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    Instrumental and Sensory Characteristics of a Baked Product Containing Barley

    Flour with Varying Amounts of Beta-Glucan and Sugar Substitute

    by

    Niti Lathia

    A Thesis submitted to the

    Graduate School-New Brunswick

    Rutgers, The State University of New Jerseyin partial fulfillment of the requirements

    for the degree of

    Master of Science

    Graduate Program in Food Science

    written under the direction of

    Dr. Henryk Daun

    Dr. Paul Takhistov

    and approved by

    ________________________

    ________________________

    ________________________

    New Brunswick, New Jersey

    October 2011

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    ABSTRACT OF THE THESIS

    Instrumental and Sensory Characteristics of Baked Product Containing Barley

    Flour with Varying Amounts of Beta-Glucan and Sugar Substitute

    By Niti Lathia

    Thesis Directors:

    Dr. Henryk DaunDr. Paul Takhistov

    The objective of this study was to determine the influence of varying levels of

    beta-glucan in barley flour on selected properties of a model baked product. Another aim

    was to reduce sugar levels in the product by incorporating a natural sweetener stevia and

    to monitor its influence using instrumental and sensory analysis. Batter rheology was

    studied using a lubricated squeezing flow technique, pasting profiles of the barley flours

    were determined with a rheometer, viscoelastic properties were evaluated using dynamic

    oscillatory rheology to measure G and G, and firmness of the baked products was

    monitored using a texture analyzer, for changes occurring due to varying -glucan levels

    in barley flour and removal of sugar. L a* b* color values of barley flour and muffins

    were obtained using a colorimeter. A descriptive sensory panel was trained to observe

    changes in product attributes when stevia was used to replace sugar in the high beta-

    glucan product.

    Water absorption index was found to be significantly higher for high -glucan

    barley flour. The color of both barley flours also had a significant difference in L*

    (lightness) and b* (yellowness) values. Similarly, muffin samples prepared without

    sugar, using stevia, were significantly lighter in surface color (higher L*), while the

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    interior colors were darker (higher b*). Low beta-glucan dough showed a lower biaxial

    extensional viscosity compared to the high beta-glucan dough, which indicates that the

    level of beta-glucan present in the barley flour has an impact on the dough viscosity. The

    pasting profiles of the flours were also found to be significantly different, where the high

    beta-glucan barley flour resulted in a significantly higher peak viscosity but lower peak

    time compared to low -glucan barley flour. Muffin firmness was found to be

    significantly higher when sugar was omitted from the formulation, but there was no

    significant difference in firmness among the two beta-glucan levels in the muffins. The

    sensory descriptive panel found significantly higher firmness, surface roughness, and

    bitterness attributes for the high -glucan muffins prepared with stevia. Additional

    efforts will be needed to mask the undesirable attributes in the model baked product

    occurring due to the removal of sugar.

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    Acknowledgement

    First and foremost, I would like to express my sincere gratitude towards both of

    my thesis advisors, Dr. Henryk Daun, who brought the project to my attention, and

    equally to Dr. Paul Takhistov whose lab I conducted my research in. Both professors

    have provided guidance, support, encouragement, and have had patience in explaining

    my numerous inquiries throughout the duration of my project research. Secondly, I

    would like to thank Dr. Kit Yam for being on my thesis defense committee, whose input

    and suggestions I value. In addition, a big thank you to my lab mates for their assistancewith learning new instrumentation as well as providing a fun learning environment.

    Also, I appreciate the efforts of the undergraduate team of students that participated in the

    sensory portion of this research as panelists; your help and cooperation was greatly

    appreciated.

    Most importantly, I would like to thank my parents and family for providing the

    financial support for my graduate studies as well as love, encouragement, moral support,

    providing comfort during the challenging times, and accepting my absence while I

    worked towards completing my degree. I have relied on them for guidance and strength

    throughout my academic career. Thank you for your confidence and unwavering support.

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    TABLE OF CONTENTS

    Page

    ABSTRACT OF THESIS ii-iii

    ACKNOWLEDGEMENTS iv

    TABLE OF CONTENTS v

    LIST OF ABBREVIATIONS vii

    LIST OF TABLES viii

    LIST OF ILLUSTRATIONS ix

    LITERATURE REVIEW CHAPTER ONE

    1.0 Introduction 1

    1.1 Health Benefits of Barley 4

    1.2 Chemical and Physical Characteristics of Barley 10

    1.3 -Glucans and Arabinoxylans 13

    1.4 Properties and Molecular Interactions among Major Food Components 17

    1.5 Beta-glucan extraction 17

    1.6 Water absorption capacity and effect on end-products 19

    1.7 Rheological Properties Influenced by barley flour beta-glucan content 21

    1.8 Stevia as a Sweetening Agent in Consumer Products 22

    1.9 Conclusions from Literature Review and Objectives for Research 24

    MATERIALS AND METHODS CHAPTER TWO

    2.1 Ingredients Used in Baking Procedures and Analytical Measurements 26

    2.2 Water Absorption Index of Low and High -glucan Barley Flour 27

    2.3 Microbakery Model Formulations for Barley Muffins 29

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    2.4 Rise and Moisture Loss of Muffins 32

    2.5 Rheological properties of barley dough using lubricatingsqueezing flow technique 33

    2.6 Pasting Properties of High -glucan and low -glucan barley flours 36

    2.7 Dynamic Rheological Properties of Muffin Batter 39

    2.8 Assessing Muffin Firmness Using a Texture Analyzer 40

    2.9 Evaluation of Colors Using a Colorimeter 42

    2.10 Nutritional Comparison of Muffins 44

    2.11 Sensory Methodologies Used to Evaluate Muffin Products 45

    RESULTS AND DISCUSSION CHAPTER THREE

    3.1 Water absorption values for barley flours 50

    3.2 Increase in Muffin Heights After Baking 51

    3.3 Muffin Moisture Loss After Baking 53

    3.4 Rheological Properties of barley flour doughs and muffin batters 55

    3.5 Pasting properties of barley flours 62

    3.6 Color values for Barley Flour Varieties 64

    3.7 Muffin Surface Color 66

    3.8 Muffin Interior Color 71

    3.9 Muffin Firmness 74

    3.10 Nutrition Facts for Muffin Formulations 77

    3.11 High -glucan Muffin Sensory Quantitative Descriptive Analysis 79

    3.13 Conclusions and Suggestions for Future Work 85

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    LIST OF ABBREVIATIONS

    United States Department of Agriculture (USDA)

    National School Lunch Program (NSLP)

    National Health and Nutrition Examination Survey (NHANES)

    Dietary Reference Intakes (DRI)

    Recommended Daily Allowances (RDA)

    Coronary Heart Disease (CHD)

    Low density lipoprotein (LDL)

    Food and Drug Administration (FDA)Code of Federal Regulations (CFR)

    Apparent Biaxial Extensional Viscosity (ABEV)

    The International Commission on Illumination (CIE)

    Rapid Visco Analyzer (RVA)

    Quantitative Descriptive Analysis (QDA)

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    LIST OF TABLES

    Table 1: Nutritional Composition of Sustagrain Barley

    Table 2: Average Nutrient Comparison for Hulless Barley, Oats, and barley variety

    Prowashonupana (Prowash) Barley

    Table 3: Formulations for Muffin Batters Used to Prepare a Model Baked Product

    Table 4: Reference standards for selected attributes used in Spectrum DescriptiveAnalysis panel

    Table 5. Descriptors used to evaluate muffin samples in the QDA panel

    Table 6: Water Absorption indices of high and low -glucan barley flour

    Table 7: Percentage Increase in Muffin Height (Rise) After Baking

    Table 8: Percentage Decrease in Muffin Weight (Moisture Loss) After Baking

    Table 9: Pasting profile for low and high -glucan barley flours

    Table 10: Consistency index and flow behavior index for muffin batter with varyingamounts of beta-glucan and sugar

    Table 11: Average L a*b* Values for low -glucan and high -glucan Barley FlourVarieties

    Table 12: Average L, a*, b* color values for surface color of muffins prepared with lowor high -glucan barley flour with 100% sugar

    Table 13: Average L, a*, b* color values for surface color of muffins prepared with lowor high -glucan barley flour with 0% sugar

    Table 14: The average L, a*, b* color values for interior color of muffins prepared withlow or high -glucan barley flour with 100% sugar

    Table 15: The average L, a*, b* color values for interior color of muffins prepared with

    low or high -glucan barley flour with 0% sugar

    Table 16: Average maximum peak force to compress muffins as a measure of firmness

    Table 17: Nutrition Facts for Muffin Formulations

    Table 18: The mean values of each attribute measured by the QDA panel for muffinsprepared with Sustagrain flour with 100% sugar and 0% sugar, sweetened with stevia

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    LIST OF ILLUSTRATIONS

    Figure 1: Molecular Structure of -(1 3)- and -(1 4)-glucan

    Figure 2: Structural elements present in arabinoxylans

    Figure 3: Extraction and purification of -glucans from barley and oats

    Figure 4: Structural Components of Stevioside, Rebaudioside A, and Steviol

    Figure 5: Nutrition Label for Bobs Red Mill Ground Flaxseed Meal and Bobs Red Millbarley flour

    Figure 6: Example of water separated from barley flour after centrifugation

    Figure 7: Apparatus and set-up of TA.XT2 Texture Analyzer for Lubricated Squeezing

    Flow Technique Analysis of Doughs Prepared Using Barley FloursFigure 8: Typical RVA pasting profile of a normal maize starch for viscosity andtemperature as a function of time

    Figure 9: Water absorption indices of high and low -glucan barley flour

    Figure 10: Percentage Increase in Muffin Height (Rise) After Baking

    Figure 11: Percentage Decrease in Muffin Weight (Moisture Loss) After Baking

    Figure 12: Biaxial Extensional Viscosity as a Function of Biaxial Strain Rate forSustagrain Dough and Bobs Red Mill Barley Flour Dough

    Figure 13: Pasting profile for low and high -glucan barley flours

    Figure 14: Strain vs. shear rate relationship of muffin batter with varying levels of beta-glucan and sugar

    Figure 15: Effect of % Strain on G of muffin batters containing varying amounts of beta-glucan

    Figure 16: Effect of % Strain on G of muffin batters containing varying amounts ofbeta-glucan

    Figure 17: Average L a*b* Values for low -glucan and high -glucan Barley FlourVarieties

    Figure 18: Visual Difference in Flour Color.

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    Figure 19: Surface color values for muffins prepared with low -glucan Bobs Red Millbrand barley flour

    Figure 20: Muffins prepared with low -glucan Barley Flour

    Figure 21: Surface color values for muffins prepared with high -glucan barley flour

    Figure 22: Muffins prepared with high -glucan Barley Flour

    Figure 23: Interior color values for muffins prepared with low -glucan barley flour

    Figure 24: Interior Surface Images of Muffins Prepared with low -glucan barley Flour.

    Figure 25: Interior color values for muffins prepared with high -glucan barley flour

    Figure 26: Interior Surface Images of Muffins Prepared with high -glucan barley flour

    Figure 27: Typical texture profile curve for high -glucan muffins prepared with 100%sugar and 0% sugar

    Figure 28: The average maximum peak force to compress muffins as a measure offirmness

    Figure 29: The mean values of attributes measured by the QDA panel for muffinsprepared with high -glucan barley flour with 100% sugar and 0% sugar, sweetened withstevia

    Figure 30: Sweet steviol glycosides from leaves of Stevia rebaudiana

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    CHAPTER 1

    LITERATURE REVIEW

    1.0 Introduction

    The history of barley usage dates back to approximately 8000 B.C. and is

    considered one of the oldest cultivated crops as it was a mainstay of ancient civilization

    and contributed to the diet of working class people up until the end of the 19 thcentury.

    Many food products such as porridges, broths, hard biscuits, and flat breads were

    prepared utilizing barley. The fermentation of the grain led to the production of varioustypes of alcoholic beverages, beer being one of the most well-known and second highest

    consumed alcoholic beverage today following wine (Jones 2009). Although barley has

    lost its place as a primary staple for modern times, mainly due to the introduction and

    proliferation of the wheat industry, the health benefits and functional food uses are being

    discovered and it is emerging as a major ingredient in current food formulations

    (Anonymous 2005).

    The concept of functional foods has been gaining much attention in the recent

    times. Although there is no legislative definition of a functional food, one of the well-

    accepted definition is Food similar in appearance to conventional food that is intended

    to be consumed as part of a normal diet, but has been modified to subserve physiological

    roles beyond the provision of simple nutrient requirements (Sir, Kpolna et al. 2008).

    Similarly, The Institute of Medicine's Food and Nutrition Board defined functional foods

    as "any food or food ingredient that may provide a health benefit beyond the traditional

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    nutrients it contains" (Hasler 1998). Thus, fiber rich foods can be considered as a

    functional food as it provides health benefits to the consumer.

    One pressing issue in the American schools has been that of childhood obesity.

    Over one-third of children ages 12 to 19 years old are overweight (Ogden, Carroll et al.

    2008) and the prevalence of childhood obesity has increased three-fold from 1980 to

    2000 according to Center of Disease Control Health data Division of Adolescent and

    School Health (2008). The health consequences and social difficulties associated with

    childhood obesity make this a tremendously problematic situation. As a result, many

    institutions and individuals are involved in trying to figure out the sources and solutionsto childhood obesity. National data show that children who participate in the National

    School Lunch Program (NSLP) obtain over half of their daily total food energy from

    school meals. Many social programs have been established at the schools, but the main

    way to target the problem is by targeting food products that are consumed by the school

    children. One such way is to introduce nutritionally-sound and wholesome food products

    into the school lunch programs, which are consumed by children and adolescents

    everyday in the public schools. It is crucial that the product shall maintain quality

    through processing, storage, preparation, and serving. Most importantly, it should

    provide sensory acceptability among children so that it will be fully consumed and thus

    provide the intended nutritional and health benefits.

    The United States Department of Agriculture (USDA) oversees a nationwide

    program called National School Lunch Program (NSLP), which provides and manages all

    breakfast and lunch meals being served to students in the public school system. The goal

    of the program is to provide healthy, nutritious and wholesome food products to school

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    children that choose to consume meals provided at the public schools. As part of the

    NSLP, the USDA sets federal nutrition requirements for the schools to follow when

    creating a menu plan. The nutritional values of the meals served to students are reported

    to the government as a weekly average. Federal nutrition requirements are that the meals

    meet one third of the Recommended Daily Allowances (RDA) for protein, calcium, iron,

    vitamin A and vitamin C as appropriate for the levels for the age group served. The

    meals should also be limited to 30% calories from fat and 10% calories from saturated

    fat. Along with these guidelines, the schools should also try to be consistent with the

    most current Dietary Guidelines of America (2005) (USDA 2009).One prevalent issue is the lack of consumption of fiber rich foods in childrens

    diets. In a study conducted by The National Health and Nutrition Examination Survey

    (NHANES), it was determined that grain based dessert products account for only 5% of

    the total fiber consumption in the diets of children and adolescents between the ages 2-18

    (2010). In 2002, the National Academy of Sciences released the Dietary Reference

    Intakes (DRI) for macronutrients and fiber, which recommended that Americans of all

    ages consume 14 g total fiber per every 1,000 kcal total energy intake, based on evidence

    for reduced cardiovascular disease risk at that level. In addition, fiber protects against

    constipation and has also been shown to have many other health benefits, including

    decreased risk of some cancers, obesity, cardiovascular disease, and diabetes. One

    striking result of this study was that the main sources of fiber in childrens diets were

    foods that were relatively low in fiber density (eg, low-fiber fruits) (Sibylle, Diane et al.

    2005). Thus, it is necessary to develop products with high fiber content which is aimed

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    toward the school lunch program and can contribute to the total daily fiber consumption

    recommendation as given in the DRI.

    1.1 Health Benefits of Barley

    Historically, barley has been used in many Asian, European, and African

    countries in various products such as soups, flat breads, and porridges. Although the use

    of barley declined through the 19thand 20thcentury as wheat and rice became more

    prominent in the global diet, the various health benefits of barley are being discoveredand it is now known to be an excellent source of whole grain. Thus, it is becoming an

    increasingly desirable product to use in formulations (Baik and Ullrich 2008). There are

    numerous health benefits of barley flour, which are predominantly attributed to the fiber

    present in the commodity.

    1.1.1 Fiber content of barley

    Dietary fiber is a major component of whole grains, which has low energy

    density, and has been shown to act as a satiating ingredient. In fact, the current

    recommendation for Americans for daily fiber consumption is between 25-35 g, with a

    quarter of that amount required as soluble fiber (Hecker, Meier et al. 1998). One group

    of dietary fibers, particularly the soluble fibers, is usually viscous or gel-forming.

    Viscous dietary fibers, present in some whole grains such as oats and barley, create

    gastric distention and delay gastric emptying. Subsequently, satiety-related hormones are

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    produced, which signal fullness. Thus, the consumption of whole grains containing

    soluble fibers has been shown to increase satiety and thus thought to reduce overall

    energy intake in a meal. Compared to other grains, barley contains a relatively high

    concentration of beta-glucan, a viscous and fermentable dietary fiber, and therefore may

    be highly satiating. Sustagrain barley, which contains 50% of the soluble fiber as beta-

    glucan, was given to human subjects to study its effect on satiety along with oatmeal and

    rice products, which contain fewer grams of fiber per serving. It was found that the

    barley product, which contained the highest amount of fiber, was the most satiating as it

    left subjects feeling not as hungry for the next meal when compared to meals where therice and oatmeal products were consumed (Schroeder, Gallaher et al. 2009).

    The arabinoxylans present in barley are a source of insoluble fiber. It has been

    found that enzymes present in the colon can specifically hydrolyze arabinoxylans,

    resulting in arabinoxylan oligosaccharides. These oligosaccharides are said to have a

    prebiotic effect, meaning that they promote the growth of beneficial bacteria in the gut.

    Soluble dietary fibers play a role in the reduction of blood cholesterol and postprandial

    blood glucose and insulin. Soluble arabinoxylans may possess these qualities as well

    (Beaver 2008).

    1.1.2 Effect of Barley on Glycemic Index and Insulin Response

    The concept of glycemic index (GI) was first introduced in 1981 as a means for

    identifying and classifying carbohydrate-rich foods based on their ability to raise

    postprandial blood glucose levels. A lower glycemic response is desirable in both

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    healthy and diabetic individuals as it prevents the spiking of blood glucose levels. In a

    study done with South Asian chapati flatbreads which contained varying percentages of

    barley flour mixed with wheat flour, it was hypothesized that high-molecular weight

    barley -glucan added to a food product subjected to mild cooking would be effective in

    lowering postprandial glycemia. As predicted, it was found that the higher levels of -

    glucan resulted in lower serum glucose levels. The fiber may affect glycemic response

    by forming a physical barrier to enzymatic hydrolysis of starch (KNUCKLES, HUDSON

    et al. 1997). This suggests that the addition of high level of -glucan containing barley

    would be beneficial those individuals and populations that have a prevalence of type-2diabetes (Thondre and Henry 2009).

    In another study done with human subjects at a higher risk for insulin resistance,

    they were fed varying amounts of -glucan containing Sustagrain barley. It was found

    that those who consumed 10 g of -glucan in their diet showed significantly lower spikes

    in blood glucose levels, which was measured over a two hour glucose tolerance test. This

    is significant for those that are at risk for developing type-2 diabetes or have insulin

    resistance, which is often seen in obese individuals and nowadays even in children.

    Although a rather larger dose of the barley fiber containing food would need to be

    consumed, it is a reasonable amount and shows that it could be very beneficial for

    individuals who are at a higher risk for developing type 2 diabetes or have insulin

    resistance (Kim, Stote et al. 2009). As this study was done in at-risk women whose

    average age was 51.6, it is possible that the amount of barley -glucan necessary for a

    similar effect on children or healthy individuals with normal blood glucose and regulated

    insulin levels would be much lower.

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    In a similar study done with 10 men who were fed bread products as a breakfast

    meal prepared with three different levels (35, 50 and 75%) of the (13;14)--glucan

    rich barley genotype Prowashonupana, 50% common barley, or 100% white wheat, and

    their postprandial blood glucose and insulin responses were measured. It was found that

    for the subjects that consumed the bread prepared with 50% and 75% Prowashonupana

    barley had a statistically significant reduction in glucose response levels. For bread that

    was prepared with a 50% common barley composition, there was not a significant

    reduction in postprandial glucose levels. This is an important indication that the higher

    percentage -glucan formulated bread had a much more beneficial effect on glucoselevels (stman, Rossi et al. 2006). There is strong evidence from numerous studies that

    high fiber, particularly from -glucan in barley, has a beneficial effect on blood glucose

    levels and thus could delay or prevent the onset of insulin resistance or diabetes.

    1.1.3 Effect of Barley Fiber on Lowering Cholesterol

    The fiber components of barley, particularly the soluble -glucan have been

    shown to have a cholesterol-reducing effect in a study conducted using rats as a model,

    where food products such as tortilla, granola bar, and pudding with added -glucan were

    fed. The soluble fraction, which contains mostly pectin, arabinoxylan, and -glucan, has

    the ability to lower blood serum cholesterol, through its tendency to increase viscosity in

    the intestine, thus affecting the bile acid-cholesterol cycle. Through this mechanism, the

    cholesterol-lowering effects occur by blocking the absorption of fat in the intestines

    (Hecker, Meier et al. 1998). In another study conducted using human subjects who were

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    fed a barley-rich diet, the low-density lipoprotein cholesterol concentration was

    significantly lower at week four in the barley group than in the standard group, who were

    not fed a diet containing barley (Li, Kaneko et al.).

    Coronary Heart Disease (CHD) is the cause of almost 500,000 deaths annually

    and the top risk factors for CHD include high total cholesterol levels and high levels of

    low density lipoprotein (LDL) cholesterol. Supporting scientific evidence shows that

    adding barley to one's diet can contribute to lowering serum cholesterol. As part of its

    continuing initiative to provide Americans with the information they need to make

    healthy nutritional choices about foods and dietary supplements, in 2005, the Food andDrug Administration (FDA) approved that whole grain barley and barley-containing

    products are allowed to carry a claim that they reduce the risk of coronary heart disease

    (CHD) under the Code of Federal Regulations (CFR 101.81). Whole barley and dry

    milled barley products such as flakes, grits, flour, meal, and barley meal are all products

    that can use this health claim. An example of the health claim that may be used on

    products is: "Soluble fiber from foods such as [name of food], as part of a diet low in

    saturated fat and cholesterol, may reduce the risk of heart disease. A serving of [name of

    food] supplies [x] grams of the soluble fiber necessary per day to have this effect." To

    qualify for the health claim, the barley-containing foods must provide at least 0.75 grams

    of soluble fiber per serving of the food, and the health claim in CFR 101.81 is based on

    the consumption of total 3 grams of beta-glucan soluble fiber daily (FDA 2005; NBFC

    2006). The addition of this health claim is very important as it is strongly supported by

    scientific evidence and will make consumers more aware of the health benefits of

    consuming barley products.

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    1.1.4 Antioxidant potential from barley

    Some recent studies have focused on demonstrating and studying the antioxidant

    properties and compounds found in barley. Antioxidants or phenolic structured

    antioxidant compounds have been detected in barley and recent studies have shown that

    cereals contain more phytochemicals than previously considered. These constituents of

    barley are considered to be the most important source of antioxidants in cereals and exist

    in both the free as well as bound form. The majority of the free phenolics are found as

    flavanol compounds, whereas the bound phenolics are mainly phenolic acids. Both ofthese groups are known to have antioxidant activity and possibly contribute health

    benefits. Cereals are therefore claimed to be good sources of natural antioxidants.

    Preliminary results suggest that these phenolic acids are absorbed in humans and that

    their antioxidant activity may reduce the risk of coronary heart diseases, cancers, and

    aging processes (Holtekjlen, Kinitz et al. 2006).

    In addition to the potential health benefits associated with phytochemicals, these

    phenolic compounds have important functional properties. Firstly, phytochemicals in

    grains contribute to product quality in terms of color, flavor, and texture. The phenolic

    acids and the flavanol polymers may be perceived as sour, bitter, and astringent.

    Secondly, they also influence bread quality by interfering with the dough formation. The

    changes in antioxidant properties were studied after baking bread containing barley. The

    most significant change was seen among the different barley varieties, but much less after

    storage or baking (Holtekjlen, Bvre et al. 2008).

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    1.2 Chemical and Physical Characteristics of Barley

    1.2.1 Sustagrain Barley Variety

    The Sustagrain barley variety is a proprietary barley variety that was developed

    by ConAgra Foods through a conventional barley breeding program at Montana State

    University in the late 1970s. This particular variety is generically known as

    Prowashonupana, which is a waxy, hulless barley variety that has a unique

    macronutrient composition. The varietys name is an acronym that represents its grain

    characteristics and lineage: PRO: high protein (high lysine); WA: waxy starch; SHO:short awned; NU: nude (hulless); and PANA: derived from the parent barley Compana.

    It is much higher in fiber and protein, but lower in starch compared to many other

    common cereal grains. Thus, this particular variety of barley can be used to formulate

    products with desirable health benefits (Arndt 2006). Sustagrain barley is available as a

    fine ground flour or and quick flakes which are tannish-brown in color (ConAgra).

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    Sustagrain Barley Nutritional Data (100g Basis)

    Calories 390 Vitamin A 0 IU

    Calories from Fat 60g Vitamin C 0 mg

    Fat 6.5g Calcium 33mg

    Saturated Fat 1.8g Iron 3.6mg

    Cholesterol 0g Vitamin B1 (Thiamin) 0.6mg

    Carbohydrates 64.3g Vitamin B2 (Riboflavin) 0.3mg

    Total Dietary Fiber 30g Vitamin B3 (Niacin) 4.6mg

    Soluble Fiber 12g Potassium 452mg

    Protein 18g Zinc 2.8mg

    Sodium 12mg

    Table 1: Nutritional Composition of Sustagrain Barley (ConAgra)

    1.2.2 Chemical Composition of Sustagrain Barley (Prowashonupana)

    The carbohydrate distribution in Prowashonupana barley is at least 30% dietary

    fiber and less than 30% starch. This unique composition of fiber to starch is about 2-3

    times the amount of fiber and about half the amount of starch compared with other

    common cereal grains. Approximately half the dietary fiber, 50%, consists of -glucan.

    This particular variety also provides other whole-grain nutrients including healthy lipids,

    vitamins, minerals, tocotrienols, and phytonutrients (Arndt 2006).

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    Table 2: Average Nutrient Comparison for Hulless Barley, Oats, and barley variety

    Prowashonupana (Prowash) Barley (Arndt 2006)

    Microscopic and chemical analyses were conducted to compare the structure,

    macronutrient distribution, and macronutrient content of Prowashonupana barley variety

    to another type of waxy, hulless (naked) barley variety: Bz 489-30. Waxy naked barleys

    have previously been reported to contain 51.760.5% starch, 12.616.6% protein, 12.6

    20.5% dietary fibre, 2.63.3% fat and 1.53.5% ash, while the content of -glucan has

    been shown to vary between 6 and 11% of dry matter. In contrast, the Prowashonupana

    variety has been shown to have a lower starch content (21-31%), while the contents of the

    dietary fiber, protein and fat have been shown to be high (33-36%, 18-22%, and 6%,

    respectively). Furthermore, the content of -glucan has been reported to be 2-3 times as

    high as in other naked waxy barleys, varying from 15-18% of dry matter. This study

    closely analyzed the association between structure and chemistry in the barley grain.

    Through chemical isolation methods it was found that the cellulose and arabinoxylan

    content was higher in the Prowashanupana variety of barley. It was hypothesized that the

    Nutrient (%)Hulless

    BarleyOats

    Prowash

    Barley

    Protein 13 15 20

    Fat 3 6 7

    Starch 60 59 21-30

    Total Dietary Fiber 13 10 30

    -Glucan 5 5 15

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    thicker cell wall indicates a high content of -glucan while a large amount of starch

    granules is associated with higher starch content. Fluorescence microscopy was used to

    study the structural characteristics of the grains and it was observed that the

    Prowashonupana barley variety had irregular endosperm cells with thicker cell walls, and

    thus the higher -glucan content is unique to the Prowashonupana variety of barley

    (Andersson, Andersson et al. 1999). These studies comparing different varieties of

    barley with the Prowashonupana (Sustagrain) barley variety show that there is a definite

    advantage to use the latter in product formulations that are geared towards providing a

    nutritional and functional advantage.

    1.3 -Glucans and Arabinoxylans

    Arabinoxylans and mixed linkage (13)(14)--D-glucans, commonly

    referred to as beta-glucans, are the major non-starch polysaccharides present in various

    tissues of barley. Depending on the genotypic or cellular origin, both polymers exhibit

    variations in their molecular structures. The molecular features of -glucans and

    arabinoxylans are important in determining their physical properties, such as water

    solubility, viscosity, and gelation properties as well as of their physiological functions in

    the gastro-intestinal tract, which most notably provides the health benefits mentioned

    previously. The potential application of -glucans as food hydrocolloids has been also

    proposed based on their rheological characteristics. In addition to enhancing solution

    viscosity, -glucans have been shown to gel under certain conditions. Arabinoxylans

    have been shown to significantly affect cereal based processes such as milling, brewing,

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    and breadmaking. Furthermore, arabinoxylans offer nutritional benefits of soluble and

    insoluble fiber, and because of the presence of phenolic moieties in their molecular

    structures, they may also have some antioxidant properties (Izydorczyk and Dexter

    2008).

    -Glucan is a trivial name for the glucose polymer found in the endosperm cell

    walls of barley and oats. The -bond is not digestible by enzymes in human

    gastrointestinal tract, resulting in the classification of -glucan as a soluble dietary fiber

    (Burkus and Temelli 2005). -glucans consist of linear unbranched polysaccharides of

    linked -(1 3)- and -(1 4)-D-glucopyranose units in a non-repeating but non-random order, as seen below:

    Figure 1: Molecular Structure of -(1 3)- and -(1 4)-glucan (Chaplin 2009)

    -glucans form 'worm'-like cylindrical molecules containing up to about 250,000

    glucose residues that may produce cross-links between regular areas containing

    consecutive cellotriose units. They form thermoreversible infinite network gels. 90% of

    the -(1 4)- links are in cellotriosyl and cellotetraosyl units joined by single -(1 3)-

    links with no single -(1 4) or double -(1 3)-links. The main use of -glucans is in

    texturizing by functioning as a fat substitute, which is made possible by the increase in

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    viscosity. One such product, Nutrim, is prepared by subjecting an aqueous suspension of

    barley flour to a high temperature mechanical shearing in the presence of thermostable -

    amylase, followed by centrifugations and drying of the supernatant. In addition to -

    glucans (515%, depending on the -glucan content in barley used for extraction), a

    Nutrim preparation contains also starch, amylodextrin, and proteins and has been used to

    make low fat cheddar cheese (Izydorczyk and Dexter 2008). In a study done using a -

    glucan fat substitute product called Trimchoice, it was found that a 35% substitution with

    fat resulted in shortbread cookies that were comparable in terms of texture, color, and

    taste with the full-fat control (Sanchez, Klopfenstein et al. 1995). High molecular weight-glucans are viscous due to labile cooperative associations whereas lower molecular

    weight -glucans can form soft gels as the chains are easier to rearrange to maximize

    linkages. Barley -glucan is highly viscous and pseudoplastic, both properties decreasing

    with increasing temperature. Although these properties cause difficulty in the brewing

    industry by negatively affecting fermentation and filtration, -glucans have important

    functionality in foods as well as physiologically (Chaplin 2009).

    Arabinoxylans are non-starch polysaccharides found in the cell walls of plants.

    They are generally classified as hemicelluloses, or more specifically pentosans, a series

    of 5 carbon sugars. Their general structure is comprised of -(1,4) linked D-

    xylopyranosyl backbone with -L-arabinofuranose units attached as side residues via -

    (1,3) and/or -(1,2) linkages (Beaver 2008). They are present in Prowashonupana barley

    at 12% dry weight basis (Andersson, Andersson et al. 1999)

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    Figure 2: Structural elements present in arabinoxylans (Izydorczyk and Dexter 2008)

    The arabinoxylan structure affects its physiochemical properties. Arabinoxylans

    have the ability to bind water which may alter the dough rheology, processing and

    finished product attributes of many baked products. The high water holding capacity of

    arabinoxylans delays starch gelatinization most likely by restricting the amount of water

    available for starch gelatinization. The arabinoxylans also protect the starch from -

    amylase enzymatic degradation which results in increased bread volume, better crumb

    elasticity and increased shelf life (Beaver 2008).

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    1.4 Properties and Molecular Interactions among Major Food Components

    Along with the many physiological benefits of barley -glucans, these compounds

    exhibit certain rheological and physical properties in the food matrix that make it a

    suitable for various food applications. Such beneficial health effects have been attributed

    to the solubility of -glucans in water and their capacity to form highly viscous solutions.

    Cereal -glucans exhibit considerable diversity in their structures, including the ratio of

    tri- to tetramers, the amount of longer cellulosic oligomers and the ratio of -(1-4):-(1-3)

    linkages. These structural features appear to be important determinants of their physicalproperties, such as water solubility, viscosity, and gelation. The potential use of -

    glucans as hydrocolloids in the food industry is based mainly on their rheological

    characteristics, i.e. their gelling capacity and ability to increase the viscosity of aqueous

    solutions. Thus, -glucans can be utilized as thickening agents to modify the texture and

    appearance of food formulations or may be used as fat mimetics in the development of

    calorie-reduced foods. -Glucan-rich fractions from cereals or purified -glucans have in

    fact been successfully incorporated into products such as breakfast cereals, pasta, noodles

    and baked goods (bread, muffins), as well as dairy and meat products (Lazaridou and

    Biliaderis 2007).

    1.5 Beta-glucan extraction

    Extraction of -glucans can be done to verify the amount present in the barley

    varieties. Subsequently, the properties of -glucans alone can be studied to model how

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    they behave when subjected to thermal, chemical, and physical changes. Extraction and

    isolation of pure -glucans from oat and barley are conducted using the procedure

    outlined in Figure 3. Pure -glucan has been added to several baked products with

    successful applications (Thondre and Henry 2009). It is to be noted that this procedure is

    widely accepted and used to confirm the amount of -glucans present in a commodity.

    Figure 3: Extraction and purification of -glucans from barley and oats (Biliaderis and

    Izydorczyk 2007).

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    1.6 Water absorption capacity and effect on end-products

    The addition of -glucans to wheat flour used for bread-baking barley has been

    shown to result in a higher water absorption capacity. The addition of -glucans to a

    dough formulation also increases the development time, the stability, the resistance to

    deformation and the extensibility of poor breadmaking quality doughs, as well as the

    specific volumes of the respective breads, exceeding even that of the good breadmaking

    cultivar. Traditionally, barley has not been used in bakery products because it lacks

    substantial gluten proteins and the end-products have poor sensory qualities (Bhatty,1999). Furthermore, studies showed that addition of fibrous materials to wheat flour

    weakens the crumb cell structure, due to the dilution and weakening of the wheat gluten

    protein network. Similarly, Dubois (1978) emphasized that especially utilization of the

    water-insoluble fractions impair the gas retention of the dough and thereby change the

    texture and appearance of the baked product. More recent studies have demonstrated that

    -glucan-enriched barley fractions, blended with wheat flour, can produce bread with

    acceptable sensory properties (Skendi, Biliaderis et al. 2010).

    In a study done by Sharma and Gujral (2010), where several barley varieties were

    used to determine differences in their water absorption, water solubility index, and oil

    holding capacity, it was found that there was some difference among the different

    varieties of barley. It was determined that the water solubility index ranged from 9.23%

    to 11.77% in different cultivars. DWR-28 and RD-2552 varieties had highest and lowest

    water solubility index respectively, however only DWR-28 showed significant difference

    in water solubility index as compared to all other cultivars. The oil holding capacity

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    significantly varied among the cultivars and ranged from 1.5 to 1.68 g/g, DWR-28 and

    RD-2052 had highest and lowest oil holding capacity among the cultivars. Water

    absorption capacity ranged from 1.38 to 1.63 g/g, the highest exhibited by PL-172 and

    RD-2052 and lowest exhibited by RD- 2503. Bhatty (1993) reported similar value for

    water holding capacity and oil absorption capacity for barley flour. The water absorption

    capacity could be attributed to -glucan content in barley flour because there was positive

    correlation (R = 0.843) between -glucan content and its water absorption capacity.

    It is known that the flow behavior and gelling properties of -glucans can largely

    vary with the concentration and the molecular size of the polysaccharide concentration.Thus, understanding the effects of barley -glucans with different molecular weights on

    the rheological properties of wheat flour doughs with different breadmaking quality is

    essential for determining both the dough handling properties during processing and the

    quality of the end products. In a recent paper published by (Skendi, Papageorgiou et al.

    2009), the effect of adding low (105Da) or high (2.03 x 105Da) molecular weight barley

    -glucans in two wheat flours of different breadmaking quality were studied. Mechanical

    spectra and creep-recovery analysis data within (low stress) and out (high stress) of the

    linear viscoelastic region were obtained and revealed that the rheological behavior of -

    glucan-enriched doughs depend on concentration and molecular weight of the

    polysaccharide as well as on the flour type used. Addition of -glucan increased the G

    values of the good breadmaking quality flour doughs, whereas decreased the G of the

    poor quality wheat cultivar. Supplementation with -glucans increased the resistance to

    deformation, flowability and elasticity of the doughs under low stress. Significant

    correlations between frequency sweep and creeprecovery parameters of optimally

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    developed doughs from both flours were found. The addition of -glucan in the dough

    recipe of the poor breadmaking wheat flour may result in similar rheological responses to

    those obtained from non-fortified good breadmaking quality wheat flour.

    Many authors have reported that due to the -glucans ability to absorb high

    quantities of water, doughs fortified with -glucans display a significant increase in the

    farinograph water absorption values. It is generally recognized that water plays the most

    important role on the viscoelastic properties of the dough during mixing; i.e. the

    distribution of the dough materials, their hydration, and the gluten protein network

    development strongly depend on the quantity of added water (Skendi, Papageorgiou et al.2010). Small deformation dynamic rheological tests and creep-recovery measurements

    are often employed for dough characterization and the derived rheological data are

    explored as predictors of breadmaking performance.

    1.7 Rheological Properties Influenced by barley flour beta-glucan content

    Rheology is concerned with how all materials respond to applied forces and

    deformations. Basic concepts of stress (force per area) and strain (deformation per

    length) are key to all rheological evaluations (Tabilo-Munizaga and Barbosa-Cnovas

    2005). Shear-thinning behavior in foods may be conceptualized by the breakdown of

    structural units in a food due to the hydrodynamic forces generated during shear. Most

    non-Newtonian foods exhibit shear-thinning behavior, including many salad dressings

    and some concentrated fruit juices. In fact, most foods fall into this non-Newtonian

    category (Rao 1999). Beta-glucans are one such material that have been shown to exhibit

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    such behavior and have demonstrated viscoelastic properties (Papageorgiou, Lakhdara et

    al. 2005). Data reported on the consistency and viscoelastic properties of batter are

    important in the development of new products (Kalinga and Mishra 2009).

    Characterizing the mechanical behavior of food materials is complicated by the

    fact that many food materials are viscoelastic, so their mechanical properties lie between

    that of a purely elastic solid and that of a viscous liquid. Using oscillatory rheology, it is

    possible to quantify both the viscous-like and the elastic-like properties of a material at

    different time scales; it is thus a valuable tool for understanding the structural and

    dynamic properties of food systems (Wyss, Larsen et al. 2007).

    1.8 Stevia as a Sweetening Agent in Consumer Products

    Stevia is a generic name for the sweetness-providing compounds, particularly the

    steviol glycosides, extracted from the herb Stevia rebaudiana(Bertoni). It is generally

    available as a mixture of steviol compounds, with the predominant sweetness compound

    being Rebaudioside A (Carakostas, Curry et al. 2008). Stevia has negligible caloric value

    for use in food products and beverages since it is used at a very low concentration. Stevia

    leaf extracts exhibit a sweetening level of 15-30 times sweeter than sucrose, dependent on

    the extract quality and raw material (SM Savita 2004). In 2008, stevia gained approval

    for mainstream food usage from a dietary supplement status. Therefore, products

    containing stevia launched thereafter, and have been gaining popularity among food

    manufacturers because of its natural label and low-calorie benefits (Lord and Sant'Angelo

    2010). Although stevia has been gaining acceptance and has increasing use in the food

    industry in the past few years, there are technical issues with the product due to the

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    presence of bitter compounds that are also incorporated into the sweetener during the

    extraction process. There is also a lingering licorice aftertaste and sweetness at a higher

    use concentration, which has been the cause for the limited acceptance of the product.

    However, there are benefits of using stevia extracts, especially in baked products

    where thermal processing occurs. Rebaudioside A has been shown to have thermal

    stability in two specific studies. One was the use in pasteurized dairy product and the

    other was in a laboratory baking study with temperatures up to 390 F. Due to its thermal

    stability, stevia is suitable for baking applications. However, stevia is not the perfect

    substitute for sugar in bakery applications. The sole function of stevia would be inproviding sweetness. Stevia lacks the ability to add texture, caramelize, feed the

    fermentation of yeast or help tenderize a batter, all properties that sugar possesses. Also,

    cakes made with stevia may not rise as well, and achieving a soft, chewy cookie may

    require additional ingredients (Jones 2006).

    Figure 4: Structural Components of Stevioside, Rebaudioside A, and Steviol

    (Carakostas, Curry et al. 2008)

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    1.9 Conclusions from Literature Review and Objectives for Research

    Appealing to the heightened awareness of health issues such as childhood obesity,

    prevalence of diabetes, and lack of fiber in the diet, more nutritious products need to be

    made available to school children. As manufacturers make the changes to their products

    to improve the nutritional value, the food quality should not diminish. Thus far, barley

    flour has limited approaches in modern-day food products despite its known health

    benefits. This may be attributed to the presence of a bitter taste and a strong whole-grain

    character as well as the requirement of high sugar content in baked products to mask thistaste. However, the addition of sugar increases the caloric value of the product. To

    address this issue, a natural sweetener called stevia will be studied by incorporating it

    into a model baked product prepared with barley flour. The essential food quality

    properties of color, taste, texture, and nutritional value will be used to assess the food

    characterization of a barley flour containing product.

    The specific objectives in this research include identifying key parameters

    responsible for processing and to monitor the effect of water absorption with varying

    levels of -glucans in barley flour, specifically noting how this difference affects some of

    the rheological properties. Another aim is to reduce sugar levels in the product by

    incorporating a natural sweetener called Stevia and to monitor its influence through

    instrumental and sensory analysis. Finally, the instrumental findings of the research will

    be compared with the sensory aspects of the model baked product to determine whether

    there exists a detectable difference though a human response.

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    CHAPTER TWO

    Materials and Methods

    A muffin type of baked product will be used as a model to assess the color,

    texture, and sensory properties as an effect of the -glucan content and reduced sugar

    levels on the final product. Most importantly, the interactions among the primary

    components of the product need to be well-understood. Thus a model system of barley

    flour, natural sweetener, water, and other minor components will be used to study the key

    processing effects through instrumental and sensory analyses. The aim is to incorporate anatural sweetener while keeping the low-calorie and low glycemic index aim in focus. At

    the same time, the functional changes occurring due to the reduction in sugar levels and

    increased beta-glucan content will also be monitored.

    Similarly, the type of fat used in the product formulation is intended to provide

    some health benefit to the consumer in addition to its role in the baked product

    formulation. Thus, flaxseed meal will be added to the product as it provides a source of

    beneficial omega-3 fatty acids and adds fiber. In a study done by (Fiscus, Harris et al.

    1999), it was found that flaxseed substituted into whole wheat flour at 25% and 50%

    levels showed similar sensory results to the control which used 100% wheat flour in the

    preparation of peanut butter cookies and banana bread. Important food quality

    characteristics such as flavor, texture, and mouthfeel were found to have no significant

    difference between that of the control and the formulation with the flaxseed added.

    Minor ingredients such as flavorings and leavening agents will present in the final

    product, but not studied in depth in the analytical parts of the project. The main objective

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    is to determine and examine the behavioral differences among the different barley flours,

    considering Sustagrain barley flour as the high -glucan and a commercially available

    brand of barley flour as the low -glucan counterpart.

    2.1 Ingredients Used in Baking Procedures and Analytical Measurements

    Two varieties of barley flour will be used to study the functional changes caused

    by the varying amounts of beta-glucan present in the two brands of barley flour. One is

    barley flour manufactured by Bobs Red Mill, a commercially available brand, which willbe considered as the low beta-glucan barley flour. The other is Sustagrain, which as

    previously mentioned, is a high beta-glucan (15%) containing barley flour sold under the

    ConAgra Mills brand. The main objective is to identify the key behavioral differences

    with other food components and compare how the beta-glucan levels affect the water

    absorption capacity, rheological properties of the batter, textural properties in a model

    baked product, and the final product color.

    Ground flaxseed meal supplied by the company Bobs Red Mill will be used in

    the muffin formulations to obtain color and textural measurements. The company

    website states the following on the product page, In a 2 Tablespoon serving size (13

    grams) the fiber content is 1.33 grams of Soluble Fiber and 2.67 grams of Insoluble Fiber.

    Ground Flaxseeds are a good source of Omega 3 Fatty Acids. In a 2 Tablespoon serving,

    there is 2400 mg of Omega 3 (2010).

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    Figure 5: Nutrition Label for Bobs Red Mill Ground Flaxseed Meal (left) and Bobs Red

    Mill barley flour

    2.2 Water Absorption Index of Low and High -glucan Barley Flours

    The amount of water used for a baked product has a great effect on the outcome

    of the final volume and texture (Osorio, Gahona et al. 2003). Therefore, it is necessary to

    determine how much water would be absorbed into the flour prior to developing product

    formulations. The water absorption index can be determined by simple methods using

    barley flour and water. Water absorption capacity of flour was measured by the ratios

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    f

    wf

    mmmWAI

    +=

    and centrifugation methods (Sharma and Gujral 2010) (Ding, Ainsworth et al. 2006).

    0.12 g Sustagrain barley flour and Bobs Red Mill Barley flour were dispersed in 1 mL of

    distilled water and placed in pre-weighed centrifuge tubes. The dispersion was stirred

    using a stir-plate for 10 min followed by centrifugation for 25 minutes at 3000 rpm. The

    supernatant was drained off by allowing the tubes to stand inverted for 10 minutes. The

    water absorption index was calculated by dividing the weight of the flour and water

    mixture obtained after draining off the supernatant by the original weight of the flour.

    The averages of triplicate measurements are recorded. It was predicted that the higher -

    glucan Sustagrain flour will have a higher water absorption index since increasingamounts of -glucans have been shown to absorb more water (Sudha, Vetrimani et al.

    2007).

    Water absorption index was calculated by:

    where mf= mass flour and mw= mass water.

    Figure 6: Example of water separated from barley flour after centrifugation

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    2.3 Microbakery Model Formulations for Barley Muffins

    A dessert type baked muffin product was used as a model to characterize the

    instrumental changes occurring in the system as a result of -glucan levels in the barley

    flour as well as study the changes occurring due to the removal of sugar from the

    formulation. Muffins were prepared using the following formulations, with process

    variations occurring in the type of barley flour utilized (high or low -glucan) and the

    reduction in sugar levels, while using stevia to maintain sweetness levels. The recipe

    ingredients were adapted from (Idzorek 2010) and modified based on preliminary baking

    experiments and the quantities were reformulated and adjusted to accommodate small-scale baking experiments. Ingredients such as eggs, banana, sugar, baking soda, vanilla

    extract, and salt were sourced from local supermarkets. Banana puree was used since

    fruit purees demonstrate humectant properties, promote tenderness and retain moistness,

    increase shelf life, and can replace some of the sugar and/or fat in muffins and cakes (Hui

    2006). The banana puree would also be helpful in providing some of the necessary

    moisture to hydrate the barley flour in the formulation. Stevia leaves extract was

    obtained from Spectrum Chemical.

    Each set of muffins were prepared with the use of 1) Bobs Red Mill Barley

    Flour as the low -glucan type or 2) Sustagrain Barley Flour as the high -glucan flour.

    Each formulation was prepared with the same ingredients, with the process variation

    occurring in the sugar:stevia use levels, as well as the addition of ground flaxseed meal in

    the 0% sugar formulations. Stevia has been shown to have a potency level perceived as

    200-300 times sweeter than sucrose (1.0). However, the sweetness potency, or the

    sweetness perception, in a product is highly dependent on the sucrose equivalency level.

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    The stevia to sugar equivalency use level has been reported as 4-8%, with 6% level being

    a reasonable average value (Prakash, DuBois et al. 2008). Initially, stevia sweetener

    levels in the product formulations were based on the previously mentioned levels. Thus,

    the amount of sucrose (9.88 grams) used in the full sugar control product required 0.59

    grams of stevia (6.0%) to have a similar sweetness equivalency.

    Preliminary baking experiments were done to test this level of sweetness using

    stevia in the formulation and it was found to be too high of a sweetness level, which also

    resulted in a strong bitter aftertaste. Stevia use level was thus reduced to 0.15 grams in

    the formulations, or approximately 0.24% of the total formulation. This stevia use levelwas employed for baking done for instrumental measurements as well as preparation of

    samples used in sensory panels.

    Table 3: Formulations for Muffin Batters Used to Prepare a Model Baked Product

    Ingredient (g)100%

    Sugar

    0%

    Sugar

    0% Sugar +

    Flaxseed Meal

    Barley Flour 16.5 16.5 16.5

    Salt 0.05 0.05 0.05

    Baking Soda 0.44 0.44 0.44

    Vegetable Oil 5 5 5

    Water 105 F 10 10 10

    Banana Puree 25 25 25

    Eggs 5.5 5.5 5.5

    Sugar 9.88 0 0

    Vanilla 0.3 0.3 0.3

    Flaxseed Meal 0 0 2.25

    Stevia 0 0.15 0.15

    Total 72.67 62.94 65.19

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    Although the total weight of the batter formulations differs, the process control

    was applied by weighing out muffin batters equally into mini-muffin tins. Equal amounts

    of batter were utilized in all baking processes for instrumental experiments and sensory

    analysis.

    Procedure for preparing muffins:

    A Frigidaire Gallery consumer electric convection oven was used for the baking

    experiments. The oven was preheated to 325F and the temperature was maintained

    throughout the preparation of the batter and baking process. A ripe banana was pureed in

    a food processor (Hamilton Beach) until it reached a smooth consistency and did notcontain any visible chunks. In a mixing bowl, the dry ingredients consisting of barley

    flour, salt, flaxseed meal and baking soda were combined and set aside. In another

    separate mixing bowl, oil, eggs, vegetable oil, and sweeteners were mixed and beaten

    with a hand-held electric mixer (Sears) on speed 3 until thoroughly blended, for

    approximately 15 seconds. Then the banana puree, vanilla, water and dry ingredients

    were added to the mixture and beat with the mixer on speed 2 for 15 additional seconds

    until all ingredients are blended and uniform in appearance. The batter was then weighed

    out to 12 1 grams in a paper muffin mold placed into a mini-muffin pan and baked for

    ten minutes. The pan was then rotated 180 and baked an additional two minutes.

    Muffins were then removed from the oven and allowed to cool in the pan at room

    temperature for two minutes. Then, the muffins were transferred to a cooling rack for an

    additional 30 minutes at room temperature. Muffins were stored in a sealed plastic zip-

    top bag at room temperature (25C) for further analysis. Texture, color measurements,

    and photographs of the surface and interior were obtained within 12 hours of preparation.

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    2.4 Rise and Moisture Loss of Muffins

    The variation for this experiment included the use of high and low -glucan

    barley flours in the model baked product as well as the complete removal of sugar. When

    sugar was omitted from the formulation, stevia was used to maintain sweetness levels.

    The heights of the muffins were evaluated by obtaining the height of the batter prior to

    baking and calculating the percentage of rise, or increase in height, after muffins were

    prepared. A toothpick was inserted into the center of the batter and the toothpick was

    marked at the level of the batter. The toothpick was then removed. The initial height of

    the batter was recorded by measuring the distance between the mark made on the

    toothpick to the end of the toothpick with a ruler, in centimeters. Final heights were

    obtained in the same manner after allowing the muffins to be cooled for 30 minutes. The

    percentage rise, or increase in muffin height, was calculated by:

    % increase in muffin height = hf hix 100%,hi

    where hfis the final height and hiis the initial height of the muffins.

    The initial weight, 12 1 grams, of the muffin batter was recorded by placing the

    batter in mini-muffin tin foil cups and weighing them on a scale. After baking, the

    muffins were allowed to cool for 30 minutes and final weights were recorded. The

    percentage change in weight, considered as moisture loss, was calculated by:

    % decrease in muffin weight = mi mf x 100%,mi

    where mfis the final mass and miis the initial mass of the muffins.

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    Average values of four replicates are reported for both the muffin rise and moisture loss

    measurements.

    2.5 Rheological properties of barley dough using lubricating squeezing flow

    technique

    The rheological properties of a dough are a great indicator of their behavior

    during baking and determining the final product texture, rise, spread, and overall quality.

    Cookie spread rate is governed by dough viscosity as doughs with lower viscosity had a

    higher spread rate compared to doughs with a higher viscosity (HadiNezhad and Butler

    2009).

    Traditional determinations of rheological properties of wheat flour dough have

    been carried out using tests and instruments such as the farinograph and alveograph,

    which provide important information on rheological characteristics for the development

    of baking products. However, it has been seen that these tests do not allow detecting

    differences of composition of the flour and addition of ingredients, nor do they provide

    detailed information on physical and characteristic properties of flow behavior (Osorio,

    Gahona et al. 2003). Thus, a technique called lubricated squeezing flow viscometry has

    been used to quantify changes in the behavior of doughs and can be used with high

    viscosity materials. This technique can detect differences among the samples that cannot

    be observed under shear conditions. It also solves two major problems that occur in food

    viscometry: 1) the slip condition in the surface and 2) the inadvertent rupture of the

    structure of the sample when introducing it in the reduced space of the conventional

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    rheometer. This technique has been previously applied to several high viscosity products

    such as peanut butter, cheese, cooked cornmeal dough, mayonnaise, tomato paste, tortilla

    dough, wheat dough, yogurt, dulce de leche (milk sweet), refried beans paste and

    mustard. The lubricated squeezing flow viscometry technique is one of the basic types of

    biaxial extensional flow, which is the type of flow behavior that occurs during the baking

    process.

    Both low and high -glucan barley dough were subjected to this type of

    compression analysis to look at the flow properties of the doughs and how they affect the

    final product texture. Dough samples were prepared using the previously determined

    water absorption index values. Water was added to the barley flours and mixed

    continuously for 3 minutes to allow the formation of a cohesive textured dough (Sudha,

    Vetrimani et al. 2007). A TA.XT2 Texture Analyzer was used for the lubricated

    squeezing flow technique. The TA-4, 38 mm cylinder probe was attached to a 25 kg load

    cell. The platform as well as the surface of the probe were well lubricated with

    commercially available food grade, edible vegetable oil to minimize any effects of

    friction (Osorio, Gahona et al. 2003) and (Stojceska, Butler et al. 2007). Pre-test and

    post-test speeds were set at 1.0 mm/sec and the test speed, or deformation rate, or

    crosshead velocity, was set to 0.1 mm/sec. Dough discs measuring 1 cm in height and 2

    cm radius were formed and placed on the platform. They were allowed to rest for 5

    minutes prior to testing in order to stabilize internal tensions. The dough was compressedto 60% of its original height, or 6.0 mm. The force, time, and distance parameters were

    recorded automatically through the Texture Expert software. Samples were prepared and

    run in triplicates with average values used for calculations of biaxial stress, biaxial strain,

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    and biaxial viscosities. Calculations to obtain biaxial viscosity vs. biaxial strain rate

    curves were obtained following detailed methodology in (Osorio, Gahona et al. 2003).

    Apparent biaxial extensional viscosity (ABEV) was calculated by the following formula:

    ABEV = 2Ftht/R2v

    where Ftis the compression force (N) at time of t; htis the height of the dough sample

    (m) at time t; R is the initial radius (m) of the dough sample; v is the crosshead speed

    (m/s). Data points were obtained at each second time interval and plotted to obtain a

    curve of the biaxial extensional viscosity versus biaxial strain rate.

    Figure 7: Apparatus and set-up of TA.XT2 Texture Analyzer for Lubricated Squeezing

    Flow Technique Analysis of Doughs Prepared Using Barley Flours

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    2.6 Pasting Properties of High -glucan and low -glucan barley flours

    Rheological properties of starch pasting are traditionally studied by an instrument

    called the Rapid Visco Analyzer. The working principle of this type of equipment is that

    the rheology is directly related to the microstructure of starch and is influenced by many

    factors such as the amylose/amylopectin ratio, the chain length of amylose and

    amylopectin molecules, the concentration of starch, shear and strain, and temperature.

    The sample is heated over a range of temperatures and the viscosity is recorded over a

    period of time. Starch granules are generally insoluble in water below 50C, so the

    viscosity of the starch mixture is low below this temperature. When the starch granules

    are heated, the granules absorb a large amount of water and swell to many times of their

    original size.

    As the instrument generates shear conditions, the viscosity increases when the

    swollen starch granules squeeze past each other. The temperature at which the rise in

    viscosity is seen is known as the pasting temperature, which indicates the minimum

    temperature required to cook a sample. As a sufficient amount of starch granules are

    heated, there is period of time where there is a rapid increase in viscosity as the

    temperature increases. The peak viscosity occurs at the equilibrium point where starch

    granules are completely swollen and just as they begin the retrogradation process. The

    peak viscosity and temperature indicates the water binding capacity of the starch. As thetemperature is held constant over a period of time, the starch granules begin to rupture

    and polymer realignment occurs, which is evident by the decrease in apparent viscosity of

    the paste and is known as the breakdown viscosity, which occurs at the beginning of the

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    cooling phase. The viscosity at this stage of heating also gives an indication of paste

    stability. As the sample is cooled down back to the starting temperature, re-association

    between the starch molecules, especially amylose, occurs to varying degrees, which

    results in an increase in viscosity once again as a gel begins to form. This phase of the

    pasting curve is referred to as the setback region, and occurs due to the retrogradation of

    the starch molecules. The final viscosity gives an indication of the stability of the cooled,

    cooked paste under low shear conditions (Cui and Liu 2005) and (Brabender 2005).

    Below is a typical pasting curve used to illustrate the specific points in the pasting profile

    determined during the duration of the run.

    Figure 8: Typical RVA pasting profile of a normal maize starch for viscosity ( - ) and

    temperature (---) as a function of time (Cui and Liu 2005)

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    The pasting properties of high and low -glucan barley flours were studied using

    a rheometer and a controlled temperature circulating water bath to emulate the Rapid

    Visco Analyzer instrument. A Brookfield Digital Rheometer Model DV-III was used in

    conjunction with a with a K10 model water bath circulator system controlled by a

    Thermo Scientific Haake DC30 temperature control system, with an accuracy of 0.1C.

    The pasting properties of the low and high -glucan barley flours was studied, as well as

    with the addition of sugar with the same ratio to that in the muffin formulation. The

    sample was prepared as a 5% wt/wt basis of barley flour in water suspension. Sugar was

    present in the muffin formulations at 13.6% of the total formulation, therefore of sugar

    was added was added to the 5% barley flour and water suspension at the same ratio. All

    samples were mixed prior to transferring into the rheometer holding cell and allowed to

    equilibrate for 3 minutes to reach an initial temperature of 50C. The shear rate was set

    to 200 rpm or 68 1/s for the duration of the run and an SC4-31 spindle was used based on

    manufacturers given maximum viscosity parameters. The temperature profile was set to

    begin at 50 C, then increase to 95 C, hold at 95 C for 5 minutes, and then cool to 50 C

    (Stojceska, Butler et al. 2007; Sharma and Gujral 2010; Sharma, Gujral et al. 2010; Sai

    Manohar, Urmila Devi et al. 2011). Time (min), temperature (C), and viscosity (mPas)

    were recorded by the Rheocalc Version 3.2 software. Peak viscosity (mPa s), breakdown

    viscosity (mPa s), final viscosity (mPa s), setback viscosity (mPa s), peak time (minutes),

    and pasting temperature (C) were obtained from the graph plotted through the durationof the sample run. Averages of triplicate values are reported.

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    2.7 Dynamic Rheological Properties of Muffin Batters

    An oscillating rheometer was used to directly study the rheological properties of

    batters to characterize the behavior of -glucan concentration on the rheological

    properties of the batter and resulting changes occurring during processing.

    Apparent Viscosity

    Muffin batters were prepared according to the same formulations used in baking

    experiments and were run through a Rheometric Scientific ARES Rheometer to measure

    the viscosity between strain rate of 1 to 398 s-1. The sample was loaded between two

    parallel plate geometry probes and the gap was adjusted to 1.0 mm. All measurements

    were made at 25C using 25 mm diameter parallel plates. Stress and strain rates were

    recorded and plotted to obtain a stress vs. strain rate curve to fit a Power-law model to

    obtain the flow behavior index (K) and the consistency index (n) from the slope of the

    line. The flow behavior was described by power law model, where shear stress (Pa) was

    related to the shear rate (1/s) and the consistency coefficient (K in Pasn), and flow

    behavior index (n) were obtained by linear regression.

    Dynamic Oscillating Rheology

    Oscillatory rheology is a standard experimental tool for studying behavior of

    foods which exhibit viscoelastic properties, essentially those foods that are between

    solids and liquids in their behavior (Wyss, Larsen et al. 2007). A Rheometric Scientific

    ARES Rheometer was used to determine the oscillating rheology properties of the muffin

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    batter. Initial experiments using barley flour solutions, based on approximate moisture

    content in the muffin batter, were used to determine the frequency parameters at which

    muffin batter samples would be tested. The frequency chosen for the duration of the

    measurements was 20 s-1. The dynamic oscillatory-shear storage and loss moduli (G and

    G) were measured and recorded as a function of % strain between 1 to 100%. All

    measurements were made at 25C using 25 mm diameter parallel plate geometry with a

    1.0 mm gap between the plates. Values reported are based on an average of 3 replicates.

    2.8 Assessing Muffin Firmness Using a Texture Analyzer

    As a measurement of food quality, texture is important for observing both

    defective and acceptable food products. Texture can be defined as a group of physical

    characteristics that arise from the structural elements of the food and are sensed primarily

    by the feeling of touch, are related to the deformation, disintegration and flow of the food

    when a force is applied (Taub and Singh 1998). A group of properties based on physical

    structure, sense of touch, and functions of mass, distance, and time compose the

    definition of texture (Bourne 2002). The classifications of this testing are puncture,

    compression-extrusion, cutting-shear, compression, tensile, torsion, bending and

    snapping and deformation. A comprehensive definition of food texture analysis and

    methods for evaluation can be found at Bourne (2002), (Rosenthal 1999), and Texture

    Technologies (2009).

    The various methods for food texture analysis depend on the properties of the

    food. A common texture instrument or universal testing machine measures force as a

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    function of time and distance. A simple test of measuring the force to push a probe into a

    food surface is used to measure texture known as a puncture test. The force to deform the

    sample is similar to the way molar teeth bite and chew. However, the puncture test

    assumes a semi-infinite geometry because of the small surface area measured. A

    compression test will measure the larger surface area of the food sample by forcing it to

    flow or fracture and deform dependent on its composition. This type of a compression

    test is widely used in the industry as a measure of food quality during shelf-life studies

    and to observe changes occurring due to ingredient modifications. When the direction of

    the force applied to the sample is parallel to the direction it is sliding this is known asshear. A food product can also be measured for the force to be divided into two sections,

    bent or pulled apart (Tabilo-Munizaga and Barbosa-Cnovas 2005). Using any test, the

    most accurate data depends on a consistent sample temperature, size, shape, speed,

    distance and direction.

    Instrumental techniques do not completely indicate textural quality of a product

    since they lack the uniqueness of consumers perception. A sensory texture analysis is

    needed to measure the quality of a food dependent on its acceptability. However, human

    experience of a trained expert can be compared to physical properties results for insight

    on the reaction of texture differences. Using the human senses to manipulate the food

    product by eating allows for many different variables to be identified. For example, in

    study of apple firmness a difference of five Newtons using instrumentation is detectable

    by human perception (Harker, Gunson et al. 2006). The process of eating can measure

    the actions of biting, chewing, swallowing, etc. and determine which sensations are

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    perceived at any given point. Since texture has a high affect on liking of the product, the

    quality of a food can depend on the description of its meeting chosen standards.

    Muffins that were prepared using the previously described formulations with low

    and high -glucan flour with varying amounts of sugar:stevia ratios and the addition of

    flaxseed meal were evaluated using the TA.XT2 Texture Analyzer. A compression test

    was conducted on all the muffin varieties to determine the firmness and how it is affected

    by each variation in the muffins and between the two types of flours. A TA-11, 1-inch

    diameter cylinder was used with a pre-test, test, and post-test speed of 2.0 mm/sec.

    Muffin samples placed on the testing platform and were compressed to 10 mm of theiroriginal height. An output of peak force (g) vs. time was obtained for each variety of

    muffin in triplicate measurements, with average values reported (Texture Technologies,

    Corp.). Differences between averages were determined by comparing muffin treatments

    according to a t-test with a significance level of 5% (p = 0.05) using the Microsoft Excel

    2003 Data Analysis ToolPak.

    2.9 Evaluation of Colors Using a Colorimeter

    Surface color is one of the important characteristics of baked products and is

    considered as a critical index for judging baking quality. Baked products develop color

    in the later stages of baking, simultaneously with crust formation and occur through

    chemical processes including the Maillard reaction and sugar caramelization. The

    Maillard reaction is responsible for color development at surface temperatures below

    150C, while caramelization reactions occur when the product surface temperature

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    exceeds 150C (Onishi M 2011). It will be important to monitor these color changes

    differences that occur due to the removal of sugar from the product, which may affect the

    caramelization reactions occurring at the higher temperature stage of the baking process.

    A Minolta CM-2500d Spectrophotometer with Spectra Match software was

    used to measure surface and interior color of the low and high -glucan barley flour

    muffin varieties in L a*b* color space. The International Commission on Illumination

    (CIE) (1976) color space measures L for the luminance or lightness component with a

    range 0 to 100 (dark to light), and a* (from green to red) and b* (from blue to yellow).

    After preparing muffin samples as described above, measurements were taken for surfacecolor on top of the muffin and the interior crumb surface color from muffins that were cut

    longitudinally from top to bottom. Measurements were taken as an average of three

    locations across the surface of the same muffin. Differences between average L, a*, and

    b* values were determined by comparing muffin treatments according to a t-test with a

    significance level of 5% (p = 0.05) using the Microsoft Excel 2003 Data Analysis

    ToolPak. It was predicted that the omission of sugar from the formulation will have a

    significant impact on the surface and interior crumb browning, thus resulting in color

    differences. The generation of brown pigments during a caramelization and Maillard

    browning reactions will be lacking when stevia is used in the muffin formulation.

    The colors for the two barley flours were also obtained by taking three

    measurements across different locations on a Petri dish containing the flours. Visual color

    differences of the barley flours and muffin surface and interiors were observed and

    recorded with digital photographs using a Sony DSC-H50 digital camera with automatic

    camera shutter speed settings and compared with L a* b* color values obtained from the

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    colorimeter to determine whether there were differences in the raw material and how they

    would translate to changes occurring in the final product color.

    2.10 Nutritional Comparison of Muffins

    Nutritional values of barley muffin prototypes were evaluated using a software

    called Recipe Calc, version 4.0 (Muller). Most ingredients nutrition facts were available

    in the software database and were used as a basis for calculation of the finished product

    nutrition facts. Ingredients that were not present in the database, such as specificproducts including the Sustagrain barley flour, Bobs Red Mill barley flour, and Bobs

    Red Mill Flaxseed Meal, were added into the software using the nutrition label provided

    on the package or supplied by the manufacturer. All ingredients were added and the

    nutrition labels were prepared based on the quantities used in the product formulations

    and calculated automatically in the software. According to the Food and Drug

    Administration, it is best to determine the values for nutrition labeling by conducting

    laboratory analyses on its products, but a manufacturer can use average values calculated

    from ingredient composition databases as long as it is confident that the values are

    accurate and accurately represent the characteristics of the product (Mermelstein 2009).

    Although the generated nutrition facts may not be an adequate tool for in labeling for

    manufactured products, they provide a clear method of comparison among the different

    formulations and varieties of the muffins for research purposes, particularly to identify

    changes in carbohydrate levels, to denote differences in sugar and fiber levels.

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    2.11 Sensory Methodologies Used to Evaluate Muffin Products

    Sensory evaluation is a scientific discipline used to evoke, measure, analyze, and

    interpret reactions to those characteristics of foods and materials as they are perceived by

    the senses of sight, smell, taste, touch, and hearing (Hui 2006). Sensory evaluation is a

    technique that food scientists use the human body and its perception of the five basic

    senses as a tool to measure differences and intensities of food characteristics. The

    objective of the sensory panels pertaining to this research included looking at key

    differences occurring due to the removal of sugar from the muffin formulations.

    2.11.1 Quantitative Descriptive Analysis (QDA) Using Spectrum Method

    Descriptive analysis methods involve the detection and the description of both the

    qualitative and quantitative sensory aspects of a product by trained panels (Meilgaard,

    Civille et al. 1999). Quantitative Descriptive Analysis (QDA) is the most sophisticated

    sensory methodology. The results of QDA are a complete sensory description of the test

    treatments (determined by the sensory panel), that provide a basis for relating specific

    ingredients to specific changes in sensory characteristics of a product. QDA, particularly

    the Spectrum Descriptive Analysis Method was chosen as the analysis tool for the study

    since it yields quantitative data from panelist scores based on perceived intensities with

    reference to pre-learned absolute intensity scales. The sensory findings may be u