EXTRUSION OF WHEAT WASHED BRAN: PHYSICOCHEMICAL AND
FUNCTIONAL PROPERTIES
By
George Nyombaire

A DISSERTATION
Submitted to
Michigan State University
in partial fulfillment of the requirements
for the degree of
DOCTOR OF PHILOSOPHY
Food Science
2012

ABSTRACT
EXTRUSION OF WASHED WHEAT BRAN: PHYSICOCHEMICAL AND
FUNCTIONAL PROPERTIES
By
George Nyombaire

Milled wheat bran contains substantial amounts of residual starchy endosperm that may
interfere with the analyses that are used to determine a bran’s composition and physicochemical
properties.

In addition, it is possible that during extrusion cooking, this residual starchy

endosperm may undergo certain chemical modifications that may affect composition,
physicochemical properties, and functionality of wheat bran. Therefore, objectives of the present
study were: (1) to develop a method that removes most of the residual starchy endosperm still
adherent to milled wheat bran, (2) to investigate the effects of particle size of non-washed and
washed wheat bran coupled with extrusion processing conditions on total dietary fiber, insoluble
dietary fiber, and soluble dietary fiber contents of non-washed and washed bran, (3) to
investigate the effects of extrusion cooking variables on the physicochemical properties of
extruded wheat bran, and (4) to evaluate the effects of treated wheat bran on the baking
properties of bread and cookies.
The washing method developed in the present study reduced starch adherent to milled
wheat bran by 76% (w/w), changed insoluble dietary fiber content in the bran from 39% to 69%
(w/w), and decreased soluble fiber content from 4.93% in NWB to 1.68% (w/w). The water
binding capacity was higher for washed bran and was not affected by bran particle size.
To investigate the effects of particle size coupled with extrusion processing conditions,
non-washed bran (NWB), ground to pass through 1000 µm (NWB1000) and 425 µm (NWB425)

screens, and washed bran (WB), ground to pass through 1000 µm (WB1000) and 425 µm
(WB425) screens, were extruded through a co-rotating and inter-meshing twin-screw extruder
under conditions of varying screw configurations (low and high shear), feed moisture (25 and
o

35%), screw speed (100 and 400 rpm), and die temperature (100 and 150 C). Extrusion
increased soluble dietary fiber in both NWB1000 and NWB425 but decreased insoluble dietary
fiber. Certain extrusion conditions increased or decreased the contents of insoluble and soluble
fiber in WB425. The insoluble dietary fiber contents were lower in extrudates made from
WB1000 than in non-extruded WB1000. Extrusion conditions increased or decreased soluble
dietary fiber in WB1000.
The effects of extrusion processing on the physicochemical properties of wheat bran
samples were studied. Extrusion increased the water binding capacity (WBC) of NWB samples.
Washing and extrusion decreased weight average molecular weight of soluble dietary fibers
extracted from NWB and WB samples. Differential Scanning Calorimetry results indicated that
thermal decomposition properties of extrudates made from NWB and WB samples were not
affected by extrusion. In vitro binding of bile acids was highest in extrudates made from
WB1000, whereas the lowest binding of bile acids was in extrudates made from NWB425.
Baking studies indicated that bread formulations containing wheat bran resulted in breads
with decreased loaf volumes. Substituting soft wheat flour with 5% or 15% of the prepared bran
samples significantly decreased cookie spread. The contents of insoluble dietary fiber in the
crumb and crusts of breads baked from hard wheat flour and hard wheat flour substituted with
bran were higher than those in the unbaked flours. The contents of soluble dietary fiber were not
significantly different from each other in the crumb and crusts of breads baked from flour
samples substituted with bran.

ACKNOWLEDGEMENTS

First and foremost, I am heartily thankful to my major advisor, Dr. Perry K.W. Ng, for
his overwhelming support throughout my Ph.D. program. This dissertation would not have been
possible without his generosity and superb guidance. His ideas have made me a better scientist. I
consider it an exceptional honor to have worked with Dr. Ng. I am truly indebted to my
committee members, Dr. Maurice R. Bennink, Dr. Gale M. Strasburg, and Dr. Irvin E. Widders,
for their wonderful contributions, encouragement, and support, all of which made this
dissertation possible.
I wish to express my thanks to the Department of Food Science and Human Nutrition for
offering me several fellowships that relieved my financial burden so I could spend more time on
my Ph.D. research. Furthermore, I would like to thank the College of Agriculture and Natural
Resources for their financial support toward the end of my Ph.D. program.
I would like to express my gratitude to Dr. Grace Lai and Dr. YuLai Jin of Kellogg
Company who allowed me to use their specialized equipment to analyze my samples. Also, I
would like to acknowledge Philip E. Hill and Richard Wolthius for their technical expertise in
fixing processing equipment in the Cereal Science laboratories.
Finally, I am grateful to my labmates, Derrick, Ken, Sarah, Ya, and Zeynep. Furthermore,
I would like to extend my thanks to my previous labmate, Rico, and former visiting scholar, Dr.
Zhong, for their friendship and support.
Finally, I am extremely grateful to my wife and children for their sacrifice and love
during the course of my Ph.D. studies.

iv

TABLE OF CONTENTS

LIST OF TABLES ......................................................................................................................... ix
LIST OF FIGURES ....................................................................................................................... xi
CHAPTER 1
INTRODUCTION .......................................................................................................................... 1
LITERATURE CITED ................................................................................................................... 6
CHAPTER 2
LITERATURE REVIEW ............................................................................................................... 9
2.1. WHEAT BRAN ................................................................................................................. 10
2.2. DIETARY FIBER .............................................................................................................. 14
2.2.1. Insoluble Dietary Fiber ................................................................................................ 16
2.2.1.1. Cellulose ............................................................................................................... 16
2.2.1.2. Hemicelluloses ...................................................................................................... 17
2.2.1.3. Lignin .................................................................................................................... 18
2.2.2. Soluble Dietary Fiber................................................................................................... 18
2.2.2.1. Arabinoxylans ....................................................................................................... 19
2.2.2.2. β-glucans ............................................................................................................... 20
2.2.2.3. Pectin..................................................................................................................... 22
2.2.3. Non-Digestible Oligosaccharides ................................................................................ 22
2.2.4. Resistant Starch ........................................................................................................... 23
2.2.5. History of Dietary Fiber .............................................................................................. 24
2.2.5. Health Benefits of Dietary Fiber ................................................................................. 25
2.3. PHYSICOCHEMICAL PROPERTIES OF HIGH FIBER INGREDIENTS .................... 27
2.3.1. Solubility ..................................................................................................................... 27
2.3.2. Water Binding Capacity .............................................................................................. 28
2.3.3. Bile Acid Binding Capacity ......................................................................................... 28
2.3.4. Molecular Weight Distribution .................................................................................... 29
2.3.5. Thermal Analysis ......................................................................................................... 30
2.4. EFFECT OF PROCESSING ON DIETARY FIBER ........................................................ 32
2.4.1. Particle Size Distribution ............................................................................................. 32
2.4.2. Extrusion Cooking ....................................................................................................... 33
2.5. WHEAT FLOUR ............................................................................................................... 36
2.6. BAKED PRODUCTS ........................................................................................................ 38
LITERATURE CITED ................................................................................................................. 40
CHAPTER 3
PHYSICOCHEMICAL PROPERTIES OF WASHED WHEAT BRAN .................................... 49
3.1. ABSTRACT ....................................................................................................................... 50
3.2. INTRODUCTION .............................................................................................................. 51
3.3. MATERIALS AND METHODS ....................................................................................... 53
3.3.1. Wheat Bran Sample ..................................................................................................... 53

v

3.3.2. Washing of wheat bran ................................................................................................ 53
3.3.3. Determination of Total Starch ..................................................................................... 56
3.3.4. Thermal Properties of Non-Washed and Washed Wheat Bran Samples..................... 57
3.3.5. Water Binding Capacity of Non-Washed and Washed Bran ...................................... 57
3.3.6. Determination of Insoluble, Soluble, and Total Dietary Fiber Content ...................... 58
3.3.7. Proximate Compositions of Non-Washed and Washed Bran ...................................... 60
3.3.8. Statistical Analysis ...................................................................................................... 60
3.4. RESULTS AND DISCUSSION ........................................................................................ 61
3.4.1. Total Starch.................................................................................................................. 61
3.4.2. Effect of Washing on the Thermal Properties of Non-Washed and Washed Wheat
Bran Starch Samples .............................................................................................................. 63
3.4.3. Water Binding Capacity of Non-Washed and Washed Bran Samples ........................ 66
3.4.4. Insoluble Dietary Fiber, Soluble Dietary Fiber and Total Dietary Fiber ..................... 68
3.4.5. Proximate Compositions (dry weight basis) of Non-Washed and Washed Wheat Bran
............................................................................................................................................... 70
3.5. CONCLUSIONS ................................................................................................................ 72
LITERATURE CITED ................................................................................................................. 73
CHAPTER 4
EFFECTS OF PARTICLE SIZE COUPLED WITH EXTRUSION PROCESSING
CONDITIONS ON DIETARY FIBER PROFILE OF NON-WASHED AND WASHED
WHEAT BRAN ............................................................................................................................ 77
4.1. ABSTRACT ....................................................................................................................... 78
4.2. INTRODUCTION .............................................................................................................. 79
4.3. MATERIALS AND METHODS ....................................................................................... 81
4.3.1. Wheat Bran Sample ..................................................................................................... 81
4.3.2. Preparation of Washed Wheat Bran ............................................................................ 81
4.3.3. Grinding and Sifting .................................................................................................... 81
4.3.4. Extrusion of Bran Samples .......................................................................................... 81
4.3.5. Determination of Soluble Dietary Fiber and Insoluble Dietary Fiber in Non-Washed
and Washed Wheat Bran ....................................................................................................... 86
4.3.7. Statistical Analysis ...................................................................................................... 86
4.4. RESULTS AND DISCUSSION ........................................................................................ 87
4.4.1. Effect of Bran Particle Size Coupled with Extrusion Processing Conditions Under .. 87
Low Shear Screw Configuration on the Contents of Insoluble Dietary Fiber (dry weight
basis) in Extrudates Made from Non-Washed and Washed Wheat Bran .............................. 87
4.4.2. Effect of Bran Particle Size and Extrusion Processing Conditions Under Low Shear
Screw Configuration on the Contents of Soluble Dietary Fiber (dry weight basis) in
Extrudates Made from Non-Washed and Washed Wheat Bran ............................................ 91
4.4.3. Effect of Bran Particle Size and Extrusion Processing Conditions Under Low and
High Shear Screw Configurations on the Contents of Insoluble and Soluble Dietary Fibers in
Extrudates Made from WB425 and WB1000........................................................................ 95
4.5. CONCLUSIONS ................................................................................................................ 98
LITERATURE CITED ................................................................................................................. 99
CHAPTER 5

vi

EFFECTS OF EXTRUSION PROCESSING ON THE PHYSICOCHEMICAL PROPERTIES
OF NON-WASHED AND WASHED WHEAT BRAN ............................................................ 103
5.1. ABSTRACT ..................................................................................................................... 104
5.2. INTRODUCTION ............................................................................................................ 105
5.3. MATERIALS AND METHODS ..................................................................................... 108
5.3.1. Raw Material ............................................................................................................. 108
5.3.2. Preparation of Washed Wheat Bran .......................................................................... 108
5.3.3. Grinding and Sifting of Wheat Bran Samples ........................................................... 108
5.3.4. Extrusion of Wheat Bran ........................................................................................... 108
5.3.5. In vitro Binding of Bile Acids ................................................................................... 108
5.3.6. Determination of Weight-Average Molecular Weight of Soluble Fiber Using HighPerformance Size Exclusion Liquid Chromatography ........................................................ 109
5.3.7. Thermal Decomposition Properties of Wheat Bran Samples ........................................... 110
5.3.8. Water Binding Capacity ............................................................................................ 110
5.3.9. Statistical Analysis .................................................................................................... 111
5.4. RESULTS AND DISCUSSION ...................................................................................... 112
5.4.1. Water Binding Capacity ............................................................................................ 112
5.4.2. Effect of Washing, Bran Particle Size and Extrusion on Weight-Average Molecular
Weights of Soluble Fibers Isolated from Wheat Bran Samples .......................................... 115
5.4.3. Thermal Decomposition of Wheat Bran Samples ..................................................... 117
5.4.4. Binding of Bile Acids by Wheat Bran Samples ........................................................ 119
5.5. CONCLUSIONS .............................................................................................................. 122
LITERATURE CITED ............................................................................................................... 123
CHAPTER 6
EFFECT OF TREATED WHEAT BRAN ON THE BAKING PROPERTIES OF BREAD AND
COOKIES ................................................................................................................................... 128
6.1. ABSTRACT ..................................................................................................................... 129
6.2. INTRODUCTION ............................................................................................................ 130
6.3. MATERIALS AND METHODS ..................................................................................... 132
6.3.1. Materials .................................................................................................................... 132
6.3.2. Preparation of Washed Bran ...................................................................................... 132
6.3.3. Grinding and Sifting .................................................................................................. 132
6.3.4. Preparation of Extruded Wheat Bran......................................................................... 132
6.3.5. Farinograph Properties .............................................................................................. 132
6.3.6. Pasting Properties ...................................................................................................... 133
6.3.7. Breadmaking .............................................................................................................. 133
6.3.8. Determination of Bread Quality Properties ............................................................... 134
6.3.9. Cookiemaking ............................................................................................................ 134
6.3.10. Determination of Cookie Quality Properties ........................................................... 135
6.3.11. Determination of Insoluble and Soluble Dietary Fibers in Bread Crumb and Crust135
6.3.12. Statistical analysis.................................................................................................... 136
6.4. RESULTS AND DISCUSSION ...................................................................................... 137
6.4.1. Farinograph ................................................................................................................ 137
6.4.2. Pasting Properties ...................................................................................................... 140
6.4.3. Bread Quality Properties ........................................................................................... 142

vii

6.4.4. Cookie Quality Properties ......................................................................................... 145
6.4.5. Effect of Baking on the Insoluble and Soluble Dietary Fibers in Bread ................... 147
6.5. CONCLUSIONS .............................................................................................................. 151
LITERATURE CITED ............................................................................................................... 152
CHAPTER 7
GENERAL CONCLUSIONS ..................................................................................................... 156
CHAPTER 8
FUTURE RECOMMENDATIONS ........................................................................................... 163

viii

LIST OF TABLES

Table 2.1. Vitamin Contents (µg/g dry basis) of Hard Red Spring Wheat and its Milling
Fractions ........................................................................................................................................ 12
Table 2.2. Effect of Fiber Supplements on Fecal Bulk ................................................................. 13
Table 2.3. Constituents of Dietary Fiber ....................................................................................... 15
Table 2.4. β-Glucan Content (dry weight basis) of Selected Cereal Grains ................................. 21
Table 3.1. Total Starch Composition of Non-Washed and Washed Wheat Bran Samples .......... 62
Table 3.2. Effect of Washing on the Thermal Properties of Non-Washed and Washed ............. 65
Table 3.3. Water Binding Capacity (WBC) of Non-Washed and Washed Wheat Bran .............. 67
Table 3.4. Dietary Fiber Composition of Non-Washed and Washed Wheat Bran samples ......... 69
Table 3.5. Proximate Composition of Non-Washed and Washed Bran Samples ......................... 71
Table 4.1. Experimental Design.................................................................................................... 83
Table 4.2. Screw Elements Used for Low and High Shear Extrusion Configurations ................. 84
Table 4.3. Extrusion Cooking Conditions for NWB425, NWB1000, WB425, and WB1000...... 85
Table 4.4. Effects of Extrusion Cooking Conditions under Low Shear Screw Configuration on
the Contents of Insoluble and Soluble Dietary Fibers of Extrudates Made from NWB425 and
NWB1000 Samples ....................................................................................................................... 90
Table 4.5. Effects of Extrusion Processing Conditions under Low Shear Screw Configuration on
the Contents of Insoluble and Soluble Dietary Fibers of Extrudates Made from WB425 and
WB1000 Samples.......................................................................................................................... 94
Table 5.1. Effects of Extrusion Conditions under Low Shear Screw Configuration on the Water
Binding Capacities of NWB425, NWB1000, WB425, and WB1000 Samples .......................... 114
Table 5.2. Effect of Washing, Particle Size, and Extrusion on the Weigh Average Molecular
Weight of Soluble Dietary Fiber in Wheat Bran Samples .......................................................... 116
Table 5.3. Thermal Decomposition Properties of Non-Washed Bran, Washed Bran, Extruded
Non-Washed Bran, and Extruded Washed Bran Samples .......................................................... 118
Table 5.4. In Vitro Binding of Bile Acids by Wheat Bran Samples ........................................... 121

ix

Table 6.1. Farinograph Properties of Hard Wheat Flour Substituted with Bran ........................ 139
Table 6.2. Pasting Properties of Hard Wheat Flour and Soft Wheat Flour Substituted with Wheat
Bran ............................................................................................................................................. 141
Table 6.3. Quality Properties of Bread ....................................................................................... 144
Table 6.4. Effect of Bran Substitution on Quality Properties of Cookies................................... 146

x

LIST OF FIGURES
Figure 3.1. Modified servodyne mixer used to wash wheat bran. For interpretation of the
references to color in this and all other figures, the reader is referred to the electronic version of
this dissertation. ............................................................................................................................ 55
Figure 4.1. Effects of low and high shear screw configurations (see Table 4.2 for details) on the
contents of soluble dietary fiber in washed wheat bran extrudates. ............................................. 96
Figure 4.2. Effect of low and high shear screw configurations (see Table 4.2 for details) on the
contents of insoluble dietary fiber in WB1000 extrudates............................................................ 97

Figure 6.6. Insoluble dietary fiber in flour samples and its quantity and distribution in subsequent
baked bread. ................................................................................................................................ 149
Figure 6.7. Insoluble dietary fiber in flour samples and its quantity and distribution in subsequent
baked bread. ................................................................................................................................ 150

xi

CHAPTER 1
INTRODUCTION

1

Wheat is one of the most cultivated and consumed food plants in the world after corn and
rice. The world annual production of wheat grain is approximately 600 million metric tons
(Wrigley 2009). Wheat is considered a unique cereal because it contains gluten proteins that when
hydrated can form dough with viscoelastic properties. This uniqueness is the main reason why
wheat, especially in the flour form, is used to manufacture bread, which is by far one of the most
common yeast-leavened products. Consumers prefer the texture of wheat-based products to any
other cereal product (Hoseney 1998).
Wheat bran is a by-product of roller milling during the milling process of wheat grain.
Wheat bran, which used to be exclusively used as animal feed, is now used by the baking industry
as a major source of dietary fiber (Vetter 1988). Wheat bran is rich in dietary fiber (USDA 2011).
In the mid-1970s, scientists hypothesized that consumption of indigestible polysaccharides
(high fiber ingredients) may prevent chronic diseases including heart disease and certain types of
cancers. Since then, sufficient research data on the effects of dietary fiber in human health have
supported the hypothesis. Because of the nutritional benefits of dietary fiber, health care officials
and nutritionists have recommended that consumption of dietary fiber can help to maintain good
health (McCleary 2007). Dietary fiber reduces the risk of cardiovascular disease, certain forms of
cancer, and constipation (Schaafsma 2004). Dietary fibers are categorized as insoluble and soluble
dietary fibers and each plays a different role in human health. Insoluble dietary fiber is important
for proper bowel function and may reduce symptoms of chronic constipation, diverticular disease,
and hemorrhoids, while soluble dietary fiber is associated with reduction in cholesterol levels and
attenuation of blood glucose (Schaafsma 2004). The physiological functions of dietary fibers are
related to their physicochemical properties such as water holding capacity, viscosity and
susceptibility to bacterial degradation in the colon (Dikeman and Fahey 2006). Because of the

2

health benefits of dietary fiber, the food industry is developing fiber-enriched food products,
including dairy products, but bakery products remain the most preferred source of dietary fiber
(Hamid and Luan 2000; Sanchez-Alonso et al 2007; Collar 2008). However, incorporation of
fiber during food processing alters the consistency, texture, rheological properties and sensory
attributes of fiber-enriched food products (Collar et al 2009). For example, addition of fiber
during the bread-making process affects the mixing properties and rheological behavior of the
dough due to the interaction of fiber ingredients with starch and protein in the bread flour (Rosell
and Foegeding 2007). Incorporation of high fiber ingredients results in loaves with reduced
volume, cookies with reduced spread and hard texture; and reduces expansion of extruded
products (Pomeranz et al 1977; Lai et al 1989; Dreher 1987; Zang and Moore 1999). The
challenge for the food industry is to develop better methods to produce fiber-enriched food
products with acceptable quality.
It is important to understand that the amount of soluble dietary fiber is lower than that of
insoluble dietary fiber in wheat bran, which implies that most of the health benefits provided by
wheat bran such as proper bowel function are attributable to insoluble dietary fiber. The amount
of soluble dietary fiber in wheat bran is 1.5 to 4.0% (w/w) whereas insoluble dietary fiber is 35 to
48% (w/w) (Kahol and Chow 2000; Nandini and Silimath 2001; Esposito et al 2005). The ability
of soluble dietary fiber to lower serum cholesterol and attenuate blood glucose has prompted
scientists to find a way of increasing soluble dietary fiber in wheat bran. Some investigators have
used extrusion cooking technology to increase the content of soluble dietary fiber in native (nontreated) wheat bran.
It is very important to note that although the major objective of roller milling is to separate
wheat bran as cleanly as possible from the adherent starchy endosperm, it is almost impossible to

3

produce pure bran during the milling process. In certain cases, some investigators (Caprez et al
1986; Camire et al 1997) have concluded that extrusion cooking causes mechanical rupture of
insoluble dietary fiber leading to an increase in soluble dietary fiber content. However, it is
crucial to investigate the role of residual starchy endosperm still adherent to bran after the milling
process on physicochemical properties and dietary fiber profile of that wheat bran. Also, it is
important to look at the role of the residual starchy endosperm on the physicochemical properties
and dietary fiber profile of extruded wheat bran. Therefore to obtain reliable dietary fiber results,
it is important to conduct extrusion cooking experiments using wheat bran that has been treated to
remove most of the starch adherent to the bran. Starch is known to undergo chemical
modifications during extrusion processing that may interfere with dietary fiber analyses (Fogliano
et al 1999).
Therefore, the present study about extrusion of washed wheat bran was designed with the
following objectives:
1. To develop a method that removes most of the residual starchy endosperm still adherent to
milled wheat bran.
2. To investigate the effects of particle size of non-washed and washed wheat bran coupled
with extrusion processing variables on total dietary fiber, insoluble dietary fiber, and
soluble dietary fiber contents.
3. To investigate the effects of extrusion cooking variables on physicochemical properties of
extruded wheat bran.
4. To investigate the effects of extrusion processing variables on in vitro binding of bile
acids by extruded washed bran.

4

5. To evaluate the effects ground extruded washed bran on the baking properties of bread and
cookies.

The purpose of the present research is to provide scientific information on how the
residual starchy endosperm still adherent to wheat bran after the process of roller-milling affects
its physicochemical properties and dietary fiber composition. The following dissertation is
divided into: (1) Literature review, (2) Physicochemical properties of washed wheat bran, (3)
Effects of particle size coupled with extrusion processing conditions on dietary fiber profile of
non-washed and washed wheat bran, (4) Effects of extrusion processing on physicochemical
properties of washed wheat bran, (5) Effect of treated wheat bran on the baking properties of
bread and cookies, (6) General conclusions, and (7) Recommendations for future studies. The
chapters of this dissertation were written according to the Cereal Chemistry journal article format
and thus some information such as introduction and laboratory procedures are very similar for
certain chapters.

5

LITERATURE CITED

6

LITERATURE CITED
Abdul-Hamid, A., and Siew Luan, Y. 2000. Functional properties of dietary fiber prepared from
defatted rice bran. Food Chem. 68:15-19.
Camire, M.E., Violette, D., Dougherty, M.P., McLaughlin, M.A. 1997. Potato peel dietary fiber
composition: effects of peeling and extrusion cooking processes. J. Agric. Food Chem. 45:
1404-1408.
Caprez, A., Arrigon, E., Amado, R., Neukom, H. 1986. Influence of different types of thermal
treatment on the chemical composition and physical properties of wheat bran. J. Cereal Sci. 4:
233-239.
Collar, C. 2008. Novel high fiber and whole grain breads. Pages 336-361 in: Technology of
Functional Cereal Products. B. Hamaker, ed. Woodhead Publishing Ltd: Cambridge, UK.
Collar, C., Rosell, C.M., Muguerza, B., and Moulay, L. 2009. Breadmaking performance and
keeping behaviour of cocoa soluble fiber enriched-white breads. Food Sci. Technol. Intern. 15.
Dikeman, C. L., and Fahey, G.C. 2006. Viscosity related to dietary fiber: A review. Crit. Rev.
Food Sci. & Nutri. 46:649-663.
Dreher, M.L. 1987. Conventional and unconventional dietary fiber components. Pages 17-52 in:
Handbook of Dietary Fiber: An Applied Approach. Marcel Decker: New York, NY.
Esposito, F., Arlotti, G., Bonifati, A.M., Napolitano, A., Vitale, D., and Fogliano, V. 2005.
Antioxidant activity and dietary fiber in durum wheat bran by-products. Food Res. Int.38:11671173.
Fogliano, V., Monti, S. M., Musella, T., Randazzo, G., and Ritieni, A. 1999. Formation of
coloured Milliard reaction products in a gluten-glucose model system. Food Chem. 66:293299.
Hoseney, R.C. 1998. Yeast-Leavened Products. Pages 229-273 in: Principles of Cereal Science
and Technology. Second Edition. AACC International, St Paul, MN.
Kahlon, T.S., and Chow, F. 2000. In vitro binding of bile acids by rice bran, oat bran and, and
corn bran. Cereal Chem. 77:518-521.
Lai, C.S., Hoseney, R.C., and Davis, A.B. 1989. Effects of wheat bran in bread baking. Cereal
Chem. 66(3):217-219.
McClearly, B.V. 2007. An integrated procedure for the measurement of total dietary fiber
(including resistant starch), non-digestible oligosaccharides and available carbohydrates.
Anal. Bional. Chem. 389:291-308.
7

Nandini, C. D., and Silimath, P.V. 2001. Carbohydrate composition of wheat, wheat bran,
sorghum, and bajra with good chapatti/roti (Indian flat bread) making quality. Food Chem.
73:197-203.
Pomeranz, Y., Shogren, M., Finney, K.F., Bechtel, D. B. 1977. Fiber in breadmaking processeffect on functional properties. Cereal Chem. 54:25-41.
Rosell, C.M., and Foegeding, E.A. 2007. Interaction of hydroxypropylmethylcellulose with
gluten proteins: small deformation properties during thermal treatment. Food Hydrocolloid.
21:1092-1100.
Sanchez-Alonso, I., Haji-Maleki, R., and Borderias, J. 2007. Wheat fiber as a functional
ingredient in restructured fish products. Food Chem. 100:1037-1043.
Schaafsma, G. 2004. Health claims, options for dietary fiber. Pages 27-28 in: Dietary fiberBioative carbohydrates for food and feed. J. W. Van der Kamp, N. G. Asp, J. Miller Jones, and
G. Schaafsma, eds. Academic Publishers: Wageningen.
United States Department of Agriculture. 2011. http://ndb.nal.usda.gov/ndb/foods/show/6378.
Vetter, J.L. 1988. Commercial available fiber ingredients and bulking agents. AIB Technical
Bulletin. 5:1-5.
Wrigley, C.W. 2009. Wheat: A Unique Grain for the World. Pages 1-17 in: Wheat Chemistry
and Technology, K. Khan and P. R. Shewry, eds. AACC International, St. Paul, MN.
Zhang, D., and Moore, W.R. 1997. Effect of wheat bran particle size on dough rheological
properties. J Sci. Food Agric. 74:490-496.

8

CHAPTER 2
LITERATURE REVIEW

9

2.1. WHEAT BRAN
World wheat production is approximately 685 million metric tons per year resulting in
nearly 140 million metric tons of wheat bran (World Agricultural Outlook Board 2012). Wheat
bran is the outer coat of the wheat kernel and comprises about 12-15% (w/w) of wheat grain (Cho
and Clark 2001). It is separated from wheat during the process of wheat milling. Wheat brans
from different wheat varieties, for example, red, white, soft, hard, and durum wheats, have
different physicochemical properties (Dreher 1987). The differences in physicochemical
properties of wheat brans could also be influenced by different environmental conditions under
which the wheats are grown.
Wheat bran contains more vitamins than any other part of a wheat kernel (Table 2.1).
Wheat bran is also rich in dietary fiber especially the insoluble type (Zook et al 1970; Kent 1974).
Nutritional studies indicate that insoluble dietary fiber can act as a bulking agent, increasing
intestinal motility and wet fecal mass (Cho and Clark 2001). Decreased intestinal residence time
and increased amount of stool are associated with reduced incidence and number of colonic
tumors (NRC 1989). Wheat bran is known for its effectiveness in the treatment of constipation
and other maladies of the colon (Burkitt 1977). The ability of wheat bran to increase stool volume
and improve laxation more than any other fiber supplement (Table 2.2) is attributed to its high
proportion of insoluble fiber (Mongeau et al 1990; Ranhotra et al 1991; Hosig et al 1996).
Although consumption of wheat bran may provide a number of health benefits, its use and
application in food processing remain a challenge as it has poor functionality and/or detrimental
effects on end use quality of food products. For example, the use of wheat bran in breadmaking
resulted in reduced loaf volume and increased crumb darkness (Lai et al 1989). Grinding of fiber
and extrusion processing with high temperature, high shear, and pressure to modify the

10

physicochemical properties of high fiber ingredients, such as wheat bran, could be an alternative
technique for improving their functionality.

11

Table 2.1. Vitamin Contents (µg/g dry basis) of Hard Red Spring Wheat and its Milling
Fractions
Vitamin
Grain
Thiamine
9.9
Riboflavin
3.1
Niacin
48.3
Biotin
0.056
Folacin
0.56
Pantothenic acid 9.1
Vitamin B6
4.7
Source: Zook et al (1970).

Flour
0.7
1.5
9.5
0.013
0.09
2.5
0.48

Shorts
10.1
1.8
23.5
0.055
0.59
7.0
5.3

12

Bran
13.2
5.5
171.4
0.162
1.59
31.7
13.0

Table 1.2. Effect of Fiber Supplements on Fecal Bulk
Fiber Supplement
Oat bran
Pectin
Guar Gum
Apple
Carrot
Cabbage
Cellulose
Wheat bran
Source: Cummings (1997)

Increase in Fecal Weight (%)
15
16-35
20
40
59
67
75
80-127

13

2.2. DIETARY FIBER
Dietary fiber is an important food component that has been found to provide positive
health benefits. It is able to improve physiological digestive functions, for example, increasing
fecal output, and normalizing blood glucose and lipid levels. The nutritional benefits have
prompted consumers to develop an interest in whole grains because they contain substantial
amounts of dietary fiber and other associated nutrients (Nelson 2001). The most common dietary
fiber constituents are depicted in Table 2.3.
According to the American Association of Cereal Chemists International (AACCI),
“dietary fiber is defined as the edible parts of plants or analogous carbohydrates that are resistant
to digestion and absorption in the human small intestine with complete or partial fermentation in
the large intestine. Dietary fiber includes polysaccharides, oligosaccharides, lignin and associated
plant substances. Dietary fibers promote beneficial physiological effects including laxation,
and/or blood cholesterol attenuation, and/or blood glucose attenuation” (AACCI 2001). This
AACCI definition is a modification of the definition developed by scientists in the mid-1970s
who defined dietary fiber as plant cell wall material that is resistant to hydrolysis by the enzymes
of the human gastrointestinal tract (Burkitt et al 1972; Trowell 1974; and Painter 1975).

14

Table 2.2. Constituents of Dietary Fiber
Non-Starch Polysaccharides and Resistant Oligosaccharides
Cellulose
Hemicellulose
Arabinoxylans
Arabinogalactans
Polyfructoses
Inulin
Oligofructans
Galactooligosaccharides
Gums
Mucilages
Pectins
Analogous carbohydrates
Indigestible dextrins
Resistant maltodextrins
Resistant potato dextrins
Synthesized carbohydrate compounds
Polydextrose
Methyl cellulose
Hydroxypropylmethyl cellulose
Resistant starches
Lignin
Substances Associated with the Non-Starch Polysaccharide and Lignin Complexes in Plants
Waxes
Phytates
Cutin
Saponins
Suberin
Tannins
Source: AACCI (2001)

15

2.2.1. Insoluble Dietary Fiber
Like soluble dietary fiber, insoluble dietary fiber is a type of fiber that cannot be
hydrolyzed by the enzymes of the alimentary canal. However, insoluble dietary fiber can be
partially fermented by colonic bacteria. Insoluble dietary fiber is commonly used to treat
constipation (Nelson 2001). The major components of insoluble dietary fiber are cellulose, certain
hemicelluloses, and lignin (Esposito et al 2005).

2.2.1.1. Cellulose
Cellulose is the major structural component of higher plant cell walls such as wheat,
barley, rye, oat straw, corn stalks and sugar cane and therefore the most common organic material
on earth (Klemm et al 2005; Nelson 2001; Izydorczyk et al 2005). It is a high molecular weight,
linear, insoluble homopolymer of D-glucopyranosyl residues linked together in β-1,4 glycosidic
linkages (French et al 1993). Its degree of polymerization ranges from 300 to 15,000 units (Donde
and Miller 1985). The size of cellulose depends on the source of cellulose and the method of
isolation. The β-1,4 linkage between glucose units enables the cellulose to form strong
intermolecular and intramolecular hydrogen bonding that result in strong and inflexible crystalline
microfibrils of up to 25 nm in diameter (McDougall et al 1996). The unique arrangement of
glucose units in cellulose makes it insoluble in hot and cold water, dilute acid, and dilute alkali
(Cho et al 1997). Because the human digestive system lacks specific enzymes to hydrolyze β-1,4
linkages, this makes cellulose a truly unavailable carbohydrate (Dreher 1987). Cellulose can be
physically and chemically modified to improve its functionality in food systems (Izydorczyk et al
2005). Modified celluloses include microcrystalline cellulose, carboxymethylcellulose, and
methylcellulose. Microcrystalline cellulose is usually prepared by treating cellulose with

16

hydrochloric acid to partially dissolve and remove its less organized amorphous regions. The
crystalline

aggregates

created

can

be

converted

to

powder

form

after

drying.

Carboxymethylcellulose which is usually used as a thickener or stabilizer is prepared by reacting
cellulose, alkali, and monochloroacetic acid. Its solubility is enhanced by presence of sodium
ions. On the other hand, methylcellulose is a non-ionic ether with thickening and film properties.
Methylcellulose is usually prepared by reacting cellulose with methyl chloride (Izydorczyk et al
2005).

2.2.1.2. Hemicelluloses
Scientists unfamiliar with fiber terminology may wrongly assume that hemicelluloses are
similar to cellulose (Cho et al 1997; McDougall et al 1996). The designation of hemi, which
means half, was used when early researchers in the area of plant polysaccharides believed that
hemicelluloses were on the way to becoming cellulose. Later, hemicelluloses were found to be
polysaccharides unrelated to cellulose (Whistler and BeMiller 1997). Both cellulose and
hemicellulose are insoluble in hot and cold water, and dilute acid. However, hemicelluloses are
distinguished from cellulose because of their solubility in dilute solutions of bases (Dreher 1987).
Hemicelluloses are not structurally related to cellulose. Unlike cellulose that consists of
polymers of only glucose units, hemicelluloses are heterosaccharides that have a much lower
degree of polymerization ranging from 50-200 residues (Cummings 1976; Theander and Aman
1979). Hemicelluloses contain a variety of monosaccharides in the backbone and side chains.
Xylose, mannose, and galactose usually make up the backbone in the beta-1,4 glycosidic
configuration. Arabinose, galactose and uronic acids predominate the side chains (Nelson 2001).
The number of sugars in the side chains varies such that some hemicelluloses are relatively linear

17

while others are highly branched. Highly branched hemicelluloses that contain acidic sugars, for
example glucuronic acid, in their side chains are actually water soluble.
The

major

groups

of

hemicelluloses

in

cereals

include

arabinoxylans,

glucuronoarabinoxylans, and β-glucans. Arabinoxylans are composed of pentoses, arabinose and
xylose (Nelson 2001).

2.2.1.3. Lignin
Although lignin is a component of insoluble dietary fiber, it is not a polysaccharide
(Nelson 2001). Lignin is a high molecular weight aromatic polymer composed of
polyphenylpropane units which are formed by condensation of the three primary alcohols during
plant growth. These alcohols are coniferyl, sinapyl and p-coumaryl (Dreher 1987).

Lignin

infiltrates the cellulose of cell walls resulting in a hard, rigid matrix of a tremendous strength.
Lignin is closely associated with cellulose and hemicellulose (Theander and Aman 1979) and is
able to form covalent bonds with them (Southgate 1995). Lignin is highly insoluble in water and
is responsible for structural adhesion of the plant cell wall components because it excludes water
when it forms networks by cross-linking with other saccharide-type molecules in plants. Lignin is
very resistant to chemical, enzymatic and bacterial breakdown. Thus, lignin resists enzymatic
breakdown in the small intestine and bacterial breakdown in the large intestine and can be
recovered almost completely intact in the feces (Cho et al 1997).

2.2.2. Soluble Dietary Fiber
Soluble dietary fiber is different from insoluble dietary fiber because of its solubility in
aqueous solutions of the digestive system. It is important to note that solubility of fibers is quite

18

different from solubility of sugars such as glucose. Fiber is never completely dissolved in aqueous
solution. Instead, it behaves as a colloid (Nelson 2001). Soluble fiber mainly consists of pectins,
certain hemicelluloses, gums, and β-glucans (Esposito et al 2005).

2.2.2.1. Arabinoxylans
In wheat grain, arabinoxylans are believed to be the major components of cell walls
(Saulinier et al 2007). Arabinoxylans are comprised of a linear backbone of β-1,4-linked Dxylopyranosyl units. α-L-arabinofuranosyl and α-D-glucuronic acids are attached to the backbone
on oxygen 2 (O-2) or oxygen 3 (O-3) as the major side chains (Dervilly et al 2000; Saulinier et al
2007). Ferulic and p-coumaric acids are esterified to the α-L-arabinofuranosyl units through
oxygen O-5 (Smith and Hartley 1983; Saulinier et al 2007).
The amount of arabinoxylans in wheat flour ranges from 0.3 to 0.7% (w/w). Although
they are not major constituents in wheat flour, arabinoxylans are known to have significant
influence on dough rheology as well as on baking properties (Biliaderis et al 1995). Unlike in cell
walls, the arabinoxylans in wheat flour (endosperm) are composed of only arabinose and xylose.
These sugars contain five carbons and are therefore referred to as pentosans. There are two types
of arabinoxylans: water-extractable (WE-AX) and water- unextractable (WU-AX). The amounts
of WE-AX and WU-AX in wheat flour are 0.5% and 1.7% (w/w), respectively (Dervilly-Pinel et
al 2001; Ordaz-Ortiz and Saulinier 2005; Saulinier et al 1995).
The structure of arabinoxylans in the residual starch (aleurone layer) adhered to wheat
bran after roller milling is different from that in the endosperm. Arabinoxylans in the aleurone
layer are not water-extractable and have a lower arabinose to xylose ratio than the arabinoxylans

19

in the endosperm (Antoine et al 2003). Also, the esterification of arabinoxylans is more
pronounced in the aleurone layer than in the endosperm (Parker et al 2005).
The functionality of arabinoxylans heavily depends on their physicochemical properties
such as viscosity and water solubility (Saulinier et al 2007). It is important to understand that the
structure and conformation of polymers, such as chain length and composition, including side
chains, are the major factors influencing the physicochemical properties of arabinoxylans
(Saulinier et al 2007).
2.2.2.2. β-glucans
Beta-glucans have gained popularity because research has shown that consumption of βglucan reduces serum cholesterol (Tapola and Sarkkinen 2007). In addition, β-glucan is of interest
because of its ability to attenuate blood glucose levels (Wood et al 1989). Beta-glucans are nonbranched polymers of glucopyranosyl units with a mixture of β-1,3 and β-1,4 linkages (Wood
1989). Grains, especially oats and barley, contain high amounts of beta-glucans (Wood et al
1989). β-glucan content (dry weight basis) in oats and wheat range from 2.5% to 6.6% (w/w) and
0.5% to 1.5% (w/w), respectively (Table 2.4). However, these values depend on environmental
factors and genetic make-up of oat cultivars (Nelson 2001). The amounts of β-glucan in barley
and rye are also listed in Table 2.4. β-glucans are often referred to as food gums or mucilages
since they hydrate easily, forming viscous solutions (Dziezak 1991). Thus, β-glucans are grouped
as hydrocolloids since they have the ability to form a gel after dispersion in water. Other types of
hydrocolloids include alginates, carrageenan, gums Arabic, karaya, tragacanth and xanthan gum
(Dziezak 1991). Beta-glucans form long cylindrical molecules containing up to 250,000 glucose
units. The presence of β-1,3 and β-1,4 linkages prevents the compact folding of β-glucans, hence
making them soluble in water (Chawla and Patil 2010).
20

Table 2.3. β-Glucan Content (dry weight basis) of Selected Cereal Grains
Grain
Barley
Oats
Rye
Wheat
Source: Cho et al (1997)

β-glucan (% w/w)
2.0 - 9.0
2.5 - 6.6
1.9 - 2.9
0.5 - 1.5

21

2.2.2.3. Pectin
Pectin is a component of higher plant cell walls where it is used for overall growth of plant
cells. It is mostly found in the middle lamella and is used for ion transport and water retention. It
is considered the largest source of soluble fiber among plant food materials and is used to
stabilize food systems due to its high viscosity properties (Endress and Mattes 2009).
The backbone of pectin is composed of α-1,4-linked galacturonic acid residues interrupted by
single α-1,2-linked rhamnose residues (Voragen et al 1995; BeMiller 1986). The side chains of
pectin consist of galactose, glucose, rhamnose, and arabinose (Cho et al 1997; McDougall et al
1996; Dreher 1987). Solubility of pectins depends on the side chain composition (Dreher 1987).
Pectins are among the primary cell wall components where they are linked to cellulose,
hemicellulose, and lignin (Lopes da Silva and Rao 2006) and are a key textural determinant in
fruits and vegetables (Voragen et al 1995; Wang et al 2002). Pectin is the major soluble fiber in
fruits and vegetables. However, in wheat, it is present in very small amounts.

2.2.3. Non-Digestible Oligosaccharides
Foods that have the ability to reduce the risk of diseases and provide better health have
become popular as consumers continue to become health conscious. In this respect, significant
attention has been paid to a group of carbohydrates commonly known as the nondigestible
oligosaccharides (NDOs) (Mussatto and Mancilla 2007). NDOs are not digestible by humans
because the human digestive system lacks the enzymes required to hydrolyze the β-links formed
among the units of some monosaccharides (Urgell and Orleans 2001). These oligosaccharides are
then fermented by beneficial bacteria in the colon where they stimulate the growth and activity of
beneficial bacteria and concomitantly repress harmful bacteria (Mussatto and Mancilla 2007). The

22

principal metabolites of fermentation are lactate and short-chain fatty acids such as acetate,
propionate and butyrate. These short-chain fatty acids are believed to: 1) increase water and
sodium absorption from digesta in the colon, 2) facilitate mucosal cell proliferation, 3) provide
energy for colonic microorganisms, and 4) acidify lumenal environment. The amounts of shortchain fatty acids produced in the colon depend on the type of NDO substrates and types of
intestinal flora (Sako et al 1999). The rate at which oligosaccharides are fermented also depends
on their degree of polymerization (Manning and Gibbson 2004).

2.2.4. Resistant Starch
Resistant starch is a type of starch that cannot be hydrolyzed by the enzymes in the
digestive system, especially the small intestine of humans. Thus, this type of starch cannot be
absorbed by humans. However, resistant starch can be fermented in the large intestine. Resistant
starch is considered a form of dietary fiber due to the fact that it is not attacked by the enzymes in
the small intestine and undergoes fermentation in the large intestine (Grooper et al 2009; Nelson
2001).
Resistant starch is categorized as RS1, RS2, RS3, and RS4. RS1 is a type of starch that is
trapped in the cell walls of plants and therefore cannot be attacked by hydrolyzing enzymes.
These starches can only be hydrolyzed by breaking the system in which they are trapped. Sources
of RS1 include partially milled grains, seeds and legumes (Gropper et al 2009; Yue and Waring
1998). RS2 encompasses starches that are intact granules and not gelatinized. These starches can
be hydrolyzed by enzymes after the process of gelatinization. Sources of RS2 starches include
raw potatoes and unripe green bananas. RS3 is a type of starch that is formed after cooking
starches under high moisture conditions. The ideal factor for the formation of RS3 is

23

gelatinization and retrogradation. Enzymes cannot hydrolyze this type of starch because of the
network formed through crystalline junction zones as a result of retrogradation.

Extrusion

cooking under high moisture is one of the popular cooking methods used to increase RS3 in
foods. Sources of RS3 include ready-to-eat cereals, and cooked and cooled potatoes and breads.
RS4 is a type of resistant starch formed by chemical modification of starches, for example,
formation of ester bonds. RS4 cannot easily be broken down by digestive enzymes because the
enzyme-attacking sites are taken up by crosslinking chemicals (Grooper et al 2009; Nelson 2001).

2.2.5. History of Dietary Fiber
For many years, dietary fiber has been known to provide health benefits. For example,
consuming foods high in indigestible carbohydrates improved bowel habits and increased fecal
volume. The term dietary fiber was later used to describe non-digestible plant components such as
cellulose, hemicelluloses and lignin (Hipsley 1953).
In the mid-1970s, there was sufficient data to suggest that consumption of foods high in
fiber could provide health benefits. Studies conducted in Africa showed that Africans who
consumed foods high in fiber had frequent and bulky stools and had fewer of the illnesses that
were common in western culture. It was eventually hypothesized that the indigestible plant
constituents were able to provide health benefits (Burkitt et al 1972; Trowell 1974; and Painter
1975).
In 1976, dietary fiber was redefined to include all indigestible polysaccharides, including
gums, modified celluloses, mucilages, oligosaccharides, and pectins. This new definition came
after new information about the chemical structures of dietary fibers was obtained. This definition
became widely accepted (AACCI 2001).

24

Understanding the chemical composition and health benefits associated with dietary fiber
prompted several researchers to develop methods to determine dietary fiber content in foods
(Southgate 1969; Baker 1979; Asp and Johanson 1981). The major purpose of the dietary fiber
content determination method was to remove all digestible components of foods from the nondigestible components using specific enzymes.
In the late 1970s, scientists sought consensus on a definition of dietary fiber that matched
with its health benefits (Prosky and Harland 1979). The consensus was achieved in 1981 during
the Spring Workshop of the Association of Official Analytical Chemists in Ottawa, Canada
(Prosky 1981).

2.2.5. Health Benefits of Dietary Fiber
Since the middle of the 1970s when Burkitt and Trowell first developed a dietary fiber
hypothesis, the role of dietary fiber has attracted significant attention in the area of health and
nutrition (Hamid and Luan 2000; Marlett et al 2002). The hypothesis put forward by Burkitt and
Trowell stated that consumption of foods low in dietary fiber was the major cause of chronic
diseases such as heart disease, diabetes, diverticulitis, and certain types of cancers that were
common in western societies. Medical data indicate that consuming diets high in dietary fiber is
associated with health benefits, especially in Western populations (McCleary 2007). Consumption
of foods high in insoluble dietary fiber improves bowel function and may reduce the symptoms of
chronic constipation, diverticular disease and hemorrhoids. Several investigators have reported
that the prevalence of Western diseases, including high cholesterol and cardiovascular disease
(Jones 2008), colon cancer (Drecker et al 2002), diabetes, obesity, and constipation, can be
reduced by consuming fiber-enriched foods (Kantor et al 2001). Although several studies have

25

shown that consumption of foods high in dietary fiber is inversely proportional to the occurrence
of chronic diseases, it is not clear if health benefits are due to dietary fiber per se or other
chemical components, for example, antioxidants that are associated with dietary fiber (Jones
2008).
Despite the above mentioned health benefits of dietary fiber, its consumption remains low,
especially in the United States. Recommended dietary fiber intake in the United States ranges
between 20 and 35 grams per day, but the average intake is between 14 and 15 grams per day
(Cho 2009). In other developed countries, such as European countries, the average consumption
of fiber is also below recommended intakes (Murakami et al 2007).
Based on published data implicating the role of dietary fiber in reducing the risks of
degenerative diseases, the Food and Drug Administration (FDA) of the United States approved a
number of claims that certain types of fiber may reduce the risk of cancer and coronary heart
disease (FDA 1993; FDA 1998).

26

2.3. PHYSICOCHEMICAL PROPERTIES OF HIGH FIBER INGREDIENTS
2.3.1. Solubility
Solubility is considered a major physicochemical property of dietary fiber (Oakenfull
2001). In fiber terminology, solubility does not mean that the fiber in question is completely
dissolved in water as in a fashion similar to glucose (Nelson 2001). Solubility rather means that
the fiber components are easily hydrated in water to form a colloidal suspension. Thus, the
solubility of high fiber ingredients refers to the state of the ingredient in water and not the
solubility of an ingredient in water.
Molecular structure is the major factor that affects solubility of polysaccharides (Morris
and Norton 1983). Branching in the backbone of a polymer can increase the polymer’s solubility
(Nelson 2001). Branching disrupts the intermolecular forces, thereby preventing the formation of
ordered crystalline structure. Examples of branched polysaccharides that are soluble include gum
acacia and arabinogalactans (Nelson 2001). On the other hand, cellulose, which is characterized
by linear and ordered crystalline structure stabilized by hydrogen bonds, is water-insoluble
(Morris 1989).
Polysaccharides with ionizing groups, for example pectins and carrageenans, are easily
dissolved in water. Electrostatic repulsion inhibits the high fiber components from close packing,
thus preventing the formation of ordered structures that tend to be insoluble in water (Patil 2008).
The solubility of a polysaccharide is also determined by the nature of linkages between its
monomers (Morris and Norton 1983). For example, the mixed β-1,3 and β-1,4 linkages of βglucans prevent the formation of linear crystalline structure, thereby enhancing hydration
(solubility). Cellulose, which consists of D-glucose entirely linked by β-1,4 glycosidic bonds, is
insoluble in water.

27

2.3.2. Water Binding Capacity
The term water binding capacity simply refers to the way in which polysaccharides
interact with water (Patil 2008). Other terminologies include water uptake, hydration, adsorption,
and absorption. The most commonly used terminologies are water binding capacity and water
holding capacity. Based on the definitions given by Rey and Rabuza (1981), water holding
capacity is the amount of water the system retains within its structures without subjection to any
given addition of pressure or stress. On the other hand, water binding capacity is the amount of
water a system retains after it has been subjected to stress, for example centrifugation.
Several factors influence the water holding capacity of high fiber ingredients, for example,
the source of the raw material, processing conditions, and the way it is measured (Patil 2008). The
ability of fiber to increase stool volume and get fermented in the large intestine depends on water
absorption properties.

2.3.3. Bile Acid Binding Capacity
Bile acids are made in the liver from cholesterol. They are conjugated with glycine or
taurine and are stored in the gallbladder. Upon eating a meal, the contents of the gallbladder are
secreted into the intestine where the bile acids serve the purpose of emulsifying fats. About 90%
of the excreted bile acids are reabsorbed by active transport in what is referred to as active
enterohepatic circulation (Hofmann 1977). Binding of bile acids and increasing their fecal
excretion has been hypothesized as a possible mechanism for lowering cholesterol by dietary fiber
(Lund et al 1989; Anderson and Siesel 1990). By binding bile acids, dietary fiber prevents
reabsorption and stimulates plasma and liver cholesterol conversion to additional bile acids
(Kritchevsky and Story 1974). Soluble fiber is known to be effective in reducing blood

28

cholesterol (FDA 1998; Seiz 2006). Previous investigators (Ralet 1990; Esposito et al 2005;
Gajula et al 2008) were able to increase the proportion of soluble fiber in wheat bran by extrusion
cooking. However, it is not known whether the soluble fiber induced by extrusion cooking has the
same cholesterol-lowering effects as naturally soluble fiber.

2.3.4. Molecular Weight Distribution
Molecular weight is one of the important characteristics that affect physical properties,
such as solubility and viscosity, of polysaccharides (Wang and Cui 2005). Polysaccharides are
made of monosaccharides of different sizes and structures and thus they are polydisperse and
polymolecular (Wang and Cui 2005; Saulnier et al 2007). In addition to different sizes and
structures, the numbers of monosaccharides that make up a polysaccharide vary, resulting in a
distribution of molecular weight for that particular type of polysaccharide (Wang and Cui 2005).
The apparent molecular weight of a polysaccharide depends on the pathway and environment of
synthesis, extraction conditions used to isolate the polysaccharide, and the method of molecular
weight measurement (Wang and Cui 2005). To better describe the molecular weight of a
polysaccharide, information on the average molecular weight is needed. The average molecular
weight is described by molecular weight averages, such as number average molecular weight
(Mn), weight average molecular weight (Mw), z-average molecular weight (Mz), and viscosity
average molecular weight (Mv). Mn is related to the chain length of a polysaccharide. It can be
physically measured by counting the number of molecules that make up a polysaccharide, for
example, by using membrane osmometry. Mn is commonly known to influence the
thermodynamic properties of polysaccharides. Mw measures the size of the polymer but by
emphasizing heavier molecules than Mn. The most common technique for measuring Mw is light

29

scattering and sedimentation equilibrium. Mz is a third power average, which is commonly
determined by sedimentation techniques. Mz, which is determined by sedimentation techniques,
refers to the melt elasticity of polysaccharides. Mv, which is the viscosity-average molecular
weight, is a measure of intrinsic viscosity of a polysaccharide and is related to molecular weight
(Wang and Cui 2005).
The effect of processing, such as extrusion cooking, may have a significant effect on the
molecular weight distribution. The mechanical shear and high temperature of extrusion can cause
degradation of a polysaccharide, thus affecting its molecular weight distribution.

2.3.5. Thermal Analysis
Thermal analysis is a family of methods that is used to monitor thermal events of a sample
as a function of temperature and time. There are several thermal analysis techniques available:
differential thermal analysis (DTA), differential scanning calorimetry (DSC), dynamic
mechanical analysis (DMA), thermogravimetric analysis (TGA), and thermomechanical analysis
(TMA) (Schenz 2003). Of these techniques, DSC is the most widely used technique to
characterize polysaccharides. Information provided by DSC measurements includes the
temperatures for glass transition (Tg) and melting (Tm). DSC also measures changes in enthalpy
(∆H) during melting. Thermal analysis techniques can provide qualitative and quantitative data
about any physical and chemical changes in a polysaccharide that involve endothermic and
exothermic processes.
There are a number of physical events that occur during heating and cooling of a
polysaccharide. For example, endothermic transition occurs when heat flows into a sample

30

causing disruption of ordered structures. Exothermic transition occurs when heat is released from
a sample system during the formation of ordered structures.

31

2.4. EFFECT OF PROCESSING ON DIETARY FIBER
Processing can alter sensory attributes, nutritional profile, safety, and stability of the fiber
containing product. Processing results in molecular, structural and functional changes which in
turn alter the physicochemical properties of fiber ingredients. Understanding how processing
affects physicochemical properties would help to understand how fibers induce physiological
effects (Guillon and Champ 2000). This can in turn provide a good link between food technology
and nutrition. Several scientists are interested in understanding how processing affects nutritional
properties of food products (Guillon and Champ 2000).
Mechanical processing, such as grinding, can cause significant changes in the
physicochemical properties of dietary fiber ingredients, for example, the hydration properties of
fiber (Zhang and Moore 1997). Wood et al (1989) reported that fibers in oats are susceptible to
degradation which also results in decreased viscosity.
Extrusion processing, which is commonly used in manufacturing breakfast cereals, can
also modify the properties of dietary fiber (Nyman et al 1994). Decrease of soluble dietary fiber
with increase in soluble dietary fiber has been reported (McDougall et al 1996). Extrusion
processing has the ability to degrade dietary fiber polysaccharides which results in lower
molecular weight and lower viscosity (Snavberg 1997). However, these changes depend on the
extrusion variables such as moisture content, screw configuration, temperature and screw speed
(Camire and Flint 1991; Ralet et al 1991).

2.4.1. Particle Size Distribution
Particle size distribution is one of the major factors that determines both fiber’s
functionality and fiber’s role in the digestive tract (transit time, fermentation, fecal excretion). The

32

shape, and consequently the size of the fibers, depends on the degree of processing and this shape
may also vary during transit in the intestine tract as a result of digestion processes (Rosell et al
2009). Heller et al (1980) found out the mean transit time of coarse wheat bran was significantly
shorter compared to that of finely ground wheat bran. Chemical composition of fibers is also
thought to affect their physiological functions such as fecal bulking: the more pentose sugars
present in the dietary fiber polymers, the greater the increase in stool weight (Cummings et al
1978).
It is known that coarse wheat bran fiber has a higher water holding capacity, which is
directly related to its physiological functions such as increasing stool bulk and decreasing mean
transit time. However, the notion that water holding capacity will explain how various fibers will
behave in the digestive tract needs to be critically analyzed. It is possible that fibers may break
down in the digestive tract.
Reduction of particle size can have varying effects on the physical structure of different
fibers. Grinding can reduce the particle diameter and collapse the fiber matrix of wheat bran
(Cadden 1987). Particle size distribution of wheat bran affects physical and sensory properties of
bread: coarse wheat bran resulted in darker crust, dull crumb and decreased loaf volume (Zhang
and Moore 1999). Panelists rated bread containing coarse bran as less acceptable.

2.4.2. Extrusion Cooking
Extrusion cooking is a thermo-mechanical process that applies high temperature, high
pressure, and shear to raw food ingredients (Heldman and Hartel 1997). Extruders have gained
popularity in recent years due to their versatility, high throughput, low-cost, product quality,
energy savings, and addressing of environmental concerns (Riaz 2001). Extrusion cooking is also

33

a high-temperature short-time technique. It is a short-time processing technique since the
materials stay in the extruder barrel for only a very short time relative to traditional cooking
methods.
The physicochemical properties of extruded products mainly depend on how extrusion
cooking variables are manipulated (Huber 2000; Ryu and Ng 2001). In extrusion, independent
variables refer to process parameters that the extruder operator can control. The major
independent variables are screw speed and screw configuration, amount of water injected into the
extruder barrel, feed rate, die size and die configuration. On the other hand, dependent variables
refer to the parameters that are influenced by independent variables. Some of the major dependent
variables include the mean residence time, pressure inside the extruder barrel, and the mechanical
energy input to the extruder (Huber 2000).
Screw speed and configuration are the major operating (independent) variables of the
extrusion cooking process. Screw speed can be used to regulate the power input to the material in
the extruder (Altimore and Ghossi 1986). In rotational systems, power is related to the torque and
angular velocity. Thus, increasing screw speed increases energy to the system and vice-versa.
Changing screw speed changes the degree of fill inside the barrel, the mean residence time, and
the shear stress on the material being extruded (Altimore and Ghossi 1986). The high shear in the
extruder causes mechanical modification of the extruded product (Riaz 2001).
Moisture is very critical during the extrusion cooking process (Huber 2000). The amount
of water injected into the extruder barrel influences the material in the extruder (Altimore and
Ghossi 1986; Heldman and Hartel 1997). Moisture in the raw materials is required during
extrusion cooking for starch gelatinization and protein denaturation. Increasing the moisture
decreases mechanical energy of the extruder (Altimore and Ghossi 1986; Huber 2000).

34

Feed rate, the amount of material that is fed into the extruder per unit time, has a
significant effect on the quality of the extruded product. Feed rate has a dramatic influence on the
mean residence time (Altimore and Ghossi 1986). The higher the feed rate, the shorter and more
uniform the residence time distribution. Using a hopper that feeds a constant uniform amount of
the dry material into the extruder allows production of a consistent and uniform extruded product
(Heldman and Hartel 1997). Inconsistent feeding may lead to inconsistent quality in the final
product.
Significant changes can take place in the polysaccharides plant cell walls during food
processing. These changes can have an impact on the physicochemical and nutritional properties
of the dietary fiber such as redistribution of soluble and insoluble fibers in total fiber with a
proportionate increase in soluble fiber. This could improve the hypocholesterolemic properties of
fiber. Extrusion cooking significantly increased the proportion of soluble fiber in extruded wheat
bran (Aoe et al 1989; Ralet et al 1990; Wang and Klopfenstein 1993; Gualberto et al 1997).
Thermo-mechanical effects during extrusion processing can have a profound effect on the
composition of dietary fiber. Applying high temperature and shear are believed to weaken and
break the glycosidic linkages holding polysaccharide monomers together. As a result, the
structure of the fiber may be altered and allow insoluble fiber to become soluble.
Contradicting results also have been published. Artz et al (1990) reported that extrusion
cooking did not result in significant changes in the amount of soluble and insoluble fibers in
wheat bran. However, it is important to note that the source of fiber, the way it is prepared before
processing, and the processing conditions themselves are all crucial when studying the effect of
extrusion cooking and other processing techniques on the physicochemical properties of dietary
fiber (Guillon and Champ 2000).

35

2.5. WHEAT FLOUR
Among food grains, wheat is one of the most extensively produced and consumed cereals.
Three common species of wheat are Triticum aestivum, T. compactum, and T. durum. There are
five classes of T. aestivum (hard red winter, hard red spring, soft red winter, hard white, and soft
white). T. compactum includes the club wheat while T. durum forms the class of durum wheat
(Artwell 2001). Wheat, mainly as flour, is utilized for the preparation of a wide range of products
such as bread (leavened and unleavened), cakes, biscuits, cookies, crackers, and others. Wheat is
a good source of vitamins B1 (thiamine), B2 (riboflavin), niacin, B6 (pyridoxine), pantothenic
acid, tocopherol, as well as iron and zinc (Hoseney 1994).
Based on 14% moisture content, wheat flour is mainly composed of starch (63-72%, w/w)
and protein (7-15%, w/w). The amounts of starch and protein also depend on wheat cultivar and
growth environments. In addition to protein and starch, wheat flour contains other
polysaccharides such as hemicellulose, pentosans, arabinose, xylose, and arabinoxylan. Wheat
flour also contains some lipids and ash.
Starch is the largest molecule in the world after cellulose. Functionality of starch depends
on its botanical source. Starch, the major polymer in wheat flour, is composed of two groups of
molecules: amyloses and amylopectins (Tester et al 2004). Amylose is believed to be a linear
polymer of glucose units joined together at carbons 1 and 4 (α-1, 4 linkage). Usually in a single
amylose molecule there are 1500-6000 glucose units (Atwell 2001) with a molecular weight of
approximately 1 x 105 to 1 x 106 (Tester et al 2004). This number depends on the wheat species
(Hoseney 1994). Starch is insoluble in cold water but dissolves in warm water (about 60oC).
Amylose is responsible for starch retrogradation. Amylose molecules tend to associate with

36

themselves after gelatinization and precipitate out of the solution. Common wheat starch contains
approximately 25% amylose (Atwell 2001).
There are two ways in which amylopectin is distinguished from amylose. First, it is a
much larger molecule compared to amylose; a single amylopectin molecule contains 300,000–
3,000,000 glucose units. Second, glucose units in amylopectin are linked together by α-1, 4 and α1, 6 glycosidic linkages. Common wheat starch contains approximately 75% amylopectin (Atwell
2001). This amount depends on the source of wheat variety.
Wheat proteins are classified based on their solubility, and approximately 80% of wheat
flour proteins are gluten proteins (Atwell 2001). In the early 1900s, Osborne classified wheat
proteins as albumins, globulins, prolamins, and glutenins. Albumins are soluble in water and
make up 15% of flour protein. Albumins are comparable to egg white protein. The functionality
of albumins is not easily affected by salt concentrations (Hoseney 1994). Globulins are the minor
proteins in wheat. They make up only 3% of the total flour protein. They are insoluble in water,
but solubility can be achieved at low salt concentrations. However, solubility of globulins is
decreased by higher salt concentrations. Globulins are the only wheat proteins that exhibit the
salting in and salting out effects. Prolamins are cereal proteins that are soluble in 70% ethanol
whereas some glutenins are soluble in dilute acids and bases.

37

2.6. BAKED PRODUCTS
The most common baked products available on the world market include breads, cakes,
pastries, cookies, crackers, muffins, and several other products. It is important to note that all the
products mentioned above have one ingredient in common: it is wheat flour. Most of the baked
products have wheat in the form of flour as a major ingredient. Wheat is one of the most
extensively produced and consumed cereals in the world (Shenoy and Prakash 2002). Couvain
and Young (2006) define baked products as “foods manufactured from recipes largely based on or
containing significant quantities of wheat or other cereal flours which are blended with other
ingredients, are formed into distinctive shapes and undergo a heat processing step which involves
the removal of moisture in the oven.”
Wheat is subdivided into hard and soft wheat. Hard wheats are high in gluten proteins and
therefore preferred for bread making. On the other hand, soft wheats are low in gluten proteins
and are thus good for non-bread products such as cakes, cookies, crackers, muffins, and other soft
wheat-based products.
Wheat bran is regarded as the major source of dietary fiber in baked products. However,
addition of wheat bran during the breadmaking process causes major changes in dough properties.
These changes can lead to detrimental effects such as reduced bread volume and increased crumb
hardness (Rosell and Foegeding 2007). Incorporation of wheat bran during cookie making results
in poor quality cookies including reduced spread during baking (Uysal et al 2007; Gujral et al
2003; Tangkanakul et al 1995).
The basic raw material for most cereal-based baked products is wheat in the form of flour.
Wheat flour obtained after a dry milling process is required as a starting point of most bakery
processes. With baked products, wheat flour has to have other raw materials added to it to form

38

doughs/batters before baking. The other ingredients combined with wheat flour in baked products
recipes impart considerable changes to the functionality of wheat flour. For example, sugar
restricts availability of water which reduces gluten-forming potential of wheat proteins and
modifies starch gelatinization as well. Sugar also acts as a substrate for yeast to produce carbon
dioxide that is responsible for the rising of the dough during proofing to produce a leavened
produt (Cauvain 2001). In addition, sugar provides sweetness to a variety of baked products
including bread, cake, muffins, and biscuits.
In general, hard wheat flour is widely used for making leavened bread whereas soft wheat
flour is suitable for non-leavened baked products such as cookies and cakes. The quality of enduse baked products depend on the physicochemical properties of flour used (Cauvain 2001).
Addition of fiber to bread or cookie formulation affects the physicochemical properties and
sensory attributes of baked products (Nelson 2001).

39

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40

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of wheat, barley, and oats. Cereal Chem. 70(6):712–5.
Wang Q., Pagan J., and Shi J. 2002. Pectin from Fruits. Page 263 in: Functional Fruits. J.Shi, G.
Mazza, and M. LeMaguer, eds. CRC Press: Boca Raton, FL.
Whister R. L and BeMiller J. N. 1997. Carbohydrate Chemistry for Food Scientists, Eagan press,
St Paul, MN.
Wood, P. J., Anderson, J. W., Braaten, J. T., Cave, N. A., Scott, F. W., Vachon, C. 1989.
Physiological effects of β-D-glucan-rich fractions from oats. Cereal Food World 34(10):878882.
World Agricultural Outlook Board. 2012. http://www.usda.gov/oce/commodity/wasde/latest.pdf
Yue, P., and Waring, S. 1998. Resistant starch in Food Applications. Cereal Food World 43:
690-695.
Zhang, D., and Moore, W. R. 1997. Effect of Wheat Bran Particle Size on Dough Rheological
Properties. J. Sci. Food Agric. 74:490-496.
Zook, E. G., Green, F. E., and Morris, E. R. 1970. Cereal Chem. 47:720-731.

48

CHAPTER 3
PHYSICOCHEMICAL PROPERTIES OF WASHED WHEAT BRAN

49

3.1. ABSTRACT
Wheat bran, a by-product of roller milling during the milling process of wheat, contains
substantial amounts of residual starch that may interfere with the analysis of bran’s
physicochemical properties. The specific objectives of this study were to develop a method that
washes away most of the starch adherent to milled wheat bran and to investigate the effects
washing has on the physicochemical properties (such as water binding capacity) and composition
(including insoluble dietary fiber, soluble dietary fiber, total dietary fiber) of washed and nonwashed wheat bran. Soft white wheat bran was washed with distilled water at room temperature
and mixed with a modified Servodyne mixer to wash residual starch away from bran. The branstarch slurry was transferred into a SoyCow presser lined with a filter cloth and rinsed to remove
as much starch as possible. The washed and non-washed bran samples were dried overnight at
60oC and ground to pass through 1000 or 425 µm screens. Washing significantly reduced starch
adherent to wheat bran by 76% (w/w), changed contents of insoluble dietary fiber and soluble
dietary fiber from 39 to 69% (w/w) and from 4.93 to 1.68% (w/w), respectively. Water binding
capacity was higher for washed bran, and was not affected by bran particle size. The transition
onset and peak temperatures of washed wheat bran samples were significantly higher than the
counterpart values of non-washed bran samples. On the other hand, transition enthalpies of
washed bran samples were lower than those of non-washed bran samples.

50

3.2. INTRODUCTION
In recent years, dietary fiber has attracted significant attention due to health care officials’
and nutritionists’ recommendations that consumption of dietary fiber can help to maintain good
health (McCleary 2007). Dietary fiber reduces the risk of cardiovascular disease, certain forms of
cancer, and constipation (Park et al 2005). Dietary fibers are categorized as insoluble and soluble
dietary fibers and each plays a different role in human health. Insoluble dietary fiber is important
for proper bowel function (Anderson 1985; Anderson and Eastwood 1986; Cummings et al 1978)
and may reduce symptoms of chronic constipation, diverticular disease, and hemorrhoids (Painter
and Burkitt 1971; Burkitt 1977; Cummings et al 1979). On the other hand, soluble dietary fiber is
associated with reduction in cholesterol levels and attenuation of blood glucose (Kantor et al
2001; Drecker et al 2002; Schaafsma 2004; Jones 2008). The physiological functions of dietary
fibers are related to their physicochemical properties such as water binding capacity and
distribution of insoluble and soluble dietary fibers (Schneeman 1999).
Wheat bran, a by-product of roller milling during the milling process, is rich in dietary
fiber and contains substantial amounts of protein, minerals and vitamins (Ortho and Shellenberger
1971). It is used by the baking industry to increase dietary fiber in baked products especially of
bread (Pomeranz et al 1977; Vetter 1988). It is important to know that in wheat bran, the amount
of soluble dietary fiber is very low compared to that of insoluble dietary fiber, which implies that
the physiological effects of wheat bran, such as proper bowel function, are attributable to the
insoluble dietary fiber. Increased stool weight and short intestinal transit times are affected by the
hydration properties of insoluble dietary fiber (Hill 1983). The ways in which fiber-containing
samples are prepared determine the hydration properties of that fiber, for example, the amount of
water a fiber sample can hold in its matrix. This, in turn, can also influence a fiber’s physiological

51

functions within the gastrointestinal tract (Rasper 1982). Wheat bran obtained after roller milling
contains significant amounts of residual starch still adherent to it. This residual starch, if not
removed, can interfere with the analyses that are used to determine the composition and
physicochemical properties of wheat bran. Therefore, the major objectives of the present study
were to develop a washing method that would remove as much residual starch as possible and
then study the effect of washing and particle size distribution on composition and
physicochemical properties of relatively pure wheat bran.

52

3.3. MATERIALS AND METHODS
3.3.1. Wheat Bran Sample
The non-washed wheat bran sample used in this study was milled from soft white wheat of
the 2009 crop and supplied by Star of the West Milling Company (Frankenmuth, MI, USA). The
o

sample was stored at 4 C until the studies were conducted.

3.3.2. Washing of wheat bran
Washing of wheat bran was performed in batches. For each batch, 500 g of non-washed
wheat bran was added to 5 L of distilled water at room temperature in a 5 gal white plastic bucket
(Paragon Molding Company, Melrose Park, IL, USA) and mixed with an electronic Servodyne
mixer (Cole-Palmer Instruments Company, Vernon Hills, IL, USA) at a speed of 150 rpm for 30
min to wash residual starch away from bran. The Servodyne mixer was modified by adding
mixing paddles as depicted in Figure 3.1. After mixing, the wheat bran and starch slurry was
transferred into a stainless steel SoyCow presser (ProSoya Inc., Ottawa, Canada) with a filter
cloth (170 µm) laid inside the presser according to the manufacturer’s manual. The tap of the
presser was opened to drain the starch slurry. Wheat bran on the filter cloth was rinsed with 5 L
distilled water a second time to remove as much residual starch as possible. The washed bran
(WB) and non-washed bran (NWB) were spread on 45 cm x 66 cm x 2.54 cm aluminum baking
o

trays (WearEver, Millville, NJ, USA) and dried overnight at 60 C in a Proctor dryer (Proctor &
Schwartz, Inc., Philadelphia, USA). Dried WB and NWB samples were ground using a hammer
mill (Model D Comminuting Machine, W.J. Fitzpatrick Company, Chicago, Illinois, USA) to
pass through a 1270 µm screen (Part Number 1532 0050, Model DAS06, W.J. Fitzpatrick
Company, Chicago, Illinois, USA). NWB and WB samples were then sifted through 1000 µm
53

(US mesh #18) or 425 μm (US mesh #40) sieves (Great Western Manufacturing Company,
Leavenworth, Kansas, USA) to obtain NWB1000, NWB425, WB1000 and WB425. The samples
were placed in one-gallon ziplock plastic bags and stored at 4ºC until required for analysis.
Hereafter, the ground samples are referred to as washed or non-washed bran samples.

54

Figure 3.1. Modified servodyne mixer used to wash wheat bran. For interpretation of the
references to color in this and all other figures, the reader is referred to the electronic
version of this dissertation.

55

3.3.3. Determination of Total Starch
Total starch content in washed and non-washed bran was determined in triplicate
according to the procedure in the Total Starch Assay Kit (Megazyme International Ireland Ltd.
Co., Wicklow, Ireland). Briefly, approximately 100 mg of bran sample were placed in 17 mL
glass test tubes followed by addition of 0.2 mL of 80% (v/v) ethanol to aid in dispersing the
sample. Three mL of thermostable α-amylase (3,000 U/mL of Ceralpha reagent) diluted (1:30) in
sodium acetate buffer (100 mM, pH 5.0) were immediately added to the samples and the tubes
were incubated in a boiling water bath (Blue M, Blue Island, IL, USA) for 6 min with vortexing at
2, 4, and 6 min. The tubes were then placed in a water bath (Blue M, Blue Island, IL, USA) at
50ºC and 0.1 mL of amyloglucosidase (3300 U/mL of soluble starch) was immediately added to
each tube. The samples were incubated for 30 min. After incubation, the contents of the test tubes
were transferred to 100 mL volumetric flasks and the volume adjusted with deionized water to
100 mL. The contents of the volumetric flasks were then transferred to 150 mL beakers and 3 mL
of the contents were placed in plastic centrifuge tubes and centrifuged at 1800 x g for 10 min at
25ºC. For each sample, a 0.1 mL aliquot of the clear supernatant was pipetted to the bottom of a
15 mL test tube, followed by addition of 3 mL of glucose oxidase-peroxidase-aminoantipyrine
o

(GOPOD) reagent. The samples were incubated in the water bath at 50 C for 20 min. A
spectrophotometer (Spectronic 5, Spectronic Instruments Inc., Rochester, NY, USA) was used to
measure the absorbance for each sample at 510 nm against the reagent blank (0.1 mL of deionized
water and 3 mL of GOPOD reagent). Total starch (%) on a dry weight basis was calculated based
on formulas outlined in the Total Starch Kit.

56

3.3.4. Thermal Properties of Non-Washed and Washed Wheat Bran Samples
The thermal properties of non-washed and washed wheat bran samples were studied by a
Differential Scanning Calorimetry (DSC Model Q100 V9.9 Build 303, Greifensee, Switzerland).
Each sample was weighed into a DSC aluminum pan and 20 µL of distilled water were added
using a micro-syringe. The sample was hermetically sealed and the moisture allowed to
o

equilibrate overnight at room temperature. Samples were heated from 20 to 200 C at the heating
o

rate of 10 C/min. A sealed empty pan was used as a reference. Transition onset temperature (To),
transition peak temperature (Tp), and transition enthalpy (∆H) were recorded and analyzed using
DSC software (Universal V4. 7A, TA Instruments, Newcastle, DE, USA).

3.3.5. Water Binding Capacity of Non-Washed and Washed Bran
Water binding capacity of non-washed and washed wheat bran samples was measured in
triplicate according to AACCI Approved Method 56-30 (AACCI 2000) with some modifications.
30 mL of deionized water were added to approximately one gram of bran sample in pre-weighed
plastic centrifuge tubes. The tubes were vortex-mixed to ensure that all the bran particles were
thoroughly wetted. The samples were allowed to hydrate for 30 min with hand shaking after 10,
20, and 30 min. The tubes were centrifuged (Model J2-21M, Beckman Instruments Inc.,
o

Fullerton, CA, USA) at 5000 x g for 30 min at 20 C. The supernatants were carefully decanted
and the tubes were inverted for 10 min to allow free drain. The tubes containing sediments were
then weighed and the difference between the dry and wet weights was calculated as the water
binding capacity.

57

3.3.6. Determination of Insoluble, Soluble, and Total Dietary Fiber Content
Insoluble, soluble, and total dietary fiber contents in washed and non-washed bran were
measured according to the procedure in the Total Dietary Fiber Assay Kit (Megazyme
International Ireland Ltd. Co., Wicklow, Ireland). The kit contained three enzymes, thermostable
α-amylase (3000 Ceralpha Units/mL), protease (350 Tyrosine Units/mL), and amyloglucosidase
(3300 Units/mL of soluble starch), that were used to hydrolyze and depolymerize starch after it
was gelatinized, solubilize and depolymerize proteins, and hydrolyze starch fragments to glucose,
respectively. MES/TRIS buffer, 0.05M, was prepared by dissolving 19.52 g 2-(N-Morpholino)
ethanesulfonic acid hydrate (MES) (M8250, Sigma-Aldrich, St. Louis, MO, USA) and 14.2 g
TRIS (hydroxymethyl) aminomethane (T1503, Sigma-Aldrich, St. Louis, MO, USA) in 1.7 L
deionized water; the pH was adjusted to pH 8.2 with 6.0 N NaOH. The buffer was then diluted to
o

2 L with deionized water and its pH adjusted to 8.3 at 20 C. Other reagent grade chemicals used
for total dietary fiber assay included ethanol (KOPTEC, King of Prussia, PA, USA), acetone
(Sigma-Aldrich, St. Louis, MO, USA) and hydrochloric acid (EMD Chemicals Inc., Gibbstown,
NJ, USA).
To determine insoluble, soluble, and total dietary fiber contents, approximately one gram
of ground bran was weighed into a 400 mL tall-form beaker followed by addition of 40 mL of
MES/TRIS buffer. The beakers were gently swirled until bran particles were completely dispersed
in the buffer solution. 50 μL of heat-stable α-amylase were added to the beakers containing
samples. The beakers were swirled, covered with aluminum foil, and incubated in a boiling water
bath (Blue M, Blue Island, IL, USA) for 35 min with continuous shaking. After incubation with
o

heat-stable α-amylase, the samples were cooled to 60 C and any rings around beakers were
scraped down with a spatula and rinsed with 10 mL of deionized water. To every sample, 100 μL
58

of protease were added before the samples were incubated for 30 min in a shaking water bath at
o

60 C. The samples were then removed from the water bath and 5 mL of 0.561 N HCl were added
to each sample to bring the pH from 4.1 to 4.8. After pH adjustment, the samples were subjected
o

to 200 μL of amyloglucosidase and incubated for 30 min in a shaking water bath at 60 C. After
amyloglucosidase treatment, the samples were filtered through cleaned [Micro-90 concentrated
cleaning solution (Z281506, Sigma-Aldrich, St. Louis, MO, USA)] Pyrex Gooch Crucibles
(CLS329450, Sigma-Aldrich, St.Louis MO, USA) containing 0.5 g celite (C8656, Sigma-Aldrich,
St. Louis, MO, USA). Each residue was washed twice with 10 mL of deionized water preheated
o

to 70 C, and the filtrate and water washings were saved for soluble dietary fiber analysis. Each
residue was then washed twice with 10 mL of 95% ethanol and 10 mL acetone. The crucibles
containing washed residues were dried overnight in the convection oven (Model 737F, Fisher
o

Scientific, Itasca, IL, USA) at 103 C. After drying, the samples were weighed and one was used
to determine protein content while the other was incinerated to determine ash content. Percent
insoluble dietary fiber was calculated using the formula stated in the Total Dietary Fiber
Megazyme Kit.
Soluble dietary fiber was determined by adding four volumes of 95% ethanol preheated to
o

60 C to filtrate and washings from the insoluble dietary fiber step and allowing soluble fiber to
precipitate for 60 min at room temperature. The soluble fiber precipitate containing enzyme digest
was filtered through crucibles containing celite. In sequence, the residue was washed twice with
15 mL of 78% ethanol, then twice with 15 mL of 95% ethanol, and finally twice with 15 ml
acetone. The crucibles containing soluble fiber residues were dried overnight in the convection
o

oven (Model 737F, Fisher Scientific, Itasca, IL, USA) at 103 C. The percentage of soluble dietary
59

fiber was calculated using the formula stated in the Total Dietary Fiber Megazyme Kit. Total
dietary fiber in washed and non-washed samples was calculated as the sum of insoluble dietary
fiber plus soluble dietary fiber.

3.3.7. Proximate Compositions of Non-Washed and Washed Bran
Ash and moisture contents of washed and non-washed bran samples were determined
according to AACCI Approved Methods 08-01 and 44-19, respectively (AACCI 2000). The
protein contents in non-washed and washed bran were determined by Leco Nitrogen Combustion
Analyzer (Model FP-2000, Leco Inc., St. Joseph, MI, USA). A factor of 5.7 was used to calculate
crude protein. Total fat contents were determined using Soxhlet extraction apparatus according to
AACCI Approved Method 30-25 with modifications. Samples were placed in 30 mm x 80 mm
thimbles and extracted for 24 hr. Following extraction, petroleum ether was removed from the
o

sample by a rotary evaporator. The fat remaining in round bottom flasks was dried at 100 C in the
convection oven (Model 737F, Fisher Scientific, Itasca, IL, USA) for 1 h.

3.3.8. Statistical Analysis
All statistical analyses were performed using SAS version 9.2 (SAS Institute Inc., Cary,
NC, USA). Analysis of variance (ANOVA) was performed using the general linear model (GLM)
procedure to determine significant differences among the samples. Means were compared using
Fisher’s Least Significant Difference (LSD) procedure. Significance was defined at the 5% level.

60

3.4. RESULTS AND DISCUSSION
3.4.1. Total Starch
Washing decreased total starch contents in NWB425 and NWB1000 by over 70% (Table
3.1). Total starch was significantly higher in NWB425 than in NWB1000, and in WB425 than in
WB1000. The higher value for total starch content in NWB425 may be caused by flour that ends
up in the bran fraction during the process of flour milling. Starchy endosperm that is loosely
attached to the bran may also easily pass through the larger sieves (e.g., 1000 µm) during sifting
and ultimately increase total starch content in NWB425. It is difficult to compare total starch
results of washed bran to published data because this is the first study to look at washed wheat
bran. Ralet et al (1990) and Xie et al (2008) measured total starch of native wheat bran and
obtained 18.60% and 17.96%, respectively, which are within the range of total starch contents of
both NWB425 and NWB1000 in the present study.

61

Table 3.1. Total Starch Composition* of Non-Washed and Washed Wheat Bran Samples
Bran Sample
Total Starch (%, w/w)
NWB425
23.41 ± 0.56a
NWB1000
14.43 ± 0.41b
WB425
5.52 ± 0.19c
WB1000
4.11 ± 0.16d
NWB425: Nonwashed bran ground to pass through a 425µm screen.
NWB1000: Nonwashed bran ground to pass through a 1000µm screen.
WB425: Washed bran ground to pass through a 425µm screen.
WB1000: Washed bran ground to pass through a 1000µm screen.
*Values followed by the same letter in the same column are not significantly different from each
other (p<0.05).
Values are means of three determinations ± standard deviation.

62

3.4.2. Effect of Washing on the Thermal Properties of Non-Washed and Washed Wheat
Bran Starch Samples
The onset transition temperatures (To) and peak transition temperatures (Tp) of washed
bran samples (WB425 and WB1000) were significantly higher than those of non-washed bran
samples (NWB425 and NWB1000), whereas the transition enthalpies (∆H) of washed bran
samples were significantly lower than those of the non-washed bran samples (Table 3.2). Goering
and DeHaas (1972) and Seib (1994) pointed out that the onset and peak transition temperatures of
starch granules are affected by the particle size distribution, with smaller particle starch granules
(B-type starch) exhibiting lower gelatinization temperatures, whereas large particle size granules
(A-type starch) exhibit higher gelatinization temperatures. However, in the present study, the
differences in the gelatinization onset temperatures and gelatinization peak temperatures of nonwashed and washed bran starch samples are thought to be caused by the differences in the water
binding capacities among the bran samples (Table 3.3). The fact that washed wheat bran samples
bind more water than non-washed bran samples may explain why the onset and peak transition
temperatures of washed wheat bran starch were higher than those of non-washed wheat bran
starch. It is possible that the ability of washed wheat bran to absorb more water reduces the water
required for gelatinization of washed wheat bran starch, thereby increasing onset and peak
transition temperatures. In the present study, the differences in the values of ∆H in non-washed
and washed bran samples may be caused by different amounts of total starch present in these
samples (Table 3.1). The higher the concentration of starch in the sample, the greater the energy
required for gelatinization to take place, thereby increasing the values of ∆H. The lower values of
total starch (Table 3.1) and transition enthalpy in washed bran samples indicate that the method

63

developed in the present study to wash away most of residual starchy endosperm from wheat bran
was effective.

64

Table 3.2. Effect of Washing on the Thermal Properties* of Non-Washed and Washed
Sample**
To (oC)
Tp (oC)
∆H (J/g)
NWB425
60.66 ± 0.14b
66.94 ± 0.13a
2.83 ± 0.25a
NWB1000
60.04 ± 0.09a
67.13 ± 0.23b
1.95 ± 0.18b
WB425
63.53 ± 0.02c
67.62 ± 0.18c
0.51 ± 0.01c
WB1000
64.22 ± 0.06d
67.63 ± 0.02c
0.53 ± 0.01c
*To: transition onset temperature; Tp: transition peak temperature; ∆H: transition enthalpy; values
followed by the same letter in the same column are not significantly different from each other
(p<0.05).
**For explanation of abbreviations, see Table 3.1.
Values are means of three determinations ± standard deviation.

65

3.4.3. Water Binding Capacity of Non-Washed and Washed Bran Samples
Water binding capacity (WBC) refers to the amount of water that a quantity of dry sample
retains after centrifugation (Nelson 2001). The WBC was significantly greater for the larger
particle size bran (Table 3.4). For the non-washed bran samples, the larger particle size (1000µm)
had a WBC 31% greater than that of the smaller particle size (425µm) bran sample. The WBC of
non-washed bran samples are consistent with those of Cadden (1987), Blackwood et al (2000),
and Zang and Moore (1997) who reported increases in WBC with increasing wheat bran particle
size. Washing increased the WBC of WB1000 by 28% and of WB425 by 51% relative to their
counterpart NWB samples. However, WBC was not significantly affected by particle size of the
washed wheat bran samples. The higher WBC of NWB1000 as compared to NWB425 was
probably related to the presence of greater amounts of soluble dietary fiber in non-washed wheat
bran (Table 3.3). Eastwood et al (1983) indicated that the manner in which various cereal fibers
bind water is very important for stool bulking effect. Because insoluble fibers absorb water in the
manner of a sponge (Oakenfull 2001), washing to remove starchy endosperm from the bran
results in the exposure of previously blocked pores and sponge-like cell structures, which may
enable the WB particles to hold more water. Robertson and Eastwood (1981) reported that fibers
that loosely bind water increased stool weight whereas those that strongly bind water had little or
no effect on stool weight.

66

Table 3. 3. Water Binding Capacity (WBC)* of Non-Washed and Washed Wheat Bran
Bran Sample**
WBC (g of water/1g of dry sample)
NWB425
2.67 ± 0.07c
NWB1000
3.90 ± 0.04b
WB425
5.47 ± 0.07a
WB1000
5.43 ± 0.20a
*Values followed by the same letter in the same column are not significantly different from each
other (p<0.05).
**For explanation of abbreviations, see Table 3.1.
Values are means of three determinations.

67

3.4.4. Insoluble Dietary Fiber, Soluble Dietary Fiber and Total Dietary Fiber
The insoluble dietary fiber (IDF), soluble dietary fiber (SDF), and total dietary fiber
(TDF) contents of NWB425, NWB1000, WB425, and WB1000 are listed in Table 3.4. The
dietary fiber contents of NWB425 and NWB1000 are in the range of those reported by Kahol et al
(2000), Nandini and Silimath (2001), Esposito et al (2005), Bilgicli et al (2007), and Claye et al
(1996). There is no published data available for comparison of dietary fiber contents of washed
bran, since this is the first study to determine dietary fiber contents of WB. The IDF content in
NWB1000 was 21.5% higher than in NWB425. Also, larger particle size (1000µm) had a 21%
higher contents of SDF than that of the smaller particle size (425µm). IDF content was
significantly higher in WB425 and WB1000 than in NWB425 and NWB1000 by 42.3% and
27.3%, respectively. SDF contents were also significantly reduced by washing. SDF was 50.5%
lower in WB425 compared to NWB425, whereas SDF was decreased by 65.9% in WB1000
relative to NWB1000. The increase in IDF contents in the washed bran samples was mainly due
to the loss of starch, water-soluble proteins, and other soluble polysaccharides during washing.
Significant decreases in SDF contents in washed wheat bran compared to non-washed bran
indicate that the residual starch still adherent to wheat bran after roller milling contains significant
amounts of soluble fiber. These results are in agreement with Dexter and Wood (1996) who
reported variations in insoluble and soluble dietary fiber contents in different layers of the wheat
kernel. Understanding how dietary fiber varies in different layers can help in developing
processing techniques that can alter physicochemical properties and the proportion of insoluble
and soluble dietary fiber in wheat bran (Dexter and Wood 1996).

68

Table 3.4. Dietary Fiber Composition* of Non-Washed and Washed Wheat Bran samples
Bran Sample**

Insoluble Dietary
Soluble Dietary Fiber
Total Dietary Fiber
Fiber (%,w/w)
(%, w/w)
(%, w/w)
NWB425
38.77 ± 0.28c
3.90 ± 0.16b
42.67 ± 0.12c
NWB1000
49.40 ± 0.14b
4.93 ± 0.01a
55.33 ± 0.20b
WB425
67.25 ± 0.21a
1.93 ± 0.05c
69.18 ± 0.23a
WB1000
68.00 ± 1.27a
1.68 ± 0.09c
69.68 ± 0.75a
*Values followed by the same letter in the same column are not significantly different from each
other (p<0.05).
**For explanation of abbreviations, see Table 3.1.
Values are means of four determinations ± standard deviation.

69

3.4.5. Proximate Compositions (dry weight basis) of Non-Washed and Washed Wheat Bran
Washing decreased protein content from 14.7% in NWB425 to 11.5% in WB425 and from
15.7% in NWB1000 to 13.6% in WB1000 (Table 3.5). It is clear from the results that washing of
milled bran resulted in a net decrease in protein. The decrease in protein content in washed bran
samples suggests that some protein was removed along with residual endosperm adherent to
wheat bran during the process of washing. However, it is important to understand that most of the
protein in washed bran comes from non-protein nitrogen. In general, fat contents were neither
significantly different for NWB425 and WB425 nor for NWB1000 and WB1000. Fat contents in
washed and non-washed bran samples are consistent with findings by other authors (Caprez et al
1986; Gualberto et al 1997). However, fat content was higher in NWB1000 and WB1000 than in
NWB425 and WB425. It is possible that large particle sized bran contains more of the germ
portion of wheat than the smaller particle sized bran. Further studies would be needed to confirm
this phenomenon.
In general, the ash contents were significantly different among the NWB425, WB425,
NWB1000, and WB1000 samples, but significantly higher in the larger particle sized bran
samples. Washing significantly affected ash contents in wheat bran samples. Ash content
increased from 4.48% in NWB425 to 5.38% in WB425 and from 6.73% in NWB1000 to 6.98%
in WB1000. The lower ash contents in NWB425 and NWB1000 may be attributed to higher
amounts of flour in these bran samples (i.e., a dilution effect). The NWB ash values in the present
study are comparable to those reported by Shenoy and Prakash (2002) and Gualberto et al (1997).

70

Table 3.5. Proximate Composition* of Non-Washed and Washed Bran Samples
Bran sample**
Moisture (%, w/w) Protein (%, w/w) Fat (%, w/w)
Ash (%, w/w)
NWB425
2.89 ± 0.04a
14.70 ± 0.02b
4.65 ± 0.04ab
4.48 ± 0.01d
NWB1000
3.15 ± 0.21b
15.67 ± 0.15a
5.19 ± 0.03a
6.73 ± 0.03b
WB425
3.00 ± 0.05b
11.50 ± 0.08c
3.78 ± 0.06b
5.38 ± 0.03c
WB1000
2.86 ± 0.05a
13.58 ± 0.08d
5.16 ± 0.08a
6.98 ± 0.02a
*Values followed by the same letter in the same column are not significantly different from each
other (p<0.05).
**For explanation of abbreviations, see Table 3.1.
Values are means of three determinations ± standard deviation.

71

3.5. CONCLUSIONS
The washing method described in this removed a significant amount of starchy endosperm
adherent to wheat bran after milling. Washing removes starchy endosperm, thereby modifying
thermal properties of the washed bran components. Washing significantly changed the contents of
IDF and SDF. Particle size was not associated with significant differences in TDF, IDF and SDF
contents of washed wheat bran. The water binding capacity of NWB425 was significantly lower
than that of NWB1000. Water binding capacities of WB425 and WB1000 were not significantly
different from each other, but significantly higher than those of their counterpart non-washed bran
samples.

Overall, the results from this study indicate that the presence of residual starchy

endosperm on milled wheat bran can easily interfere with analyses to determine the bran samples’
physicochemical properties and composition.

72

LITERATURE CITED

73

LITERATURE CITED
AACC International. 2000. Approved Methods of the AACC, 10th ed. AACCI, St. Paul, MN.
Anderson, J.W. 1985. Dietary Fiber in nutrition and management of diabetes. Pages 343-361 in:
Dietary Fiber: Basic and Clinical Aspects. G.V. Vahouny and D. Kritcheveky, eds. Plenum
Press: NY.
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Dexter, J. E., and Wood, P. J. 1996. Recent Publications of debranning of wheat before milling.
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Drecker, E., Beecher, G., Slavin, J., Miller, H. E., and Marquart, L. 2002. Whole grains as source
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Eastwood, M. A., Robertson, J. A., Brydon, W. G., and MacDonald, D. 1983. Measurement of
water-holding capacities of fiber and their fecal bulking ability in man. Brit. J. Nutr. 50:539.
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granules-starch from Colocasia esculenta. AACCI: St. Paul, MN.
Gualberto, D. G., Bergman, C. J., Kazemzadeh, M., and Weber, C. W. 1997. Effect of extrusion
processing on the soluble and insoluble fiber, and phytic acid contents of cereal brans. Plant
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Guillon, F., and Champ, M. 2000. Structural and physical properties of dietary fibers, and
consequences of processing on human physiology. Food Res. Int. 33:233-245.
Hill, M. J. 1983. Bacteria Dietary Fibre and chronic intestinal disease. Page 255 in: Dietary Fiber.
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variety of grains daily, especially whole grains: a challenge for consumers. J. Nutr. 131:473486.
Mcclearly, B. V. 2007. An integrated procedure for the measurement of total dietary fiber
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Anal. Bional. Chem. 389:291-308.
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sorghum, and bajra with good chapatti/roti (Indian flat bread) making quality. Food
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Park, Y., Hunter, D. J., Spiegelman, D., Bergkvist, L., Berrino, F., Van Den Brandt, P.A.,
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physicochemical properties of wheat bran. J. Cereal Sci. 11:249-59.
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Schneeman, B. O. 1999. Building scientific consensus: the importance of dietary fiber. Am. J.
Clin. Nutr. 25:691-699.
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properties. J Sci. Food Agric. 74:490-496.

76

CHAPTER 4
EFFECTS OF PARTICLE SIZE COUPLED WITH EXTRUSION PROCESSING
CONDITIONS ON DIETARY FIBER PROFILE OF NON-WASHED AND WASHED
WHEAT BRAN

77

4.1. ABSTRACT
Wheat bran, a by-product of roller milling during the milling of wheat, contains
substantial amounts of residual starch that may interfere with the analysis of its dietary fiber
profile. Therefore, the specific objective of the present study was to develop a method (washing)
of removing most of the starch adherent to milled wheat bran and to investigate the effects of
extrusion processing conditions on the contents of soluble dietary fiber and insoluble dietary fiber
in non-washed bran (NWB) and washed bran (WB). NWB and WB samples obtained from
milled soft white wheat were ground to pass through 1000 or 425 µm screens and extruded
through a co-rotating and inter-meshing twin-screw extruder under conditions of varying screw
configurations (low and high shear), feed moisture (25 and 35%), screw speed (100 and 400 rpm),
o

and die temperature (100 and 150 C). The results of the present study indicate that for NWB of
both particle sizes, extrusion processing conditions slightly increased soluble dietary fiber
content, but decreased insoluble dietary fiber content. Certain extrusion processing conditions
slightly increased soluble dietary fiber in both washed bran samples. The insoluble dietary fiber
contents of extrudates made from the larger particle size WB were significantly lower than those
of the counterpart non-extruded WB.

78

4.2. INTRODUCTION
The role of dietary fiber in human nutrition cannot be overlooked. In recent years, dietary
fiber has attracted significant attention due to health care officials’ and nutritionists’
recommendendations that consumption of dietary fiber can help to maintain good health
(McCleary 2007). Dietary fiber reduces the risk of cardiovascular disease, certain forms of cancer,
and alleviates constipation (Park et al 2005). Dietary fibers are grouped into insoluble and soluble
dietary fibers and each plays a different role in human health. Insoluble dietary fiber is important
for proper bowel function and may reduce symptoms of chronic constipation, diverticular disease,
and hemorrhoids, while soluble dietary fiber is associated with the reduction of cholesterol levels
and attenuation of blood glucose (Schaafsma 2004).
Wheat bran, which comprises approximately 15% of the whole wheat kernel, is rich in
dietary fiber. Like any other plant cell wall, wheat bran is composed of cellulose, hemicelluloses,
pectin, and other associated components such as lignin, cutin, and suberin (Devries et al 1999).
Wheat bran is obtained as a by-product of roller milling during the milling process of wheat grain.
It is used to make “all bran” extruded breakfast cereal (Eastman et al 2001) besides being used by
the baking industry as the major source of dietary fiber (Vetter 1988). Although wheat bran is rich
in dietary fiber, the soluble dietary fiber content is lower than that of insoluble dietary fiber. The
range of soluble dietary fiber content in wheat bran is 1.5 to 4.0% (w/w) whereas that of insoluble
dietary fiber is 35 to 48% (w/w) (Kahol and Chow 2000; Nandini and Silimath 2001; Esposito et
al 2005). The dietary fiber profile of wheat bran depends on the source of wheat and the milling
technique used.
Extrusion cooking technology, a common processing technique to produce breakfast
cereals and a variety of ready-to-eat foods including breakfast cereals, snacks, baby foods, and

79

meat analogs (Vasanthan et al 2002), has been used by a number of investigators to study its
effects on the dietary fiber profiles of high-fiber ingredients including wheat bran (Asp 1986;
Theander and Westerlund 1987; Aloe et al 1989; Camire et al 1990). The high temperature, shear
and other variables employed during extrusion processing are believed to alter the architecture of
cell wall polysaccharides (Selvendra and Robertson 1994; Nyman et al 1994). The effect of
extrusion cooking conditions on the dietary fiber profile in wheat bran has been investigated by a
number of researchers. Ralet et al (1990), Wang and Klopfenstein (1993), and Gaulberto et al
(1997) showed that extrusion processing conditions cause disintegration of insoluble dietary fiber
linkages, leading to an increase in soluble dietary fiber. On the other hand, Varo et al (1983)
reported that heat treatment had no effect on dietary fiber distribution in wheat flour.
Previous studies have used non-washed (native) wheat bran to study the effects of
extrusion cooking variables on its dietary fiber profile. However, wheat bran obtained upon roller
milling of wheat has a substantial amount of residual endosperm still adherent to it. During
extrusion cooking, it is possible that the residual endosperm adherent to wheat bran undergoes
certain chemical modifications and interferes with the analyses that are subsequently used to
determine the dietary fiber profile in extruded wheat bran. Therefore, the objective of the present
study was to investigate the effects of bran particle size coupled with extrusion processing
conditions on the contents of soluble dietary fiber and insoluble dietary fiber in both non-washed
and washed wheat bran.

80

4.3. MATERIALS AND METHODS
4.3.1. Wheat Bran Sample
The non-washed wheat bran sample used in this study was milled from soft white wheat of
the 2009 crop and supplied by Star of the West Milling Company (Frankenmuth, MI, USA). The
o

sample was stored at 4 C until the studies were conducted.

4.3.2. Preparation of Washed Wheat Bran
Washing of wheat bran was performed according to section 3.3.2 of this dissertation.

4.3.3. Grinding and Sifting
Dried Non-Washed Bran (NWB) and Washed Bran (WB) samples were ground according to
section 3.3.3 of this dissertation.

4.3.4. Extrusion of Bran Samples
Washed and non-washed wheat bran samples, sifted and collected sequentially through
1000µm and 425µm screens, were extruded in duplicate on a co-rotating and inter-meshing twinscrew extruder (Model MP 19T2-25, APV Baker, Grand Rapids, MI, U.S.A.) with a 19-mm
barrel diameter and a barrel length to diameter ratio (L/D) of 25:1. A twin-screw extruder feeder
system (K-TRON, Pittman, NJ, U.S.A.) was used to feed bran samples at 2 kg/hr through the
hopper into the extruder barrel. The extruder barrel was divided into five heating zones with zone
1 closest to the feed port and zone 5 at the die exit end. The extruder and the feeder were
equipped with control panels that displayed all the processing conditions, such as temperature for
each of the five heating zones, screw speed, pressure, torque, and feed rate. The Brook Crompton
81

E2 Metripump (Hudders Field, England) was used to inject water inside the extruder barrel that
resulted in the desired moisture content during extrusion processing of wheat bran samples.
The effect of extrusion processing variables on the contents of soluble dietary fiber and insoluble
dietary fiber was investigated using a 2 x 2 x 2 x 2 x 2 complete factorial design (Table 4.1), with
two levels of bran particle size (425 µm and 1000 μm), two screw speeds (100 and 400 rpm), two
o

temperatures (100 and 150 C), two moisture levels (25 and 35%), and two screw configurations
(low and high). The low and high shear screw configurations used in the present study are
illustrated in Table 4.2 and extrusion processing conditions (moisture, screw speed, and barrel
temperature profile) are depicted in Table 4.3. The low shear screw configuration consists of more
conveying screw elements whereas the high shear screw configuration consists of more mixing
o

screw elements. Extruded samples were dried in a convection oven overnight at 60 C. Grinding
of extruded samples was accomplished using a Udy Cyclone mill (Udy Corp., Fort Collins, CO)
equipped with a 0.5 mm screen. The ground extruded samples were stored in “Ziplock” bags at
o

4 C until further analysis.

82

Table 4. 1 . Experimental Design
Independent Variables
Bran Treatment
Bran Particle Size (µm)
Extruder Shear Configuration*
Extruder Temperature (ºC)
Feed Moisture Content (%)
Extruder Screw Speed (rpm)
*See Table 4.2.

Factors
washed, non-washed
1000, 425
low, high
100, 150
25, 35
100, 400

83

Table 4.2. Screw Elements Used for Low and High Shear Extrusion Configurations
Low Shear Configuration

High Shear Configuration

8 D Twin Lead Screws
o
7 x 30 Forward Kneading Elements
8 D Twin Lead Screws
o
3 x 60 Forward Kneading Elements

8 D Twin Lead Screws
o
7 x 30 Forward Kneading Elements
8 D Twin Lead Screws
o
4 x 60 Forward Kneading Elements

o

3 x 30 Reverse Kneading Elements
2 D Single Lead Screws
o
4 x 60 Forward Kneading Elements
o

3 x 30 Reverse Kneading Elements
2 D Single Lead Screws

o

4 x 30 Reverse Kneading Elements
2 D Twin Lead Screws
o
6 x 60 Forward Kneading Elements
4 x 30 Reverse Kneading Elements
1 D Single Lead Screws
o
7 x 90 Kneading Elements
2 D Single Lead Screws

D: screw diameter (= 19 mm).
One kneading element = 0.25 D.

84

Table 4.3. Extrusion Cooking Conditions* for NWB425, NWB1000, WB425, and WB1000
Feed Moisture (% wb)

Screw Speed (rpm)

Barrel Temperature (ºC) for
Zones 1-5**
25
100
40/60/80/100/100
25
400
40/60/80/100/100
35
100
40/60/80/100/100
35
400
40/60/80/100/100
25
100
40/60/100/140/150
25
400
40/60/100/140/150
35
100
40/60/100/140/150
35
400
40/60/100/140/150
*Same for low and high shear screw configurations (see Table 4.2).
**Zone 5 is nearest the exit die.
NWB425: Nonwashed bran ground to pass through a 425µm screen.
NWB1000: Nonwashed bran ground to pass through a 1000µm screen.
WB425: Washed bran ground to pass through a 425µm screen.
WB1000: Washed bran ground to pass through a 1000µm screen.

85

4.3.5. Determination of Soluble Dietary Fiber and Insoluble Dietary Fiber in Non-Washed
and Washed Wheat Bran
Total dietary fiber content in washed and non-washed bran was measured according to the
procedure in the Total Dietary Fiber Assay Kit (Megazyme International Ireland Ltd. Co.,
Wicklow, Ireland) and modified as stated in section 3.3.5 of this dissertation.

4.3.7. Statistical Analysis
All statistical analyses were performed using SAS version 9.2 (SAS Institute Inc., Cary,
NC, USA.). Analysis of variance (ANOVA) was performed using the general linear model
(GLM) procedure to determine significant differences among the samples. Means were compared
using Fisher’s Least Significant Difference (LSD) procedure. Significance was defined at the 5%
level.

86

4.4. RESULTS AND DISCUSSION
4.4.1. Effect of Bran Particle Size Coupled with Extrusion Processing Conditions Under
Low Shear Screw Configuration on the Contents of Insoluble Dietary Fiber (dry weight
basis) in Extrudates Made from Non-Washed and Washed Wheat Bran
The insoluble dietary fiber contents in non-extruded NWB425 and NWB1000 were 38.8%
(w/w) and 49.4% (w/w), respectively (Table 4.4). The lower value of insoluble dietary fiber in the
NWB425 before extrusion may be attributed to the presence of proportionately more starch
compared with that in NWB1000 (Table 3.1). Extrusion processing slightly decreased the
contents of insoluble dietary fiber in extrudates made from NWB425 for most extruded conditions
studied, except for the sample extruded at 150-25-400 (Table 4.4). The insoluble fiber contents in
all extruded NWB1000 samples were lower than that in the non-extruded NWB1000 sample. The
contents of insoluble dietary fiber in extrudates made from NWB1000 were not statistically
significantly different from each other, except for the sample extruded at 150-35-400, which had
the lowest amount of insoluble dietary fiber (44.4%, w/w) (Table 4.4). The slight decreases in the
insoluble dietary fiber contents in the present study are in agreement with results of previous
investigators (Ralet et al 1990; Lue et al 1991; Wang et al 1993; Quaglia et al 1995; Gualberto et
al 1997) who reported loss of insoluble dietary fiber in wheat bran, upon extrusion processing.
These authors suggested that the decrease in the insoluble dietary fiber during extrusion was
caused by high shear of the rotating screws. However, contradictory results have also been
reported. Siljestrom et al (1986), Asp and Bjorck (1989), Wang et al (1993) and Artz et al (1990)
reported that extrusion processing did not have an effect on the contents of insoluble and soluble
dietary fibers in wheat bran. In the present study, the largest decreases in insoluble dietary fiber
were from 38.8% for NWB425 to 37.3% in ENWB425 (a 4% decrease) and from 49.4% for

87

NWB1000 to 44.4% in ENWB1000 (a 10% decrease) under extrusion conditions listed in Table
4.4.
The effects of extrusion processing conditions on the contents of insoluble dietary fiber in
extrudates made from WB425 and WB1000 are shown in Table 4.5. The insoluble dietary fiber
contents in WB425 were minimally affected by extrusion processing conditions. However, certain
extrusion conditions resulted in very minor increases in the contents of insoluble dietary fiber in
WB425 extrudates. Björck (1984) and Camire et al (1990) reported that one of the possible
mechanisms by which insoluble dietary fiber increases during extrusion is the formation of
polysaccharide-lipid complexes which could not be hydrolyzed by the enzymes used in the
enzymatic-gravimetric procedure of dietary fiber analysis, and thus measure as insoluble dietary
fiber. Vitagoliane et al (2008) suggested three ways through which extrusion processing may
cause increases in insoluble dietary fiber: by (1) starch retrogradation, (2) formation of proteinpolysaccharide complexes, and (3) oxidation of dietary fiber and phenolic compounds. Some of
these mechanisms were suggested for samples containing substantial amounts of starch. It is not
clear if the same mechanisms are exactly applicable to WB425, which had a total starch content
of not more than 5.52%, w/w (Table 3.1, Chapter 3). The increases in insoluble dietary fiber
contents in WB425 extrudates were very small; hence, it is difficult to conclude that these
increases were due to chemical reactions as described by previous authors. Unlike WB425, which
was virtually unaffected by extrusion conditions, the contents of insoluble dietary fiber in
extrudates from WB1000 were significantly lower compared to non-extruded WB1000 (Table
4.5). However, the insoluble dietary fiber contents were not significantly different among the
extruded samples. Since an extruder is considered a form of a grinder, it is possible that larger
bran particles are easily mechanically broken by screw shear during extrusion, thereby decreasing

88

the contents of insoluble dietary fiber. On the other hand, the small bran particles in WB425 may
not be as easily affected by extrusion shear. This probably explains why the contents of the
insoluble dietary fiber in extrudates from WB425 were not significantly decreased by low shear
extrusion processing.

89

Table 4.4. Effects of Extrusion Cooking Conditions under Low Shear Screw Configuration
on the Contents of Insoluble and Soluble Dietary Fibers* of Extrudates Made from
NWB425 and NWB1000 Samples**
NWB425
NWB1000
Extrusion Conditions***
IDF (% w/w) SDF (% w/w) IDF (% w/w) SDF (% w/w)
Non-extruded
38.8 ± 0.3b 3.9 ± 0.2a
49.4 ± 0.1b 4.9 ± 0.1a
100-25-100
37.3 ± 0.7a
5.0 ± 0.3bc
45.6 ± 1.2ab 5.9 ± 0.1bc
100-25-400
37.5 ± 0.3a
4.6 ± 0.2abc 46.1 ± 1.9ab 6.1 ± 0.4c
100-35-100
37.3 ± 0.5a
4.7 ± 0.2abc 45.7 ± 1.0ab 5.8 ± 0.6abc
100-35-400
38.3 ± 0.3a
5.5 ± 0.6c
46.1 ± 1.0ab 5.9 ± 0.2bc
150-25-100
37.3 ± 0.3a
4.8 ± 0.2ab
46.7 ± 1.1ab 6.3 ± 0.1c
150-25-400
39.1 ± 0.3b 4.2 ± 0.5b
44.7 ± 1.6ab 6.0 ± 0.2c
150-35-100
37.3 ± 0.0a
4.5 ± 0.4abc 45.4 ± 1.6ab 5.7 ± 0.7abc
150-35-400
37.3 ± 0.0a
5.0 ± 0.6bc
44.4 ± 1.1a
5.1 ± 0.4ab
*Values followed by the same letter in the same column are not significantly different from each
other (p<0.05).
**IDF: insoluble dietary fiber; SDF: soluble dietary fiber; for NWB425 and NWB1000, see Table
4.3.
***100-25-100 (temperature, degrees C – moisture, % wet basis – screw speed, rpm).
Values are means of four determinations ± standard deviation.

90

4.4.2. Effect of Bran Particle Size and Extrusion Processing Conditions Under Low Shear
Screw Configuration on the Contents of Soluble Dietary Fiber (dry weight basis) in
Extrudates Made from Non-Washed and Washed Wheat Bran
The contents of soluble dietary fiber in NWB425 and NWB1000 before extrusion were
3.90% and 4.94%, respectively (Table 4.4). The lower value of soluble dietary fiber in NWB425
compared with that in NWB1000 may be attributed to the presence of more residual starchy
endosperm in NWB425, which was susceptible to hydrolysis by the α-amylase used in the total
dietary fiber determination protocol. The physical interaction between residual endosperm and the
bran may explain the differences in soluble dietary fiber contents in NWB425 and NWB1000.
Most of the residual endosperm in NWB425 was in the form of discrete flour particles whereas
that in NWB1000 was adhered to bran tissue. It is possible that the residual endosperm adherent
to NWB1000 is higher in soluble dietary fiber than that in the NWB425 sample. Dexter and
Wood (1996) reported that soluble dietary fiber content distribution varies in different layers of
the wheat kernel. The results of the present study indicated that soluble dietary fiber content was
higher in the residual starchy endosperm adherent to the bran of milled wheat.
The soluble dietary fiber contents in most of the extruded NWB425 and NWB1000
samples were higher than in their respective non-extruded samples. The largest percent increase in
soluble dietary fiber with low shear extrusion was from 3.90% to 5.50% (a 29% increase) for
NWB425, and from 4.9% to 6.3% (a 22% increase) for NWB1000; the largest increase in soluble
dietary fiber in NWB425 was obtained when the sample was extruded at the highest temperature
o

(150 C), lowest moisture (25%), and lowest screw speed (100 rpm), whereas that in NWB1000
o

was obtained when the sample was extruded at the lowest temperature (100 C), highest moisture
o

(35 C), and highest screw speed (400 rpm). These samples were both extruded under low shear
91

screw configuration. The results of soluble dietary fiber in the NWB425 and NWB1000 upon low
shear extrusion are comparable to those reported in literature (Caprez et al 1986; Aoe et al 1989;
Ralet et al 1990; Wang et al 1993; Gaulberto et al 1997). Several investigators have suggested
that the increase in the contents of soluble dietary fiber in extruded samples may be caused by
breakage of insoluble dietary fiber, thereby forming small fragments that are soluble in the
aqueous ethanol that is used in the total dietary fiber determination procedure (Camire et al 1990;
Gaulberto et al 1997; Guillon and Champ 2000; Vasanthan et al 2002; Singh et al 2007;
Vitagoliane et al 2008). According to the results of soluble dietary fiber obtained from the present
study, the amount (on a weight per weight basis) by which soluble dietary fiber increased was
higher than the amount of insoluble dietary fiber lost during extrusion (Table 4.4.). It is also
important to note that even for the extrusion condition (150-25-400) that did not change the
amount of insoluble dietary fiber in NWB425 and NWB1000 samples, soluble dietary fiber was
still increased by 7.4%. Therefore, the notion should be questioned that increased soluble dietary
fiber is brought about by breakdown of insoluble dietary fiber. These results suggest that
insoluble dietary fiber breakdown is not the only mechanism by which soluble dietary fiber
increases during extrusion processing. It is possible that the residual endosperm, still adherent to
wheat bran after the milling process, undergoes certain chemical modifications during extrusion
that result in chemical components that are resistant to hydrolysis by enzymes used in the dietary
fiber quantification procedure. This is in agreement with Theander and Westerlund (1987) who
pointed out that extrusion cooking generates 1, 6-anhydrosaccharides that react with starch and
starch-containing products to form new branched glucans that are resistant to α-amylase
hydrolysis.

92

Unlike NWB425 and NWB1000, for which all studied low shear extrusion processing
conditions increased soluble dietary fiber, low shear extrusion of WB425 and WB1000 samples
resulted in either increases or decreases in the quantity of soluble dietary fiber (Tables 4.5).
Although the contents of insoluble dietary fiber in all extruded WB1000 samples were
significantly lower than in non-extruded WB1000, the contents of soluble dietary fiber in
WB1000 extrudates were either higher or lower than non-extruded WB1000. Again, this implies
that the increase in soluble dietary fiber during extrusion processing is not caused merely by
redistribution of insoluble dietary fiber. It is difficult to compare results from the washed bran
samples in this study with those in published literature which mainly focus on non-washed wheat
bran. It is unknown if the cause of the variation in soluble dietary fiber contents of extruded
washed bran samples was due to extrusion processing conditions or whether this reflects
limitations of the current procedure for the determination of dietary fiber, i.e., that the procedure
may not be sensitive enough for samples containing very small amounts of soluble dietary fiber.

93

Table 4.5. Effects of Extrusion Processing Conditions under Low Shear Screw
Configuration on the Contents of Insoluble and Soluble Dietary* Fibers of Extrudates Made
from WB425 and WB1000 Samples**
WB425
WB1000
Extrusion conditions***
IDF (% w/w) SDF (% w/w) IDF (% w/w) SDF (% w/w)
Non-extruded
67.3 ± 0.2a
1.93 ± 0.01de 68.0 ± 1.3a
1.68 ± 0.10bcd
100-25-100
68.6 ± 0.5ab 1.96 ± 0.04de 62.4 ± 0.6b 2.16 ± 0.21e
100-25-400
68.4 ± 0.8ab 2.28 ± 0.04f 63.5 ± 1.7b 1.72 ± 0.16cd
100-35-100
67.6 ± 0.2a
1.73 ± 0.21cd 63.3 ± 0.5b 1.39 ± 0.16abc
100-35-400
68.5 ± 0.3ab 1.92 ± 0.20de 64.0 ± 0.9b 1.91 ± 0.01de
150-25-100
69.7 ± 0.1b 1.60 ± 0.04bc 63.6 ± 1.7b 1.22 ± 0.16a
150-25-400
69.3 ± 1.1b 2.05 ± 0.04ef 63.4 ± 0.3b 1.37 ± 0.11ab
150-35-100
67.5 ± 0.4a
1.28 ± 0.11a 63.5 ± 0.8b 1.28 ± 0.18a
150-35-400
67.5 ± 0.4ab 1.42 ± 0.13ab 63.0 ± 0.2a
1.92 ± 0.12de
*Values followed by the same letter in the same column are not significantly different from each
other (p<0.05).
**See Table 4.4.
***For explanations of abbreviations, see Tables 4.3 and 4.4.
Values are means of four determinations ± standard deviation.

94

4.4.3. Effect of Bran Particle Size and Extrusion Processing Conditions Under Low and
High Shear Screw Configurations on the Contents of Insoluble and Soluble Dietary Fibers
in Extrudates Made from WB425 and WB1000
No relationship was found between either low or high shear screw configuration and the
contents of insoluble or soluble dietary fiber of the extruded samples studied (Figs. 4.1 and 4.2).
Certain extrusion conditions with high shear configurations, increased or decreased the soluble
dietary fiber and insoluble dietary fiber in both non-washed and washed wheat bran. Wang et al
(1993) studied the effect of screw speed during extrusion of non-washed wheat bran milled from
hard red winter wheat. In that study, wheat bran was extruded at low screw speed (100 rpm),
medium screw speed (300 rpm), and high screw speed (400 rpm). They reported that the insoluble
dietary fiber content was significantly decreased from 53.7% (non-extruded wheat bran) to 50.2%
in extrudates made from wheat bran extruded at the highest screw speed (400 rpm). Under the
same condition, the soluble dietary fiber content significantly increased from 1.72% to 4.25%. In
another study, Gualberto et al (1997) investigated the effect of screw speed (50, 70, and 100 rpm)
on the contents of insoluble and soluble dietary fiber in non-washed wheat bran milled from a
Canadian hard red wheat. They reported a slight decrease in insoluble dietary fiber in bran
extrudates extruded at 100 rpm. However, they reported a significant increase in soluble dietary
fiber under the same extrusion condition.

95

Soluble fiber (% db)

3.00
2.50
2.00
1.50
1.00

0.50

Low shear

0.00

High shear

Extrusion conditions*
Figure 4.1. Effects of low and high shear screw configurations (see Table 4.2 for details) on
the contents of soluble dietary fiber in washed wheat bran extrudates.
*For abbreviations, see Tables 4.3 and 4.4.

96

Insoluble fiber (% db)

75.0
60.0
45.0
30.0
15.0
0.0

Low Shear
High Shear

Extrusion conditions*
Figure 4.2. Effect of low and high shear screw configurations (see Table 4.2 for details) on
the contents of insoluble dietary fiber in WB1000 extrudates.
*For abbreviations, see Tables 4.3 and 4.4.

97

4.5. CONCLUSIONS
Extrusion increased the quantity of soluble dietary fiber in both NWB425 and NWB1000
samples. Most of the extrusion cooking conditions studied demonstrated slight decreases in
insoluble dietary fiber of both NWB425 and NWB1000. Certain extrusion processing conditions
slightly increased the soluble dietary fiber in both WB425 and WB1000. The current protocol for
determining dietary fiber content may not be sensitive enough to detect very small amounts of
dietary fiber.

98

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99

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Guillon, F., and Champ, M. 2000. Structural and physical properties of dietary fibers, and
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Kahlon, T. S., and Chow, F. 2000. In vitro binding of bile acids by rice bran, oat bran and, and
corn bran. Cereal Chem. 77: 518-521.
Lambo, A. M., Oste, R., and Nyman, M. E. 2005. Dietary fiber in fermented oat and barley betaglucan rich concentrates. Food Chem. 89:283-293.
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Wang, W. M., Klopfenstein, C. F., and Ponte, J. G. 1993. Effects of twin-screw extrusion on the
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102

CHAPTER 5
EFFECTS OF EXTRUSION PROCESSING ON THE PHYSICOCHEMICAL
PROPERTIES OF NON-WASHED AND WASHED WHEAT BRAN

103

5.1. ABSTRACT
The effects of extrusion processing on water binding capacity of bran, molecular weight of
soluble fiber, thermal degradation properties of bran samples, and in vitro binding of bile acids of
non-washed and washed wheat bran samples were studied. Extrusion processing increased the
water binding capacity of non-washed bran samples but did not have an effect on the water
binding capacity of washed bran samples. The molecular weights of soluble dietary fibers
extracted from non-washed bran samples were significantly higher than those of soluble dietary
fibers extracted from washed bran samples. Extrusion further decreased the molecular weight of
soluble fiber in all wheat bran samples. The onset degradation temperature and peak degradation
temperature of non-washed bran were higher than those of washed bran samples. However,
degradation enthalpy of washed bran was significantly higher than that of non-washed bran.
Extrusion did not affect degradation properties of wheat bran samples. In vitro binding of bile
acids was slightly higher in washed bran samples compared to non-washed bran samples. The
increased bile acid binding by samples with very low soluble dietary fiber suggests that binding of
bile acids in wheat bran may be due to mechanisms other than binding by soluble dietary fiber.

104

5.2. INTRODUCTION
Extrusion cooking is a high-temperature short-time (HTST) processing technique that is
used to produce a variety of food products such as ready-to-eat breakfast cereals, texturized
vegetable protein, pet foods, instant powders, baby foods, expanded products and pasta products
(Ficarella et al 2006; Singh and Singh 2004; Jin et al 1995; Vasanthan 2002). During extrusion,
the sample ingredients are mixed and sheared by rotating screws inside a heated barrel. There is a
possibility that extrusion cooking can result in molecular and structural changes of extruded food
ingredients including wheat bran. These extrusion-induced changes may affect the
physicochemical properties of extruded high-fiber ingredients as well as physiological properties
(Margareta and Nyman 2003).
The high demand for healthy foods by consumers has prompted the food industry to
increase the amount of insoluble and soluble dietary fibers in processed food products (Chawla
and Patil 2010). Consumption of foods high in dietary fibers from different sources has been
shown to reduce risks of cardiovascular disease, obesity (Alfieri et al 1995; Haworth et al 2005),
diverticulitis and hemorrhoids (Gordon 1989; Brown et al 1999; Park et al 2005; Gutkoski et al
2007), and result in effective stool bulking and reduced transit time (Cummings 1997).
Wheat bran is used by the baking industry to increase the amount of dietary fiber in baked
products. It is very important to know that the ratio of soluble dietary fiber to insoluble dietary
fiber in native wheat bran is very low. The risk of cardiovascular disease is decreased with highly
viscous soluble dietary fiber due to its ability to reduce cholesterol in the serum (Park et al 2005).
Ralet et al (1990), Wang and Klopfenstein (1993), and Gaulberto et al (1997) reported that
extrusion cooking increased the amounts of soluble dietary in wheat bran. However, it is not clear

105

if the soluble dietary fiber created by this process has the same bile acid binding properties as the
original soluble dietary fiber.
Bile acids are synthesized from cholesterol in the liver and used in the digestion of lipids.
After aiding in lipid digestion, more than 90% of bile acids are reabsorbed back to the liver by
enterohepatic circulation (Hoffman 1977), where they are reutilized. Binding of bile acids by
cholesterol-lowering drugs and soluble fibers in the duodenum are some of the ways that may
lead to reduction of blood cholesterol, especially of low-density lipoproteins, the type known to
increase the risk of cardiovascular disease (Eastwood and Hamilton 1968; Kritchevsky and Story
1974).
To understand the role of processing on the physiological properties of fibers, it is
important to evaluate the physicochemical properties, such as molecular weight, water binding
capacity, and binding of bile acids. Functional properties of polymers including dietary fibers are
affected by their molecular weights. For example, polymers that are more viscous, in general have
higher molecular weights (Shibanuma et al 1996). High performance size exclusion liquid
chromatography equipped with multi-angle laser-light scattering and refractive index is a
powerful tool that has been widely used to determine absolute molecular weight of soluble
polymers (Aberle et al 1994; Fishman et al 1996).
Processing that involves cooking has been reported to decrease molecular weight of
soluble polysaccharides. Snavberg et al (1995) reported that molecular weight of soluble fiber in
carrots was significantly lower following different types of processing. It is possible that
processing may result in the de-polymerization of fiber polysaccharides. Bjork et al (1984) and
Nyman et al (1987) pointed out that breakage of glycosidic linkages may facilitate fermentation
by the beneficial microorganisms in the colon. However, de-polymerization of soluble fiber

106

polysaccharides may negatively affect their nutritional properties (Albersheim et al 1960).
Cleavage of fiber polysaccharides results in a decreased viscosity that may lower their ability to
alter lipid metabolism. Jenkins et al (1978) pointed out that cleavage of glycosidic linkages of
soluble polysaccharides may decrease their ability to regulate glucose metabolism. Ralet et al
(1991) investigated the effects of extrusion cooking conditions on the water binding capacity of
wheat bran. They reported that extrusion cooking increased the ability of wheat bran to bind more
water compared to non-extruded wheat bran. The ability of fiber to improve bowel movement and
to undergo fermentation in the colon mainly depends on their water binding properties.
No research has been conducted to understand how particle size and extrusion cooking
conditions affect physicochemical properties of wheat bran that has been washed to remove most
of the residual starchy endosperm. Therefore, the overall objective of the present study was to
evaluate the effects of particle size coupled with extrusion cooking conditions on water binding
capacity, thermal degradation, molecular weight, and binding of bile acids of extruded nonwashed and washed wheat bran.

107

5.3. MATERIALS AND METHODS
5.3.1. Raw Material
The wheat bran used in this study was milled from soft white wheat of the 2009 crop and
supplied by the Star of the West Milling Company (Frankenmuth, MI, U.S.A).

5.3.2. Preparation of Washed Wheat Bran
Preparation of washed wheat bran is described in section 3.3.2 of this dissertation. Nono

washed and washed wheat bran samples were stored at 4 C until extrusion studies were
conducted.

5.3.3. Grinding and Sifting of Wheat Bran Samples
Dried non-washed bran (NWB) and washed bran (WB) samples were ground and sifted
according to section 3.3.3 of this dissertation.

5.3.4. Extrusion of Wheat Bran
Extrusion of wheat bran samples is described in section 4.3.4 of this dissertation.

5.3.5. In vitro Binding of Bile Acids
In vitro binding of bile acids in wheat bran samples was determined according to the
method of Camire et al (1993) as modified by Kahlon and Chow (2000). Briefly, conjugated bile
acids:

glycochenodeoxycholic

acid,

glycocholic

acid,

taurochenodeoxycholic

acid,

taurodeoxycholic acid, and taurocholic acid (CBA-1KT, Sigma-Aldrich, St. Louis, MO, USA)
were formulated based on composition of human bile to form a stock solution. To simulate acidic
108

gastric digestion, 100 mg of wheat bran samples were weighed into 17 mL-screw capped tubes
and 1 mL of 0.01N HCl was added to each test tube. The samples were incubated for 1 hr in a
o

shaking water bath maintained at 37 C. After this acidic digestion, the pH was adjusted to 6.3 by
addition of 0.1 mL 0f 0.1N NaOH. To simulate conditions in the small intestine, 4 mL of bile acid
mixture and 5 mL of pancreatin solution (providing amylase, protease and lipase for digestion of
o

samples) were added to each sample and incubated for 1 h at 37 C under continuous agitation.
Samples were transferred to 10- mL Oak Ridge centrifuge tubes and centrifuged (Model J2-21M,
o

Beckman Instruments Inc., Fullerton, CA, USA) at 33,000 x g for 20 min at 25 C. Concentration
of bile acids in the supernatant was determined enzymatically according to the Bile Acid Kit No.
450-A (Trinity Biotech, Bray, Wicklow, Ireland).

5.3.6. Determination of Weight-Average Molecular Weight of Soluble Fiber Using HighPerformance Size Exclusion Liquid Chromatography
The soluble fiber powder used for determining weight-average molecular weight was
prepared according to the procedure described in section 3.3.4 of this dissertation. The weightaverage molecular weights of soluble fibers from non-washed, washed, extruded non-washed and
extruded washed bran samples were determined using high performance size exclusion liquid
chromatography (HPSELC, Agilent 1200, Agilent Technologies, Santa Clara, CA, USA).
HPSELC analyses were conducted using an Ohpak SB-804 HQ column (Shoko America Inc.,
CO, USA) with packing materials having pore diameters of 10 µm. A Wyatt miniDawn TREOS
multi-angle laser light scattering detector (Wyatt Tech. Corp., Santa Barbara, CA, USA) and a
Wyatt Optilab refractive index detector (Wyatt Tech. Corp., Santa Barbara, CA, USA) were used

109

to measure light scattering and refractive index values, respectively. The laser sources of these
detectors were operated at 658 nm. Each sample of soluble fiber powders was mixed with
deionized water (2.5 mg/mL) and filtered through a 0.2 µm syringe filter prior to injection. The
o

HPSELC analyses of the soluble fiber isolated from bran samples were performed at 70 C for 30
min, with sample concentrations of 2.5 mg/mL and eluent flow rates of 0.5 mL/ min. De-ionized
water was used as the solvent. The data processing was performed using the ASTRA 5.3.4.18
software package (Wyatt Tech. Corp., Santa Barbara, CA, USA)

5.3.7. Thermal Decomposition Properties of Wheat Bran Samples
Thermal decomposition properties studies of non-washed, washed, extruded non-washed,
and extruded washed wheat bran samples were studied by a Differential Scanning Calorimetry
(DSC) (Model Q100 V9.9 Build 303, Greifensee, Switzerland). Each sample was weighed into a
o

o

DSC aluminum pan and heated and from 20 to 400 C at the heating rate of 10 C/min. A sealed
empty pan was used as a reference. Onset decomposition temperature (To), peak decomposition
temperature (Tp), and decomposition enthalpy (∆H) were recorded and analyzed using DSC
software (Universal V4. 7A, TA Instruments, Newcastle, DE, USA).

5.3.8. Water Binding Capacity
Water binding capacity was determined according to section 3.3.5 of this dissertation.

110

5.3.9. Statistical Analysis
All statistical analyses were performed using SAS version 9.2 (SAS Institute Inc., Cary,
NC, USA.). Analysis of variance (ANOVA) was performed using the general linear model
(GLM) procedure to determine significant differences among the samples. Means were compared
using Fisher’s Least Significant Difference (LSD) procedure. Significance was defined at the 5%
level.

111

5.4. RESULTS AND DISCUSSION
5.4.1. Water Binding Capacity
Table 5.1 shows the effects of particle size coupled with extrusion cooking conditions
(low shear screw configuration, temperature, moisture, and screw speed) on the water binding
capacity of washed and non-washed bran samples. Among the non-extruded wheat bran samples,
the water binding capacity of NWB1000 was significantly higher than that of NWB425. The
results of water binding capacities of NWB425 and NWB1000 samples from the present study are
consistent with those reported by others (Kirwani 1974; Connell 1976; Heller et al 1977; Kimura
1977), who reported higher water binding capacity for the large particle size non-washed bran.
They reported that the inability of fine wheat bran to bind more water, as compared to large
particle size bran, may be attributed to lower contents of hydrophilic hemicelluloses in small
particle size fibers. Ralet et al (1990) reported that water binding capacity of wheat bran increases
with increases in insoluble fiber. In the present study, the insoluble dietary content in NWB1000
was higher than that in NWB425 (Table 3.3). The water binding capacities of washed bran
samples were significantly higher than those of non-washed bran samples before extrusion (Table
3.4). Contrary to non-washed bran, water binding capacities of washed bran samples were not
significantly affected by particle size. The possible explanation for this phenomenon is that
removal of starch by washing aids in the exposure of wheat bran interstices, which are easily
filled up with water. The water binding capacity of washed bran samples cannot be compared to
published literature, since this is the first study to look at water binding capacity of washed bran.
Extrusion cooking conditions significantly increased the water binding capacity in
extruded NWB425 and NWB1000. However, there were no significant differences among the
individual extrusion conditions. The increases in water binding capacities in extrudates from

112

NWB425 and NWB1000 may be due to the presence of partially gelatinized starch. Anderson and
Ng (2003) pointed out that the ability of starch to bind water was influenced by extrusion
processing conditions. The water binding capacities of extruded WB425 and WB1000, which
were lower in starch than the NWB samples, were not affected by the extrusion cooking
conditions studied.
The water binding capacity of a particular fiber is an indicator of its impact in the
gastrointestinal tract. Water binding can be affected by the chemical composition and physical
properties of fibers (Guillon and Champ 2000). Thibault et al (1992) indicated that water binding
capacity of fibers can be influenced by the way fibers are processed, for example, by grinding and
extrusion cooking. In the present study, the water binding capacity of the bran samples was
affected by particle size, washing, and extrusion.

113

Table 5. 1. Effects of Extrusion Conditions under Low Shear Screw Configuration on the
Water Binding Capacities of NWB425, NWB1000, WB425, and WB1000 Samples*
Extrusion Conditions**

Water Binding Capacity
NWB425
NWB1000
WB425
WB1000
Non-extruded
2.67 ± 0.07a
3.89 ± 0.04a
5.47 ± 0.07abc 5.43 ± 0.02b
100-25-100
5.51 ± 0.07bc
4.89 ± 0.03e
5.66 ± 0.13bc
5.48 ± 0.04b
100-25-400
5.61 ± 0.16d
4.38 ± 0.04b
5.83 ± 0.02c
5.39 ± 0.03ab
100-35-100
5.50 ± 0.14bc
4.71 ± 0.09d
5.76 ± 0.07c
5.40 ± 0.18b
100-35-400
5.61 ± 0.01d
4.45 ± 0.11bc 5.84 ± 0.10c
5.27 ± 0.02ab
150-25-100
5.54 ± 0.01bc
4.56 ± 0.07cd 5.38 ± 0.04ab
5.86 ± 0.06c
150-25-400
5.34 ± 0.04b
4.46 ± 0.04bc 5.64 ± 0.03bc
5.33 ± 0.14ab
150-25-100
5.48 ± 0.08bc
4.94 ± 0.10e
5.25 ± 0.05a
5.45 ± 0.01b
100-25-400
5.43 ± 0.06bc
4.34 ± 0.08b
5.20 ± 0.02a
5.50 ± 0.05b
*Values followed by the same letter in the same column are not significantly different from each
other (p<0.05).
NWB425: Nonwashed bran ground to pass through a 425µm screen.
NWB1000: Nonwashed bran ground to pass through a 1000µm screen.
WB425: Washed bran ground to pass through a 425µm screen.
WB1000: Washed bran ground to pass through a 1000µm screen.
**100-25-100 (temperature, degrees C – moisture, % wet basis – screw speed, rpm).
Values are means of three determinations ± standard deviation.

114

5.4.2. Effect of Washing, Bran Particle Size and Extrusion on Weight-Average Molecular
Weights of Soluble Fibers Isolated from Wheat Bran Samples
The weight average molecular weights of soluble fibers isolated from NWB425 and
NWB1000 were significantly higher than those of soluble fibers isolated from WB425 and
WB1000 (Table 5.2). The weight average molecular weights of soluble fibers isolated from
extruded NWB425 (ENWB425), ENWB1000, EWB425, and EWB1000 were lower than those of
soluble fibers isolated from their respective counterpart non-washed bran samples (Table 5.2).
The lower weight average molecular weights of soluble fibers isolated from washed bran samples
may be due to the loss of soluble polymers, such as starch, during the washing process. This is
confirmed by comparison of the total starch contents in the non-washed and washed bran samples
(Table 3.1). Snavberg et al (1997) studied the effect of microwave cooking on soluble fiber in
green beans and found that the weight average molecular weight decreased after microwave
treatment. It is possible the extrusion processing conditions break down fiber glycosidic linkages,
resulting in de-polymerization of fiber (Margaretta and Nyman 2003). In another study, Svanberg
et al (1995) reported a decrease in weight average molecular weight of soluble fiber in carrots
following blanching and boiling. The lower weight average molecular weights of soluble fibers
isolated from extruded non-washed and extruded washed bran samples may be due to extrusion
conditions, such as high temperature and shear, which may have led to depolymerization of
soluble polymers. Decrease in weight average molecular weight has nutritional significance.
Lowering the weight average molecular weight of soluble polymers, such as soluble dietary fiber,
may impair their ability to modulate glucose and hormonal responses in the blood (Gustafsson et
al 1995).

115

Table 5. 2. Effect of Washing, Particle Size, and Extrusion on the Weigh Average Molecular
Weight* of Soluble Dietary Fiber in Wheat Bran Samples
Sample*

Weight Average Molecular
5
Weight (x10 )
NWB425
751.0
NWB1000
390.0
WB425
28.3
WB1000
23.6
ENWB425
70.6
ENWB1000
13.7
EWB425
2.2
EWB1000
2.2
*E: Extruded; for all other abbreviations, see Table 5.1.

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5.4.3. Thermal Decomposition of Wheat Bran Samples
Differential Scanning Calorimetry (DSC) is a very important technique that is used to
study thermal properties of polymers. The onset decomposition temperature (To), peak
decomposition temperature (Tp), and decomposition enthalpy values (∆H) of non-washed,
washed, extruded non-washed and extruded washed bran samples are depicted in Table 5.3. From
the results, it is clear that the onset decomposition temperature (To) of NWB1000 was
significantly higher than that of WB1000. Extrusion cooking did not have an effect on the To of
the samples in this study. The Tp values of NWB and ENWB were not significantly different
from each other. Likewise, the Tp values of WB and EWB did not significantly differ from each
other. However, Tp values of WB and EWB were significantly higher than those of NWB and
ENWB, respectively. The decomposition enthalpies of NWB and ENWB were significantly lower
than those of WB and EWB. The higher decomposition enthalpy values for WB and EWB
indicate that more energy is required to decompose washed wheat bran. Based on the fact that the
amount of insoluble dietary fiber is significantly higher in WB samples than in NWB samples
(Table 3.3), it could be speculated that there is proportionately more cellulose in washed wheat
bran than in non-washed wheat bran (weight per weight); cellulose requires substantial amounts
of energy to decompose. The thermal properties of non-washed, washed, extruded non-washed
and extruded washed bran samples cannot easily be compared to previous studies because of
limited reports in the literature on this topic.

117

Table 5.3. Thermal Decomposition Properties* of Non-Washed Bran, Washed Bran,
Extruded Non-Washed Bran, and Extruded Washed Bran Samples
o
o
∆H (w/g)
To ( C)
Tp ( C)
NWB1000
257.8 ± 0.2b
285.8 ± 1.4b
102.8 ± 0.3a
WB1000
250.9 ± 2.8a
279.5 ± 0.3a
140.6 ± 0.1b
ENWB1000
257.2 ± 0.1b
287.3 ± 0.1b
102.6 ± 0.3a
EWB1000
250.2 ± 1.4a
277.2 ± 0.1a
131.3 ± 1.4c
*To: onset decomposition temperature; Tp: peak decomposition temperature; ∆H: decomposition
enthalpy; values followed by the same letter in the same column are not significantly different
from each other (p<0.05).
**For explanation of abbreviations, see Tables 5.1 and 5.2.
Values are means of three determinations ± standard deviation.

Sample**

118

5.4.4. Binding of Bile Acids by Wheat Bran Samples
Cholestyramine significantly bound more bile acids than any of the wheat bran samples
(Table 5.4). Assigning a bile acid binding of 100% to cholestyramine, the relative binding of bile
acids by NWB425, WB425, NWB1000, WB1000, ENWB425, EWB425, ENWB1000, and
EWB1000 were 18.09, 20.31, 19.20, 23.53, 17.09, 22.42, 18.65, and 25.64%, respectively. It is
clear from the results that bile acid binding was higher in washed wheat bran compared to nonwashed bran. Particle size did not have an effect on the binding of bile acids. Extrusion did not
significantly affect binding of bile acids either.
Two mechanisms have been proposed to explain the ability of dietary fibers to decrease
cholesterol in the blood. The first mechanism is that increased amounts of propionate are
produced during microbial fermentation of these fibers and that the propionate inhibits cholesterol
biosynthesis (Anderson et al 1990). The second proposed mechanism is that the presence of
highly viscous fibers in the ileum interferes with bile acid re-absorption (Marlett 1997). Zhang et
al (1992) and Lund et al (1989) reported that increased loss of bile acids from the body may be
attributed to the higher intestinal bacterial count with diets high in fermentable fiber, because
bacteria may bind bile acids in the colon. Marcus and Heaton (1986) and Alberts et al (1996)
reported that wheat bran reduces intestinal transit time and binds bile acids, thereby decreasing
the risks of colon cancer.
According to the results from the present study, washed bran samples bound more bile
acids than non-washed bran samples. The increased bile acid binding by washed bran samples,
which are lower in soluble dietary fiber but higher insoluble dietary fiber (Table 3.3), indicate that
binding of bile acids by wheat bran may be due to mechanisms other than binding by soluble
dietary fiber. These results are consistent with those reported by Kahlon and Chow (2000). They

119

studied in vitro binding of bile acids by rice bran, oat bran, wheat bran, and corn bran and
reported that rice and wheat brans significantly bound more bile acids than oat fiber, even though
oat bran is higher in soluble dietary fiber than rice bran and wheat bran. The soluble dietary fiber
in oat bran has been linked to reduction in serum cholesterol.

120

Table 5.4. In Vitro Binding of Bile Acids by Wheat Bran Samples
Sample*

Bile acid binding**

Cholestyramine
9.01 ± 0.07c
NWB425
1.63 ± 0.14ab
NWB1000
1.73 ± 0.14ab
WB425
1.83 ± 0.27ab
WB1000
2.12 ± 0.27ab
ENWB425
1.54 ± 0.27a
EWB425
1.68 ± 0.14ab
ENWB1000
2.02 ± 0.11ab
EWB1000
2.31 ± 0.22b
*For explanation of abbreviations, see Tables 5.1 and 5.2.
**Values are means of three determinations ± standard deviation.

121

Binding relative to
cholestyramine (%)
100.00c
18.09ab
19.20ab
20.31ab
23.53ab
17.09a
18.65ab
22.42ab
25.64b

5.5. CONCLUSIONS
The extrusion cooking conditions utilized in the present study increased the water binding
capacities of NWB425 and NWB1000. Extrusion cooking had no effect on water-binding
capacities of washed bran samples. The weight average molecular weights of WB samples were
lower than those of NWB samples. Extrusion processing decreased weight average molecular
weights of soluble fiber in both non-washed and washed bran samples. Thermal decomposition
properties of wheat bran samples were not affected by extrusion cooking. Washing did affect
thermal decomposition of wheat bran samples. In vitro binding of bile acids was higher in washed
bran than in non-washed bran. Binding of bile acids was found to be higher in extrudates from
WB425 and WB1000. Overall, in vitro binding of bile acids was higher for the larger bran
particle size samples, for both non-extruded and extruded samples.

122

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123

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CHAPTER 6
EFFECT OF TREATED WHEAT BRAN ON THE BAKING PROPERTIES OF BREAD
AND COOKIES

128

6.1. ABSTRACT
The ability of dietary fibers to provide health benefits has prompted the food industry to
develop a variety of fiber-enriched foods including baked products. The purpose of this study was
to investigate the effects of adding non-washed, washed, extruded non-washed, and extruded
washed wheat bran samples on (1) the Farinograph results and pasting properties of a hard wheat
flour and a soft wheat flour, (2) the quality properties of bread and cookies substituted with 5% or
15% bran sample, and (3) the dietary fiber profile in bread samples. Farinograph water
absorptions of each of the flour samples substituted with wheat bran samples were higher than
that of the respective hard wheat flour, and increased with the increase in wheat bran substitution
level. Pasting properties obtained from the Rapid Visco Analyzer indicated that peak, trough,
breakdown, and final viscosities of hard and soft wheat flour samples containing wheat bran were
lower than those of the respective hard and soft wheat flours. Bread formulations containing
wheat bran resulted in breads with increased loaf weights and decreased loaf volumes.
Substituting soft wheat flour with 5% or 15% bran significantly decreased cookie spread.
Insoluble and soluble dietary fiber distributions were affected by baking conditions. The contents
of insoluble dietary fiber in the crumb and crusts of breads baked from hard wheat flour and hard
wheat flour substituted with various bran samples were significantly higher than those in the
control hard flour. On the other hand, the contents of soluble dietary fiber in the crumb and crusts
of breads baked with hard wheat flour substituted with wheat bran were not significantly different
from each other.

129

6.2. INTRODUCTION
Baked products are a major source of revenue in the United States. Fresh bread and
cookies alone generated 11 billion dollars in 2010 (AIB 2011). Bread, a commonly consumed
baked product all over the world, is made from hard wheat flour in addition to yeast, salt, and
water. Other ingredients that may be included in the formula are fat, sugar, milk, oxidants,
enzymes, surfactants, and other additives to increase the taste, quality, and shelf life of the bread.
It is important to know that flour is the major ingredient in the bread formulation. The proteins in
the flour, when water is added, form a visco-elastic dough that retains gas and results in the
formation of bread. Yeast is also an important ingredient in the bread formulation. It converts
fermentable carbohydrates into carbon dioxide and ethanol, and these gases are responsible for the
production of a leavened loaf of bread. Salt is usually added to bread formulations as a source of
taste as well as an improver of dough rheological properties. Another important ingredient is
water, which acts as a plasticizer and solvent. Without water, there would be no dough and
therefore no viscous flow properties and many of the reactions that normally take place during
fermentation could not occur.
Cookies are different from bread because they are made from soft wheat flour.

In

addition, cookie formulations are high in sugar and fat and low in water. Cookies are popular
baked products because they are convenient and have extended shelf life (Singh and Mohamed
2007).
Wheat is a major ingredient of staple foods, such as breads and noodles for people all over
the world (Posner 2000) and is a good source of protein, vitamins, and minerals besides being an
excellent source of dietary fiber. The positive impact of dietary fiber on human health has
prompted the food industry to develop foods enriched with dietary fiber. Dietary fiber is

130

incorporated in a variety of foods, including baked products, to improve their nutritional benefit.
Wheat bran is one of the major sources of dietary fiber incorporated into baked products. Wheat
bran has the ability to provide numerous health benefits in the gastrointestinal tract (Anderson et
al 1994). Insoluble dietary fiber in wheat bran is more effective in preventing constipation than
that found in fruits and vegetables (Anderson et al 1994). Consumption of wheat bran
significantly increases stool weight and volume (Balasubramanian et al 1987; Jenkins 1987;
Lampe et al 1993) and may possibly reduce the risk of colon cancer (National Research Council
1989; Kroon et al 1997; Cho and Clark 2001).
The challenge encountered by food technologists is that incorporation of wheat bran into
food systems deteriorates overall quality properties of end-use products (Guillon and Champ
2000). Extrusion may modify physicochemical properties of wheat bran and reduce its
deteriorative effects and result in improved quality attributes in processed foods, for example of
baked products, which could ultimately increase human fiber consumption. The main purposes of
the present study were to investigate the effects of adding non-washed, washed, extruded nonwashed and extruded washed wheat bran samples on (1) the Farinograph results and pasting
properties of a hard wheat flour and a soft wheat flour, (2) the quality properties of bread and
cookies substituted with 5% or 15% of the bran samples, and (3) the amounts of insoluble dietary
fiber and soluble dietary fiber in wheat flour and in the crust and crumb of baked bread.

131

6.3. MATERIALS AND METHODS
6.3.1. Materials
Hard wheat flour was obtained from the Mennel Milling Co. (Fostoria, OH, U.S.A). Soft
wheat flour and wheat bran were obtained from the Star of the West Milling Co. (Frankenmuth,
MI, U.S.A).

6.3.2. Preparation of Washed Bran
Wheat bran was washed according to the procedure described in section 3.3.2 in this
dissertation.

6.3.3. Grinding and Sifting
Dried Non-washed Bran (NWB) and Washed Bran (WB) samples were ground according
to section 3.3.3 of this dissertation.

6.3.4. Preparation of Extruded Wheat Bran
Extruded wheat bran samples were extruded according to section 4.3.4 of this dissertation.

6.3.5. Farinograph Properties
Farinograph properties of hard wheat flour samples containing 0%, 5% (w/w) or 15%
(w/w) of non-washed, washed, extruded non-washed, or extruded washed wheat bran were
determined using a 50-g mixing bowl Farinograph according to the AACCI Approved Method
54-21 (AACCI 2000). Values of water absorption, dough development time, stability, and mixing
tolerance index were obtained from the Farinograms.

132

6.3.6. Pasting Properties
The pasting properties of hard and soft wheat flour samples containing 0%, 5% (w/w) or
15% (w/w) of non-washed, washed, extruded non-washed, or extruded washed bran were
determined using a Rapid Visco Analyzer (RVA Model 4, Newport Scientific Inc., Warriewood,
Australia). Three grams (at a 14% moisture basis) of each sample were mixed with 25 mL of
distilled water in an RVA cup and mixed in the RVA heating block. Pasting properties of
samples were analyzed using Standard 1 profile according to the AACCI Approved Method 76-21
(AACCI 2000). Pasting data such as peak viscosity (highest viscosity during heating), trough
viscosity (lowest viscosity following peak viscosity), breakdown viscosity (peak viscosity minus
trough viscosity), setback (final viscosity minus peak viscosity), and final viscosity (viscosity at
the end of heating cycle) were processed and obtained by the Thermocline version 1.2 Software
(Newport Scientific Inc., Warriewood, Australia).

6.3.7. Breadmaking
Hard wheat flour (100 g at 14% moisture) was used to make control bread. Breads were
also prepared from wheat flour substituted with 5% (w/w) or 15% (w/w) of non-washed bran,
washed bran, extruded non-washed or extruded washed bran samples. Breadmaking was
performed according to the optimized straight-dough breadmaking procedure (AACCI Approved
Method 10-10B; AACCI 2000) with some modifications. In addition to flour and bran, the bread
formulation contained sugar (6.0 g), salt (1.5 g), shortening (3.0 g), yeast (1.96 g, Safmex,
Milwaukee, WI, USA), ascorbic acid (5.0 mg), and water. The amount of water in the formulation
was calculated based on the Farinograph water absorption. Breads were baked in a rotary oven

133

o

(National MFG Co., Lincoln, NE, USA) at 215 C for 24 min. After baking, breads were removed
from the baking pans and allowed to cool at room temperature for 2 hr before quality properties
were determined.

6.3.8. Determination of Bread Quality Properties
After cooling, the breads were weighed and their volumes determined by the rapeseed
displacement method. Loaves of bread were then stored in sealed Ziplock bags at room
temperature for 24 hr prior to bread firmness determination. Using a slicer (Model FF-3199,
Hobart MFG. Co. Troy, OH, USA), the loaves were cut to obtain two bread slices (each 12.5 mm
thick) from the center of the loaf which were used for firmness analysis. Bread firmness was
determined by compression using a texture analyzer (Model TA-XT2, Texture Technologies,
Corp., Scarsdale, NY, USA) equipped with a 25 kg load cell. The force (N) required to compress
25% (6.25 mm) of two stacked slices of bread was determined with an acryl probe (38 mm
diameter) at a compression rate of 1.77 mm/s. After determining bread firmness, the same two
o

slices were stored in sealed 1 gal Ziplock bags at -20 C until dietary fiber analysis.

6.3.9. Cookiemaking
Cookies from soft wheat flour and soft wheat flour substituted with 5% (w/w) or 15%
(w/w) of non-washed bran, washed bran, extruded non-washed or extruded washed bran samples
were prepared according to the AACC Method 10-54 (AACCI 2000) for Micro Wire-Cut
Cookies. Cookie doughs were baked in a rotary oven (National MFG Co., Lincoln, NE, USA) for

134

o

11 min at 215 C. Baked cookies were placed on a wire rack and cooled for 30 min at room
temperature before determination of quality properties.

6.3.10. Determination of Cookie Quality Properties
The cookie quality properties evaluated after 30 min cooling time included weight,
diameter, and thickness. Cookies were then placed in Ziplock bags and stored for 24 hr before
cookie hardness was determined. Cookie hardness was determined using a texture analyzer
(Model TA-XT2, Texture Technologies Corp., Scarsdale, NY, USA) equipped with a 25 kg load
cell. The cookie was cut into two pieces by a blade (3 mm thickness) at a compression rate of 2.0
mm/s, and the maximum force (N) was recorded.

6.3.11. Determination of Insoluble and Soluble Dietary Fibers in Bread Crumb and Crust
The frozen slices of bread were thawed for 12 hr before crumb and crust samples were
collected. The crumb that was used in this study was taken 2 cm from the edge of the bread slice.
The crust used was 1 cm from the edge of the bread slice. The crumb and crust were torn into
o

pieces by hand, placed in aluminum bowls and dried at 50 C for 18 hr in the conventional oven
(Model 737F, Fisher Scientific, Itasca, IL, USA). The dried pieces of crumb and crust were then
crushed by hand prior to grinding in a Udy Cyclone Mill (Udy Corp., Fort Collins, CO, U.S.A)
o

equipped with a 0.5 mm screen. The ground samples were stored in “Ziplock” bags at 4 C until
insoluble and soluble dietary fibers determination.
Insoluble and soluble dietary fibers in bread crumb and crust were determined according
to section 3.3.4 of this dissertation with a modification. The weights of the dried insoluble dietary

135

fiber and soluble dietary fiber residues were designated as the weights of insoluble dietary fiber
and soluble dietary fiber, respectively. No corrections for ash and protein were made due to the
small amounts of each fiber in the samples.

6.3.12. Statistical analysis
All statistical analyses were performed using SAS version 9.2 (SAS Institute Inc., Cary,
NC, USA.). Analysis of variance (ANOVA) was performed using the general linear model
(GLM) procedure to determine significant differences among the samples. Means were compared
using Fisher’s Least Significant Difference (LSD) procedure. Significance was defined at the 5%
level.

136

6.4. RESULTS AND DISCUSSION
6.4.1. Farinograph
The Farinograph properties of hard wheat flour (control) and hard wheat flour samples
substituted with 5% (w/w) or 15% (w/w) bran are listed in Table 6.1. All the hard wheat samples
that substituted with wheat bran had higher water absorption values than that of hard wheat flour
(control). Farinograph water absorption increased significantly when bran substitution was
increased to 15% (w/w). Water absorption was slightly higher for all samples containing washed
bran as compared to the counterpart non-washed bran. The higher values of water absorption in
hard wheat flour substituted with washed bran may be due to the fact that washed bran samples
bind more water than non-washed bran samples (see water binding capacities of non-washed and
washed bran samples listed in Table 3.3, Chapter 3). These results are consistent with those of
Lang et al (1990) who reported that increasing the amount of high-fiber ingredients in wheat flour
increases Farinograph water absorption. The dough development times of wheat flour samples
substituted with 5% wheat bran samples were not significantly different from that of the control
flour. However, with 15% bran-substitution, the dough development times were significantly
different from that of the control flour and those of the flour samples substituted with 5% bran. It
is possible that the higher amounts of fiber particles in the 15% bran-substituted samples
interfered to greater extents with the hydration of flour proteins and starch by competing for water
(Nelson 2001). The Farinograph stabilities of all flour samples substituted with 5% or 15% wheat
bran samples were significantly lower than that of the control flour (Table 6.1). The lowest
stability was found with flour sample substituted with 15% washed wheat bran. Stability is one of
the dough mixing properties that indicates dough strength, with higher stabilities indicating
stronger doughs and vice-versa. The decrease in Farinograph stability upon addition of cereal

137

brans has been reported by other investigators. Laurikainen et al (1998) reported a decrease in
stability of the dough made from hard wheat flour substituted with 5% rye bran. The highest
mixing tolerance index was displayed by hard wheat flour substituted with 15% washed bran. The
increase in mixing tolerance indices for samples substituted with wheat bran was also reported by
Zhang and Moore (1997) and Shenoy and Prakash (2002), although these authors utilized nonwashed bran.

138

Table 6.1. Farinograph Properties of Hard Wheat Flour Substituted with Bran*
Sample

Water
Dough
Stability (min)
Mixing
Absorption
Development
Tolerance
(%)
Time (min)
Index (BU)
HWF
59.0 ± 0.3a
5.5 ± 0.0b
16.0 ± 0.7f
20.0 ± 0.0a
HWF-5NWB
61.2 ± 0.3b
5.5 ± 0.1b
13.5 ± 0.3d
20.0 ± 0.0a
HWF-5WB
62.2 ± 0.1c
5.0 ± 0.0a
10.0 ± 0.1b
30.0 ± 10.0b
HWF-5ENWB
61.0 ± 0.6b
5.0 ± 0.3a
15.0 ± 0.3e
20.0 ± 10.0a
HWF-5EWB
62.8 ± 0.1d
5.5 ± 0.1b
12.0 ± 0.3c
30.0 ± 0.0b
HWF-15NWB
66.2 ± 0.2e
6.0 ± 0.1c
12.0 ± 0.3c
20.0 ± 0.0a
HWF-15WB
69.9 ± 0.1g
6.5 ± 0.3d
9.0 ± 0.4a
50.0 ± 10.0c
HWF-15ENWB
67.4 ± 0.3f
6.5 ± 0.7d
12.0 ± 0.1c
20.0 ± 0.0a
HWF-15EWB
70.2 ± 0.3h
7.5 ± 0.1e
12.0 ± 0.3c
30.0 ± 0.0b
*Values followed by the same letter in the same column are not significantly different (p<0.05).
HWF: hard wheat flour; NWB: non-washed bran; WB: washed bran; ENWB: extruded nonwashed bran; EWB: extruded washed bran; 5 and 15 are bran substitution levels (%, w/w).
Values are means of two determinations ± standard deviation.

139

6.4.2. Pasting Properties
The pasting data indicate that soft wheat flour had higher peak, breakdown, trough,
setback and final viscosities than hard wheat flour (Table 6.2). These results are consistent with
those of Ragaee and Abdel-Aal (2006). Both soft wheat flour and hard wheat flour samples
containing 5% or 15% wheat bran had lower peak viscosities than the respective control flour
samples. The decrease in peak viscosity in bran-substituted samples is likely due to a starch
diluting effect.
Understanding how high-fiber ingredients such as wheat bran affect pasting properties of
hard and soft wheat flours is very important since starch plays important roles during the
manufacture of baked products, e.g., bread and/or other processes in which starch is heated in the
presence of sufficient amounts of water. During heating, starch granules imbibe water, which
increases viscosity of the food system, a process known as gelatinization. The increase in
viscosity during heating of starch and other processes, such as cooling, are responsible for product
structure, e.g., bread crumb (Artwell 2001; Couvain and Young 2006).

140

Table 6.2. Pasting Properties of Hard Wheat Flour and Soft Wheat Flour Substituted with
Wheat Bran*
Sample

Peak
Trough
Breakdown
Final
Viscosity
Viscosity
Viscosity
Viscosity
(RVU)
(RVU)
(RVU)
(RVU)
HWF
184.4
98.8
67.2
224.4
HWF-5NWB
151.5
101.1
50.4
218.8
HWF-15NWB
121.9
78.9
43.0
193.8
HWF-5WB
165.4
105.6
59.8
221.5
HWF-15WB
119.2
78.7
41.2
173.2
HWF-5ENWB
145.7
95.7
50.0
196.1
HWF-15ENWB
115.2
79.6
35.6
172.3
HWF-5EWB
117.6
82.2
35.3
168.5
HWF-15EWB
113.8
77.0
36.8
160.8
SWF
196.0
127.8
85.6
270.8
SWF-5NWB
109.2
56.3
52.8
143.3
SWF-15NWB
108.3
60.3
48.1
156.8
SWF-5WB
125.8
64.5
61.3
156.3
SWF-15WB
86.7
45.7
41.0
115.5
SWF-5ENWB
140.5
72.5
60.5
172.2
SWF-15ENWB
98.5
53.8
44.8
135.7
SWF-5EWB
130.8
74.9
55.8
165.2
SWF-15EWB
105.8
54.3
51.4
136.6
*SWF: Soft Wheat Flour; for all other abbreviations refer to Table 6.1.

141

Setback
(RVU)
125.6
117.8
114.9
115.9
94.6
100.4
92.7
86.2
83.8
142.0
87.0
96.6
91.8
69.8
99.8
81.9
90.2
82.5

6.4.3. Bread Quality Properties
Bread formulations containing 5% or 15% bran samples resulted in significantly lower
loaf volumes than bread made from control flour (Table 6.3). There were no significant
differences in bread volumes among breads made from flour samples substituted with 5% of nonwashed, washed, extruded non-washed, or extruded washed bran. The loaf volumes of bread
samples made from flour samples substituted with 15% WB were significantly lower than those
substituted with 15% NWB. The bread volumes of bread samples made from flour samples
substituted with 15% ENWB were significantly lower than those made from flour samples
substituted 15% NWB. Breads made from hard wheat flour substituted with 15% EWB had the
lowest bread volumes.
Addition of various high-fiber ingredients in bread formulations has been reported to
decrease loaf volume (Hamid and Luan 2000; Pomeranz et al 1977; Dalgetty and Baik 2006;
Hung et al 2007). Results obtained in the present study are consistent with those of Dalgetty and
Baik (2006). Pomeranz et al (1977) reported that reduction in bread loaf volume in formulations
with added fiber ingredients was caused by gluten dilution that ultimately reduces gas retention.
Mujoo and Ng (2003) reported that addition of immature wheat meal (i.e., proportionately high in
fiber) to the bread formulation decreased loaf volume. They suggested that reduction in loaf
volume was probably due to reduced gas retention. In the present study, it can be speculated that
the decrease in loaf volumes of breads made from flour substituted with 15% WB and 15% EWB,
as compared to their counterpart NWB and ENWB samples, is due to their higher water binding
capacities, (Table 5.1, Chapter 5 of this dissertation). It is possible that the presence of washed
and extruded-washed wheat bran in the dough samples compete for water with flour proteins
gliadins and glutenins, and impair formation of gluten network, which is responsible for gas

142

retention during fermentation and baking. In addition to gluten dilution caused by direct
replacement of flour, the presence of wheat bran in the system may interfere with the
development of the gluten network, thus impairing the ability of the dough to retain gas.
Increasing the concentration of wheat bran in bread formulation increased bread weight. This
could be caused by the ability of wheat bran to bind more water than wheat flour on a weight by
weight basis.
Crumb firmness values of all breads containing 5% or 15% added bran were significantly
higher than that of the control. There were significant differences in crumb firmness among all the
samples supplemented with 5% or 15% wheat bran. Crumb firmness values of all breads
containing 15% added bran were significantly higher than of the control and the breads baked
from formulations containing 5% bran. The effect of high-fiber ingredients on crumb firmness
was also studied by Hamid and Luan (2000). They reported that crumb firmness of bread baked
from hard wheat flour was significantly lower than that of hard wheat flour containing 5%
FIBREX, a commercial fiber.

143

Table 6.3. Quality Properties of Bread*
Sample

Bread Weight (g)

Firmness (N)

Bread Volume
(cm3)
HWF
127.5 ± 4.7a
2.92 ± 0.01a
840.0 ± 14.1f
HWF-5NWB
137.9 ± 0.7b
4.19 ± 0.16d
697.5 ± 3.5e
HWF-5WB
139.4 ± 2.2c
3.70 ± 0.08c
695.0 ± 7.1e
HWF-5ENWB
139.5 ± 2.0c
3.12 ± 0.10b
692.0 ± 3.5e
HWF-5EWB
140.7 ± 2.0d
4.49 ± 0.08e
695.0 ± 7.1e
HWF-15NWB
144.7 ± 1.6e
4.62 ± 0.19f
645.0 ± 7.1d
HWF-15WB
151.7 ± 2.4f
5.97 ± 0.08g
630.0 ± 7.1c
HWF-15ENWB
147.9 ± 1.5g
6.57 ± 0.25h
617.5 ± 3.5b
HWF-15EWB
151.9 ± 2.0h
9.36 ± 0.11i
610.0 ± 14.1a
*Values followed by the same letter in the same column are not significantly different (p<0.05);
for abbreviations, refer to Table 6.1.
Values are means of two determinations ± standard deviation.

144

6.4.4. Cookie Quality Properties
Diameter, height, and hardness of cookies baked from soft wheat flour substituted with
5% or 15% of non-washed, washed, extruded non-washed and extruded washed bran samples are
listed in Table 6.4. Cookie weights were not significantly affected by adding bran samples to
cookie formulations. Cookie diameter decreased whereas cookie height increased with increasing
bran substitution. Cookie hardness was higher for cookies made from soft wheat flour containing
5% or 15% bran samples compared to the cookies made from control flour. The results of the
present study are in agreement with those obtained by Uysal et al (2007), Gujral et al (2003), and
Tangkanakul et al (1995). They reported that increasing the concentration of apple fiber in the
cookie formulation results in decreased cookie spread. The diameters of cookies with 4, 8, or 12
% (w/w) added apple fiber were significantly smaller than the control (Chen et al 1988). Chen et
al (1988) reported that addition of 12% wheat bran decreased cookie spread. James et al (1989)
found that adding rice bran to cookies reduced their crispness and their diameters.
The effect of wheat bran to decrease cookie spread may be related to its water binding
properties (Vratania and Zabik 1978; Gorczyca and Zabik 1979; Jeltema et al 1983; Sievert et al
1990; Artz et al 1990; Chen et al 1988). In the present study, the reduced diameters of cookies
baked from soft wheat flour containing wheat bran may be caused by the higher water binding
capacity of wheat bran. It is possible that during baking, binding of water by wheat bran leaves
less water in the system, water that would otherwise be used to dissolve sucrose and increase
viscosity, which is responsible for cookie spread.

145

Table 6.4. Effect of Bran Substitution on Quality Properties of Cookies*
Sample
Weight (g)
Diameter (cm) Height (cm)
Hardness (N)
SWF
20.82 ± 0.03ab
8.09 ± 0.02f
0.91 ± 0.01ab
27.6 ± 0.8a
SWF-5NWB
21.56 ± 0.04ab
7.68 ± 0.05d
0.93 ± 0.01bc
32.3 ± 1.3abc
SWF-5WB
20.18 ± 0.03a
7.46 ± 0.02c
1.01 ± 0.01d
29.0 ± 1.2ab
SWF-5ENWB
20.85 ± 0.03ab
7.74 ± 0.02e
0.94 ± 0.02c
32.3 ± 1.3abc
SWF-5EWB
21.76 ± 0.02b
7.29 ± 0.06b
1.01 ± 0.02d
30.4 ± 0.8abc
SWF-15NWB
21.65 ± 0.04b
7.76 ± 0.02e
0.89 ± 0.01a
29.5 ± 1.6ab
SWF-15WB
21.36 ± 0.02ab
7.14 ± 0.05a
1.03 ± 0.02d
32.4 ± 0.0abc
SWF-15ENWB
21.47 ± 0.02ab
7.68 ± 0.03d
0.94 ± 0.02c
35.8 ± 1.9c
SWF-15EWB
21.74 ± 0.02b
7.11 ± 0.03a
1.08 ± 0.02e
34.0 ± 1.30bc
*Values followed by the same letter in the same column are not significantly different (p<0.05);
SWF: Soft
Wheat Flour; for all other abbreviations refer to Table 6.1.
Values are means of two determinations ± standard deviation.

146

6.4.5. Effect of Baking on the Insoluble and Soluble Dietary Fibers in Bread
The contents of insoluble and soluble dietary fiber presented in Figures 6.1 and 6.2 were
calculated based on equal weight of flour on a dry weight basis. According to the results
presented in Figure 6.1, the contents of insoluble dietary fiber in crumb and crust of bread made
from hard wheat flour were not significantly different from each other but significantly higher
than insoluble dietary fiber in hard wheat flour (HWF). The contents of insoluble dietary fiber in
crumb and crust of bread made from wheat flour substituted with 15% wheat bran (HWF-15B)
were significantly higher than the control unbaked flour substituted with 15% bran (HWF-15). On
the other hand, baking significantly increased the contents of soluble dietary fiber with higher
increases reported in crust than in crumb (Fig. 6.2). For the hard wheat flour substituted with 15%
wheat bran, baking significantly increased soluble dietary fiber in the bread crust whereas the
soluble dietary fiber was not significantly different from that in the control (HWF-15, Fig. 6.2).
Vitaglione et al 2008 reported that starch gelatinization and retrogradation together with
the formation of protein-polysaccharide complexes through Maillard reactions could be
responsible for the increases in insoluble dietary fiber in processed foods, as long as long as
processing conditions are favorable for chemical reactions to take place. Wang et al (1993)
reported that high processing temperature could disrupt covalent and non-covalent bonds in the
carbohydrate-protein complexes resulting in smaller fragments with increased solubility. The
results from this study reveal that dietary fiber distribution varies within a loaf of bread with the
crust containing more soluble fiber than the crumb. Substituting a certain portion of wheat flour
with wheat bran is one way to increase insoluble dietary fiber in food products. However, addition
of wheat bran to hard wheat flour is not the best way to increase soluble dietary fiber in bread.

147

This may be due to the limited amount of soluble dietary fiber present in wheat bran. Overall,
substituting wheat flour with wheat bran increases total dietary fiber in bread.

148

% insoluble fiber (w/w)

16.50
15.00
13.50
12.00
10.50
9.00
7.50
6.00
4.50
3.00
1.50
0.00

15.19
13.46
10.00
crust
5.72

crumb

5.19

2.38

HWF

HWFB

HWF-15 HWF-15 B

Figure 6.6. Insoluble dietary fiber in flour samples and its quantity and distribution in
subsequent baked bread.
HWF: hard wheat flour; HWFB: bread baked from hard wheat flour; HWF-15: hard wheat flour
substituted with 15% bran (w/w); HWF-15B: bread baked from hard wheat flour substituted with
15% bran (w/w).

149

4.50
% soluble fiber (w/w)

3.84

3.77

4.00
3.50
3.00
2.50
2.00

2.32

2.18 2.07

crust
crumb

1.50

1.50
1.00
0.50
0.00
HWF

HWFB

HWF-15 HWF-15 B

Figure 6.7. Insoluble dietary fiber in flour samples and its quantity and distribution in
subsequent baked bread.
For abbreviations, refer to Fig. 6.1.

150

6.5. CONCLUSIONS
The results from the present study show that addition of wheat bran to bread and cookie
formulations affect the physicochemical properties of the doughs and endproducts. Addition of
wheat bran increased Farinograph water absorption of hard wheat flour. Also, addition of bran to
hard and soft wheat flours decreased the pasting viscosities, as analyzed by the Rapid Visco
Analyzer. Bread and cookie formulations containing bran resulted in lower loaf volume and
cookie spread, respectively. Breads containing 15% bran had lower volumes compared to those
containing 5% bran. Increasing the concentration of wheat bran to 15% decreased loaf volume
and increased bread firmness. Increasing wheat bran in the cookie formula did not significantly
affect cookie hardness. The contents of insoluble and soluble dietary fiber were higher in bread
than in control flours.

151

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155

CHAPTER 7
GENERAL CONCLUSIONS

156

Wheat bran, a by-product of dry milling of wheat, contains substantial amounts of
minerals and vitamins and is rich in dietary fiber than any other part of a wheat kernel. It is
technically impossible to obtain bran free of endosperm during the process of roller milling of
wheat as there is always residual starchy endosperm still adherent to the bran. Previous studies on
the effect of processing, such as particle size distribution and extrusion processing conditions on
the composition, physicochemical, and physiological properties of wheat bran, have been
conducted using non-washed wheat bran. The neglect by previous investigators of the residual
starchy endosperm adherent to wheat bran motivated the present author to investigate the effect of
this adherent starchy endosperm on dietary fiber composition, physicochemical properties, and
functionality of wheat bran. Therefore, the specific objectives of the present study were: (1) to
develop a method that removes most of the residual starchy endosperm still adherent to the milled
wheat bran, (2) to investigate the effects of particle size of non-washed and washed wheat bran
coupled with extrusion processing variables on insoluble and soluble dietary fiber contents of the
bran, (3) to investigate the effects of extrusion cooking conditions on the physicochemical
properties of extruded non-washed and washed wheat bran samples, and (4) to evaluate the effects
of treated wheat bran on the baking properties of bread and cookie products.
A washing method was developed to remove as much as possible of the residual starchy
endosperm still adherent to wheat bran. Washed bran was then ground to pass through 1000 µm
(WB1000) and 425 µm (WB425) screens. Non-washed bran ground to pass through 1000 µm
(NWB1000) and 425 µm (NWB425) screens was used as controls. The washing method
developed in the present study decreased total starch content in washed bran samples by over
70%. Transition enthalpies of washed bran samples, as measured by Differential Scanning
Calorimetry, were much smaller compared to those obtained from non-washed bran samples,

157

which indicated that the washing method removed significant amounts of residual starchy
endosperm. The insoluble dietary fiber contents in WB1000 and WB425 were significantly higher
than those in NWB1000 and NWB425. On the other hand, soluble dietary fiber contents were
significantly lower in washed bran samples than in non-washed bran samples. Total dietary fiber
in NWB1000 was significantly higher than that it NWB425. However, the insoluble, soluble, and
total dietary fibers in WB1000 and WB425 were not significantly different from each other. The
water binding capacity was significantly higher in NWB1000 than in NWB425. On the other
hand, the water binding capacities of the washed wheat bran samples were not significantly
affected by particle size of the bran. Based on the findings of the present study, it is evident that
the presence of residual starchy endosperm on milled wheat bran can easily interfere with
analyses that are used to determine a bran sample’s composition and physicochemical properties.
An extrusion study was conducted to investigate the effects of extrusion cooking on the
contents of insoluble and soluble dietary fibers in non-washed and washed bran samples. Most of
the extrusion cooking conditions decreased insoluble dietary fiber and increased soluble dietary
fiber contents in NWB425 and NWB1000. On the other hand, extrusion of WB425 did not have a
defined trend with regard to insoluble and soluble dietary fiber contents. Insoluble dietary fiber
contents in extruded WB1000 samples were significantly lower than in the non-extruded
WB1000, but soluble dietary fiber contents either increased or decreased during extrusion. It is
not clear if the fluctuation in soluble dietary fiber contents in washed bran samples was due to the
effects of extrusion conditions or if the level of dietary fiber content present was below the
sensitivity level of the assay used.
The present study also examined the effects of extrusion cooking conditions on the
physicochemical properties, such as water binding capacity, molecular weight, binding of bile

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acids, and thermal degradation, of extruded non-washed and extruded washed wheat bran
samples. The results of the present study indicate that the water binding capacity values of
extruded non-washed bran samples were significantly higher than those of non-extruded
NWB1000 and NWB425. Extrusion conditions increased the values of water binding capacity in
non-washed wheat bran samples. However, certain extrusion conditions increased or decreased
the water binding capacities of washed bran samples. Washing and extrusion cooking decreased
molecular weights of soluble dietary fiber in both non-washed and washed bran samples. Thermal
decomposition properties of wheat bran samples were not affected by extrusion. The onset
decomposition temperature and peak decomposition temperature of washed bran were lower than
those of non-washed bran. On the other hand, decomposition enthalpies of non-extruded and
extruded washed bran were significantly higher than those of non-extruded and extruded nonwashed bran. Extrusion increased in vitro binding of bile acids in extruded NWB1000 and
WB1000.
The results of baking studies indicated that addition of non-washed, washed, extruded
non-washed, and extruded washed wheat bran samples affected the quality properties of baked
bread and cookies. Substituting hard wheat flour with 5% or 15% of wheat bran sample increased
its Farinograph water absorption. Wheat bran was found to reduce the pasting viscosities of hard
and soft wheat flour samples. Breads containing wheat bran had decreased bread volume
compared to the wheat flour-only control. Cookie formulations containing bran produced cookies
with reduced spread. Quality properties of bread and cookies deteriorated with an increase in bran
substitution.
Overall, the findings obtained from the present study indicate that the residual starchy
endosperm still adherent to milled wheat bran affects analyses of the insoluble and soluble dietary

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fiber profile as well as other physicochemical properties of the bran sample. The lower values of
soluble dietary fiber in washed bran indicate that residual starchy endosperm adherent to milled
wheat bran is high in soluble dietary fiber. Expanding upon these findings, it may be possible for
the soluble dietary fiber content in milled wheat bran to be increased by altering the mill settings
as well as by treatment of wheat grain before milling to obtain wheat bran with substantial
amounts of residual endosperm. In addition, this scientific information could be used by wheat
breeders to develop varieties that contain residual endosperm that cannot easily be peeled off
during the milling process. This would increase the amount of soluble dietary fiber content in
high bran-containing cereals, such as “All Bran” cereal. On the other hand, the washing method
developed in this study could be used by the food industry to make pure wheat bran, as
ingredients for novel products.
The findings of this study indicate that the bran’s residual starchy endosperm affects the
dietary fiber profile and physicochemical properties, such as water binding capacity of milled
wheat bran. Wheat bran that was treated to remove most of the residual starchy endosperm
(washed wheat bran) contained substantial amounts of insoluble dietary fiber and had a higher
water binding capacity as compared to non-washed wheat bran. Insoluble dietary fiber content
and water binding capacity greatly affect physiological properties of wheat bran. Therefore,
washed wheat bran could be used as a functional ingredient in products intended to improve
intestinal regulation.
Extrusion processing conditions decreased the contents of insoluble dietary fiber and
increased the contents of soluble dietary fiber in all extrudates made from non-washed bran. On
the other hand, extrusion decreased insoluble dietary fiber but did not increase soluble dietary
fiber in all extrudates made from washed wheat bran. This indicates that the residual starchy

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endosperm still adherent to milled wheat bran undergoes certain chemical modifications during
extrusion cooking to form new components that have properties similar to those of soluble dietary
fiber. This phenomenon contradicts previous investigators who suggested that extrusion shear
results in breakage of the glycosidic bonds of insoluble dietary fiber, thereby increasing soluble
dietary fiber in non-washed bran.
The fact that binding of bile acids was not affected by the amount of soluble dietary fiber
present in the sample does not mean that soluble dietary fiber is unable to decrease serum
cholesterol. There may be other mechanisms by which soluble dietary fiber decreases cholesterol
in the blood. Unlike cholestyramine, which decreases cholesterol in the blood through ionic
binding of bile acids in the intestine, the mechanism by which soluble dietary fiber removes
cholesterol from the body may be different. Because soluble dietary fiber binds water to form a
viscous gel, it is possible that bile acids are physically trapped by the gel and eventually end up in
the feces.
The ability of wheat bran to provide physiological benefits such as fecal bulking is
affected by their its binding capacity. Previous studies have indicated that the fecal bulking
capacity of large particle size bran is higher than that of small particle size bran. The explanation
for this phenomenon is that the water binding capacity of large particle size bran is higher than
that of small particle size. The results of the present study showed that large particle size bran is
higher in insoluble dietary fiber than is small particle size bran. Therefore, it can be speculated
that the increased fecal bulking by large particle size wheat bran is due to its higher insoluble
dietary fiber content. In addition, the results of the present study indicated that particle size of
washed wheat bran had no effect on water binding capacity and the insoluble dietary fiber content
of that bran sample. If fecal bulking depends on the water binding capacity and insoluble dietary

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fiber content, it can be suggested that fecal bulking is not affected by particle size of washed
wheat bran. However, it is important to note that the smaller particle size bran increases surface
area available for bacterial fermentation in the large intestine. Increased fermentation impairs the
ability of fiber to increase fecal bulking.

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CHAPTER 8
FUTURE RECOMMENDATIONS

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1. Injecting water into the extruder barrel during extrusion does not provide enough time for
the water to penetrate the bran. It is recommended that future studies investigate how
extrusion of wheat bran that has been pre-soaked may affect its physicochemical
properties. Extruding soaked wheat bran may increase heat transfer and facilitate cooking
during extrusion, and change the soluble and insoluble fiber distribution.
2. Generally, extrusion conditions changed dietary fiber distribution in non-washed and
washed bran samples. It is recommended that further studies examine the effects of
extrusion conditions on the composition of dietary fiber monosaccharides, such as
rhamnose, arabinose, xylose, mannose, galactose, and glucose.
3. Extrusion processing increased or decreased soluble dietary fiber in washed bran samples.
It is not clear if these were actual changes due to extrusion conditions, or if the protocol
used to determine soluble dietary fiber was not sensitive enough to accurately measure
low levels of soluble dietary fiber. More studies are recommended to examine this
phenomenon.
4. Addition of wheat bran to bread formulation decreased loaf volume. Future studies should
examine effects of bran substitution together with addition of dough enhancers, such as
vital gluten, on quality properties of bread.
5. Nutritional studies have indicated that large particle size non-washed wheat bran increases
stool volume to a greater degree than small particle size non-washed wheat bran. In these
studies, water binding capacity of bran has been identified as an important factor that
affects physiological functions of fibers. The results of the present study indicate that the
water binding capacities of small and large particle size washed brans were not

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significantly different from each other. Therefore, it is recommended that further studies
investigate the effect of particle size of washed wheat bran on fecal bulking.

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