JFS: Sensory and Nutritive Qualities of Food

Optimization of Gluten-Free Bread Prepared

from Cornstarch, Rice Flour, and Cassava Starch
H.D. SANCHEZ, C.A. OSELLA, M.A. DE LA TORRE

ABSTRACT: The proportions of cornstarch, cassava starch, and rice flour were optimized for production of gluten-
free bread (with 0% and 0.5% soy flour) to maximize specific volume (Y1,Y1’), crumb-grain score (Y2,Y2’), and bread
score (Y3,Y3’). A central composite design involving cornstarch/cassava starch ratio (X1) and rice flour/cassava
starch ratio (X2) was used, and 2nd-order models for Yi and Yi’ were employed to generate response surfaces. The
maxima of response surfaces for crumb-grain score and bread score indicate that optimal gluten-free bread can be
prepared from 74.2% cornstarch, 17.2% rice flour, and 8.6% cassava starch. Addition of soy flour at the 0.5% level
also improved bread texture.
Key Words: celiac disease, gluten-free bread, cornstarch, cassava starch, rice flour

Introduction were obtained when these raw materials were used together

A LTHOUGH BREAD PRODUCTS MADE FROM WHEAT FLOUR ARE

consumed worldwide, some individuals are intolerant to
prolamins of wheat and the related cereals oats, rye, and
barley. This intolerance, celiac disease, seriously impairs in-
testinal absorption and can lead to severe malnutrition
(Davison and Bridges 1987; Ciclitira and Ellis 1987). The only
certain remedy to this pathology is total omission of gluten
from the diet. That is very difficult, however, since many
foods contain gluten (Baldo and Wrigley 1984).
Many investigators have attempted to develop gluten-free
yeast breads. Wheat starch has often been used as a wheat
flour replacement (Kulp and others 1974; Ranhotra and oth-
ers 1975). Proteases have been used to hydrolyze the small
percentage of protein left attached to starch (Sánchez and
others 1996). However, many individuals are so sensitive to
gluten’s gliadin fraction that they cannot tolerate even very
small amounts of gliadin (Hamilton and McNeill 1972). The
World Health Organization states that gluten-free foods
should contain less than 1 mg gliadin per 100-g product.
Non-wheat cereal products may also be used in bread-
making. Rice flour is useful since it lacks gluten and contains
low levels of sodium and a high amount of easily digested
carbohydrates, making it desirable in celiac diets (Nishita and
others 1976; Ylimaki and others 1988; Ylimaki and others
1991). Corn and cassava starches may be consumed by those
with celiac disease, but they cause technological difficulties
in breadmaking and impart unusual taste to bread. Gums,
replacing gluten in bread formulations, offer substantial
technological improvements (Sánchez and others 1996;
Christianson and others 1974).
Soy flours and soy protein concentrates were used by Ra-
Sensory and Nutritive Qualities of Food
in the mixture.
The aim of the present study was to use rice flour, corn-
starch, and cassava starch in gluten-free breadmaking and to
statistically establish optimal amounts of each ingredient.
Addition of soy flour was also tested to improve internal
bread characteristics.
Materials and Methods
Materials
PERLA cornstarch [12.4% moisture, 0.16% protein (%
N 3 6.25), 70 8C gelatinization temperature] was from In-
dustrias de Maíz S.A. (Buenos Aires, Argentina). ALDEMAN
cassava starch [14% moisture, 0.06% protein (% N 3 6.25),
63 8C gelatinization temperature] was from Cooperativa
Agrícola Industrial San Alberto Ltda. (Misiones, Argentina).
TRIMACER rice flour [13.7% moisture, 6.0% protein (% N 3
6.25), 70 8C gelatinization temperature] was a product of
Atilio Betella y Cía (Santa Fe, Argentina). TRIMACER soy
flour [9% moisture, 35.1% protein (% N 3 6.25), 18% fat]
was also from Atilio Betella y Cía. The fat used was OPTIMA
oleomargarine (melting point 36 8C) from Molinos Río de la
Plata S.A. (Buenos Aires, Argentina). The premix (contain-
ing gum, sucrose, and salt) was developed by ITA-FIQ-Uni-
versidad Nacional del Litoral in Santa Fe, Argentina, and is
in the patent process.
The following equipment was used: The General Electric
mixer had 2 stainless-steel whisks with speeds of 400 and
600 rpm. The DALVO proofing box with temperature and
humidity control was a product of Ojalvo S.A. (Santa Fe, Ar-
gentina). An apparatus that was used to control proofing

nhotra and others (1975) to prepare soy-fortified, gluten-free
bread. Grain and texture showed a marked improvement
from a rough, crumbly, open-faced interior to a more ten-
der, close-grain, even texture. Soy could not be used at high
levels without severely decreasing bread quality, however.
At previous trials, rice flour, cornstarch, and cassava
starch were used separately in preparing bread because
these raw materials are normally used by celiac people in
food preparation. Nevertheless, better technological results
consisted of a glass cylinder (75 mm in height, 45 mm i.d.)
with a tight-fitting plastic piston that rises during proofing.
The DALVO oven (Ojalvo S.A.) had electrical heating and
temperature control up to 300 ºC. A SETA cone penetrome-
ter (Stanhope-Seta Limited, Surrey, England) was fitted with
a flat-nosed cone (greater dia: 3.2 cm; smaller dia: 1.1 cm;
length: 2.5 cm; weight: 55.0 g). Displacement in the mixture
was measured at 3 s (range 0 to 400 units, equivalent to 0 to
40 mm of penetration).

416 JOURNAL OF FOOD SCIENCE—Vol. 67, Nr. 1, 2002 © 2002 Institute of Food Technologists
Gluten-Free Breadmaking . . .
Breadmaking
Solid ingredients (cornstarch 1 rice flour 1 cassava
Figure 1—Contour plots for X1 against X2 at constant val-
ues of Y1 = specific volume (A), Y2 = crumb-grain score (B),
and Y 3 = bread score (C) for gluten-free breads containing
no soy flour
starch, 300 g; soy flour, 0 and 1.5 g; fat, 30 g; premix, 31 g;
yeast, 30 g) were mixed with water (250 to 300 mL) at 400
rpm for 1 min and 600 rpm for 2 min. Water was adjusted so
mixtures reached 250 units on the cone penetrometer.
Doughs (250 g) were then placed in greased bread pans and
proofed at 27 8C and 80% humidity, controlling raising with
the push-meter (Sánchez and others 1996) until samples
reached 30 mm. Breads were baked at 210 8C for 40 min
without steam.
Volume Measurement and Sensory Evaluation
Specific volume (cc/g of bread) was determined 60 min
after baking by millet-seed displacement, then breads were
evaluated by an expert panel of 3 individuals with more than
20 years of experience in baking. The samples, including the
center points, were served as slices, at the same time. Each
expert scored each sample once. As recommended by Pyler
(1973), the parameters evaluated and the maximum scores
were: volume 15, crust 15, texture 15, crumb color 10, crumb
grain 10, aroma 15, and taste 20.
Experimental Design
Three responses were measured for runs without soy
flour (Yi) and with 0.5% soy flour (Y i’): specific volume
(Y1,Y1’), crumb-grain score (Y2,Y2’), and bread score (Y3,Y3’).
The variables chosen, in order to be independent, were X 1
(%cornstarch/%cassava starch) and X2 (%rice flour/%cassa-
va starch). The sum of these 3 raw materials must always
equal 100. Each variable to be optimized was coded at 5 lev-
els: -1.41421, -1, 0, 1, and 1.41421. Table 1 shows the vari-
ables, their symbols, and levels. The selection of extreme lev-
els was based on previous studies by the authors. A central
composite design, shown in Table 2, was arranged to allow
for fitting of a 2nd-order model. The star points were added
to the factorial design to provide for estimation of curvature
of the model (Montgomery 1991). Six replicates (runs 9, 10,
11, 12, 13, 14) at the center of the design were used to allow
for estimation of the pure error at sum of the square.
Statistical Analysis
A software package (STATGRAPHICS, Manugistics, Inc.,
Rockville, M.D., U.S.A.) was used to fit the 2nd-order models
and generate response surface plots. The model proposed
Sensory and Nutritive Qualities of Food
Figure 2—Gluten-free breads made with 0% and 0.5% soy
flour and optimal proportions of cornstarch (74.2%), rice
flour (17.2%), and cassava starch (8.6%)
Vol. 67, Nr. 1, 2002—JOURNAL OF FOOD SCIENCE 417
 Gluten-Free Breadmaking . . .

Table 1—Variables and their levels for central composite Table 2—Central composite design arrangement and re-
design sponses
Coded Variable Levels Coded Responsesa
Variable Symbol –1.41421 –1 0 1 1.41421 Variable Levels Soy Flour 0 % Soy Flour 0.5 %
%cornstarch/ X1 2 3.4 7 10.6 12 Run X1 X2 Y1 Y2 Y3 Y1’ Y2’ Y3’
%cassava starch 1 1 –1 4.75 7 80.0 4.25 7 79.0
%rice flour/ X2 0.5 0.9 2 3.1 3.5 2 –1 –1 4.80 5 75.0 4.00 6 66.0
%cassava starch 3 1.41421 0 5.00 5 78.5 4.55 8 82.5
4 –1.41421 0 3.80 8 84.0 4.55 5 63.0
5 0 –1.41421 4.30 5 71.0 4.50 8 72.5
6 1 1 4.45 6 75.5 4.50 9 85.0
for each response was: 7 –1 1 3.60 8 83.5 4.35 4 68.0
8 0 1.41421 4.00 8 82.5 4.35 4 68.0
Y = b0 1 b1 X1 1 b2 X2 1 b11 X1 2 1 b22 X2 2 1 b12 X1 X2 9 0 0 4.85 7 73.0 4.30 10 88.5
10 0 0 5.00 6 73.5 4.25 10 91.0
11 0 0 4.85 7 73.0 4.30 9 89.0
where b0 is the value of the fitted response at the center point 12 0 0 4.90 6 73.5 4.25 10 88.5
of the design, that is, point (0,0); b1 and b2 are the linear regres- 13 0 0 4.85 7 73.0 4.65 9 88.0
sion terms; b11 and b22 are the quadratic regression terms; and 14 0 0 4.90 6 73.5 4.60 9 89.0
b12 is the cross-product regression term (Montgomery 1991). a Y and Y ’ = specific volume (cc/g); Y and Y ’ = crumb-grain score (max.
1 1 2 2
10); Y 3 and Y 3’ = bread score (max. 100)
Sensory and Nutritive Qualities of Food

Figure 3—Response surfaces for Y2’ = crumb-grain score Figure 4—Contour plots of X1 against X2 at constant val-
(A) and Y3’ = bread score (B) as a function of X1 and X2 for ues of Y 2’ = crumb-grain score (A) and Y3’ = bread score
0.5% soy flour formulation (B) for gluten-free breads containing 0.5 % soy flour

418 JOURNAL OF FOOD SCIENCE—Vol. 67, Nr. 1, 2002

Gluten-Free Breadmaking . . .

Table 3—Results of analysis of variancea Table 4—Regression coefficients of polynomialsa

F F Soy Flour 0% Soy Flour 0.5%
Degree Soy Flour 0% Soy Flour 0.5% Coefficient Y1 Y2 Y3 Y1’ Y2’ Y3’
Variable of Freedom Y1 Y2 Y3 Y1’ Y2’ Y3’
b0 4.888 6.5 73.3 4.388 9,501 89.0
X1 1 20.33** 4.11 5.22 0.35 17.5** 67.1** b1 0.304 –0.530 –1.360 0.044 1.280 7.204
X2 1 12.69** 8.90* 18.55** 0.38 5.3* 0.04 b2 –0.240 0.780 2.563 0.046 –0.707 0.179
X12 1 9.11* 0.0 38.14** 0.13 20.3** 63.7** b11 –0.212 0.0 3.825 0.028 –1.437 –7.306
X22 1 22.03** 0.0 6.26* 0.26 28.0** 89.0** b22 –0.329 0.0 1.55 –0.040 –1.687 –8.631
X1.X2 1 5.21 7.31* 14.69** 0.01 5.3* 0.6 b12 0.217 –1.00 –3.225 –0.010 1.000 0.950
Total Error 8 0.29 4.38 22.66 0.35 6.01 49.5 a Y and Y ’ = specific volume; Y and Y ’ = crumb-grain score; Y and Y ’ =
(sum of squares) 1 1 2 2 3 3
bread score
Coefficient R2 0.89 0.72 0.91 0.13 0.90 0.96
Statistical significance: *P # 0.05 **P # 0.01
a Y and Y ’ = specific volume; Y and Y ’ = crumb-grain score; Y and Y ’ =
1 1 2 2 3 3
bread score
A verification experiment at the optimum conditions (us-
ing 0.5% soy flour), consisting of 6 runs, was performed. The
result was: Y1’ = 4.32, Y2’ = 9.5, and Y3’ = 89.8. By using the hy-
pothesis-testing technique (Montgomery 1991), differences
Results and Discussion between responses from models and from verification ex-
T ABLE 2 SHOWS THE CENTRAL COMPOSITE DESIGN ARRANGE -
ment and responses for 0% and 0.5% soy flour formula-
tions. Analysis of variance for each response (Table 3) shows
periment were shown to not be significant at the 5% level.

that a significant effect is found for specific volume, in the
absence of soy flour (Y1), with regard to X1, X2, X12, and X22,
whereas no significant effect appears in the presence of 0.5%
soy flour (Y1’). For crumb grain, a highly significant
(P # 0.01) effect is exhibited by all variables when soy flour is
in the formulation (Y2’); without soy flour (Y2), however, only
variables X 2 and interaction X1.X2 are significant and at the
5% level. ANOVA analyses of bread scores (Y 3 and Y 3’) indi-
cate marked effects for almost all variables. These results in-
dicate that, in samples containing 0.5% soy flour, both corn/
cassava (X 1) and rice/cassava (X2) ratios significantly influ-
ence crumb-grain score and bread score but not specific
volume. In contrast, in soy-free formulations, the effect of
variables X 1 and X2 is mainly on specific volume and bread
score, suggesting that high values for specific volume are due
to large holes that impair crumb structure and, consequent-
ly, bread score. This abnormality was corrected by using soy
flour in the recipe, thus avoiding coalescence of bubbles in
the crumb. This effect is noticeable in the presence of soy
proteins with water-binding and stabilizing properties, which
appeared to be resistant to disruption at baking tempera-
tures. Polynomial coefficients for the 2nd-order equation
noted above are given in Table 4, showing the relative impor-
tance of each variable.
For samples containing no soy flour, contour plots of re-
sponse surfaces are shown in Figure 1. Low crumb-grain
score ( Y2 = 6.1) and medium bread score (Y 3 = 74.6) values
are obtained for the maximum specific volume (Y1 = 4.99).
Samples of gluten-free breads with this internal defect and of
bread without holes (containing 0.5% soy flour) are shown in
Figure 2.
Response surfaces of crumb-grain score ( Y2’) and bread
score (Y3’) in terms of X1 and X 2 for samples containing soy
flour are shown in Figure 3. Maxima of both surfaces, indica-
Conclusions
O UR RESULTS SHOW , THEREFORE, THAT HIGH-QUALITY, GLU-
ten-free bread with good taste and appearance, suitable
for celiac patients, can be made from a mixture of 3 raw ma-
terials normally consumed in other foods: cornstarch, rice
flour, and cassava starch. The indicative optimal formulation
for this bread was cornstarch 74.2%, rice flour 17.2%, and
cassava starch 8.6%, and its validation will be made with a
larger panel and ultimately with appropriately qualified con-
sumers. In addition, inclusion of 0.5% soy flour improves
crumb-structure quality.
References
Baldo BA, Wrigley CW. 1984. Allergies to cereals. In: Pomeranz Y. Advances in
cereal science and technology. Vol. VI. St. Paul, Minn.: AACC. p 331-344.
Christianson D, Gardner HW, Warner K, Boundy BK, Inglett GE. 1974. Xanthan
gum in protein-fortified starch bread. Food Tech 28:23-29.
Ciclitira PJ, Ellis HJ. 1987. Investigation of cereal toxicity in coeliac disease.
Postgrad Med J 63:767-775.
Davison AGF, Bridges M. 1987. Coeliac disease. Clin Chim Acta 163:1-40.
Hamilton JR, McNeill LK. 1972. Chilhood coeliac disease: response of treated
patients to a small uniform daily dose of wheat gluten. J Pedia 81,855.
Kulp K, Hepburn FN, Lehmann TA. 1974. Preparation of bread without gluten.
The Baker’s Dig 48(3):34-37.
Montgomery D. 1991. Diseño y Análisis de Experimentos. 3rd Ed. Mexico: Grupo
Editorial Iberoamérica. 589 p.
Nishita KD, Roberts RL, Bean MM. 1976. Development of a yeast leavened rice
bread formula. Cereal Chem 53(5):626-635.
Pyler EJ. 1973. Baking science and technology. Chicago: Siebel Publishing Com-
pany. P 891-895.
Ranhotra GS, Loewe RJ, Puyat LU. 1975. Preparation and evaluation of soy for-
tified gluten free bread. J Food Sci 40(1):62-64.
Sánchez HD, Osella CA, de la Torre MA. 1996. Desarrollo de una fórmula para
Pan sin gluten. Información Tecnológica 7(2):35-42.
Ylimaki G, Hawrysh ZJ, Hardin RT, Thomson BR. 1988. Application of response
surface methodology to the development of rice flour yeast breads: objective
measurements. J Food Sci 53(6):1800-1805.
Ylimaki G, Hawrysh ZJ, Hardin RT, Thomson BR. 1991. Application of response
surface methodology to the development of rice flour yeast breads: sensory
evaluation. J Food Sci 53(3):751-759.
MS 20000851 Submitted 8/29/00, Accepted 10/5/01, Received 10/19/01
We thank Julio C. Mancuello for his collaboration in breadmaking, and we also thank to
Jerold A. Bietz for critically reviewing the manuscript.
Sensory and Nutritive Qualities of Food

tive of optimal gluten-free breads, are near the center point;

the coded values X 1 = 0.48 and X2 = 0 correspond to X1 =
%cornstarch/%cassava starch = 8.7 and X 2 = %rice flour/
%cassava starch = 2.0 (Figure 4); using the models, the re-
sponse values were: Y1’ = 4.4, Y2’ = 9.7, and Y3’ = 90.7.
Authors are with the Instituto De Tecnologia De Alimentos, Facultad de
Ingeniería Química, Universidad Nacional del Litoral, C.C. 266, (3000)
Santa Fe, Argentina. Direct correspondence to Hugo Sanchez (E-mail:
hsanchez@fiqus.unl.edu.ar).

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