VOL. 111, NO.

2, 2005 137
Estimation of Ethanol Yield
in Corn Mash Fermentations
Using Mass of Ash as a Marker
R. Soto
13
, I. Russell
2
, N. Narendranath
1
, R. Power
1
and K. Dawson
1

ABSTRACT
J. Inst. Brew. 111(2), 137143, 2005
An equation to estimate the ethanol yield by mass of ash in fer-
mentation was generated. The equation was based on the as-
sumption that mass of ash does not change during the course of
fermentation. Ethanol yield estimated by the mass of ash equa-
tion was validated by contrasting it to ethanol yield obtained by
the distillation method. Results showed no difference (P = 0.925)
in yields estimated by the aforementioned methods. Further-
more, this innovative method was tested in three fuel ethanol
plants located in the American Midwest. The results from each
of these plants showed that the ethanol yield estimated by mass
of ash is valid, consistent and can be used to compare individual
fermentations and/or treatments.
Key words: Ash, ethanol, fermentation, fermenter, yield.
INTRODUCTION
The fuel ethanol industry lacks an accurate method to
measure the ethanol yield in a particular fermenter. This is
due to the problem of not being able to measure the quan-
tity of grain that goes into each fermenter. Not having
better methodology, most of the industry relies on yield
measurements based on averages of the total amount of
grain received per month, and the total volume of alcohol
pumped from the distillation equipment into the storage
tanks
10
. This information gives an average yield for the
month, but there is no information pertaining to a specific
fermenter, or to a specific period of time i.e. a week.
Therefore, there was a need for a better way to estimate
the ethanol yield per fermenter.
Ash content measurement is a simple method and it is
easy to determine in a fuel ethanol plant laboratory. This
led to the use of the concept of mass of ash to estimate
ethanol yield. The mass of ash in an ethanol fermentation
is given by the ash content of a stream multiplied by the
total mass of the stream. Therefore, the initial mass of
grain in a fermenter can be estimated by the use of a ma-
terials balance for the mass of ash (Fig. 1). If the amount
of grain in the fermenter can be estimated, then the etha-
nol yield for that particular fermenter can be estimated.
Yield is an important measurement for determining the
efficiency of a plant to convert grain, such as corn, into
ethanol. Yield is defined in the fuel ethanol industry as the
volume units of ethanol obtained from a mass unit of grain
via fermentation, usually expressed as gallons per bushel
in the U.S.A., however for research purposes usually ex-
pressed as litres per tonne (LPT).
Although there are other important measurements to
evaluate ethanol yield, such as theoretical yield and starch
utilization, to address these was not the objective. The aim
of the present work was to estimate ethanol yield using
the mass of ash in a fermenter as a marker in a materials
balance. This estimation needed to be validated against a
well trusted method, it needed to be consistent, and an
evaluation needed to be made to see if it was useful for
monitoring and evaluating the success of process changes.
MATERIALS AND METHODS
Corn (Purina brand, U.S.A.) used for the experiments
was ground using a hammer mill (U.S. #4 screen), to pro-
duce a particle size distribution typical of the fuel ethanol
industry
10
. The ground corn was sampled accordingly and
analyzed
17
to determine homogeneity in its composition,
and vacuum packed for later use. This was carried out to
ensure a standardized corn composition, to reduce corn
effects on ethanol yield across fermentations.
Mash preparation
Five-hundred grams of mash per flask was prepared at
30% dry solids
10
(30 g dry corn/100 g mash). Ground corn
(175 g) was added to a 1000 mL round bottom flask, fol-
lowed by 325 g hot water (90C). The flask was heated
using a heating mantle (Glas-Col, Terre Haute, IN) coupled
to a rheostat (Staco Energy Products Co., Dayton, OH)
and maintained at 85C. Alpha-amylase (High T DS, All-
tech Inc., Nicholasville, U.S.A.), a 0.072 g aliquot, was
added to the flask. The mash was continuously mixed at
400 rpm for 20 min using a mixer (Cole-Palmer Instru-
ment Co., Barrington, IL) attached to a controller (Cole-
Palmer Instrument Co., Barrington, IL). The contents of
the flask were autoclaved for 15 min at 121C. Once auto-
claved, the mash was immediately cooled to 85C in cold
water. A second aliquot of -amylase (0.048 g) was added,
1
Alltech Bio-science Centre, Alltech Inc., Nicholasville, Kentucky
40356, USA.
2
International Centre for Brewing and Distilling, School of Life Sci-
ences, Heriot Watt University, Riccarton, Edinburgh EH14 4AS, UK.
3
Corresponding author. E-mail: rsoto@alltech.com
Publication no. G-2005-0718-268
2005 The Institute of Brewing & Distilling
138 JOURNAL OF THE INSTITUTE OF BREWING
and the mash was maintained at 85C for 60 min with con-
tinuous mixing.
Fermentation
A simultaneous saccharification and fermentation was
conducted once the mash had cooled to 30C. Glucoamyl-
ase (Allcoholase II-400, Alltech Inc., Nicholasville, U.S.A.)
was added (0.3 g). Each flask was inoculated to have an
initial viable yeast count of 3 10
7
cells/g mash using
active dry yeast (Superstart

, Alltech Inc. Nicholasville,

KY). Active dry yeast had a viable count of 2.7 10
10

cells/g determined by the methylene blue method
6
. The
flasks were placed in a circulating water bath maintained
at 30C. The mash was fermented over a period of 48 h.
Validation
A series of laboratory fermentations were designed to
evaluate the use of mass of ash in the estimation of etha-
nol yield. These were compared to the ethanol yield deter-
mined using the distillation method, to validate the mass
of ash method. Ethanol yields were estimated from nine
replications for the mass of ash method, and seven repli-
cations for the distillation method. All the fermentations
were conducted under equal controlled conditions. From
the first set of flasks (mass of ash method) samples were
withdrawn, and analysed for ash content in the mash be-
fore and after fermentation, as well as ethanol in the fer-
mented mash. The information obtained was then used to
estimate ethanol yield. The second set of seven flasks was
used for ethanol determination by the distillation method
(no sampling).
Samples
Procedures. It was found that sampling was of para-
mount importance in describing the fermentation system
in an industrial environment. The front end is a continu-
ous process subject to changes in the amount of corn (sol-
ids) supplied to the mash tank. These changes occur as the
fermenter is being filled. The filling process takes be-
tween 6 and 12 hours depending on the fermenter work-
ing volume. Therefore collecting several samples at equal
time intervals depicts the mash in a more realistic way. At
predetermined periods, samples of 250 mL were collected,
and mixed to prepare the composite sample from each
sampling point.
Sample pre-treatment for ash content determina-
tion. When carrying out the ash content determination in
the various samples, it was clear that the mash solids had
a tendency to sediment quickly. This was observed to have
an effect on the ash determination, yielding high variabil-
ity among the replicates. Several different approaches
were taken to address this problem. Freeze drying a 200 g
portion of the sample resulted in a very low coefficient of
variability amongst the replicates and helped produce esti-
mated yields very close to the distillation counterparts.
Industrial sampling and sample handling. Samples
were collected from the plant, preserved in dry ice, and air
freighted next day to the Alltech Bio-science Centre
(Nicholasville, KY) facilities to be analysed.
Development of the equation to estimate
ethanol yield
In estimating the yield, it is important to take into ac-
count the peculiarities of the process (Fig. 1), such as the
use of backset (recycled stillage) and changes in the mash
density
10
. In developing the corresponding equation, the
most important assumption was that the mass of ash was
not increased or reduced during the fermentation process.
The main change during the process was that carbon di-
oxide was lost due to its nature and this modified the mash
volume in the fermenter, and additional losses included
volatile compounds and water vapour loss. Due to these
Fig. 1. Simplified diagram of the sampling points and parameters used in the mass of ash estimation of ethanol yield.
VOL. 111, NO. 2, 2005 139
losses in volume, the ash content was modified i.e. it was
the same mass of ash in less volume. The mash prepared
for fuel ethanol production, due to the variations in the
process, was ~2530% solids prior to fermentation and
~1015% solids towards the end of fermentation
7
. The
change in density was considerable, usually progressing
from ~1.28 g/mL to a value closer to 1.00 g/mL. This
change was not only the result of a reduction in solids, but
was also due to the presence of ethanol The ethanol con-
tent of the fermented mash in the fuel ethanol production
process was typically ~15% v/v. Consequently, the etha-
nol content could be as much as one-sixth of the sample.
Ethanol density
14
is 0.78924 g/mL, thus the effect could
easily be assessed.
The mass of ash that goes into a fermenter in an indus-
trial operation has two main sources: the corn and the
backset (Fig. 1). The former typically is added to make a
30% solids mash. The latter can make up to 50% of the
total volume of mash. The test plants all had methods to
calculate the volume of the backset that went into the
preparation of the mash; therefore, this was a known
value. Starting from this value (B), and knowing the total
volume of the fermenter (V
i
):
V
b
= V
i
* B (1)
Where:
V
b
= Backset volume in mash [=] litre of backset
V
i
= Fermenter volume at fill [=] litre of mash
B = Percentage of backset used in mash preparation
From equation (1) the mass of ash in backset can be
calculated using the following equation:
M
b
(Kg of Ash in backset) = V
b
* A
b
*
b
(2)
Where:
A
b
= Ash content of the backset [=] kg of ash per 100
kg of backset

b
= Backset density [=] kg of backset per litre of
backset
Knowing the mass of ash coming from the backset,
then the mass of ash corresponding to corn can be calcu-
lated from the total mass of ash in the fermenter:
M
i
(kg of Ash in fermenter) = V
i
* A
i
*
i
(3)
Where:
A
i
= Ash content of the fermenter at fill [=] kg of ash
per 100 kg of mash

i
= Mash density of fermenter at fill [=] kg of mash
per litre of mash
Therefore, the mass of ash coming from corn (M
a
) can
be calculated:
M
a
= M
i
M
b
(4)
Substituting the initial and backset mass of ash, we ob-
tain
M
a
= (V
i
* A
i
*
i
) (V
b
* A
b
*
b
) (5)
Since backset volume can be calculated from equation
(1), we substitute this in equation (5) to obtain:
M
a
= (V
i
* A
i
*
i
) (V
i
* B * A
b
*
b
) (6)
Equation (6) can be also expressed as:
M
a
= V
i
[(A
i
*
i
) (B * A
b
*
b
)] (7)
In order to calculate the mass of corn in the fermenter
(M
m
) we need to know the ash content of corn (A
m
) and
use it in the equation:
M
m
= M
a
A
m
(8)
Substituting equation (7) in (8):
M
m
= V
i
[(A
i
*
i
) (B * A
b
*
b
)] /A
c
(9)
As for the volume of ethanol (V
E
) produced during fer-
mentation, it can be calculated from the following:
V
E
= V
f
* E/
E
(10)
Where:
V
f
= Final fermenter volume [=] litres of mash
E = Ethanol content in mash [=] kg of ethanol per 100
litre of mash

E
= Ethanol density [=] 0.78924 kg of ethanol per litre
of ethanol
10

Due to the losses during fermentation i.e. carbon di-
oxide, the final volume is difficult to determine, however,
the initial volume is known. Using the principle for mass
of ash conservation through fermentation, the final vol-
ume can be easily obtained from:
V
i
* A
i
*
i
= V
f
* A
f
*
f
(11)
Solving for final volume:
V
f
= V
i
* A
i
*
i
/A
f
*
f
(12)
Combining equations (10) and (12) to obtain volume
of ethanol produced:
V
E
= V
i
* A
i
*
i
* E/A
f
*
f
*
E
(13)
By definition ethanol yield () is litres of ethanol pro-
duced per tonne of corn, therefore, if equation (13) is di-
vided by equation (9) the resulting value will be ethanol
yield:
= V
E
/M
m
(14)
Substituting equations (13) and (9) in (14):
= [V
i
* A
i
*
i
* E/A
f
*
f
*
E
]
/{V
i
[(A
i
*
i
) (B * A
b
*
b
)] /A
c
} (15)
Since V
i
is in the numerator as well as in the denomi-
nator, it can be cancelled to simplify equation (15) to its
final form:
= {[(
i
A
i
E)/(0.78924 A
f

f
)] /[((
i
A
i
)(
b
A
b
B)) /A
c
]}
* (16)
Where:
= Ethanol yield [=] litres of ethanol per tonne of corn

i
= Density of mash at set point [=] kg per litre
140 JOURNAL OF THE INSTITUTE OF BREWING

f
= Density of fermented mash [=] kg per litre

b
= Density of backset [=] kg per litre
A
i
= Ash content of mash at set point [=] kg ash per
100 kg mash
A
f
= Ash content of fermented mash [=] kg ash per 100
kg mash
A
b
= Ash content of backset [=] kg ash per 100 kg
backset
A
c
= Ash content of corn in dry basis [=] kg ash per
100 kg corn
E = Ethanol content in the fermented mash
[=] kg ethanol per 100 litre of mash
B = Percentage of backset in the fermenter
[=] litres of backset per 100 litre of mash
0.78924 = Ethanol density in kg per litre at 20C
10

= Conversion factor to litres per tonne = 10
This equation is also useful in the laboratory when corn
only fermentations are carried out i.e. no backset added.
Then it takes the simpler form:
= {[(
i
A
i
E)/(0.78924 A
f

f
)] /[(
i
A
i
) /A
c
]}*
As can be observed, the resultant equations do not in-
volve the volume of mash in the calculation of yield. All
the other components of the equation are easily obtained
by sampling the fermenter. Correct sampling technique,
especially during fermenter fill, is of paramount impor-
tance.
Testing of the equation in the fuel ethanol
industry
Industrial fermentations, from three different fuel etha-
nol plants located in the American Midwest, were sampled
to test the equation. The first two plants were used to as-
sess the validity and consistency of the yield estimation.
The third plant was used to evaluate a change in the pro-
cess.
First and second industrial trials. Plants A and B
agreed to allow sampling of the required streams in their
fermentations (Fig. 1). From these the following parame-
ters were determined: percent of backset added to mash,
grain moisture content, mash density (initial and final),
backset density, mash ash content (initial and final),
ground corn ash content, and backset ash content.
Third industrial trial. Plant C trial evaluated the use
of the equation to estimate ethanol yield by mass of ash
when changes were made in the process. A solid state fer-
mentation enzyme complex, Rhizozyme

(Alltech, KY),
was used, at two levels of addition, to examine the effect
on fermentation yield. All other parameters were as in
Plant Trial A and B.
Ethanol determination
Distillation method. The fermented mash was distilled
in a glass distillation apparatus
5
. (Pyrex, 24/40 No. 2540,
UK). The collected distillate was weighed and analysed us-
ing an Anton Paar Alcolyzer (Anton Paar, Ashland, VA).
HPLC. High Pressure Liquid Chromatography (HPLC)
was used to determine alcohol content in the fermented
mash. Samples were centrifuged at 4000 g for 15 min.
Supernatant was subsequently filtered through a 0.2 m
nylon filter into a glass vial. Vials were placed in a 717
plus Autosampler (Waters, Milford, MA,). Injection sam-
ples were passed through an Aminex HPX-87H column at
65C (BIO-RAD, CA) for 25 min, with a Refractive Index
Detector (Waters 2410, Milford, MA) using sulphuric acid
0.0025 M as a mobile phase and a sample injection vol-
ume of 20 L. Eluent flow was set to 0.6 mL/min (515
HPLC pump, Waters, Milford, MA). The results were col-
lected and analysed using the Millenium software (Waters,
Milford, MA), and expressed as ethanol content (weight
basis).
Ash determination
The ash content of the corn, initial and fermented mash
samples was determined using the official British Stan-
dard BS 4603:1970 as described by James
8
.
Yield calculation for distillation method
The yield was calculated in litres of ethanol per tonne
of corn. For the calculation, the grams of ethanol recov-
ered in the distillation were obtained from the weight of
the distillate and multiplied by the percentage of ethanol
(weight basis) reported by the Alcolyzer according to the
standards of the O.I.M.L. alcohol tables
14
.
Corn Tonne
g 10 1
mL 1000
Ethanol L
Ethanol g 0.78924
Ethanol mL
Corn g
Ethanol g
(LPT) Yield
6

=

Statistical analysis of data
Analysis of variance (ANOVA) and t-test assuming un-
equal variances were carried out for laboratory results
2
.
For plant C, a complete randomised experiment ANOVA
was used in statistics software
15
to analyse ethanol yield
using main and linear effect of enzyme addition as con-
trasts.
RESULTS AND DISCUSSION
Laboratory fermentations
In the laboratory trials, the variance of the mass of ash
method was higher than when the distillation method was
used, however, there was no statistical difference (P =
0.92) in yield of ethanol (LPT) calculated using either
method (Table I). This result suggested that the ash con-
tained in the grain, in terms of the total mass of ash being
added into the flasks, had no changes throughout the fer-
mentation. This confirmation indicated that the generated
equation using ash content is a sound way to estimate the
amount of grain added to a fermenter and therefore for
estimation of ethanol yield.
Mash preparation for the fermentation process has an
effect primarily on starch and protein. These compounds
are hydrolyzed, the former into primarily glucose, and the
latter into peptides and amino acids. Yeast can utilize
VOL. 111, NO. 2, 2005 141
these compounds for growth and reproductive functions.
Micronutrients such as calcium, potassium and phospho-
rus are essential to yeast metabolism
16,18
. The main source
for these micronutrients in these ethanol fermentations
was corn. The other source for these elements was the
backset which originated from the corn mash fermenta-
tion (see Fig. 1).
Water and other compounds (i.e. aqueous ammonia,
enzymes, etc) required during the preparation of the mash
do not contain or contribute significant amounts of these
micronutrients. Based on the amount and the ash content
of each of these compounds, their contribution to the mass
of ash was calculated as negligible. As the fermentation
proceeded and the yeast utilized the micronutrients, these
micronutrients passed from the mash to the yeast. Since
ash determination was conducted by incinerating the or-
ganic matter, the elements contained in the yeast cells and
corn were liberated. No loss of micronutrients occurred.
The carbon dioxide and water vapor that escaped from the
fermenter were essentially free of these elements. To cal-
culate the mass of ash it was important to know the ash
content of a representative sample and the total mass of
mash at the time of the sampling. The mass of ash was
therefore the total mass of the mash times the ash content
of the mash. Therefore, at any point or at the end of the
fermentation, the mass of ash calculated remained un-
changed.
Ash is one of the most widely used markers in animal
science with numerous applications in the field of nutri-
tion
1,3,4,1113
. The ash content of a feedstock is the residue
remaining after all the water has been removed and all the
proteins, fats, carbohydrates, vitamins, organic acids, etc.
have been burnt away. The resulting residue is considered
to be a measure of the mineral content of the feedstock
8
.
The ash content determined by the igniting procedure is
consistent in value and is characteristic for each feed-
stock. In animal feed, the inorganic elements (i.e. calcium
and magnesium) in the grain suffer no alteration during
digestion. Similarly, during fermentation the total mass of
ash coming from corn and later contained in the mash,
remains constant throughout the process. Using this unique
situation, it is possible to estimate the original amount of
grain used in any given fermentation.
The use of the mash of ash estimation showed a good
correlation to the distillation method. Due to the success-
ful laboratory trials, the method was next tested on large
scale plant industrial fermentations.
Plant trials
Plant A. The results of the determinations are shown in
Table II. The value obtained by the ash method was
361.56 LPT. The plant estimation was 394.50 LPT and
this yield value provided by the plant was based on the
average yield for the month.
Table I. Yields (LPT) obtained using the equation to estimate yield by
mass of ash and the distillation method in laboratory fermentations.
Replicate
Ethanol yield (LPT)
sets Mass of ash Distillation
1 332.93 336.73
2 386.45 327.71
3 330.57 323.35
4 324.03 326.65
5 327.79 331.58
6 268.03 291.25
7 293.28 303.85
8 297.85 n/a
9 332.30 n/a
Average = 321.47 320.16
n/a = not available
Table II. Use of the equation to estimate yield by mass of ash in Plant A.

Backset %EtOH
Ash content
Fermentor
Ferm
ID
to mash
(%of Total)
final
(%w/v)
A
i

(%w/w)
A
f

(%w/w)
A
b

(%w/w)
A
c

(%w/w)
yield (LPT)
as is basis
246 34.4 11.99 0.7198 0.8017 0.9211 1.2773 361.56
Table III. Estimation of ethanol yield by mass of ash method in plant B.
Fermenter ID
174 177 187 190 Average SEM CV
Backset to mash (B) (% of total) 28.16 28.43 23.53 25.20 26.3 1.19 4.5%
Final ethanol content (E) (% w/v) 11.72 11.81 11.94 11.58 11.8 0.08 0.6%
Grain moisture (%) 13.40 13.70 13.60 14.70 13.9 0.29 2.1%
Density

i
(kg/L) 1.015 1.063 1.074 1.124 1.069 0.022 2.1%

f
(kg/L) 1.011 0.988 1.021 1.001 1.005 0.007 0.7%

b
(kg/L) 1.008 0.993 0.995 0.998 0.998 0.003 0.3%
Ash content
A
i
(%) 0.4235 0.6240 0.6365 0.6129 0.5742 0.0505 8.8%
A
f
(%) 0.5648 0.5583 0.5837 0.5379 0.5611 0.0094 1.7%
A
b
(%) 0.5465 0.6594 0.6869 0.5466 0.6098 0.0370 6.1%
A
m
(%) 1.1450 1.1797 1.3017 1.3232 1.2374 0.0441 3.6%
Yield
(LPT) 403.42 383.84 373.28 384.15 386.17 6.28 1.6%
SEM = Standard Error of the Mean, CV = Coefficient of Variation
142 JOURNAL OF THE INSTITUTE OF BREWING
The main issue in terms of accuracy of the measure-
ment is related to the size of the plant and the amount
of grain being processed. A typical working volume of a
fermenter is 1,700 m
3
and for a 30% dry solids mash,
700,000 kg of corn are required per fermenter
9
. The in-
accuracies in determining the amount of grain ground per
day due to the limitations of the weighing belts are high
9
.
Plant B. Four fermenters were monitored and sampled
according to the previous methodology. The results of
plant B fermentations are shown in Table III. The average
value estimated for the yield by the ash method was
386.17 LPT. The plant estimated average yield for the
month was 394.52 LPT. As in plant A, the yield estimated
by the plant was based on inventory practices, not on the
direct measurement of corn going to the fermenters. These
results validated the use of the ash method in an industrial
environment, and reinforced that sampling and the pre-
treatment of the sample are of paramount importance to
obtain reliable information.
Plant C. The results are shown in Fig. 2. Although 5
fermenters of each treatment were sampled, some samples
were damaged during transportation, and thus no tests
were conducted on those samples. The data generated was
analyzed using ANOVA with SAS
15
. The analysis showed
that the use of the Rhizozyme

enzyme complex im-

proved (P = 0.02) ethanol yield. The effect showed linear-
ity (P = 0.03) as the Rhizozyme

dosage increased. The

plant was able to observe a difference in total ethanol vol-
ume from the distillation column to the storage tank i.e.
the ash method was able to show the plant manager differ-
ences amongst process treatments. This information is
what a plant manager requires to make decisions in terms
of improvement in the economics of the process.
The use of a method such as the ash estimation of etha-
nol yield can help the fuel ethanol industry improve their
understanding of how different factors affect fermenta-
tion. For this purpose, individual fermenters can be used
to test the variables, instead of long trials that require use
of all the fermenters. The latter is an expensive option and
due to the problems discussed previously is not as effec-
tive a way to determine differences in treatments to justify
process changes. However, to determine if the ash estima-
tion was more or less accurate than the current estimation
method used in the industry was not the purpose of this
study, but rather it was to provide a valuable tool for the
ethanol industry to evaluate changes in the process in or-
der to optimise ethanol production. Other ethanol indus-
tries i.e. whisky and spirits could benefit from this method,
especially when individual fermenters require monitoring
for treatment or change of feedstock effect. The informa-
tion generated will help to identify the parameters that
need to be modified to improve ethanol yield. The latter
combined with organoleptic testing of batches could pro-
vide answers to faster fermentations without risking prod-
uct quality.
Method alternatives
The content of selected minerals was determined for
the composite samples from Plant A (data not shown). The
possibility of using some of these micronutrients as a
marker was explored. Mineral analysis was carried out by
inductively coupled plasma source (ICP) mass spectrome-
try. The use of ICP mass spectrometry was rejected as a
feasible tool currently due to the cost of the equipment
and/or the need to depend on an external laboratory, as
this would make the estimation of yield a cumbersome,
lengthy and expensive process. Other alternatives to deter-
mine minerals are available, but due to the simplicity and
relative inexpensiveness of the ash determination, the
other methods were not explored further.
CONCLUSIONS
The results obtained validated the use of the estimation
of ethanol yield by the mass of ash method. This method
offers a simple solution for the fuel ethanol industry that
delivers consistent results. It also demonstrated that such a
method can be used to compare fermenter performance or
to monitor improvements in yield due to modification of
the current practices or use of new products, such as in the
example presented here. The current cost structure of fuel
ethanol plants in America leaves minimum operational
profit. Therefore, these companies are in constant search
of new ways to reduce ethanol production costs. At the
same time, suppliers are constantly offering new and in-
novative products to this industry. The historical plant
method of estimating ethanol yield is not capable of pro-
viding sound information on the performance of these
new products. The method described in this paper can
help the fuel ethanol industry to optimise fermentation
procedures and aid in the evaluation of materials such as
improved enzymes and novel feedstocks.
ACKNOWLEDGEMENTS
The authors wish to thank Alltech Inc. for funding the pres-
ent work. Special acknowledgement to Dave Kelsall for organiz-
ing the trials with the plants and coordinating the sampling and
shipping and for sharing his valuable knowledge of the operation
of fuel ethanol plants. Thanks are due to Dr. Juan Tricarico for
his help with statistical analyses and to Janna Norton for provid-
ing the requested papers, to Dr. Joe Power, and Tara Graves for
the valuable discussions and help, and to Dr. Colm Moran for
his suggestions and corrections in the writing of this paper. Spe-
cial thanks to the managers and staff of ethanol plants A, B, and
C for their support and encouragement while carrying out these
trials.
Fig. 2. Ethanol yield by mass of ash estimated for the fermenta-
tions using Rhizozyme

against control in Plant C.

VOL. 111, NO. 2, 2005 143
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(Manuscript accepted for publication June 2005)