 mg (Mettler Instruments, Hightstown, NJ).
(e) Glass weighing spoon.—With opening for dis-
pensing sample into the titration flask through the
septum stopper (Brinkmann 6.2412.000 or equivalent)
(Brinkman Instruments, Inc., Westbury, NY).
Safety
Hydranal Composite 5 contains 5 hazardous compo-
nents—iodine, sulfur dioxide, imidazole, diethylene
glycol monoethyl ether, and hydriodic acid—and should
be handled with care. Methanol: Highly flammable;
toxic by inhalation, in contact with skin, and if swal-
lowed. Formamide: Moderately irritating to skin, mu-
cous membranes.
Sample Preparation
Grind samples to pass a 1 mm, or smaller, opening
to ensure homogeneity.
Drying or Conditioning the Cell
Dry titration cell by completing a pretitration as
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follows: Dispense 80 mL methanol-formamide solvent
into titration vessel. Close cell to minimize addition of
atmospheric moisture. Heat to 50° ± 1°C. Dry cell (in-
cluding solvent, cell walls, electrode walls, generator,
cell atmosphere) by performing a blank run, including
homogenization and titration. Homogenize at
24 000 RPM for 60 s. Begin titration immediately. Be-
fore reaching end point, homogenize momentarily to
rinse untitrated methanol that may have been sitting
under the cap into solution. The end point is reached
when no change in potential is observed for 10 s (titra-
tion system programmed for stop criterion: time; delay:
10 s). A dried titration cell has a maximum drift con-
sumption of 5-10 uX Karl Fischer reagent/min.
Standardization
Heat cell to 50° ± 1°C. Dry cell as described above.
Immediately after drying the cell, quickly weigh
150-250 mg sodium tartrate dihydrate standard into
glass weighing spoon and record weight of spoon and
standard to nearest 0.1 mg (S). Quickly transfer
weighed sample into the titration flask through septum
stopper. Reweigh empty sample spoon to obtain tare
weight (T). Obtain weight of standard material added
by subtracting tare weight (T) from weight of spoon
plus standard (S). Record weight of standard material
(S — T) in mg to the nearest 0.1 mg. Homogenize at
24 000 RPM for 60 s. Begin titration immediately. Be-
fore reaching end point of titration, homogenize mo-
mentarily to rinse down moisture that may have been
sitting under the cap. This ensures that moisture of
particles sitting under the cap or hanging on the glass
walls above the liquid surface is titrated. Titrate to
same endpoint as described in the conditioning step,
recording volume of reagent required for titration (mL
reagent) to the nearest 0.001 mL. Repeat a total of
5 times. Calculate titer and then average the 5 values.
The relative standard deviation should be < 2%.
mL reagent
mg N a 2 C 4 H 4 0 6 • 2 H 2 0 X 0.1566
mL reagent
where mg N a 2 C 4 H 4 0 6 • 2 H 2 0 is S — T, in mg.
 VAN EREM ET AL.: JOURNAL OF AOAC INTERNATIONAL VOL. 81, No. 1,1998
Determination
Heat cell to 50° + 1°C. Dry cell as described above
before running a sample or each time solvent is replen-
ished in titration cell. Check drift in titration cell. A
dried titration cell has a maximum drift consumption of
5-10 |JLL Karl Fischer reagent/min.
Analyze sample as follows: Immediately after drying
cell, quickly weigh ca 0.5 g sample (to contain ca 2 5 -
50 mg water) into glass weighing spoon and record
weight of spoon plus sample (W). Quickly add weighed
sample into titration flask through septum stopper.
Reweigh empty sample spoon and record tare weight
(T). Obtain sample weight by subtracting tare weight
(T) from weight of spoon plus sample (W). Record
sample weight (W — T) in grams to the nearest 0.1 mg.
Homogenize at 24 000 RPM for 60 s. Begin titration
immediately. Before reaching end point of titration,
homogenize momentarily to rinse down moisture that
may have been sitting under the cap. This ensures that
moisture of particles sitting under cap or hanging on
glass walls above the liquid surface is titrated. The end
point is reached when no change in potential is ob-
served for 10 s (stop criterion: time, delay: 10 s). Re-
peat determination in triplicate. Relative standard de-
viation of replicates should be < 5 % . The cell need not
be emptied between each sample. Usually about
3 titrations can be performed before the cell needs to
be emptied and replenished.
Calculations
H 2 0 , m g = VX titer
V X titer
H
20, % = — : —
10 X sample weight
where V is volume of titrant (mL) and sample weight is
W- T(g).
Method Repeatability, Reproducibility, and
Comparability
Five samples representing various types of forages,
animal feeds, and pet food were selected for repeatabil-
ity testing, reproducibility testing, and for comparisons
with oven methods. Samples included corn silage, tex-
turized feed, baked dog food, total mixed ration (TMR),
and legume hay. Corn silage, TMR, texturized feed,
and legume hay were dried at 60°C overnight. Samples
were ground to pass a 0.75 mm screen in a Brinkmann
Retch Mill Model ZM-1. Samples were split and placed
in air-tight, heat-sealed plastic containers to minimize
moisture change. Analysts were instructed to open bags
on the day of analysis.
To test method repeatability, 5 samples were ana-
lyzed at South Dakota State University (SDSU) on 3
27
different days. To test method comparability, the same
5 samples were analyzed in triplicate by 4 oven meth-
ods on the same 3 days as the repeatability tests. The
oven methods chosen were selected because of their
frequent use in laboratories analyzing these types of
samples: Oven Method I (7), drying in a forced draft
oven at 135°C for 2 h; Oven Method II (8), drying in a
forced draft oven at 104°C for 3 h; Oven Method III
(9), drying in a vacuum oven at 95°C for 5 h; and Oven
Method IV (10), drying in a forced draft oven at 105°C
for 16 h. Methods I and III are AOAC official methods
for animals feeds and plants, Method II is an AOAC
official method for malt, and Method IV is an AOAC
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official method for peat. To test method reproducibil-
ity, the 5 samples were distributed to 3 different labo-
ratories for analysis.
Oven Methods
Apparatus
(a) Drying oven, forced draft convection.—Capable of
maintaining temperature at ±1°C. Oven should be
equipped with a metal rod shelf to allow circulation of
air. Exhaust vent on top of oven should be wide open.
Gallen Kamp Model Plus II, Leicester, UK.
(b) Drying oven, vacuum oven.—Equipped with
pressure gauge and needle-type air valve. Oven should
be equipped with system to dry input air. Precision-
Thelco Model 19, Chicago, IL.
(c) Vacuum pump.—Connected to vacuum oven via
vacuum tubing, Welch Model 1404, Skokie, IL.
(d) Moisture pans.—Aluminum, 50 id X 20 mm
deep; lids are optional, Fisher Scientific, Itasca, NY.
(e) Analytical balance.—Electronic, sensitive to
0.1 mg. (Mettler Instruments).
(f) Desiccator. (Corning, Corning, NY).
Determination
Dry moisture pans at > 100°C for at least 2 h. Trans-
fer moisture pan to a desiccator and cool. (A desiccator
full of containers should cool for at least 1 h but not
more than 2-3 h.) Weigh moisture pan; record tare
weight (T) in g to nearest 0.1 mg. Tare pan to zero
while on balance. Add 1.5-2 g well-mixed sample to
pan. Record sample weight (S) in g to nearest 0.1 mg.
Repeat each analysis in triplicate. Gently shake pan to
uniformly distribute sample and expose maximum area
for drying. Place samples in preheated oven. Leave
space around each pan to allow free air circulation.
Dry at the temperatures and times indicated in Table 1.
Oven should come to temperature ca 5 min after sam-
ples are put into it.
Transfer dried samples to desiccator and cool.
Weigh; record weight of pan and dried sample (D) in g
to nearest 0.1 mg. Moisture pans can be cleaned by
blowing with air. Note: Official moisture methods call
for use of a lid with moisture pan. Place lid on bottom
of pan. Dry and weigh empty pan and lid together. As
28 V A N E R E M E T A L . : JOURNAL OF A O A C INTERNATIONAL V O L . 81, No. 1, 1998

Table 1. Oven conditions

Oven
method Temperature, °C Drying time, h Oven type

135 ± 2 2 Forced draft

104 + 1 3 Forced draft
95-100 5 Vacuum oven
(<100 mm Hg)
IV 105 ± 1 16 Forced draft

dried sample is being removed from oven, close mois- Results and Discussion

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ture pan with its lid.
Method Repeatability
Calculations
Five samples representing various types of forages,
(D - T) X 100 animal feeds, and pet food were analyzed at SDSU on
Dry matter, % 3 different days. Means, standard deviations (SDs), and
coefficients of variation (CVs) for each day are re-
Moisture, % = 100 - % dry matter ported by product in Table 2. Combining results from
3 days, intralaboratory CVs ranged from 1.41 for dog
food to 5.09 for TMR. There were no statistical differ-
where D is weight of pan and dried sample (g), T is ences between days (method repeatability) for corn
tare weight of pan (g), and S is sample weight (g). silage, TMR, dog food, or legume hay. Only the mean
Data were statistically analyzed by using PROC for day 1 differed from those of day 2 and day 3 for
ANOVA in SAS (11). texturized feed (P = 0.012). On the basis of these

Table 2. Results of repeatability studies 2

Water found

Feed Day n Mean, % SD CV

Corn silage 1 3 4.63 b 0.05 1.08

2 3 4.72b 0.13 2.75
3 4 4.74 5 0.21 4.40

Combined days 10 4.70 0.16 3.31

b
Texturized feed 1 4 7.21 0.29 4.05
2 3 7.65 c 0.07 0.87
3 3 7.82 c 0.08 1.02

Combined days 10 7.52 0.20 2.64

b
Dog food 1 3 6.94 0.07 1.04
2 3 6.94 fa 0.09 1.30
3 3 7.05 b 0.13 1.78

Combined days 9 6.97 0.10 1.41

b
Total mixed ration 1 4 4.83 0.27 5.60
2 7 5.24 fa 0.29 5.50
3 3 5.08 b 0.09 1.83

Combined days 14 5.09 0.26 5.09

b
Legume hay 1 4 6.89 0.26 3.83
2 6 6.93" 0.31 4.51
3 3 6.58 fa 0.08 1.14

Combined days 13 6.84 0.27 3.90

a
Means within feeds with the same superscript letter are not statistically different (P = 0.05).
 V A N E R E M E T A L . : JOURNAL OF A O A C INTERNATIONAL V O L . 81, No. 1,1998 29

results, method precision is adequate, and the method
is considered to be repeatable.
Method Reproducibility
Five samples were distributed to 3 different labora-
tories for analysis. Means, SDs, and CVs for each
laboratory are reported by product in Table 3. Combin-
ing results from 3 laboratories, interlaboratory CVs
ranged from 1.64 for dog food to 4.46 for TMR. There
were no statistical differences between laboratories
(method reproducibility) for TMR or legume hay.
practical differences and were attributed to differences
in intralaboratory variability. All laboratories matched
on 2 of 5 samples, and 2 of 3 laboratories matched on
the other 3 samples. Interlaboratory variability was ac-
ceptable, and the method was sufficiently reproducible.
Comparison with Oven Methods
Five samples were analyzed in triplicate by each of
4 oven methods on the same 3 days as repeatability
tests were conducted. Means, SDs, and CVs are re-
ported Table 4. For corn silage, Karl Fischer and oven
method II with respective means of 4.70 and 4.55%

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Means for laboratory 3 were different from those of
laboratories 1 and 2 for corn silage (P = 0.0015) and
texturized feed (P = 0.0001). Means for laboratory 1
differed from those of Laboratories 2 and 3 for dog
food (P = 0.0002). Differences between laboratory 3
and laboratories 1 and 2 for corn silage and texturized
feed were apparently due to differences in precision
between the laboratories. Generally, laboratory 3 had
the least experience with Karl Fischer methods and
also had more variability on all samples. Laboratories 1
and 2 generally had extremely low SDs. Single-day,
intralaboratory CVs for laboratories 1 and 2 were ex-
cellent, ranging from 0.66 to 1.79. Therefore, although
some statistical differences were found between 2 labo-
ratories on 3 of 5 samples, they were not considered
were similar. All other methods were different (P =
0.05). For corn silage, oven methods I and IV gave high
means of 6.33 and 5.57%, respectively. Oven method III
gave the lowest mean, 4.00%.
Karl Fischer, oven method II, and oven meth-
od III gave similar means of 7.52, 7.08, and 7.45% for
texturized feed and 5.09, 4.85, and 4.95%, respectively,
for TMR. All other methods were different (P = 0.05).
The means for texturized feed and TMR for oven
method I and oven method IV of 9.17 and 8.22%, and
6.57 and 5.86%, respectively, were considerably higher
than those from the other 3 methods.
For dog food, Karl Fischer and oven method I gave
similar means of 6.97 and 7.03%. All other methods
were different (P = 0.05). Means for oven methods II,

Table 3. Results of reproducibility studies 9

Water found
Feed Laboratory n Mean, % SD CV

Corn silage 10 4.85 b 0.03 0.66

3 4.98fa 0.07 1.51
9 4.70 c 0.15 3.24
All 21 4.81 0.10 2.17
b
Texturized feed 1 5 8.37 0.12 1.45
2 3 8.06" 0.09 1.15
3 10 7.52 c 0.33 4.38
All 18 7.85 0.27 3.38
Dog food 1 10 7.25' 0.13 1.79
2 3 7.00c 0.10 1.48
3 9 6.97c 0.10 1.47
All 22 7.10 0.12 1.64
fa
Total mixed 1 10 5.11 0.08 1.50
ration 2 3 5.05 b 0.04 0.73
3 14 5.09 b 0.30 5.92
All 27 5.09 0.23 4.46
b
Legume hay 1 10 7.01 0.08 1.13
2 3 6.79 6 0.04 0.66
3 13 6.84" 0.28 4.17
All 26 6.90 0.21 3.08
Means within feeds with the same superscript letter are not statistically different (P - 0.05).
30 VAN EREM ET AL.: JOURNAL OF AOAC INTERNATIONAL VOL. 81, No. 1, 1998

Table 4. Comparison of proposed Karl Fischer (KF) method with various oven methods*

Water found
Feed Method n Mean, % SD CV

b
Corn silage Proposed KF 10 4.70 0.15 3.10
Oven method 1 9 6.33° 0.34 5.30
Oven method II 9 4.55b 0.45 9.83
Oven method III 9 4.00cf 0.58 14.42
Oven method IV 9 5.57e 0.28 5.05

Texturized Proposed KF 10 7.52" 0.33 4.38

feed Oven method 1 9 9.17c 0.44 4.73
Oven method II 9 7.08b 0.36 5.08

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Oven method III 9 7.45fa 0.84 11.34
Oven method IV 9 8.22d 0.26 3.16

Dog food Proposed KF 9 6.97" 0.10 1.47

Oven method 1 9 7.03fa 0.12 1.71
Oven method II 9 6.17c 0.31 5.09
Oven method III 9 6.38 d 0.11 1.77
Oven method IV 9 6.61e 0.13 1.89

Total mixed Proposed KF 14 5.09fa 0.30 5.92

ration Oven method 1 9 6.57c 0.34 5.22
Oven method II 9 4.85b 0.21 4.25
Oven method III 9 4.95b 0.41 8.33
Oven method IV 9 5.86d 0.14 2.39

Legume hay Proposed KF 13 6.84b 0.29 4.17

Oven method 1 9 7.81c 0.20 2.54
Oven method II 9 6.00 d 0.16 2.61
Oven method III 9 6.21e 0.20 3.28
Oven method IV 9 6.69b 0.22 3.32

Means within feeds with the same superscript letter are not statistically different (P = 0.05).

Ill, and IV of 6.17, 6.38, and 6.61% were all lower than
the 2 other methods.
For legume hay, Karl Fischer and oven method IV
gave similar means of 6.84 and 6.69%. All other meth-
ods were different (P = 0.05). The mean for oven
method I of 7.81% was considerably higher than those
for 2 other methods, and the means for oven methods
II and III of 6.00 and 6.21%, respectively, were lower
than those for other methods.
A summary of CVs and variances for the Karl Fis-
cher and various oven methods is provided in Table 5.
CVs averaged for all feeds were 3.81, 3.90, 5.37, 7.83,
and 3.16 for Karl Fischer and oven methods I, II, III,
and IV, respectively. Compared with oven methods, the

Table 5. Comparison of variability of proposed Karl Fischer method with various oven methods

Corn Texturized Dog Total mixed Legume

Method silage feed food ration hay Mean

Proposed Karl CV 3.10 4.38 1.47 5.92 4.17 3.81

Fischer Variance 0.021 0.109 0.010 0.091 0.081

Oven method 1 CV 5.30 4.73 1.71 5.22 2.54 3.90

Variance 0.112 0.190 0.014 0.118 0.040

Oven method II CV 9.83 5.08 5.09 4.25 2.61 5.37

Variance 0.200 0.130 0.099 0.042 0.024

Oven method III CV 14.42 11.34 1.77 8.33 3.28 7.83

Variance 0.321 0.712 0.013 0.171 0.042

Oven method IV CV 5.05 3.16 1.89 2.39 3.32 3.16

Variance 0.079 0.068 0.016 0.020 0.049
 VAN EREM ET AL.: JOURNAL OF AOAC INTERNATIONAL VOL. 81, No. 1,1998 31

proposed Karl Fischer method performed well. Be-
cause sample size (about 0.5 g) for the proposed method
is small relative to sample sizes (about 2 g) for oven
methods, the precision obtained with the proposed
method would be most affected by sample heterogene-
ity. Despite the smaller sample size, CVs for the pro-
posed method were lower than those for all oven
methods for corn silage and dog food. For texturized
feed and TMR, CVs for the proposed method were
comparable with those of oven methods. The highest
CV observed for legume hay was for the proposed Karl
Fischer method. This sample type was probably the
most heterogeneous; thus the smaller sample size would
Oven Method IV overestimated water in corn silage,
texturized feed, and TMR and underestimated water in
dog food. For all feeds, the bias for oven method IV
ranged from -0.37 to 0.87%, which is 95-120% of the
Karl Fischer result.
Conclusions
The proposed Karl Fischer method is acceptable for
determining water in animal feeds and forages on the
basis of repeatability, reproducibility, and comparison
with results of 4 oven methods. Results of the method
are not influenced by the presence of other volatile

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have the greatest effect on precision for this sample.
Average CV for the proposed Karl Fischer method was
3.81%, and the range was from 1.47 to 5.92%. Only
oven method IV had a lower average CV of 3.17%.
The differences between Karl Fischer means and
oven method means are given in Table 6. In this table,
the difference is also expressed as a percentage of the
Karl Fischer mean. For all feeds except dog food, oven
method I overestimated water content. For all feeds,
the bias expressed as % H 2 0 for oven method I ranged
from 0.06 to 1.63, which is 101-135% of the Karl
Fischer result. These results are consistent with those
of Windham et al. (4,12) who reported biases of 151,
121, 121, and 119% for silages, legumes, rations, and all
forages averaged, respectively.
Oven method II underestimated water content in
dog food, and legume hay. For all feeds, the bias for
oven method II ranges from -0.15 to -0.84%, which is
88-97% of the Karl Fischer result.
Oven method III underestimated water content in
corn silage, dog food, and legume hay. For all feeds,
the bias for oven method III ranged from -0.14 to
-0.70%, which is 91-99% of the Karl Fischer result.
components of feed samples, as are results of oven
methods. Throughput for the proposed method is good,
relative to other Karl Fischer methods used in the
author's laboratory. About 25-30 samples can be ex-
tracted easily and titrated in an 8 h day after instru-
ment standardization.
Comparison of the Karl Fischer method to oven
methods illustrates the inconsistency of oven methods.
No one oven method is appropriate for all types of
samples. Further study is needed to evaluate the appro-
priateness of oven methods for specific types of feed.
Acknowledgments
We thank Paul D. Evenson, professor of plant sci-
ence and experiment station statistician, College of
Agriculture and Biological Sciences, SDSU, for assis-
tance with statistical analysis.
References
(1) Mitchell, J., Jr, & Smith, D.M. (1980) Aquametry, Part
III. The Karl Fischer Reagent, John Wiley & Sons, New
York, NY

Table 6. Method bias—difference between oven methods and proposed Karl Fischer (KR) method

Corn Textured Dog Total Mixed Legume

Method Parameter silage feed food ration hay Mean

Proposed KF % H20 4.70 7.52 6.97 5.09 6.84 6.22

Oven method 1 % H20 6.33 9.17 7.03 6.57 7.81 7.38

Bias from KFa 1.63 1.64 0.06 1.48 0.97 1.16
% of KFb 135 122 101 129 114 119

Oven method II % H20 4.55 7.08 6.17 4.85 6.00 5.73

Bias from KFa -0.15 -0.44 -0.81 -0.23 -0.84 -0.49
% of KFb 97 94 89 95 88 92

Oven method III % H20 4.00 7.45 6.38 4.95 6.21 5.80
Bias from KFa -0.70 -0.08 -0.60 -0.14 -0.62 -0.43
% of KFb 85 99 92 97 91 93
Oven method IV % H20 5.57 8.22 6.61 5.86 6.69 6.59
Bias from KFa 0.87 0.70 -0.37 0.77 -0.15 0.36
% of KFb 119 109 95 115 98 106
a
Difference between oven method mean and Karl Fischer mean.
b
Difference between means divided by Karl Fisher mean times 100, or percent H 2 0 recovered based on Karl Fischer.
32 VAN EREM ET AL.: JOURNAL OF AOAC INTERNATIONAL VOL. 81, No. 1, 1998
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