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EFFECTS OF EXTRUSION AND BAKING PROCESSES ON
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EFFECTS OF EXTRUSION AND BAKING PROCESSES ON
GIN SENOSIDES IN WHEAT FLOUR-GINSENG POWDER
BLENDS

By

Yoon Hyuk Chang

A DISSERTATION

Submitted to
Michigan State University
in partial fulﬁllment of the requirements
for the degree of

DOCTOR OF PHILOOSPHY
Department of Food Science and Human Nutrition

2008

ABSTRACT

EFFECTS or EXTRUSION AND BAKING PROCESSES ON GINSENOSIDES
IN WHEAT FLOUR-GINSENG POWDER BLENDS
By

Yoon Hyuk Chang

The speciﬁc objectives of the present study were: (1) to explore optimum
extraction conditions of ginsenosides (G, ginseng saponins) from a blend of wheat ﬂour
(WF)-ginseng powder (GP), (2) to investigate effects of extrusion process variables on
extractable G in wheat-ginseng extrudates (WGE), (3) to measure quality properties of
WGE extruded under different conditions, and (4) to evaluate physicochemical
properties of bread and cookies baked from hard wheat ﬂour and soﬂ wheat ﬂour,
respectively, blended with GP and effects of baking on their extractable G.

Through studies on optimization of extraction conditions for G from WF-GP,
it was observed that interactions between a starch fraction and G as well as between a
gluten fraction and G did occur in WF-GP heated at 90°C. It was also found that the
interactions between WF components and G could be completely disrupted by
increasing ultrasonic extraction time up to 90 min, for the maximum extraction.

To evaluate effects of extrusion process variables on the extractable G in and
quality properties of WGE samples, WF-GP (10% GP, w/w) was extruded in a twin-
screw extruder with full factorial combinations of feed moisture (25, 30, and 35%),
screw speed (200 and 300 rpm), and zone 5 barrel temperature (110, 120, 130, and
140°C). The quantities of G (Rbl, Re, and Rd) extracted from WGE samples extruded

at a zone 5 barrel temperature of 2 120°C were signiﬁcantly higher than those extracted

from the non-extruded WF-GP blend. New G compounds (Rg2 and Rg3), which were
not present in raw non-extruded WF-GP, were found in certain WGE samples,
indicating that high temperature extrusion cooking could lead to the chemical
modiﬁcation of thermolabile protopanaxatriol type G and protopanaxadiol type G into G
Rg2 and G Rg3, respectively. The amounts of G (Rg2 and Rg3) were dependent on
each of the three extrusion process variables studied. The physical (expansion ratio,
water absorption index, and water solubility index), pasting, and thermal properties of
WGE samples were signiﬁcantly affected by changes in extrusion process variables
studied.

From studies on effects of baking processes (bread-baking and cookie-
baking) on G of WF-GP blends, it was found that the temperatures used for bread- and
cookie-baking could increase the amounts of extractable G Rbl, G Re, and G Rd in the
hard wheat-ginseng bread and soﬁ wheat-ginseng cookie samples. Ginsenosides Rg2
and Rg3 were present only in the hard wheat-ginseng bread sample, indicating that not
only the higher baking temperature (215°C) but also the longer baking time (24 min)
used for bread-baking could cause the production of ginsenosides Rg2 and Rg3, as

compared to cookie-baking conditions (205°C and 11 min).

ACKNOWLEDGEMENTS

Many people have helped me in the completion of this dissertation, and I
would like to acknowledge and thank them here. First of all, I would like to thank Dr.
Perry K.W. Ng, my major advisor, for being a wonderful mentor and for his constant
support throughout my study and research. His expert guidance and generosity with
his time and ideas have enriched my graduate school experience and have contributed
greatly to my personal and professional development over the years. I feel very
fortunate to have had an advisor whose help, kindness, patience, and concern for my
well being helped make my doctoral program an enjoyable and rewarding experience.
I also would like to thank my committee members, Dr. James F. Steffe, Dr. Gale M.
Strasburg, and Dr. Mark A. Uebersax, for the valuable contributions they have made to
help improve this work and for the support and encouragement they have all given me

along the way.

Thanks are extended to previous labmates, Ed, Hilda, and Ritu, and previous
visiting scholars, Dr. Ahn, Dr. Kim, Dr. Koh, and Dr. Rhim. Furthermore, I would like

to thank my current labmates, George and Kevser.

Finally, I am extremely grateful to my parents, parents-in-law, wife, and son
for all of their love, support, encouragement, and sacriﬁce throughout the course of this

study.

iv

TABLE OF CONTENTS

Page
LIST OF TABLES ................................................................................ ix
LIST OF FIGURES ................................................................................ xi
CHAPTER 1
INTRODUCTION .................................................................................. 1
CHAPTER 2
LITERATURE REVIEW ......................................................................... 9
2.1. EXTRUSION ............................................................................. 10
2.1.1. Independent Variables ............................................................. 12
2.1.2. Dependent Variables ............................................................... 13
2.1.2.1. Expansion Ratio and Bulk Density ..................................... 14
2.1.2.2. Water Absorption Index and Water Solubility Index ............... 16
2.1.2.3. Pasting Properties ........................................................ 16
2.1.2.4. Thermal Properties ...................................................... 17
2.1.2.5 Breaking Strength ......................................................... 18
2.1.3. Changes in Starch, Proteins, and Heat-Labile Components during
Extrusion Cooking ................................................................. 18
2.1.3.1. Starch ...................................................................... 18
2.1.3.2. Proteins .................................................................... 19
2.1.3.3. Heat-Labile Components ................................................ 20
2.2. WHEAT FLOUR .......................................................................... 22
2.2.1. Main Components of Wheat Flour: Starch and Protein ..................... 22
2.2.2. Baked Products .................................................................... 24
2.2.3. Cereal-Based Snack Foods from Extruders ................................... 25
2.3. GINSENG ................................................................................. 27
2.3.1. Processing Methods of Ginseng ................................................ 27
2.3.2. Major Components of Ginseng ................................................. 28
2.3.3. Detection of Ginsenosides ...................................................... 30
2.3.4. Medicinal Effects of Ginseng ................................................... 30
2.4. LITERATURE CITED ................................................................... 32
CHAPTER 3
EXTRACTION OF GINSENOSIDES FROM A BLEND OF WHEAT FLOUR
AND GINSENG POWDER ..................................................................... 44
3.1. ABSTRACT ............................................................................... 45
3.2. INTRODUCTION ........................................................................ 47
3.3. MATERIALS AND METHODS ...................................................... 49
3.3.1. Materials ............................................................................ 49
3.3.2. Fractionation of Wheat Flour .................................................... 49
3.3.3. Chemical Analysis ................................................................. 50
3.3.4. Optimization of Sample-to-Solvent Ratio for Extraction of
Ginsenosides ........................................................................ 50
3.3.5. Temperature Effects on Extractable Ginsenosides ............................ 50
3.3.6. Optimization of Ultrasonic Extraction Time for Ginsenosides from
Heat-Treated Samples ............................................................. 51

3.3.7. Fractionation and Identiﬁcation of Ginsenosides by RP-HPLC. . . . . ........52

3.3.8. Statistical Analysis ................................................................. 52
3.4. RESULTS AND DISCUSSION ........................................................ 54
3.4.1. Chemical Analysis ................................................................. 54
3.4.2. Optimization of Sample-to-Solvent Ratio for Extraction of
Ginsenosides ........................................................................ 54
3.4.3. Temperature Effects on Extractable Ginsenosides ............................. 56
3.4.4. Validation of Interactions between Wheat Flour Components and
Ginsenosides in a Blend of Wheat Flour and Ginseng Powder .............. 59
3.4.5. Optimization of Ultrasonic Extraction Time for Ginsenosides from
Heat-Treated Samples ............................................................. 60
3.5. SUMMARY ............................................................................... 63
3.6. LITERATURE CITED .................................................................. 64
CHAPTER4
EFFECTS OF EXTRUSION PROCESS VARIABLES ON EXTRACTABLE
GIN SENOSIDES IN WHEAT-GINSENG EXTRUDATES ............................. 75
4.1. ABSTRACT ............................................................................... 76
4.2. INTRODUCTION ........................................................................ 77
4.3. MATERIALS AND METHODS ....................................................... 80
4.3.1. Materials ............................................................................ 80
4.3.2. Extrusion Cooking ................................................................. 80
4.3.3. Speciﬁc Mechanical Energy ...................................................... 81
4.3.4. Optimization of Ultrasonic Extraction Time for Ginsenosides from
Wheat-Ginseng Extrudates ....................................................... 82
4.3.5. Fractionation and Identiﬁcation of Ginsenosides by RP-HPLC. . . . 8.2
4.3.6. Statistical Analysis ................................................................. 83
4.4. RESULTS AND DISCUSSION ........................................................ 84
4.4.1. Optimization of Ultrasonic Extraction Time for Ginsenosides from
Wheat-Ginseng Extrudates ........................................................ 84
4.4.2. Changes in Ginsenosides during Extrusion Cooking .......................... 84
4.4.3. Correlations among Ginsenosides and Product Temperature ................ 90
4.5. SUMMARY ............................................................................... 92
4.6. LITERATURE CITED ................................................................... 93
CHAPTER 5
EFFECTS OF EXTRUSION PROCESS VARIABLES ON QUALITY
PROPERTIES OF WHEAT-GINSENG EXTRUDATES .............................. 104
5.1. ABSTRACT ............................................................................. 105
5.2. INTRODUCTION ...................................................................... 106
5.3. MATERIALS AND METHODS ..................................................... 109
5.3.1. Materials ........................................................................... 109
5.3.2. Extrusion Cooking ............................................................... 109
5.3.3. Speciﬁc Mechanical Energy .................................................... 109
5.3.4. Expansion Ratio .................................................................. 109
5.3.5. Water Absorption Index and Water Solubility Index ........................ 109
5.3.6. Pasting Properties ................................................................ 110
5.3.7. Thermal Properties ............................................................... 110

vi

5.3.8. Statistical Analysis ............................................................... 111

5.4. RESULTS AND DISCUSSION ...................................................... 112
5.4.1. Expansion Ratio .................................................................. 112
5.4.2. Water Absorption Index and Water Solubility Index ........................ 113
5.4.3. Pasting Properties ................................................................. 115
5.4.4. Thermal Properties ............................................................... 117
5.4.5. Correlations among Extrudate Quality Properties and
System Variables ................................................................. 119
5.5. SUMMARY ............................................................................. 121
5.6. LITERATURE CITED ................................................................. 122
CHAPTER 6
PHYSICOCHEMICAL PROPERTIES OF BREAD AND COOKIES BAKED
FROM WHEAT FLOUR BLENDED WITH GINSENG POWDER ................ 129
6.1. ABSTRACT ............................................................................. 130
6.2. INTRODUCTION ...................................................................... 132
6.3. MATERIALS AND METHODS ..................................................... 134
6.3.1. Materials ........................................................................... 134
6.3.2. Chemical Analysis ................................................................ 134
6.3.3. Farinograph Properties .......................................................... 134
6.3.4. Thermal Properties ............................................................... 134
6.3.5. Pasting Properties ................................................................. 135
6.3.6. Breadmaking ...................................................................... 135
6.3.7. Bread Quality ...................................................................... 136
6.3.8. Cookiemaking ..................................................................... 136
6.3.9. Cookie Quality .................................................................... 137
6.3.10. Sample Preparation for RP-HPLC ............................................ 137
6.3.11. Extraction of Ginsenosides .................................................... 138
6.3.12. Fractionation and Identiﬁcation of Ginsenosides by RP-HPLC ......... 138
6.3.13. Statistical Analysis ............................................................. 139
6.4. RESULTS AND DISCUSSION ....................................................... 140
6.4.1. Farinograph Properties .......................................................... 140
6.4.2. Thermal Properties ............................................................... 140
6.4.3. Pasting Properties ................................................................ 141
6.4.4. Bread Quality Properties ........................................................ 142
6.4.5. Cookie Quality Properties ...................................................... 144
6.4.6. Changes in Ginsenosides during Bread-Baking and Cookie-Baking. . 144
6.5. SUMMARY ............................................................................. 148
6.6. LITERATURE CITED ................................................................. 149
CHAPTER 7
GENERAL CONCLUSION ................................................................... 159
CHAPTER 8
FUTURE RECOMMENDATIONS ......................................................... 164
APPENDIX 1

Effects of Sample Heating of Wheat Flour (WF, 0.9 g), Ginseng Powder

vii

(GP, 0.1 g), and a Blend of WF-GP (0.9 g WF + 0.1 g GP) on Subsequent

Extractable Ginsenosides (Rbl, Re, and Rd) ............................................. 166
APPENDIX 2

Moisture Contents of Wheat-Ginseng Extrudates Extruded at Different

Conditions ........................................................................................ 167
APPENDIX 3.

Effects of Ultrasonic Extraction (U E) Time on Extractability of Ginsenosides
(Rbl, Rc, Rd, Rg2, and Rg3) from Wheat-Ginseng Extrudates Extruded
at Different Conditions .......................................................................... 168

APPENDIX 4.
Die Pressure Related to Different Extrusion Process Variables of
Wheat-Ginseng Extrudates .................................................................... 172

APPENDIX 5.
Residence Time Related to Different Extrusion Process Variables of
Wheat-Ginseng Extrudates .................................................................. 173

APPENDIX 6.
Pasting Properties of Wheat-Ginseng Extrudates Extruded at
Different Conditions ............................................................................ 174

APPENDIX 7.

Thermal Properties' of Hard Wheat Flour (HWF), Ginseng Powder (GP),

Blends of HWF-GP (10, 20, and 30% GP, w/w), Soft Wheat Flour (SWF),

and Blends of SWF-GP (10, 20, and 30% GP, w/w) ..................................... 175

APPENDIX 8.

Pasting Properties of Hard Wheat Flour (HWF), Ginseng Powder (GP),

Blends of HWF-GP (10, 20, and 30% GP, w/w), Soft Wheat Flour (SWF),

and Blends of SWF-GP (10, 20, and 30% GP, w/w) ...................................... 176

viii

LIST OF TABLES

Page
CHAPTER 2
2.1. Comparison between White Ginseng and Red Ginseng
(ﬁ'om Cho et a1 1995) ..................................................................... 40
CHAPTER 3
3.1. Moisture and Protein Contents of Wheat Flour, Ginseng Powder,
Starch Fraction, and Gluten Fraction Samples ........................................ 68

3.2. Quantities of Ginsenosides (Rbl, Re, and Rd) Ultrasonically Extracted

for 30 min at Room Temperature from Wheat Flour (WF, 0.9 g),

Ginseng Powder (GP, 0.1 g), and a Blend of WF-GP (0.9 g WF + 0.1 g GP)

with Different Sample-to-Solvent Ratios .............................................. 69
3.3. Effects of Ultrasonic Extraction Time on Extractability of

Ginsenosides (Rbl, Re, and Rd) ﬁ'om a Heated (90°C) Starch Fraction (SF),

a Gluten Fraction (GF), a Blend of SF-Ginseng Powder (GP), and

a Blend of GF-GP ......................................................................... 70
3.4. Effects of Ultrasonic Extraction Time on Extractability of

Ginsenosides (Rbl, Re, and Rd) ﬁ'om Heated (90°C) Wheat Flour (WF),

Ginseng Powder (GP), and a WF-GP Blend .......................................... 71
CHAPTER 4
4.1. Extrusion Conditions for a Wheat Flour-Ginseng Powder (GP) Blend
( 10% GP, w/w) ............................................................................ 96
4.2. Screw Conﬁguration for Twin-Screw Extruder ...................................... 97

4.3. Effects of Ultrasonic Extraction Time of Wheat Flour (WF, 0.9 g),
Ginseng Powder (GP, 0.1 g), and a WF-GP (0.9 g WF + 0.1 g GP) Blend
on Extractable Ginsenosides (Rbl, Re, and Rd) ..................................... 98
4.4. Quantities of Ginsenosides (Rbl, Rc, Rd, Rg2, and Rg3) Ultrasonically
Extracted for 150 min from a Wheat Flour-Ginseng Powder (GP)

Blend (10% GP, w/w) and Wheat-Ginseng Extrudates ............................... 99
4.5. System Variables Related to Different Extrusion Process Variables of
Wheat-Ginseng Extrudates ............................................................. 100
4.6. Pearson’s Coefﬁcients of Correlation (r) among Ginsenosides (Rbl, Rc, Rd,
and Rg2) and Product Temperature ...................................................... 101
CHAPTER 5

5.1. Effects of Extrusion Process Variables on the Expansion Ratio,

Water Absorption Index, and Water Solubility Index

of Wheat-Ginseng Extrudates .......................................................... 124
5.2. Effects of Extrusion Process Variables on Peak Viscosity,

Transition Peak Temperature, and Transition Enthalpy

of Wheat-Ginseng Extrudates .......................................................... 125
5.3. Pearson’s Coefﬁcients of Correlation (r) among Quality Properties

(Expansion ratio, Water Absorption Index, Water Solubility Index,

Peak Viscosity, Transition Peak Temperature, and Transition Enthalpy) of

Wheat-Ginseng Extrudates and System Variables (Speciﬁc Mechanical Energy

and Product Temperature) ............................................................. 126

ix

CHAPTER 6
6.1. Moisture and Protein Contents of Hard Wheat Flour, Soﬁ Wheat Flour,
and Ginseng Powder Samples ......................................................... 152
6.2. Farinograph Properties of Hard Wheat Flour (HWF), a Blend
of HWF-Ginseng Powder (GP), Soft Wheat Flour (SWF),

and SWF-GP .............................................................................. 153
6.3. Thermal Properties of Hard Wheat Flour (HWF), Ginseng Powder (GP),

a Blend of HWF-GP, Soft Wheat Flour (SWF), and SWF-GP .................... 154
6.4. Pasting Properties of Hard Wheat Flour (HWF), Ginseng Powder (GP),

a Blend of HWF-GP, Soft Wheat Flour (SWF), and SWF-GP .................... 155
6.5. Quality Properties of Breads Baked from Hard Wheat Flour (HWF)

and a Blend of HWF—Ginseng Powder ................................................ 156
6.6. Quality Properties of Cookies Baked from Soﬁ Wheat Flour (SWF)

and a Blend of SWF-Ginseng Powder ................................................ 157

6.7. Quantities of Ginsenosides (Rbl, Rc, Rd, Rg2, and Rg3) Ultrasonically
Extracted for 150 min from Ginseng Powder (GP), Bread Crusts
and Bread Crumbs Baked from Hard Wheat Flour (HWF) and
a Blend of HWF-GP, and Cookies Baked from Soﬁ Wheat Flour (SWF)
and a Blend of SWF-GP ................................................................ 158

LIST OF FIGURES

Page
CHPATER 2
2.1. A traditional processing method for making red ginseng ........................... 41
2.2. Structures of the three main types of aglycon moieties of ginsenosides
(from Shibata 2001) ....................................................................... 42
CHAPTER 3

3.1. RP-HPLC chromatograms of ginsenosides ultrasonically extracted

for 30 min from pure ginseng powder (GP), wheat ﬂour (WP) and a blend of

WF-GP in 70% (v/v) aq. methanol ...................................................... 72
3.2. Effects of sample heating of pure ginseng powder (GP) and

a wheat ﬂour-GP blend on subsequent extractable

ginsenosides (Rbl, Re, and Rd) ........................................................ 73
3.3. Effects of sample heating of a starch fraction-ginseng powder (GP) blend

and a gluten fraction-GP blend on subsequent extractable

ginsenosides (Rbl, Re, and Rd) ......................................................... 74

CHAPTER 4

4.1. Effects of ultrasonic extraction time of wheat-ginseng extrudates
extruded at 30% feed moisture, 200 rpm screw speed, and 120°C zone 5 barrel
temperature on extractable ginsenosides (Rbl, Rc, Rd, and Rg2) ................ 102

4.2. RP-HPLC chromatograms of ginsenosides ultrasonically extracted for 150 min
from a non-extruded blend of wheat ﬂour-ginseng powder
and a wheat-ginseng extrudate extruded at 25% feed moisture,
300 rpm screw speed, and 140°C zone 5 barrel temperature ...................... 103

CHAPTER 5

5.1. Rapid Visco Analyzer pasting curves of a non-extruded wheat ﬂour-
ginseng powder (GP) blend (10% GP, w/w), and a wheat-ginseng extrudate
extruded at 30% feed moisture, 200 rpm screw speed,
and 110°C zone 5 barrel temperature ................................................. 127

5.2. Differential scanning thermograms of a wheat-ginseng extrudate
extruded at 30% feed moisture, 300 rpm screw speed, and 140°C zone 5 barrel
temperature, and a non—extruded wheat ﬂour and ginseng powder (GP) blend
(10% GP, w/w) ........................................................................... 128

xi

CHAPTER 1

INTRODUCTION

Wheat is one of the most important cereal crops to humankind. Common
wheat is divided into soft wheat and hard wheat varieties, depending on its kernel texture.
In addition, hard wheat has higher protein content and stronger protein strength as
compared to soft wheat (Atwell 2001; Posner 2000). Hard wheat ﬂour (HWF) is used
mainly for the production of bread products, while soﬁ wheat ﬂour (SWF) is usually
used to produce snacks, cookies, cakes, pastries, crackers, and pretzels.

Extrusion cooking is a versatile, low cost, and very efﬁcient technology in
food processing, and has been used in the production of breakfast cereals, baby foods,
snack foods, pasta products, pet foods, instant powders, modiﬁed starches, etc. (F icarella
et al 2006; Jin et al 1995; Singh and Singh 2004). During extrusion cooking, a variety
of steps including feed transport, mixing, heating, forming, and drying occur within a
short time (Harper 1998). The food components in the extruder barrel experience high
temperature, shear, and pressure during extrusion cooking (Guy 2001; Harper 1981).
The addition of diverse ingredients to extrusion materials has been done with the
objective of improving the nutritional quality of ﬁnal extruded products and imparting
desirable functional properties (Jin et a1 1995; Schmid et a1 2005; Zazueta-Morales et al
2002)

Bread can be produced by many different ingredients and procedures
depending on tradition, types of ﬂour, and types of bread desired (Pyler 1988). Water
and HWF are the most important ingredients in breadmaking, and yeast, salt, shortening,
and sugar are also added according to manufacturer’s choice. Bread can be produced
by three basic processing steps: mixing or dough formation, fermentation, and baking
(Hosney 1998). Recently, new ingredients such as various ﬁbers, proteins, and

vitamins have been introduced in bread formulations not only to improve the texture but

also to make nutritionally superior products (Dalgetty and Baik 2006; Filipovic et al
2007; Ranhotra et a1 2000).

The formulation for making cookies basically includes SWF, sugar,
shortening, and water. The addition of nutraceutical ingredients to the formulation of
cookies could have advantages, including health beneﬁts. Many recent studies have
reported on nutraceutical cookies, such as high-protein cookies (Singh and Mohamed
2007) and high-dietary ﬁber cookies (Bilgicli et a1 2007).

Ginseng has long been used in Asian countries as a traditional medicine
(Fuzzati 2004). Ginsenosides (G, ginseng saponins) have been known as the main
active components in ginseng root and have been typically used as marker compounds
for quality properties (Chuang et al 1995; Lau et a1 2004; Li et al 1996). It has been
known that the extraction efﬁciency of G from raw plant materials, extracts, and
commercial products is dependent on extraction conditions used, such as extraction
solvents (pure methanol, aq. methanol, pure ethanol, or aq. ethanol), sample-to-solvent
ratio, extraction time, extraction temperature, and extraction methods (heating or
ultrasonication) (Court et al 1996; Fuzzati 2004; Lau et al 2004).

Raw ginseng is processed into two kinds of ginseng, white ginseng and red
ginseng, to improve its preservation and efﬁcacy (Kim et al 2000; Wang et a1 2006).
White ginseng is usually air-dried raw ginseng, while red ginseng is usually made by
ﬁrst steaming raw ginseng at 90-100’C for 2-3 hr and then air-drying (Ha et a1 2005).
Red ginseng is known to be more pharmaceutically active than white ginseng, because
the steaming process causes changes in the chemical composition of G, thereby
enhancing biological activities of ginseng (Ha et a1 2007; Shibata 2001). The

ginsenosides Rg3, Rg5, Rg6, Rh2, Rh3, Rh4, Rs3, Rkl, and F4 are not found in raw and

white ginseng, whereas red ginseng contains these new G generated during the steaming
process (Ha et al 2007; Kim et a1 2000; Kwon et a1 2001). In particular, G Rg3 have
been found to exhibit various biological activities, including anticancer activity (Shibata
2001; Wang et al 2006; Yun et al 2001) and radical scavenging activity (Kang et a1
2006). In addition, it has been reported that G Rg3 inhibit the growth of Helicobacter
pylori (Bae et al 2002).

Healthy foods, especially those with nutraceutical properties, are in great
demand in our health-conscious society. Nutraceutical snacks, bread, and cookies
could be good vehicles in this respect if a portion of the wheat ﬂour (WF) were to be
replaced with nutraceutical ingredients, such as ginseng. Extrusion cooking, bread-
baking, and cookie-baking normally involve the use of high temperature to affect and
alter the nature of many food constituents. Thus, there is a possibility of using WF in
combination with ginseng to produce extruded snack foods, and baked bread and
cookies that contain the ginsenosides found in red ginseng.

In order to pursue these possibilities, the present study was set up with the
following objectives:

1. To explore optimum extraction conditions of G from a blend of WF-GP.

2. To investigate effects of extrusion process variables on extractable G in wheat-
ginseng extrudates (WGE).

3. To evaluate quality properties of WGE extruded under different processing
conditions.

4. To evaluate effects of adding GP on Farinograph, thermal, and pasting properties
of HWF and SWF.

5. To study quality properties of hard wheat-ginseng bread and soft wheat-ginseng

cookies.
6. To investigate changes in G components in hard wheat-ginseng bread and soft
wheat-ginseng cookies upon baking.

The information generated through the present study is aimed at providing
information on a novel processing method of ginseng, and aiding production of new
nutraceutical snack foods, bread, and cookies that incorporate ginseng components.
The following dissertation entails: (1) Literature review, (2) Extraction of ginsenosides
from a blend of wheat ﬂour and ginseng powder, (3) Effects of extrusion process
variables on extractable ginsenosides in wheat-ginseng extrudates (4) Effects of
extrusion process variables on quality properties of wheat-ginseng extrudates, and (5)
physicochemical properties of bread and cookies baked from wheat flour blended with
ginseng powder. The chapters in this dissertation were written in the journal paper

format and thus some of the information from chapter to chapter might appear repetitive.

1.]. LITERATURE CITED

Atwell, W. A. 2001. Composition of commercial ﬂour. Pages 1-14 in: Wheat. W. A.
Atwell, ed. AACC International: St. Paul, MN.

Bae, E. A., Han, M. J., Choo, M. K., Park, S. Y., and Kim, D. H. 2002. Metabolism of
20(8)- and 20(R)-ginsenoside Rg3 by human intestinal bacteria and its relation to in
Vitro biological activities. Biol. Pharm. Bull. 25:58-63.

Bilgicli, N., Ibanoglu, S., and Herken, E. N. 2007. Effect of dietary ﬁbre addition on the
selected nutritional properties of cookies. J. Food Eng. 78:86-89.

Chuang, W. C., Wu, H. K., Sheu, S. J., Chiou, S. H., Chang, H. C., and Chen, Y. P. 1995.
A comparative study on commercial samples of ginseng radix. Planta. Med. 61:459-
465.

Court, W. A., Hendel, J. G., and Elmi, J. 1996. Reversed-phase high-performance liquid
chromatographic determination of ginsenosides of Panax quinquefolium. J.
Chromatogr. A 755:11-17.

Dalgetty, D. D., and Baik, B. 2006. Fortiﬁcation of bread with hulls and cotyledon ﬁbers
isolated from peas, lentils, and chickpeas. Cereal Chem. 83:269-274.

Ficarella, A., Milanese, M., and Laforgia, D. 2006. Numerical study of the extrusion
process in cereals production: Part II. Analysis of variance. J. Food Eng. 72:179-
188.

Filipovic, N., Djuric, M., and Gyura, J. 2007. The effect of the type and quantity of
sugar-beet ﬁbers on bread characteristics. J. Food Eng. 78: 1047-1053.

Fuzzati, N. 2004. Analysis methods of ginsenosides. J. Chromatogr. B 812:119-133.

Guy, R. 2001. Raw materials for extrusion cooking. Pages 5-28 in: Extrusion cooking. R.
Guy, ed. Woodhead Publishing Limited: Cambridge, UK.

Ha, D. C., Lee, J. W., and Ryu, G. H. 2005. Changes in ginsenosides and maltol in dried
raw ginseng during extrusion process. Food Sci. Biotechnol. 14:363-367.

Ha, Y. W., Lim, S. 8., Ha, I. J., Na, Y. C., Seo, J. J., Shin, H. 8., Son, S. H., and Kim, Y.
S. 2007. Preparative isolation of four ginsenosides from Korean red ginseng (steam-
treated Panax ginseng C. A. Meyer), by high-speed counter-current chromatography
coupled with evaporative light scattering detection. J. Chromatogr. A 1151:37-44.

Harper, J. M. 1981. The food extruder. Pages 7-19 in: Extrusion of foods. J. M. Harper,
ed. AACC International: St. Paul, MN.

Harper J. M. 1998. Food extruders and their applications. Pages 1-15 in: Extrusion
cooking. C. Mercier, P. Linko, and J. M. Harper, eds. AACC International: St. Paul,

6

MN.

Hosney, R. C. 1998. Yeast-leavened products. Pages 229-305 in: Principles of cereal
science and technology. R. C. Hosney, ed. AACC International: St. Paul, MN.

Jin, J., Hsieh, F., and Huff, H. E. 1995. Effects of soy ﬁber, salt, sugar, and screw speed
on physical properties and rnicrostructure of corn meal extrudate. J. Cereal Sci.
22:185-194.

Kang, K. S., Kim, H. Y., Yamabe, N., and Yokozawa, T. 2006. Stereospeciﬁcity in
hydroxyl radical scavenging activities of four ginsenosides produced by heat
processing. Bioorg. Med. Chem. Lett. 16:5028-5031.

Kim, W. Y., Kim, J. M., Han, S. B., Lee, S. K, Kim, N. D., Park, M. K., Kim, C. K.,
and Park, J. H. 2000. Steaming of ginseng at high temperature enhances biological
activity. J. Nat. Prod. 63: 1702-1704.

Kwon, S. W., Han, S. 3., Park, I. H., Kim, J. M., Park, M. K, and Park, J. H. 2001.
Liquid chromatographic determination of less polar ginsenosides in processed
ginseng. J. Chromatogr. A 921:335-339.

Lau, A. J., Seo, B. H., Woo, S. 0., and Koh, H. L. 2004. High-performance liquid
chromatographic method with quantitative comparisons of whole chromatograms of
raw and steamed Panax notoginseng. J. Chromatogr. A 1057: 141-149.

Li, T. S. C., Mazza, G., Cottrell, A. C., and Gao, L. 1996. Ginsenosides in roots and
leaves of American ginseng. J. Agric. Food Chem. 44:717-720.

Posner, E. S. 2000. Wheat. Pages 1-29 in: Handbook of cereal science and technology.
K Kulp, and J. G. Ponte, eds. Marcel Dekker, Inc.: New York, NY.

Pyler, E. J. 1988. Wheat and wheat ﬂour. Pages 300-377 in: Baking science and
technology. E. J. Pyler, ed. Sosland Publishing Company: Merriam, KS.

Ranhotra, G. S., Gelroth, J. A., and Okot-Kotber, B. M. 2000. Stability and dietary
contribution of vitamin E added to bread. Cereal Chem. 77: 159-162.

Schmid, A. H., Dolan, K. D., and Ng, P. K. W. 2005. Effect of extruding wheat ﬂour at
lower temperatures on physical attributes of extrudates and on thiamin loss when

using carbon dioxide gas as a pufﬁng agent. Cereal Chem. 82:305-313.

Shibata, S. 2001. Chemistry and cancer activities of ginseng saponins and some related
triterpenoid compounds. J. Korean Med. Sci. 16:828—37.

Singh, M., and Mohamed, A. 2007. Inﬂuence of gluten-soy protein blends on the quality
of reduced carbohydrates cookies. LWT. 40:353-360.

Singh, J., and Singh, N. 2004. Effect of process variables and sodium alginate on

extrusion behavior of nixtamalized corn grit. Int. J. Food Prop. 7:329-340.

Wang, C. 2., Zhang, B., Song, W. X., Wang, A., Ni, M., Luo, X., Aung, H. H., Xie, J. T.
Tong, R., He, T. C., and Yuan, C. S. 2006. Steamed American ginseng berry:
ginsenoside analyses and anticancer activities. J. Agric. Food Chem. 54:9936-9942.

Yun, T. K, Lee, Y. S., Lee, Y. H., Kim, S. I., and Yun. H. J. 2001. Anticarcinogenic
effect of Panax ginseng C. A. Meyer and identiﬁcation of active compounds. J.
Korean Med Sci. 16:86-18.

Zazueta-Morales, J., Martinez-Bustos, F., Jacobo-Valenzuela, N., Ordorica-Falomir, C.,
and Paredes-Lopez, O. 2002. Effects of calcium hydroxide and screw speed on
physicochemical characteristics of extruded blue maize. J. Food Sci. 67:3350-3358.

CHAPTER 2

LITERATURE REVIEW

2.1. Extrusion

A food extruder can be described as a device that transforms a variety of raw
ingredients into intermediate and/or ﬁnal extruded products. Extrusion processes
involve operations of feed transport, mixing, heating, and forming at the same time
(Anderson and Ng 2003; Carnire et al 1990; Ding et a1 2006; Harper 1979). The
cooking temperature can be relatively high during extrusion cooking, but the residence
time (time materials actually spend within the barrel) is usually relatively short, hence
the extrusion cooking process is often referred to as a high temperature short time
(HTST) process. Extrusion cooking can produce ﬁnal extruded products with high
quality, including high digestibility and nutritional value (Cauvain 2001), all
characteristics with appeal.

In general, food extruders can be classiﬁed into two types: single-screw and
twin-screw extruders (Harper 1979). Single-screw extruders are manufactured with
and without grooved barrels and with either single-piece screws or individual slip-on
screw elements of various designs (Van Zuilichem and Stolp 1984). Single-screw
extruders are typically known to include three ﬁrndamental zones: a primary feed zone, a
melt zone, and a pump zone. On the other hand, twin-screw extruders can be classiﬁed
as co-rotating or counter-rotating and as intermeshing or non-intermeshing. The twin
parallel screws ﬁt into a horizontal barrel housing all the ingredients. The most popular
type of twin-screw extruder is those with co-rotating and intermeshing screws, which
have been found to be suitable for many extrusion processes due to the wider control of
the residence time distribution, good mixing intensity, superior heat transfer, and self-
wiping operation (Moore 1994; Van Zuilichem and Stolp 1984).

Via its heat treatment, extrusion cooking can affect and change the nature of

10

many food constituents, including starches and proteins, by changing physical, chemical
and nutritional properties, such as starch gelatinization, protein denaturation, complex
formation between amylose and lipids, and degradation reactions of vitamins, pigments,
etc. (Camire et al 1990; Harper 1979; Iwe et a1 2004; Sgaramella and Ames 1993). In
addition, the moisture content in the ﬁnal product largely determines the end product’s
physical state (glass, rubber, and crystalline).

In recent years, there has been an increasing interest in the production of
extruded foods, such as texturized vegetable proteins, ready-to-eat (RTE) breakfast
cereals, expanded snacks, baby foods, pet foods, instant powders, and pasta products
(Anderson and Ng 2003; Ficarella et a1 2006; Jin et a1 1995; Singh and Singh 2004).
An extruder essentially consists of one or two screws, mixing (kneading) paddles, an
external barrel and an exit die (Ficarella et a1 2006). During extrusion cooking, the
ingredients are mixed, sheared and subjected to elevated temperatures and pressures in
the barrel. The dough exhibits some degree of plasticity that enables the expansion of
the ﬁnal product upon exiting the die (Schmid et a1 2005). In fact, the sudden drop in
pressure at the die results in the release of gases entrapped in the product, determining
the pufﬁng of extrudates (Camire et al 1990).

With extrusion processing, a wide variety of foods can be produced using an
economical, energy-efﬁcient, rapid, and continuous system with numerous ingredients
and processing conditions (Ficarella et al 2006). On the other hand, the extrusion
process presents its own challenges as it is fundamentally extremely complex since there
is a great deal of process variability. Not only process variables, but also product
quality, can be inﬂuenced by small changes in processing conditions (Desrumaux et a1

1999). Based on the extruder type, screw conﬁguration and speed, feed moisture,

ll

temperature proﬁle in the barrels, feed rate, and die type, the ﬁnal product quality can be

changed remarkably (Unlu and Faller 2002).

2.1.1. Independent Variables

Extrusion process variables that directly control product quality properties
are referred to as independent variables (Ryu and Ng 2001). The stability of an
extrusion system and product quality is an optimization of extrusion process variables
and feed ingredients. The major extrusion process variables, which include feed rate,
feed moisture, screw designs and conﬁgurations, screw speed, die design and size, barrel
temperatures, and the addition of other ingredients, can be directly controlled by the
extruder operator (Harper 1981). Ryu and Ng (2001) evaluated the effects of extrusion
process variables such as screw speed, feed moisture, and barrel temperature on the
expansion ratio and mechanical properties of wheat ﬂour and whole cornmeal extrudates.
They observed that the expansion ratio, speciﬁc length, and breaking strength of these
extrudates are signiﬁcantly affected by changes in feed moisture and in barrel
temperature.

The primary crucial extrusion process variable is the stable and consistent
supply of feed ingredients into the extruder. Inconsistent introduction of feed
ingredients is often the cause of inconsistent expansion ratios, poor shape, and varied
textures of ﬁnal extrudates (Moore 1994).

The screw is the key element of an extruder since its conﬁguration and speed
can inﬂuence the unit operation of the extruder. Screw speed directly affects the
degree of the barrel ﬁll, the residence time distribution, and the shear stress on the

material being extruded. Moreover, die pressure during extrusion cooking by the

12

action of both single-screw and twin-screw extruders can be signiﬁcantly affected by the
conﬁguration of the screw and its rotational speed (Harper 1981). Mechanical energy
dissipation in the screws causes the temperature of the ingredients to increase rapidly,
transforming the mass from a granular state into a continuous plasticized mass. The
shear occurring during extrusion cooking is very high and causes mechanical
modiﬁcation of extruded ingredients (Ha et a1 2005). In particular, damage to starch or
protein molecules can reduce their ability to form an elastic matrix capable of expanding
and retaining texture upon passing through the die (Harper 1998).

Most extruders have the ability to control temperature, and the degree of
indirect heating or cooling depends on how the extruder is operated. Barrel heating
generates conductive and convective heat in ﬁlled and partially-ﬁlled zones, and the
temperatures of the barrel zones depend on the physical and rheological properties of the

feed, as well as the chosen barrel temperature proﬁle and screw speed (Moore 1994).

2.1.2. Dependent Variables

Dependent variables change as a result of changes made to the independent
variables, and they include melt temperature, residence time in the extruder, pressure in
the extruder, speciﬁc mechanical energy (SME) and quality properties of ﬁnal extruded
products (Ryu and Ng 2001). Melt temperature corresponds to the disappearance of
native starch crystallinity and is a function of the feed material, mechanical energy input,
and heat transfer (Harper 1981). The SME is a measure of the amount of work being
input 'to the material during extrusion cooking and is affected by the feed rate, feed
moisture, screw conﬁguration, screw speed, barrel temperature proﬁle, and rheological

properties of the extruded material (Onwulata et a1 2001). The SME can also control

13

target-dependent variables such as the expansion ratio, water absorption index (WAI)
and water solubility index (WSI), density of extrudates, and many other measurable
parameters (Ryu and Ng 2001).

The pressure at the ﬁnal die of an extruder attains a value which balances the
output of the extruder with the ﬂow through the die (Harper 1981). The viscosity is
considerably affected by composition and moisture of the feed materials, residence time,
shear rate, and temperature. The rheological properties of a food dough in the extruder
are quite complex and are known to be non-Newtonian (Hagenimana et al 2007).

Residence time is deﬁned as the length of time which the process ingredients
spend in the barrel of the extruder during extrusion cooking (Unlu and Faller 2002).
This has commonly been used in evaluating the performance of an extruder. Analysis
of residence time provides information about the degree of mixing, cooking, and
shearing, all of which play signiﬁcant roles in the quality of the ﬁnal products. The
residence time data are also used for scale-up and improving equipment design (Van
Zuilichem and Stolp 1984).

The most important dependent variables are the quality properties, such as
physicochemical properties [expansion ratio, density, water absorption index (WAI),
water solubility index (W SI), texture, gelatinization, ﬂavor, color, appearance, etc.],
pasting properties (peak, trough, breakdown, ﬁnal viscosity, and setback), and thermal
properties [transition onset temperature (To), transition peak temperature (Tp), transition
enthalpy (AH)] of the ﬁnal extruded products. The following will review the effects of

extrusion process variables on the major quality properties of extrudates.

2.1.2.1. Expansion Ratio and Bulk Density

l4

Harper (1979) reported a couple of elements affecting the expansion ratio of
an extrudate: dough viscosity and elasticity. Feed moisture, however, is known to be
the main factor in controlling both extrudate density and expansion of ﬁnal extruded
products (Ding et al 2006; Fletcher et a1 1985; Liu et a1 2000). A reduction in
gelatinization resulted in a decrease in the expansion ratio of and an increase in the bulk
density of wheat ﬂour extrudates (Ding et al 2006). In addition, Rinaldi et al (2000)
showed that a decreasing trend in the expansion ratio of wheat ﬂour extrudates enriched
with wet okara, and an increasing trend in the bulk density of the extrudates were
observed as the extrusion conditions became increasingly more severe (i.e., high shear
screw conﬁguration). They reported that the decreased expansion and increased bulk
density may be because of the breakdown of starch granules into smaller particles,
which may interfere with bubble expansion upon exiting the die.

The expansion ratio of an extrudate can also be affected by feed materials
and addition of other ingredients. Anderson and Ng (2003) evaluated the effects of
feed protein content levels on extrudate expansion of wheat ﬂour. They reported that
the expansion ratio of wheat ﬂour extrudates was negatively affected by increasing feed
protein content of raw feed material; therefore, higher protein content of raw materials
may not be beneﬁcial to expansion in the extrusion processing of wheat ﬂour.
According to Park et a1 (1993), increasing protein content levels in the feed for the
extrusion of soy ﬂour, corn starch, and raw beef blends reduced the expansion ratio of
the extrudates and increased the density of the extrudates.

Calcium is one ingredient that has been reported to affect extrudate

expansion ratio. In work done by Zazueta-Morales et a1 (2002), it was shown that the

15

addition of calcium hydroxide (0.0 to 0.2%, w/w) to blue maize ﬂour decreased the

expansion ratio of the extrudates ﬁom 3.4 to 1.4.

2.1.2.2. Water Absorption Index (WAI) and Water Solubility Index (W SI)

The WAI measures the amount of water absorbed, mainly by starch
(Anderson et al 1969). According to Ding et al (2006), increasing feed moisture
signiﬁcantly increased values of WAI of wheat ﬂour extrudates; however, an increase in
barrel temperature was observed to cause a signiﬁcant decrease in values of WAI of the
extrudates. Hagenimana et al (2006) reported that high feed moisture during extrusion
cooking of rice ﬂour causes higher values of WAI of the extrudates due to reduced
amounts of degradation of starch granules. On the other hand, values of WAI
decreased with increasing barrel temperature and screw speed, because of an increase in
starch degradation or fragmentation (J in et a1 1995).

The WSI measures the amount of soluble components released from the
starch upon extrusion cooking (Kirby et a1 1988) and has been used as an indicator of
degradation of molecular components (Ding et a1 2005). Hagenimana et a1 (2006)
showed that values of WSI of rice ﬂour extrudates increased with increasing both the
severity of the screw proﬁle in the extruder and the processing temperature. Tanhehco
and Ng (2005) reported that decreasing extrusion feed moisture content or increasing

extruder screw speed led to an increase in WSI values of wheat ﬂour extrudates.

2.1.2.3. Pasting Properties
Extrusion cooking of starch leads to gelatinization, partial or complete

destruction of the crystalline structure, and molecular fragmentation of starch

l6

(McPherson et a1 2000). The degree of gelatinization of starch granules and the extent
of their molecular breakdown are known to affect the viscosity of ﬁnal extruded
products (E-Dash et al 1983).

McPherson et al (2000) investigated the pasting properties of extruded corn
starch using a Rapid Visco Analyzer (RVA), and they reported that starches extruded at
different conditions had differences in hot paste viscosity and ﬁnal viscosity values.
For instance, increasing feed moisture during extrusion cooking increased the starch
viscosity, while increasing extrusion temperature and shear decreased the starch
viscosity. High peak and ﬁnal viscosity values were observed for the corn starch
extrudates prepared at high moisture content and low screw speed (Blanche and Sun
2004); the researchers assumed that the degree of starch transformation was lower

under these extrusion conditions.

2.1.2.4. Thermal Properties

Extrusion cooking causes physical and structural conversions of starches in
wheat ﬂour (Anderson and Ng 2001; Brent et al 1997). The phase transitions
(including Tp and AH) of the extrudates have been studied using differential scanning
calorimetry (DSC). Anderson and Ng (2001) reported that the value of Tp is
particularly associated with the heat stability of non-extruded and extruded samples.
Blanche and Sun (2004) found no gelatinization peak on DSC thermograms obtained for
corn starch extrudates. They speculated that starch granules were completely
gelatinized as a result of extrusion cooking or that starch molecules might be broken

down or depolyrnerized due to heat and shear forces during extrusion cooking.

17

2.1.2.5. Breaking Strength

The breaking strength (hardness) is the maximum force required for a probe
to snap an extrudate into two pieces. As already mentioned, feed moisture, screw
speed, and barrel temperature are the main factors affecting the density and expansion of
extrudates. Hardness is considerably inﬂuenced by the density and expansion of
extrudates. Rinaldi et a1 (2000) observed that breaking strength increased as expansion
ratio decreased and as bulk density increased; they postulated this was due to thickening
of the cell walls and decreasing air cell size, which enhanced the resistance of the
extrudates to fracttu'ing. Onwulata et al (2001) reported that the addition of proteins to
starches increases the sites available for cross-linking and affects texture properties of
the extrudates. Ryu and Ng (2001) observed that breaking strength of wheat ﬂour
extrudates extruded at higher SME input was lower than those made at lower SME input,
and they suggested that extrudates produced under higher SME conditions had a softer

texture than those extruded under lower SME conditions.

2.1.3. Changes in Starch, Proteins, and Heat-Labile Components during Extrusion
Cooking
2.1.3.1. Starch

It has been reported that starch is the predominant functional ingredient in
extruded snacks, RTE cereals, and other extruded products (Mercier and Feillet 1975).
Extrusion cooking of starch granules causes changes in their morphological and
molecular structure depending on several factors including feed moisture, cooking
temperature, and mechanical and thermal energy input (Barres et a1 1990). Ozcan et al

(2005) showed that extrusion cooking of corn starch at high temperature (165°C) and

18

low moisture content (8.72%) with moderate shear resulted in changing both gelatinized
and melted starches into fully cooked amorphous extrudates whose starches crystallize
after being cooled. Moreover, it has been known that increasing extrusion temperature
causes a higher degree of gelatinization, while increasing screw speed and increasing die
diameter each lead to a reduction in gelatinization (Blanche and Sun 2004; McPherson et

al 2000).

2.1.3.2. Proteins

Heat and shear during extrusion cooking can induce the denaturation of
proteins and weaken the forces that stabilize the tertiary and quaternary structures of
proteins. The result is the formation of a new molecular aggregate structure (Guy
2001). According to Anderson and Ng (2003), during extrusion cooking of wheat ﬂour,
gluten is signiﬁcantly involved in the microstructural and textural formation of the
extrudates. Furthermore, intermolecular disulﬁde bonding is the main cause of protein
aggregation in the extrusion process (Anderson and Ng 2000; Guy 2001 ).

During extrusion cooking, a reducing sugar such as glucose, fructose, lactose,
or maltose may react with free amino groups in proteins following the so-called Maillard
reaction (Harper 1979). This is a complex reaction that is often desired for the
development of golden brown color and caramel aroma in extruded products, but it is
also generally responsible for a reduction in the nutritional value (Harper 1979).
According to Guy (2001), during extrusion cooking, the Maillard reaction was favored
by conditions of high temperature (more than 180°C) and screw speed (more than 100

rpm) in combination with low feed moisture content (less than 15%).

19

2.1.3.3. Heat-Labile Components

A disadvantage of the extrusion process may be related to the relatively high
temperature, shear, and pressure conditions. High temperatures in barrels during
extrusion cooking can cause the loss of some heat-labile or unstable ingredients such as
vitamins, antioxidants, and ﬂavors (Camire et a1 1990). Camire et a1 (2002) showed
that the amounts of grape and blueberry anthocyanins in RTE breakfast cereals were
signiﬁcantly reduced during the extrusion process. According to Bhandari et al (2001),
the addition of ﬂavoring is sometimes required as a ﬁnal step of extrusion due to the
destruction of a great deal of ﬂavor during the extrusion process. Schmid et a1 (2005)
reported that vitamins were destroyed or decreased during high temperature extrusion
cooking, and they reported that the loss of vitamins results from breakdown of chemical
bonds because of shearing (mixing) or the reduction in vitamin stability during heating.
After extrusion cooking, some extruded foods are fortiﬁed with vitamins by spraying
onto the surface of the extrudate because of the critical loss of vitamins by high
temperature extrusion cooking (Burns et a1 2000).

According to Bhandari et a1 (2001), feed moisture is the most important
factor affecting ﬂavor loss during extrusion cooking, and they reported that increasing
the feed moisture decreases the losses in ﬂavor. Killeit (1994) showed that the
reduction in thiamin loss at higher feed moisture contents was related to the lower dough
viscosities, since the higher feed moisture extrusion conditions can lead to reductions in
degree of shear and energy input on the extruded product.

Results from effects of different screw speeds on nutrient loss during
extrusion cooking are still controversial. There have been two main opposing results:

Guzman-Tello and Cheftel (1987) showed that elevating the screw Speed caused more

20

thiamin loss at high temperatures, even though the extrusion residence time was shorter.
Schmid et al (2005) also found similar results. They reported that increasing the screw
speed resulted in more thiamin loss, indicating that shear effects played the more
dominant role in thiamin degradation than did temperature effects. Conversely, in
other studies, reduction in vitamin content was lessened with increasing screw speed,

and was reported to be due to decreased residence time in the barrel (Killeit 1994).

21

2.2. Wheat Flour

Wheat is one of the most important cereal crops in the world. It has been
used for human food since it has a higher nutritive proﬁle and is relatively easily
harvested, stored, transported and processed, as compared to other grains (Dobraszczyk
2001; Posner 2000). There are three main wheat species: Triticum aestivum, which is
also called common wheat, T. durum, or durum wheat, and T. compactum, or club wheat
(Pyler 1988; Yamazaki et a1 1981).

Wheat ﬂour consists mainly of starch (about 70-75%), water (about 12-14%),
protein (about 8-16%), and minor constituents such as non-starch polysaccharides (about
2-3%), lipids (about 2%), and ash (about 1%). Quality of wheat ﬂour depends on the
quantity and characteristics of its constituents that are different ﬁom other wheat

cultivars.

2.2.1. Main Components of Wheat Flour: Starch and Protein

Starch, the most important storage polysaccharide and the most abundant
component of many plant seeds and tubers, is a major component of wheat ﬂour.
Wheat starch, in particular, is used in the food industry to provide the desired
rheological properties or to improve the physical stability of different products, such as
salad dressings, sauces, soups, extruded products, and baked goods (Dobraszczyk 2001).

The major components of starch are the glucose polymers, amylose and
arnylopectin (Heinemann et al 2003; Tester et a1 2004). Amylose is a fundamentally
linear molecule, comprised of a-1,4 linked glucose polymers. In general, a single
amylose molecule has 1500-6000 glucose units (Atwell 2001) and a molecular weight of

about 1 x 105 to l x 10" (Tester et al 2004). In recent years, it has also been reported

22

that a fraction of the amylose molecules has a low degree of branching by a-l,6 linkages
(Bail et al 2005). In contrast, amylopectin is a very large and highly branched chain
molecule, consisting of both (rt-1,6 and a-1,4 linked glucose polymers. In general, a
single amylopectin molecule has 3 x 105 to 3 x 106 glucose units (Atwell 2001) and a
molecular weight of about 1 x 107 to 1 x 109 (Tester et a1 2004). The
amylose/amylopectin ratio differs among starches, but typical levels of amylose and
amylopectin are 25-28% and 72-75%, respectively (Atwell 2001; Tester et al 2004).

According to Osborne (1907), wheat proteins can be classiﬁed into albumins,
globulins, gliadins, and glutenins. Albumins are extractable in water, globulins in salt
solutions, gliadins in aqueous alcohol, and glutenins in dilute acid or alkali. Among
these four groups of proteins, the gluten proteins, which include the gliadins and
glutenins, are the major storage proteins of wheat.

The gluten proteins are known to make up 80 to 85% of total wheat proteins
(Atwell 2001). Gluten proteins can be easily separated from other components
including starch and water-soluble constituents in wheat ﬂour because they are insoluble
in water. Therefore, gluten can also be deﬁned as the rubbery mass that remains when
wheat dough is washed to remove starch granules and water-soluble constituents
(Wieser 2007). Gliadins represent a highly polymorphic group of monomeric gluten
proteins with molecular weights about 40,000 and are very sticky when hydrated
(Lindsay and Skerritt 1999). Gliadins are known to be associated with the
cohesiveness of wheat dough (Hoseney 1998). Gliadins can be divided into the a, B, y,
and (1)-type in terms of their different primary structures (Wieser 2007).

In contrast, glutenins are a heterogeneous mixture of polymers with

molecular weights varying from about 100,000 to several millions (Atwell 2001).

23

Generally speaking, glutenins comprise 55-70% of the gluten, and glutenin molecules
are larger than gliadin molecules. Glutenins appear to be responsible for the resistance
of wheat dough to extension (Hoseney 1998). It is known that glutenin is built of
glutenin subunits that are linked via disulﬁde bonds (Wieser 2007). These glutenin
subunits can be separated by reduction of these disulﬁde bonds with reducing agents
such as B-mercaptoethanol or dithiothreitol. Four different groups of glutenin subunits
can be distinguished: high molecular weight glutenin subunits (HMW-GS) with
molecular weights between 65,000 and 90,000, and B-, C-, and D-type low molecular
weight glutenin subunits (LMW-GS) with molecular weights between 30,000 and

60,000 (Hou et a1 1996; Lindsay and Skerritt 1999).

2.2.2. Baked Products

Common wheat is divided into soft wheat and hard wheat, based on its kernel
texture. In particular, soft wheat has lower protein content and weaker protein strength
as compared to hard wheat (Posner 2000). Hard wheat ﬂour is used mainly for the
production of bread products, while soft wheat ﬂour is usually used to produce
chemically-leavened products such as cakes, cookies, crackers, and pretzels (Pyler 198 8).

A Visco-elastic dough can be made when wheat ﬂour is combined with water.
It is known that the dough can retain the gas produced during fermentation, thus yielding
a leavened product, especially bread (Cauvain 2001). In general, the major ingredient
for making bread is hard wheat ﬂour with other ingredients being yeast, salt, and water
(Hoseney 1998), whereas the major ingredient for baking cookies is soft wheat ﬂour.
In particular, the cookie formulation has higher sugar, higher shortening, and lower

water contents as compared to the bread formulation (Hoseney 1998). In addition,

24

other ingredients such as non-fat dry milk, salt, sodium bicarbonate, high ﬁuctose corn syrup,

and ammonium bicarbonate are added to make cookies (T ownsend 2001; Yamazaki 1962).

2.2.3. Cereal-Based Snack Foods from Extruders

Snack foods contribute a signiﬁcant portion to many consumers’ daily food
and caloric intake. They consist essentially of a cereal blend extruded with a certain
amount of water. Wheat ﬂour and corn meal have usually been used as ingredients in
snack foods (Matz 1993). Wheat ﬂour has been widely used in the extrusion industry
and the effects of process variables on wheat extrudate properties have been studied (Ali
et a1 1996; Anderson and Ng 2001; Ding et a1 2006; Harper 1979; Mercier and Feillet
1975; Ryu and Ng 2001; Schmid et a1 2005).

Today’s consumers are expecting a wide variety of selection of snack foods.
Extrusion has imparted one manufacturing method for producing novel snack products
and has modernized many traditional snack manufacturing processes (Huber and Rokey
1990). Currently, there is an expanding number of extrusion equipment types and
suppliers available in the market for snack producers. Although this may challenge the
snack producers in their choice of equipment, it should also encourage a wider use of
extrusion cooking and provide the possibility of producing low cost and functional snack
foods (Huber 2001; Huber and Rockey 1990).

During the extrusion cooking of snack foods, ﬂour and other ingredients are
combined and cooked simultaneously in barrels. The resulting mass is forced through
an extrusion die. Upon exiting the die, moisture in the extrudate ﬂashes off, leading to

expansion and cooling of the product (Killeit 1994). The shaped products are cut into

25

individual pieces, and the newly produced snack food is then ready for packaging and
distribution.

A snack food can be made of many diverse ingredients including those from
plant, animal, and chemical sources. A usual formula for making snack foods is
composed fundamentally of carbohydrates (starch), plus smaller amounts of proteins,
fats and seasonings (Moore 1994). Starch is a main food constituent, present in most
extruded snack food products at about a 70% level, and it ﬁrnctions in extruded products
as a binding agent, viscosity builder, and emulsiﬁer. In addition, it enhances the
textural characteristics of foods, such as the expansion ratio, bulk density, mouth feel,

and the aesthetic characteristics, such as color (Guy 2001).

26

2.3. Ginseng

Ginseng is one of the most popular medicinal plants in the world, and has long
been conventionally consumed for strengthening immunity and relieving fatigue (Kim et a1
2005). Many current researchers have conﬁrmed the pharmaceutical effects of ginseng (Bae
et a1 2002; Kang et al 2006; Shibata 2001; Yun et al 2001). Ginseng is referred to as being in
a group of “adaptogenic” herbs in most of the alternative medicine literature, because ginseng
has many beneﬁcial effects, including increased resistance to physical, chemical, and
biological stresses and improved general vitality (Fuzzati et a1 1999; Fuzzati 2004). In recent
years, ginseng products have become increasingly popular and readily available in pharmacies
and health stores around the world.

Its name, ginseng, originated in the Chinese language from the similarity of its root
shape to the shape of the human body (Kwon et a1 1990). The genus name Panax means
“cure all” in Greek (Cho et a] 1995; Ha et a1 2005). The most widely used Panax species
are Panax ginseng (Korean or Asian ginseng), Panax quinquefolius (American ginseng),
Panax notoginseng (Tienchi or Sanchi), Panax vietnamensis (Vietnamese ginseng) and
Panax japom'cus (Japanese ginseng). Korean ginseng has been studied frequently by

many researchers, and is known as a tonic and a panacea in Asia (Fuzzati 2004).

2.3.1. Processing Methods of Ginseng

In general, 4—6 year old Korean ginseng plants are used for medicinal purposes.
“Susam”, which is freshly harvested ginseng root, is also called fresh or raw ginseng because
it is not yet dried. It contains about 70-75% water and easily spoils, so that it is very hard to
store Susarn for more than one week Raw ginseng root is processed to make white and red

ginseng. White ginseng is another name for dried ginseng (about 12-16% moisture content,

27

wb), and has a much longer storage stability than Susarn (Cho et a1 1995). Sun or hot-air
drying is required to produce white ginseng. The color of white ginseng is either milky white
or light yellow, depending on the variety (Ha et a1 2005). Red ginseng is a unique ginseng
product produced by the steaming and then drying of raw ginseng. It preserves the original
root shape but the color changes to a light reddish brown or deep maroon and it can be stored
for a long period of time (Cho et a1 1995).

The traditional processing of red ginseng is depicted in Figure 2.1. During the
heating or steaming process to produce red ginseng, the ginsenosides (G, ginseng saponins),
polyacetylene, acidic polysaccharides, and amino acids go through chemical changes and are
transformed to active substances that do not exist in raw ginseng or white ginseng (Ha et a1

2005; Kwon et al 2001). Table 2.1 shows the compositional differences between white and

red ginseng.

2.3.2. Major Components of Ginseng

In general, herbal medicines include very intricate chemical compositions
which may differ signiﬁcantly based on factors that include growing location, climate,
and species used (Ji et al 2001; Lau et al 2004; Li et al 1996). Ginseng is one of the
most popular herbal medicines in the world and it has particularly complex compositions
since many products from different genera and species can be described as ‘ginseng’ (Ji
et a1 2001).

Traditionally, the dried roots of the plants belonging to the genus Panax are
called ginseng The dried root contains various pharmaceutical components, including
ginsenosides (G, ginseng saponins), polyacetylenes, polyphenolic compounds, and

acidic polysaccharides (Kim et al 2005). The various Panax species are used slightly

28

differently for traditional medicines and contain different components, but all species
contain 2G, which are the most widely studied chemical components of ginseng. In
particular, they are believed to be a main active constituent (Attele et a1 1999), and are
typically used as marker compounds for quality properties (Chuang et a1 1995; Lau et a1
2004; Li et a1 1996). About 30 different G [Ra], Ra2, Ra3, Rbl, Rb2, Rb3, Rc, Rd,
20(S)-Rg3, 20(R)-Rg3, Rh2, R31, R32, Quinquenoside-Rl , Notoginsenoside-R4, malonyl
Rbl, malonyl Rb2, malonyl Rc, malonyl Rd, Re, Rf, Rgl, Rg2, Rbl, 20Glc-Rf,
Notoginsenoside-Rl, 20(R)-Rg2, 20(R)-Rhl, Rh4, and Re] have so far been separated from
various ginseng species, including ﬁve major G (G Rbl, G Rb2, G Rc, G Re, and G
Rgl) that constitute more than 80% of the total G content (Kim et a1 2005).

Aglycons of G are of the dammarane-type triterpenes. The characteristic
dammarane G can be divided into three major groups, depending on their characteristic
aglycon: the protopanaxadiol, protopanaxatriol, and oleanoic acid (Li et a1 2006).
Figure 2.2 shows the structures of the three main types of aglycon moieties of G. The
only oleanolic acid-type G is ginsenoside Ro. Four malonyl derivatives (malonyl G
Rbl, malonyl G Rb2, malonyl G Re, and malonyl G Rd) of G Rbl, G Rb2, G Re, and G
Rd, respectively, have also been isolated and characterized (Fuzzati et a1 1999). In
addition, the G R0 and the malonyl G are regarded as “acidic” G; however, the others
are typically called “neutral” G (Fuzzati 2004). The protopanaxadiols include the G
Rbl, G Rb2, G Rc, G Rd, and G Rg3 and the protopanaxatriols include G Re, G Rf, G
Rgl, G Rg2, and G Rh (Kwon et a1 1990). G (Rg3, Rg5, Rg6, Rh2, Rh3, Rh4, RS3,
Rkl , and F4) are known to be unique to red ginseng, while malonyl G Rbl, malonyl G Rb2,
malonyl G Re, and malonyl G Rd are known to be unique to white ginseng (Cho et a1 1995;

Ha et a1 2007).

29

2.3.3. Detection of Ginsenosides

Many studies have been done since the 1970’s with the purpose of
developing analytical methods for the extraction, identiﬁcation, quantiﬁcation and
quality control of G in raw plant materials (Corbit et al 2005; Court et a1 1996; Fuzzati et
al 1999; Ji et al 2001; Li et al 1996; Wang et a1 2006), extracts (Choi et al 1982; K0 et a1
2005), and marketed products (Choi et a1 1984; Harkey et a1 2001). Those researchers
attempted to differentiate the G composition among the various Panax species.
Furthermore, studies regarding changes in G composition due to different traditional
processing of white and red ginseng have been undertaken (Fuzzati 2004; Kim et al
2000; Kwon et a1 2001; Lau et al 2003; Lau et al 2004).

There are many methods for the analysis of G, including thin layer
chromatography (TLC), gas chromatography (GC), high performance liquid
chromatography (HPLC), GC-mass spectrometry (MS), HPLC-MS and most recently
HPLC-MS/MS. HPLC has been used for the identiﬁcation and fractionation of
ginsenosides in raw ginseng or ginseng products, but generally does not offer structural
information. The advantage of GC-MS is to identify the structural information, but on
the other hand, the broad fractionation that occurs during GC-MS could make
identiﬁcation of speciﬁc G challenging. Since HPLC-MS and HPLC-MS/MS
procedures are somewhat expensive, they can be impractical for typical commercial

analysis (Ji et al 2001).

2.3.4. Medicinal Effects of Ginseng

G Rg3, one of the most important components of the protopanaxadiols and

30

major components of red ginseng, have been found to show various biological activities
including anticancer activities (Shibata 2001; Yun et al 2001), and radical scavenging
activities (Kang et al 2006). Furthermore, it has been reported that G Rg3 inhibit the
growth of Helicobacter pylori (Bae et a1 2002). G Rb] and G Rd (the
protopanaxadiols) are reported to inﬂuence the overall activity of the CNS (central
nervous system, Cho et a1 1995). G Rgl and G Rg2 (the protopanaxatriols) are known to
possess properties of CNS excitation and anti-fatigue (Cho et a1 1995; Li et a1 2005).
In addition to G, ginseng also contains polyacetylenes, which have anticancer activities
(Matsunaga et a1 1989), phenolic compounds, which have antioxidant activities (Suh et
al 1996), and acidic polysaccharides, which have immune-system-controlling activities
(Jie et a1 1984). Na et a1 (2004) reported that ginseng polysaccharides also have
antitumor, anticomplementary, antiulcer, and immunopotentiating effects even though G
have been regarded as the major component for explanation of ginseng’s medicinal

effects.

31

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39

Table 2.1.

Comparison between White Ginseng and Red Ginseng (from Cho et a1 1995)

 

 

White Ginseng Red Ginseng
Processing Methods Sun or air drying Steaming and drying
Color Milky white or light Light reddish brown or
yellow deep maroon

Unique Ginsenosides

Acidic Polysaccharides

Enzymes

Starch
Polyacetylene Compounds

Plgments

Malony ginsenosides

(Rbl, Rb2, Re, and Rd)

2-3%

Mostly remain active
(Amylase, invertase)
Intact
0.1-0.2 mg/g

Natural pigments

Ginsenosides Rg3, Rg5,
Rg6, Rh2, Rh3, Rh4,
RS3, Rkl, and F4
7-8%

All inactivated

Gelatinized
0.6-1.0 mg/g

Melanoidin pigments

 

40

 

Washing of freshly harvested raw ginseng

 

 

 

ﬂ

 

Steaming (90 to 100°C, for 2-3hr)

 

 

 

ﬂ

 

First drying at 50-55°C (to a moisture content of 35-40%)

 

 

 

[l

 

Aging for 24—48 hr

 

 

 

ll

 

Second drying at 25-50°C (to a moisture content of 16%)

 

 

 

ﬂ

 

Red ginseng

 

 

 

Fig. 2.1. A traditional processing method for making red ginseng.

41

<Protopanaxadiols>

R10

 

 

 

Ginsenosides R1 R2
Rbl Glcz-Glc Glc6-Glc
sz Glcz-Glc Glc”-Ara(p)
Rc Glcz-Glc GIc‘-Ara(f)
Rd Glcz-Glc Glc
Malonyl-Rbl Glcz-Glc6-Ma Glc6-Glc
Malonyl-Rb2 Glcz-Glc°-Ma Glc°-Ara(p)
Malonyl-Rc Glcz-Glc6-Ma Glc°-Ara(f)
Malonyl-Rd Glcz-Glc6-Ma Glc
Rg3 Glcz-Glc H
Rh2 Glc H

 

(p): pyranosyl; (f): furanosyl; Ma: HOOC-CHz-COOH.
Fig. 2.2. Structures of the three main types of aglycon moieties of ginsenosides

(from Shibata 2001).

42

<Protopanaxatriols>

 

 

 

R20
/” “NR
.W,”
’i
I
Ginsenosides R1 R2
Rg2 Glcz-Rha H
Re Glcz-Rha Glc
Rh] Glc H
Rf Glcz-Glc H

 

<Oleanolic acid>

COOGk

 

 

 

 

R0
Ginsenosides R
R0 Glcz-Glc
Fig. 2.2. (Continued).

43

CHAPTER 3
Extraction of Ginsenosides from a Blend of Wheat

Flour and Ginseng Powder

3.]. ABSTRACT

This study explored conditions for maximum extraction of ginsenosides (G,
ginseng saponins) from a blend of wheat ﬂour (WF)-ginseng powder (GP). The weight
of each sample (WP: 0.9 g, GP: 0.] g, or WF-GP: 0.9 g WF + 0.1 g GP) was ﬁxed and
different amounts of 70% (v/v) aq. methanol (1-14 mL) were added to each sample.
Samples were then ultrasonically extracted for G for 30 min. Individual ginsenosides
Rbl, Re, and Rd were fractionated and identiﬁed by RP-HPLC. The maximum
extraction of the G were obtained by the sample-to-solvent ratio of 0.025;] (mg/uL) for
GP alone and the sample-to-solvent ratio of 0.071 (mg/uL) for WF-GP. To investigate
temperature effects on extractable G in a WF-GP blend, WP (0.9 g), GP (0.] g), or WF-
GP (0.9 g WF + 0.1 g GP) was mixed with distilled water (4.5, 0.5, or 5.0 mL,
respectively) and heated at temperatures from 25 to 90°C prior to extraction. The
quantities of G extracted from GP were constant at all the temperatures studied, while
the quantities of G extracted ﬁom WF-GP decreased with increasing heating
temperature. Results suggested that interactions between WF components and G (Rbl ,
Re, and Rd) existed, and that the interactions increased with increasing temperature.
WF was fractionated into two components: a starch fraction (SF) and a gluten fraction
(GF) to examine the interactions between WF components and G in a heated blend of
WF-GP. Extractable G (Rbl, Re, and Rd) decreased with increasing heating
temperature for both the SF-GP blend and the GF-GP blend. These ﬁndings conﬁrmed
that interactions occurred not only between the SF and G but also between the GF and G
in a blend of WF-GP at the higher temperature (90°C). In addition, the quantities of
extractable G ultrasonically extracted for 30 or 60 min from GF-GP heated at 90°C were

signiﬁcantly higher than those of extractable G ultrasonically extracted for 30 min from

45

SF-GP heated at 90°C, indicating that the SF has more interactions with G than did the
GF. To disrupt the interactions, ultrasonic extraction (UE) time was increased to 60, 90,
or 120 min for each sample. The quantities of G extracted ﬁom GP were not affected
by increasing UE time. However, the quantities of G extracted from WF-GP increased
with increasing UE time, and similar quantities of Rbl, Re, and Rd (0.87, 0.53, and 0.26
mg per 100 mg of GP, respectively) were obtained after 90 min of UE as were extracted
from pure GP at 30 min. Thus, the interactions between WF components and G could

be disrupted by increasing UE time to 90 min for maximum extraction.

46

3.2. INTRODUCTION

Saponins are known as glycosides with distinctive foaming and emulsifying
characteristics and are composed of steroid or triterpenoid aglycons and various sugar
moieties (Ikedo et al 1996). Saponins have been found in many plant species,
including ginseng, spinach, asparagus, sugarbeets, oats, and legumes. Ginsenosides (G,
ginseng saponins) are the major active components of ginseng, one of the most popular
herbal medicines worldwide, contributing to pharmaceutical activity. Their basic
structure contains triterpenoid aglycon and various sugar moieties (Fuzzati 2004;
Shibata, 2001). Over 30 G have been identiﬁed; among these, Rbl, Rb2, Rc, Rd, and
Re are the most abundant (Corbit et al 2005; Kim et a1 2005).

Numerous studies have reported that proteins interact with relatively small
molecules, such as saponins and ﬂavor compounds (Heng et al 2004; Ikedo et a1 1996;
Potter et a1 1993; Samthein-Graf and Mesa 2004). Potter et a1 (1993) studied the
interactions of isolated soy proteins and caseins with quillaja saponin using gel
electrophoresis. They noted that the saponins interacted with soy proteins much more
quickly than with caseins. In addition, Ikedo et a1 (1996) reported that soyasaponin
interacts with bovine serum albumin (BSA). They hypothesized that the interactions
between BSA and soyasaponin include electrostatic and hydrophobic interactions.

It has been shown that starch interacts with various components of food
systems, such as lipids, proteins, iodine, organic alcohols or acids, and ﬂavor
compounds, including terpenes, aldehydes, and lactones (Arvisenet et al 2002; Derycke
et a1 2005; Heinemann et al 2001; Jouquand et a1 2006; Le Bail et a1 2005; Raphaelides

and Georgiadis 2006; Toro-Vazquez et a1 2003). Arvisenet et a] (2002) reported that

47

the interactions of gelatinized starch with its interaction partners occur during heating
process of a starch-based food system.

Wheat ﬂour (WF) primarily consists of proteins and starch. Depending on
the wheat variety, the contents of protein and starch in ﬂour can vary from 7-15% and
63-72%, respectively, at a 14% moisture basis (Atwell 2001). Therefore, in the present
study, it was hypothesized that interactions between WF components and G would take
place during the heating process required to make a cereal product, if ginseng is to be
used as one of the ingredients. It was also hypothesized that the extraction efﬁciency
of G from the cereal product would be related to the degree of interaction between WF
components and the G. For more than 30 years, numerous studies on the extraction of
G have been performed on raw plant materials (Corbit et al 2005; Court et a1 1996;
Fuzzati et a1 1999; Ji et a1 2001; Kwon et a1 2001; Li et al 1996; Park et a1 1996; Wang
et al 2006), extracts (Choi et a1 1982; K0 et a1 2005), and commercial products (Choi et
a1 1984; Harkey et a1 2001). Studies have employed various extraction solvents (pure
methanol, aq. methanol, pure ethanol, or aq. ethanol) and extraction methods (heating or
ultrasonication) to facilitate the extraction of G from various sources. However, the
optimum extraction conditions, such as solvent, time, temperature, and sample-to-
solvent ratio, for G that have interacted with other components (such as proteins and
starch) in a food system have not been reported. Therefore, the objective of the present
study was to explore optimum extraction conditions of G (Rbl, Re, and Rd) from a

blend of WF and ginseng powder (GP).

48

3.3. MATERIALS AND METHODS

3.3.]. Materials

Soft WF was obtained from the Mennel Milling Co. in Fostoria, OH, USA.
The raw ginseng root (Panax ginseng C. A. Meyer), harvested after four years growth
and milled into powder (particle size of S 167 um), was cultivated in Geumsan, Korea,
and purchased at a ginseng market. Standards for ginsenosides Rbl, Re, and Rd were

provided by ILHWA Co. (Kuri, Korea).

3.3.2. Fractionation of Wheat Flour

Wheat ﬂour was ﬁactionated into two components: a starch ﬁ'action (SF) and
a gluten fraction (GF) using the procedure described by MacRitchie (1985) with some
modiﬁcations. WF (100 g) was mixed into dough with an amount of water according
to its Farinograph water absorption; the dough was mixed for 30 see, just enough to give
a cohesive mass that was easy to handle. The SF was isolated by hand kneading this
dough in small aliquots of distilled water (total of 800 mL), which were collected, and
the GF was obtained as the residual viscoelastic mass. The distilled water-starchy
Slurry was centrifuged at 5,000 x g for 10 min using a Sorvall RC-SB centriﬁrge (Dupont,
Wilmington, DE, USA) and the supernatant was decanted. The SF (sediment) was
spread in aluminum pans and allowed to air-dry at room temperature for 72 hr. The
wet GF was divided into six parts, frozen, and freeze-dried for 48 hr. The air-dried SF
and freeze-dried GF were initially each ground using a mortar and pestle and then ﬁnally
ground to a particle size of S 330 um (54 mesh screen) using a coffee grinder (Braun
1110., Wobum, MA, USA). All ground fraction samples were placed in ziplock bags,

sealed, and stored at -20°C until needed.

49

3.3.3. Chemical Analysis

WF, GP, SF, and GF samples were analyzed for their moisture and protein
contents using AACCI Approved Methods 44-15A and 46-13 (AACCI, 2000),
respectively. Protein contents were calculated by multiplying the nitrogen content by

the conversion factor 5.7, and reported on a 14% moisture basis.

3.3.4. Optimization of Sample-to-Solvent Ratio for Extraction of Ginsenosides
Different amounts of extraction solvent (70%, v/v, aq. methanol) were added
to each sample (GP, WF, or a blend of WF-GP): l, 2, 3, 4, 5, 6, 8, or 10 mL of 70%
(v/v) aq. methanol was added to GP (0.] g); 14 mL of 70% (v/v) aq. methanol to WF
(0.9 g); and 12 or 14 mL of 70% (v/v) aq. methanol to WF-GP (0.9 g WF + 0.1 g GP).
These tested amounts of extraction solvents gave sample-to-solvent ratios that ranged
from 0.07:] to 0.10:] (mg/uL). The suspension was ultrasonically extracted for 30 min
using an Ultrasonicator (Model FS 14H, Fisher Scientiﬁc, Pittsburgh, PA, USA, Power:
155W) at room temperature and then centrifuged at 11,200 x g for 20 min using a
centrifuge (Model 12-21M, Beckrnan Instruments Inc., Fullerton, CA, USA) at 25°C.
The supernatant was evaporated using a Micro Rotary Evaporator (Model 421-4000,
Labconco, Kansas City, MO, USA) at 50°C. Prior to RP-HPLC analysis for G, the
residue was dissolved in 2 mL of 70% (v/v) aq. methanol and ﬁltered through a 0.45 um
nylon ﬁlter membrane (Millipore, Ireland). Each sample was extracted in duplicate and

two chromatographic runs were performed for each extract.

3.3.5. Temperature Effects on Extractable Ginsenosides

50

WP (0.9 g), GP (0.] g), or WF-GP (0.9 g WP + 0.1 g GP) was mixed with distilled
water (4.5, 0.5, or 5.0 mL, respectively) and heated at 25, 50, 70, or 90°C using a water bath
for 30 min. The SF (0.9 g), GF (0.9 g), a blend of SF-GP (0.9 g SF + 0.1 g GP), or a
blend of GF-GP (0.9 g GF + 0.1 g GP) was mixed with distilled water (4.5, 4.5, 5.0, or
5.0 mL, respectively) and heated at 25 or 90°C using a water bath for 30 min. The
mixture was frozen and dried in a ﬁ'eeze-dryer for 24 hr. To prevent the loss of G from the
ﬁeezedried samples, the samples were not ground An aliquot of 14, 4, or 14 mL of 70%
(v/v) aq. methanol was added to the prepared ﬁ'eeze-dried WF sample, GP sample, or
WF-GP sample, respectively. Fourteen mL of 70% (v/v) aq. methanol were added to
each sample of freeze-dried SF, GF, SF-GP, or GF-GP. The suspension was
ultrasonically extracted for G for 30 min, then centrifuged, evaporated, dissolved, and ﬁltered
as described in 3.3.4. Each sample was extracted in duplicate and two chromatographic

runs were performed for each extract.

3.3.6. Optimization of Ultrasonic Extraction Time for Ginsenosides from Heat-
Treated Samples

WP (0.9 g), GP (0.] g), or WF-GP (0.9 g WF + 0.1 g GP) was mixed with distilled
water (4.5, 0.5, or 5.0 mL, respectively) and heated at 90°C using a water bath for 30 min.
SF (0.9 g), GF (0.9 g), SF-GP (0.9 g SF + 0.1 g GP), or GF-GP (0.9 g GF + 0.1 g GP) was
mixed with distilled water (4.5, 4.5, 5.0, or 5.0 mL, respectively) and heated at 90°C using a
waterbathfor30 min. Eachmixturewas frozenanddriedinaﬁeezedryerfor24hr. To
prevent the loss of G from the freeze-dried samples, the samples were not ground Fourteen,
4, or 14 mL of 70% (v/v) aq. methanol were added to the prepared sample of freeze-dried

WF, GP, or WF-GP, respectively. Fourteen mL of 70% (v/v) aq. methanol were added

51

to each prepared sample of freeze-dried SF, GF, SF-GP, or GF-GP. The suspension
was ultrasonically extracted for G for 30, 60, 90, or 120 min, then centrifuged,
evaporated, dissolved, and ﬁltered as described in 3.3.4. Each sample was extracted in

duplicate and two chromatographic runs were performed for each extract.

3.3.7. Fractionation and Identification of Ginsenosides by RP-HPLC

RP-HPLC was carried out on a Millenium 2010 HPLC Workstation,
consisting of a Waters 600E multi-solvent delivery system (Waters, Milford, MA, USA),
a temperature control module, and a 996 photodiode array detector. Separation of G
was performed at 30°C on a Phenomenex 300 RP, Jupiter 018 column (4.6 x 250 mm, 5
um particle diameter; Phenomenex, Torrance, CA, USA). A Phenomenex security
guard with the same packing material served as the guard column.

The solvents were A: water (distilled water ﬁltered through Milli-Q-Plus,
Millipore, Bedford, MA, USA), and B: HPLC-grade acetonitrile. Separation was
achieved using the following gradient: 0-5 min, 20-30% B; 5-25 min, 30-40% B; 25-30
min, 40-20% B. The solvent ﬂow rate was 1 mL/min and the injection volume was 20
[IL The solvents were purged with helium at the rate of 20 mL/min. The column was
equilibrated for 10 min with 20% acetonitrile prior to injection. The UV detection
wavelength was set at 203 mn (Lau et a1 2004), and the detector output was transmitted

simultaneously to the computer for data storage and graphic representation.

3.3.8. Statistical Analysis
All statistical analyses were performed using SAS version 9.1 (SAS

Institute Inc., Cary, NC, USA). Analysis of variance (ANOVA) was performed using

52

the general linear models (GLM) procedure to determine signiﬁcant differences among
the samples. Means were compared by using Fisher’s least signiﬁcant difference

(LSD) procedure. Signiﬁcance was deﬁned at the 5% level.

53

3.4. RESULTS AND DISCUSSION

3.4.]. Chemical Analysis

Moisture and protein contents of WF, GP, SF, and GF samples are reported
in Table 3.1. The moisture contents of the samples ranged from 1.9 to 11.2% (wb).
Only a very low protein content (2.1%) in the SF was obtained, indicating that good
fractionation had been achieved. The protein content of the WP (8.3%) represents a

typical soft wheat variety.

3.4.2. Optimization of Sample-to-Solvent Ratio for Extraction of Ginsenosides

The representative RP-HPLC chromatograms of G ultrasonically extracted
for 30 min ﬁom pure GP and a blend of WF-GP are illustrated in Figure 3.1. The RP-
HPLC chromatograms of G extracted from WF-GP were nearly identical to the
chromatograms of G extracted from pure GP. This result indicated that adding WF to
GP led to no signiﬁcant impact on the extractable chemical proﬁle of GP. Rbl, Re,
and Rd were the most abundant among all the G and, based on information in the
literature, they are associated with the pharmaceutical activity (Ji et a1 2001).
Therefore, these three G were ﬁactionated and identiﬁed in the present study.
. Ginsenosides Rbl, Re, and Rd were eluted in that order during RP-HPLC ﬁactionation.
This ﬁnding was in good agreement with reports in the literature (Ji et a1 2001; Lau et al
2003; Lau eta12004; Li et al 1996).

Previous studies by others have shown that different solvents extract different
amounts of individual G and the amounts of G change depending on the amounts of the
solvents used (Court et a1 1996; Fuzzati 2004; Lau et a1 2003; Lau et a1 2004). In

addition, Corbit et a1 (2005) noted that the quantities of G are affected by the physical

54

and chemical extraction procedures and extraction time used. F uzzati (2004) reported
that most extraction methods of G employ pure methanol or aq. methanol. Lau et a1
(2003) showed that the amounts of G (Rl, Rbl, Rc, Rd, Re, and Rgl) extracted from
raw Panax notoginseng in 70% (v/v) aq. methanol are higher than those of the G
extracted in 100% methanol. Therefore, they concluded that 70% (v/v) aq. methane] is
the optimum solvent for the extraction and quantiﬁcation of the G. Corbit et a1 (2005)
reported similar results from the study using Panax quinquefolius, i.e., that 100%
methanol was less effective than 70% (v/v) aq. methanol when each solvent was used in
combination with the ultrasonic extraction (UE) method. Court et al (1996) compared
UE and the Soxhlet method for evaluating the quantities of G (Rbl, Rb2, Rc, Rd, Re,
and Rgl) extracted from Panax quinquefolius. They found that UE yields higher
quantities of the G than the Soxhlet method. Furthermore, Lau et a1 (2003) noted that
UE is a rapid and efﬁcient extraction method for G, as compared to the Soxhlet method.
Based on the studies mentioned above, 70% (v/v) aq. methanol (extraction solvent) and
the UE method were chosen for extraction of ginsenosides in the present study.

A preliminary study was performed to identify the most appropriate range of
sample-to-solvent ratios for the extraction of G from WF, GP, and WF-GP. The weight
of each sample (WP: 0.9 g, GP: 0.] g, or WF-GP: 0.9 g WF + 0.1 g GP) was ﬁxed and
different amounts of 70% (v/v) aq. methanol were added to each sample. Solvent
functions as a medium to solubilize target compounds, such as G; thus, the sample-to-
solvent ratio is one of several critical parameters to be considered. Table 3.2 lists the
quantities of G (Rbl, Re, and Rd; on a mg per 100 mg GP basis) ultrasonically extracted
for 30 min from WF, GP, and WF-GP with different sample-to-solvent ratios. The

quantities of G (Rbl, Re, and Rd) extracted from pure GP increased with decreases in

55

the sample-to-solvent ratio from 0.100:l to 0.025;] (mg/uL), but did not increase
signiﬁcantly with further decreases in the ratio. This observation established that the
maximum extraction of G Rbl, G Re, and G Rd from pure GP could be obtained by the
sample-to-solvent ratio of 0025:] (mg/uL). The quantities of G (Rbl, Re, and Rd)
extracted from WF-GP with the sample-to-solvent ratio of 0071:] (mg/uL) were not
signiﬁcantly different ﬁom the quantities of the G extracted from pure GP with the
sample-to-solvent ratios of less than 0.025;] (mg/uL). However, no measurable G
(Rbl, Re, and Rd) in pure WF were found. Based on these results, the sample-to-
solvent ratio of 0025:] (mg/uL) for pure GP and the sample-to-solvent ratio of 0071:]

(mg/uL) for WF-GP were chosen as one of the UE conditions for subsequent studies.

3.4.3. Temperature Effects on Extractable Ginsenosides

Effects of heating of pure GP and a blend of WF-GP on subsequent
extractable G (Rbl, Re, and Rd) are shown in Figure 3.2. Extractable G (Rbl, Rc, and
Rd) decreased with increasing heating temperature for the WF-GP-water blend, but
remained essentially the same for pure GP. No measurable G (Rbl, Re, and Rd) in pure
WF, with distilled water, were found at any of the temperatures studied (data not shown).
These ﬁndings together indicate that when blends of WF-GP-water were heated,
increased interactions between WF components and G took place at the higher
temperatures (70-90’C), causing more difﬁculty in the extraction of G from these blends.
The decreased amounts of extractable G in the heated blends can be related to the
interactions of G with WF components. Furthermore, the interactions between WF
components and G increased with increasing heating temperature. It is possible that

the G are interacting with wheat proteins or with starch during the heating process.

56

To examine the interactions between WF components and G in a heated
blend of WF-GP, WF was fractionated into a SF and a GP. A blend of SF-GP or a
blend of GF-GP was mixed with distilled water and heated at 25 or 90°C for 30 min.
Figure 3.3 illustrates effects of heating these SF-GP and GF-GP samples on subsequent
extractable G (Rbl, Rc, and Rd). Extractable G (Rbl, Re, and Rd) decreased with
increasing heating temperature for both the SF -GP-water blend and the GF-GP-water
blend. No measurable G (Rbl, Re, and Rd) in pure SF and GF, with distilled water, were
found at any of the temperatures studied (data not shown). These ﬁndings conﬁrmed that
interactions between the SF and G and between the GF and G in a blend of WF-GP-
water heated at 90°C did exist. Furthermore, the quantities of extractable G
ultrasonically extracted for 30 or 60 min ﬁ'om GF-GP heated at 90°C were signiﬁcantly
higher (on a g/g basis) than those of extractable G ultrasonically extracted for 30 min
from SF-GP heated at 90°C (Table 3.3). This result could indicate that the SF had a
higher degree of interactions with G than did the GF, and/or that interactions between
the SF and G were stronger than those between the GF and G, thereby resulting in more
difﬁculty in the extraction of G from SF-GP under the given extraction conditions.

As mentioned earlier, G have a complex chemical structure consisting of
triterpenoid aglycon and various sugar moieties. Because the aglycon is highly
hydrophobic and the sugar is hydrophilic, G can be considered as amphiphilic molecules
(Kim et al 1985). According to Ikedo et al (1996), saponins show excellent
emulsifying characteristics due to the presence of the hydrophilic and hydrophobic
groups. These groups can participate in interactions with proteins. Shimoramada et al
(2000) observed interactions between soybean saponin and whey proteins. Potter et a1

(1993) reported that interactions between quillaja saponin and casein occur via the sugar

57

moieties of the saponin with the free amino groups of the proteins (e.g., Maillard
reaction). In the present study, it is suggested that interactions between hydrophilic
groups (sugar moieties) of G and the free amino groups of wheat proteins could take
place during the heating process of the WF-GP-water blend. Moreover, upon
unfolding of wheat protein molecules during the heating process, the previously buried
hydrophobic groups in wheat proteins become accessible for interactions with
hydrophobic groups (aglycons) in G via hydrophobic interactions.

In general, starch undergoes gelatinization upon heating of its granules in the
presence of water. During gelatinization, amylose leaches out of the starch granules
and forms a helix. According to Conde-Petit et al (2006), the outside surface of the
helix is hydrophilic due to the oxygen and hydrogen atoms, while the inner surface of
the helix is hydr0phobic because of the carbon atoms. They also reported that the helix
can interact with hydrophilic and hydrophobic molecules. According to Heinemann et
a1 (2005), various compounds in food system such as terpenes, alcohols, aldehydes, and
lactones can form complexes with amylose during the heating process, and these
complexes display a V type pattern as assessed by X-ray difﬁaction. When amylose is
not forming a complex with other molecules, it is in the form of a double helix (A and B
type, Zobel et a1 1988). Conde-Petit et a1 (2006) reported that higher temperatures are
necessary to plasticize amylose in order to enable the formation of complexes with its
interaction partners. Heinemann et a1 (2003) evaluated the structural features of potato
starch and lactone complexes and they explained interactions between amylopectin and
lactone, as well as interactions between amylose and lactone. Because the external
branches of amylopectin have the same structure as amylose, amylopectin can also

interact in the same way with lactone. In the present study, it is assumed that the outer

58

and inner groups of gelatinized amylose helix could interact with the hydrophilic and
hydrophobic groups, respectively, of G during the heating process. Furthermore, the
leaching of amylose from starch granules could be increased at higher temperatures,

improving the interactions between gelatinized amylose helix and G.

3.4.4. Validation of Interactions between Wheat Flour Components and
Ginsenosides in a Blend of Wheat Flour and Ginseng Powder

The amounts of G Rbl ultrasonically extracted for 30 min from GP and
blends of WF-GP, SF-GP and GF-GP, each heated at 90°C for 30 min, were used to
validate the interactions between WF components and G Rb] in the WF-GP blend.
The amounts of G Rbl ultrasonically extracted for 30 min from pure GP (100 mg)
heated at 90°C and a blend of WF (900 mg)-GP (100 mg) heated at 90°C were 0.88
mg/ 100 mg GP and 0.72 mg/ 100 mg GP, respectively (Table 3.4). The amounts of G
Rb] ultrasonically extracted for 30 min from a blend of SF (900 mg)-GP (100 mg)
heated at 90°C and a blend of OF (900 mg)-GP (100 mg) heated at 90°C were 0.71
mg/ 100 mg GP and 0.77 mg/ 100 mg GP, respectively (Table 3.3).

Based on the amounts of G Rbl ultrasonically extracted from heated GP and
WF-GP, the amount of G Rb] which could interact with WF components in the heated
WF-GP blend was calculated as:
0.88 mg/ 100 mg GP (the amount of G Rb] extracted ﬁom heated GP) - 0.72 mg/ 100 mg
GP (the amount of G Rb] extracted from heated WF-GP) = 0.16 mg/ 100mg GP (the
amount of G Rb] which could interact with WF components in heated WF-GP)

The amounts of G Rb] which could interact with the SF or with the GP in

each blend were calculated in the same way; the amounts of G Rbl calculated were 0.17

59

mg/100 mg GP for the SF and 0.11 mg/IOO mg GP for the GF. For purposes of
comparison, it was assumed that WF (based on dry weight) grossly consists of about
85% SF, 10% GF, and 5% nonstarch polysaccharides and lipids. Based on this
assumption, the following WF composition was obtained for a 900 mg sample:

900 mg (WF) = 765 mg (SF) + 90 mg (GF) + 45 mg (nonstarch polysaccharides and
lipids)

Based on results in the present study of the amounts of G Rbl which could
interact with the SF (900 mg) or the GF (900 mg), calculated values were determined for
the total amounts of G Rb] which could interact with the SF (765mg) and the GF (90
mg) in a blend of WF (900 mg)-GP (100 mg) heated at 90°C. The amounts of G Rbl
calculated were 0.14 mg/ 100 mg GP for the SF and 0.0] mg/ 100 mg GP for the CF.
The sum (0.15 mg/ 100 mg GP) of those two calculated values is close to the measured
amount (0.16 mg/ 100 mg GP) of G Rb] which appeared to interact with WF
components in the WF-GP blend heated at 90°C. Moreover, G Re and G Rd showed
the same trends with their respective calculated values (data not shown). Therefore,
based on the data obtained and the calculations, the degree of interactions of G (Rbl, Re,
and Rd) with the SF in a heated blend of SF-GP and the G with the GP in a heated blend
of GF-GP could explain the total interactions between WF components and the G in a
heated blend of WF-GP. Furthermore, it is postulated that, quantitatively, higher
amounts of G are interacting with the SF than with the GP in a blend of WF-GP heated

at 90°C.

3.4.5. Optimization of Ultrasonic Extraction Time for Ginsenosides from Heat-

Treated Samples

60

Effects of UE time of heated (90°C) samples of WF, GP, SF, GF, and blends
of WF-GP, SF-GP, and GF-GP on extractable G (Rbl, Re, and Rd) are listed in Tables
3.3 and 3.4. No measurable G (Rbl, Re, and Rd) in pure WF, SF, or GF, with distilled
water, were found at any of the UE times studied. The quantities of G (Rbl, Re, and
Rd) extracted from pure GP were not affected by increasing UE time. However, the
quantities of G (Rbl, Re, and Rd) extracted from WF-GP increased with increasing UE
time, and amounts of the three individual G obtained after 90 min of UE were similar to
their counterpart amounts extracted from pure GP at 30 min. Thus, the interactions
between WF components and G could be disrupted by increasing UE time up to 90 min,
for the maximum extraction.

The quantities of G (Rbl, Re, and Rd) extracted from the blend of SF-GP
increased with increasing UE time, and similar amounts of G Rbl, G Re, and G Rd were
obtained after 90 min of UE time as were extracted from pure GP at 30 min. However,
similar amounts of G (Rbl, Re, and Rd) extracted from GF-GP were obtained after 60
min of UE time as were extracted ﬁ'om pure GP at 30 min. Thus, the interactions
between the SF and G as well as between the GF and G could be disrupted by increasing
UE time up to 90 min and 60 min, respectively, for the maximum extractions, indicating
a difference in the strength of the interactions of ginsenosides with starch and protein.

According to Onoﬁ'e and Hettiarachchy (2007), ultrasonication can increase
the extraction efﬁciency of targeted compounds. They also reported that the UE
efficiency is associated with the rate of diffusivity of the compounds from the solid to
the solvent, and is signiﬁcantly affected by the UE conditions such as UE temperature
and UE time. Ultrasound breaks down the cell walls and matrix of a sample

mechanically, exposing targeted compounds to the solvent and subsequently enhancing

61

the rate of mass transfer of compounds to the extractrant (Capelo and Mota 2005;
Onofre and Hettiarachchy 2007). It is plausible in the present study that the number of
interactions disrupted increases with increasing UE time, facilitating the access of the
solvent (70% aq. methanol) to the G, resulting in an increase in the quantities of
individual G until achieving the maximum extraction of the G. Based on the results
obtained in the present study, optimization of UE time appears to be an important
parameter for accomplishing the maximum extraction of G from a cereal product

containing ginseng.

62

3.5. SUMMARY

It is necessary to quantify G in a cereal product if ginseng is to be used as one
of the ingredients, and to be able to do so to an accurate degree despite any interactions
with components of the ingredients. In the present study, optimum extraction
conditions of G (Rbl, Re, and Rd) from a blend of WF-GP was investigated. The
amounts of extractable G (Rbl, Re, and Rd) decreased with increasing heating
temperature of the blends of WF-GP, SF-GP, or GF-GP, indicating interactions between
the SF and G as well as between the GF and G in a blend of WF-GP heated at 90°C. In
addition, it was revealed that G had more interactions with the SF than with the GP in
the heated WF-GP, due to proportionally greater amounts of starch in the WP. The
interactions between WF components and G in WF-GP blend heated at 90°C were
increasingly disrupted with increasing UE time, up to 90 min for the maximum
extraction of G. Based on the ﬁndings in the present study, optimized extraction
conditions for G from a WF-GP blend were a sample-to-solvent ratio of 0071:] (mg/uL)
followed by ultrasonication for 90 min for as maximum an extraction of G as from pure

GP.

63

3.6. LITERATURE CITED

AACC International. 2000. Approved Methods of the American Association of Cereal
Chemists. 10th Ed. Method 44-15A and 46-13. The Association: St. Paul, W.

Arvisenet, G., Bail, P. L., Voilley, A., and Cayot, N. 2002. Inﬂuence of physicochemical
interactions between amylose and aroma compounds on the retention of aroma in
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Atwell, W. A. 2001. Composition of commercial ﬂour. Pages 27-45 in: Wheat ﬂour. W.
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Capelo, J. L., and Mota, A. M. 2005. Ultrasonication for analytical chemistry. Curr.
Anal. Chem. 1:193-201.

Choi, J. H., Kim, D. H., Sung, H. S., Kim, W. J., and Oh, S. K. 1982. Kinetic studies on
the thermal degradation of ginsenosides in ginseng extract. Korean J. Food Sci.
Technol. 14:197-202.

Choi, J. H., Byun, D. S., and Park, K. D. 1984. Studies on stability for the quality of
ginseng products. 1. Quality characteristics of freeze and spray dried red ginseng
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Conde-Petit, B., Escher, F. and Nuessli, J. 2006. Structural features of starch-ﬂavor
complexation in food model systems. Trends Food Sci. Technol. 17:227-235.

Court, W. A., Hendel, J. G., and Ehni, J. 1996. Reversed-phase high-performance liquid
chromatographic determination of ginsenosides of Panax quinquefolium. J.
Chromatogr. A 755:11-17.

Corbit, R. M., Ferreira, J. F. S., Ebbs, S. D., and Murphy, L. L. 2005. Simpliﬁed
extraction of ginsenosides ﬁ'om American ginseng (Panax quinquefolius L.) for
high-performance liquid chromatography-Ultraviolet analysis. J. Agric. Food Chem.
53:9867-9873. ‘

Derycke, V., Vandeputte, G. E., Vermeylen, R., De Man, W., Goderis, B., Koch, M. H.
J., and Delcour, J. A. 2005. Starch gelatinization and arnylose-lipid interactions
during rice parboiling investigated by temperature resolved wide angle X-ray
scattering and differential scanning calorimetry. J. Cereal Sci. 42:334-343.

Fuzzati, N., Gabetta, B., Jayakar, K, Pace, R., and Peterlongo, F. 1999. Liquid
chromatography-electrospray mass spectrometric identiﬁcation of ginsenosides in
Panax ginseng roots. J. Chromatogr. A 854:69-79.

Fuzzati, N. 2004. Analysis methods of ginsenosides. J. Chromatogr. B 812:119-133.

Harkey, M. R., Henderson, G. L., Gershwin, M. E., Stern, J. S., and Hackman, R. M.
2001. Variability in commercial ginseng products: an analysis of 25 preparations.

54

Am. J. Clin. Nutr. 73:1101-1106.

Heinemann, C., Conde-Petit, B., Nuessli, J., and Escher, F. 2001. Evidence of starch
inclusion complexation with lactones. J. Agric. Food Chem. 49:1370-1376.

Heinemann, C., Escher, F., and Conde-Petit, B. 2003. Structural features of starch-
lactone inclusion complexes in aqueous potato starch dispersions: the role of
amylose and amylopectin. Carbohyd. Polym. 51:159-168.

Heinemann, C., Zinxli, M., Renggli, A., Escher, F., and Conde-Petit, B. 2005. Inﬂuence
of amylose-ﬂavor complexation on build-up and breakdown of starch structures in
aqueous food model systems. LWT. 38:885-894.

Heng, L., van Koningsveld, G. A., Gruppen, H., van Boekel, M. A. J. S., Vincken, J. P.,
Roozen, J. P. and Voragen, A. G. J. 2004. Protein-ﬂavour interactions in relation to
development of novel protein foods. Trends Food Sci. Technol. 15:217-224.

Ikedo, S., Shimoyamada, M., and Watanabe, K. 1996. Interaction between bovine serum
albumin and saponin as studied by heat stability and protease digestion. J. Agric.
Food Chem. 44:792-795.

Ji, Q. C., Harkey, M. R., Henderson, G. L., Gershwin, M. E., Stern, J. S., and Hackman,
R. M. 2001. Quantitative determination of ginsenosides by high-performance liquid
chromatography-tandem mass spectrometry. Phytochem. Anal. 12:320-326.

Jouquand, C., Ducruet, V., and Le Bail, P. 2006. Formation of arnylose complexes with
C6-aroma compounds in starch dispersions and its impact on retention. Food Chem.
96:461-470.

Kim, J. W., Lee, H. B., and Joe, C. N. 1985. Biochemical studies on ginseng saponin
(XXV) ginseng saponin-protein interaction. Korean Biochem. J. 18:453-45 8.

Kim, M. K, Lee, J. W., Lee, K. Y., and Yang, D. C. 2005. Microbial conversion of
major ginsenoside Rb] to pharmaceutically active minor ginsenoside Rd. J.
Microbiol. 43:456-462.

K0, S. K, Lee, K. H., Hong, J. K, Kang, S. A., Sohn, U. D., Im, B. 0., Han, S. T., Yang,
B. W., Chung, S. H., and Lee, B. Y. 2005. Change of ginsenoside composition in
ginseng extract by vinegar process. Food Sci. Biotechnol. 14:509-513.

Kwon, S. W., Han, S. 3., Park, I. H., Kim, J. M. Park, M. K, and Park, J. H. 2001.
Liquid chromatographic detemrination of less polar ginsenosides in processed
ginseng. J. Chromatogr. A 921:335-339.

Lau, A. J., Woo, S. O., and Koh, H. L. 2003. Analysis of saponins in raw and steamed

Panax notoginseng using high-performance liquid chromatography with diode array
detection. J. Chromatogr. A 1011:77-87.

65

Lau, A. J., Seo, B. H., Woo, S. 0., and Koh, H. L. 2004. High-performance liquid
chromatographic method with quantitative comparisons of whole chromatograms of
raw and steamed Panax notoginseng. J. Chromatogr. A 1057:141-149.

Le Bail, P., Rondeau, C., and Buleon, A. 2005. Structural investigation of amylose
complexes with small ligands: helical conformation, crystalline structure and
therrnostability. Int. J. Biol. Mactomol. 35:1-7.

Li, T. S. C., Mazza, G., Cottrell, A. C., and Gao, L. 1996. Ginsenosides in roots and
leaves of American ginseng. J. Agric. Food Chem. 44:717-720.

MacRitchie, F. 1985. Studies of the methodology for fractionation and reconstitution of
wheat ﬂours J. Cereal Sci. 3:221-230.

Onofre, F. O., and Hettiarachchy, N. S. 2007. Extraction, quantiﬁcation, and
characterization of phenolics extracted with the aid of sonication ﬁ'om rice bran.
Cereal Chem. 84:337-342.

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1993. Protein-saponin interaction and its inﬂuence on blood lipids. J. Agric. Food
Chem. 41:1287-1291.

Raphaelides, S. N., and Georgiadis, N. 2006. Effect of fatty acids on the rheological
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Samthein-Graf, C., and Mesa, C. L. 2004. Association of saponins in water and water-
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Shibata, S. 2001. Chemistry and cancer activities of ginseng saponins and some related
triterpenoid compounds. J. Korean Med. Sci. 16:828-37.

Shinmoyamada, M., Ootsubo, R., Naruse, T., and Watanabe, K. 2000. Effects of
soybean saponin on protease hydrolyses of B-lactoglobulin and a-lactalbumin. Biosci.
Biotechnol. Biochem. 64:891-893.

Toro-Vazques, J. F., Gomez-Aldapa, C. A., Aragon-Pina, A., Brito-de la Fuente, E.,
Dibildox-Alvarado, E., and Charo-Alonso, M. 2003. Interacion of granular maize
starch with lysophosphatidylcholine evaluated by calorimetry, mechanical and
microscopy analysis. J. Cereal Sce. 38:269-279.

Wang, C. Z., Wu, J. A., Mcentee, E., and Yuan, C. S. 2006. Saponins composition in

American ginseng leaf and berry assayed by high-performance liquid
chromatography. J. Agric. Food Chem. 54:2261-2266.

66

Zobel, H. E, Young, S. N., and Rocca, L. A. 1988. Starch gelatinization: An X-ray
diffraction study. Cereal Chem. 65:443-446.

67

Table 3.].
Moisture and Protein Contents of Wheat Flour (WF), Ginseng Powder (GP),

Starch Fraction (SF), and Gluten Fraction (GF) Samples

 

 

Sample Moisture Content (%, wb) Protein Content (%)"
WF 11.2 8.3
GP 8.3 11.2
SF 1.9 2.]
GF 2.6 62.3

 

I'Protein percentages are on a 14% moisture basis.

68

Table 3.2.
Quantities of Ginsenosides (Rbl, Re, and Rd) Ultrasonically Extracted for 30 min
at Room Temperature from Wheat Flour (WF, 0.9 g), Ginseng Powder (GP, 0.] g),
and a Blend of WF-GP (0.9 g WF + 0.] g GP) with Different Sample-to-Solvent'

Ratios

 

Sample Solvent Sample-to-Solvent Ginsenosides (mg/100 mg GP)

 

 

(FL) Ratio (mg/11L) Rb] Rc Rd
GP 1000 0.100:1 0.530 0.390 0.18d
2000 0.050:1 0.70c 0.47c 0.21c
3000 0.030:1 0.83b 0.49b 0.23b
4000 0.025:1 0.89a 0.52a 0.25a
5000 0.020:1 0.88a 0.53a 0.25a
6000 0.017:1 0.88a 0.52a 0.25a
8000 0.013:1 0.88a 0.52a 0.25a
10000 0.010:1 0.89a 0.52a 0.25a
WF 14000 0.064;] ND ND ND
WF-GP 12000 0083:] 0.82b 0.500 0.230
14000 0.071:1 0.88a 0.53a 0.25a

 

Values with different letters within the same column differ significantly (P<0.05).

ND: not detectable.

'70% (v/v) aq. methanol

Table 3.3.
Effects of Ultrasonic Extraction (U E) Time on Extractability of Ginsenosides (Rbl,
Re, and Rd) from a Heated (90°C) Starch Fraction (SF), a Gluten Fraction (GF), a

Blend of SF-Ginseng Powder (GP), and a Blend of GF-GP

 

Sample UE Time (min) Ginsenosides (mg/100 mg GP)

 

 

Rbl Rc Rd
SF 30 ND ND ND
60 ND ND ND
90 ND ND ND
120 ND ND ND
SF-GP 30 0.7 1d 0.46c 0.20d
60 0.82b 0.50b 0.24b
90 0.87a 0.54a 0.271!
120 0.88a 0.54a 0.26a
GF 30 ND ND ND
60 ND ND ND
90 ND ND ND
120 ND ND ND
GF-GP 30 0.77c 0.49b 0.23c
60 0.88a 0.54a 0.2711
90 0.87a 0.53a 0.2711
120 0.88a 0.54a 0.27a

 

Values with different letters within the same column differ significantly (P<0.05).
ND: not detectable.

70

Table 3.4.
Effects of Ultrasonic Extraction (U E) Time on Extractability of Ginsenosides (Rbl,
Re, and Rd) from Heated (90°C) Wheat Flour (WF), Ginseng Powder (GP), and a

WF-GP Blend

 

Sample UE Time (min) Ginsenosides (mg/100 mg. GP)

 

 

Rbl Rc Rd
GP 30 0.88a 0.54a 0.27a
60 0.86a 0.53a 0.26a
90 0.87a 0.53a 0.26a
120 0.87a 0.53a 0.26a
WF 30 ND ND ND
60 ND ND ND
90 ND ND ND
120 ND ND ND
WF-GP 30 0.72c 0.47c 0.21c
60 0.82b 0.51b 0.24b
90 0.87a 0.53a 0.26a
120 0.87a 0.54a 0.26a

 

Values with different letters within the same column differ significantly (P<0.05).

ND: not detectable.

71

 

GP

 

 

 

0 5 10 15 20 25 30
Time (min)

 

 

 

 

0.00 I I I I I
0 5 10 15 20 25 30

Time (min)

 

WF-GP

 

 

 

0 5 10 l 5 20 25 30
Time (min)

Fig. 3.1. RP-HPLC chromatograms of ginsenosides ultrasonically extracted for 30
min from pure ginseng powder (GP), wheat flour (WF), and a blend of WF-GP in

70% (v/v) aq. methanol. 1: Rh]; 2: Re; 3: Rd.

72

Ginsenosides (mg/100 mg GP)
.° P P P 1"
N h as ea 9

P
e

0.4

0.2

Ginsenosides (mg/100 mg GP)

0.0

 

 

GP

<>————<>~<>—<>

HM

[—o—Rbr -<>—Rc —A—Rdl

 

 

 

 

20 40 60 80 100
Temperature (°C)

 

 

WF-GP

HR

o—W
AM

I—D—Rbl -0-Rc —a—Ra|

 

 

 

l L I l

 

20 40 60 80 100
Temperature (°C)

Fig. 3.2. Effects of sample heating of pure ginseng powder (GP) and a wheat ﬂour

(WF)-GP blend on subsequent extractable ginsenosides (Rbl, Re, and Rd).

73

 

l"
c

SF-GP

 

 

 

 

 

 

E?
U
M
E
G
2 0.6 -
”'50
E <>\<>
8 0.4 -
:2
3
a:
g 0.2 - AM
0 I—CI—Rbl -<>—Rc —A—Rd]

0.0 l l L I

0 20 40 60 80 100
Temperature (°C)
1'“ GF-GP

0.3 - D\D

<>\<>
0.2 - A\A

l—D—Rbl —<>-Rc —A—Rd|
0.0 ' ' ‘ '

0 20 40 60 80 100
Temperature (°C)

 

Ginsenosides (mg/100 mg GP)

 

 

 

 

Fig. 3.3. Effects of sample heating of a starch fraction (SF)-ginseng powder (GP)
blend and a gluten fraction (GF)-GP blend on subsequent extractable ginsenosides

(Rbl, Rc, and Rd).

74

CHAPTER 4
Effects of Extrusion Process Variables on Extratable

Ginsenosides in Wheat-Ginseng Extrudates

75

4.1. ABSTRACT

Effects of extrusion process variables (feed moisture, screw speed, and barrel
temperature) on extractable ginsenosides (G, ginseng saponins) in wheat-ginseng
extrudates (WGE) produced under different extrusion conditions were investigated. A
wheat ﬂour (WF)-ginseng powder (GP) blend (10% GP, w/w) was extruded in a twin-
screw extruder (L/D ratio of 25:1) with full factorial combinations of feed moisture (25,
30, and 35%), screw speed (200 and 300 rpm), and zone 5 barrel temperature (110, 120,
130, and 140°C). Samples of WF, GP, WF-GP, and WGE extruded at different
conditions were ultrasonically extracted for G from 60 to 150 min. Individual G (Rbl,
Re, Rd, Rg2, and Rg3) were fractionated and identiﬁed by RP-HPLC from each sample.
The quantities of G (Rbl, Re, and Rd) extracted from WGE samples extruded at a zone
5 barrel temperature of more than 120°C were signiﬁcantly higher than those extracted
from the control blend, explaining the conversion of malonyl G (Rbl, Re, and Rd) into
G (Rbl, Re, and Rd, respectively) during extrusion cooking. Neither G Rg2 nor G Rg3
were found in pure WF, GP, nor in the WF-GP blend. G Rg2 were found in all the
WGE samples studied. Increasing feed moisture caused a decrease in the quantities of
G Rg2 in WGE samples. In contrast, increasing screw speed and barrel temperature
each led to an increase in the quantities of G Rg2 in WGE samples. G Rg3 were
present in WGE samples produced under only the following three extrusion conditions:
(1) 25% feed moisture, 300 rpm screw speed, and 130°C zone 5 barrel temperature, (2)
25% feed moisture, 300 rpm screw speed, and 140°C zone 5 barrel temperature, and (3)
30% feed moisture, 300 rpm screw speed, and 140°C zone 5 barrel temperature. Thus,
it was suggested that the production of new G Rg3 was dependent upon the extrusion

process conditions used.

76

4.2. INTRODUCTION

Extrusion cooking is a widely used technology to process cereals or starches
into food products, such as snack foods, ready-to-eat breakfast cereals, infant formulas,
etc. (Harper 1979). This cooking processing, carried out using a high temperature and
short time (HTST) treatment, yields ﬁnished products with desirable qualities, such as
high digestibility and nutritional value (Harper 1998).

Wheat ﬂour (WF) is a commonly used material in the extrusion industry and
the effects of extrusion process variables on the properties of WF extrudates have been
studied (Ali et al 1996; Anderson and Ng 2001, 2003; Ding et a1 2006; Harper 1979;
Mercier and Feillet 1975; Ryu and Ng 2001; Schmid et a1 2005). Based on these
studies, extrusion process variables (including screw conﬁguration, screw speed,
temperature proﬁle of the barrel, residence time, feed rate, feed moisture, diameter of
exit die, and others) have been found to alter the physical and chemical properties of
ﬁnal products. The addition of diverse ingredients to extrusion materials has been done
with the objective of improving the nutritional quality of ﬁnal extruded products and
imparting desirable functional properties (Jin et al 1995; Rinald et a1 2000; Schmid et a1
2005; Singh & Singh 2004; Zazueta-Morales et al 2002).

Ginseng has been frequently used in Asian countries as a traditional medicine
(Fuzzati 2004). Ginsenosides (G, ginseng saponins) have been known as the main
active components and have been typically used as marker compounds for quality
properties (Chuang et al 1995; Lau et a1 2004; Li et al 1996). G are dammarane-type
triterpene glycosides, and can be classiﬁed into three groups: the protopanaxadiols,

protopanaxatriols, and oleanolic acid (Li et al 2006). Four malonyl G, including

77

malonyl G Rbl, malonyl G Rb2, malonyl G Re, and malonyl G Rd, are also found in
ginseng (Fuzzati et al 1999).

Raw ginseng has been processed into two kinds of ginseng, white ginseng
and red ginseng, to improve its preservation and efﬁcacy (Kim et al 2000; Wang et a1
2006). White ginseng is usually air-dried raw ginseng, while red ginseng is usually
made by ﬁrst steaming raw ginseng at 90-100’C for 2-3 hr and then air-drying (Ha et al
2005). Red ginseng is known to be more pharmaceutically active than white ginseng,
because the steaming process causes changes in the chemical compositions of G, thereby
enhancing the biological activity of the ginseng (Ha et a1 2007; Shibata 2001). The
ginsenosides Rg3, Rg5, Rg6, Rh2, Rh3, Rh4, RS3, Rkl, and F4 are not found in raw and
White ginseng, whereas red ginseng contains these new G generated during the steaming
process (Ha et al 2007; Kim et al 2000; Kwon et a1 2001). In particular, G Rg3 have
been found to show various biological activities, including anticancer activity (Shibata
2001; Wang et al 2006; Yun et al 2001) and radical scavenging activity (Kang et al
2006). Moreover, it has been reported that G Rg3 inhibit the growth of Helicobacter
pylori (Bae et a1 2002).

In spite of medical beneﬁts of red ginseng and the processing efﬁciency (e. g.,
high temperature and short time) of extrusion cooking for making red ginseng products,
Ha et a1 (2005), who studied changes in G in ﬁnal extruded Panax ginseng, only applied
the extrusion processing technique to produce pure red ginseng products. They
reported that the application of extrusion cooking to produce red ginseng can
considerably reduce the processing time, as compared to the traditional red ginseng
process. However, no studies have apparently reported the production of extruded WF

snack products with ginseng, nor have studies been found that investigate changes in G

78

in snack products extruded under different conditions. Extrusion cooking has been
employed to make nutraceutical snack foods and there is a possibility of using WF in
combination with ginseng to provide highly nutritious snack food products. Thus, the
objective of this study was to investigate the effects of extrusion process variables (feed
moisture, screw speed, and barrel temperature) on extractable G in wheat-ginseng

extrudates (WGE).

79

4.3. MATERIALS AND METHODS

4.3.]. Materials

Soft WF was obtained from the Mennel Milling Co. in Fostoria, OH, USA.
The raw ginseng root (Panax ginseng C. A. Meyer), harvested after four years’ growth
and milled into powder (particle size of S 167 um), was cultivated in Geumsan, Korea,
and purchased at a ginseng market. WF and GP were analyzed for their moisture
contents using AACCI Approved Method 44-15A (AACCI, 2000) and the moisture
contents of WP and GP were 11.2% (wb) and 8.3% (wb), respectively. Standards for

ginsenosides Rbl, Rc, Rd, Rg2, and Rg3 were provided by ILHWA Co. (Kuri, Korea).

4.3.2. Extrusion Cooking

WP and ginseng powder (GP) were mixed in a weight ratio of 9:] using a
mixer (Model A-200, Hobart MFG. Co., Troy, OH, USA) for 20 min; hereafter, this
blend is referred to as WF-GP. Extrusion cooking was done with an APV co-rotating,
twin screw extruder, with a barrel diameter of 19 mm and a barrel length to diameter
(L/D) ratio of 25:] (Model MP 19T2-25, APV Baker, Grand Rapids, MI, USA).
Heating of the barrel was controlled by electric heating elements jacketing the barrel and
thermal probes. Heating was divided into ﬁve zones along the length of the barrel,
with zone 1 nearest the feed section and zone 5 nearest the exit die. Feed moisture was
adjusted to the predetermined dough moisture content during extrusion cooking by
injection of water into the barrel using a Brook Crompton E2 Metripump (Hudders Field,
England). Dry materials were fed using a K-TRON K2M twin-screw volumetric feeder
(K-TRON, Pittman, NJ, USA). Product temperature and Die pressure was measured

by a thermocouple (Model TB422J, Dynisco, Sharon, MA, USA) and by a pressure

80

transducer (Model ERP3-3M, Dynisco, Sharon, MA, USA), respectively, inserted before
the die. Torque was measured as % of the total motor power (2000W).

A full factorial arrangement was used for the present study. Extrusion
process variables were: (1) feed moisture (25, 30, and 35%), (2) screw speed (200 and
300 rpm), and (3) zone 5 barrel temperature (110, 120, 130, and 140°C), for a total of 24
different combinations (Table 4.1). Zone 1/2/3/4 barrel temperatures were constant
(40/70/90/110°C, respectively) for all the extrusion process conditions studied (Table
4.1). The dry material feed rate was kept constant (2.0 kg/hr) along with the screw
conﬁguration (Table 4.2). Extrusion cooking of WF-GP was repeated on a different
day.

The WGE samples produced at different extrusion conditions were collected

and dried in an air oven (45°C) overnight. After drying, portions of each WGE sample

were ground to a particle size of S 330 um (54 mesh screen) using a coffee grinder
(Braun Inc., Woburn, MA, USA). After grinding, moisture contents of ground WGE
samples were measured by AACCI Approved Method 44-15A (AACCI, 2000). Both
ground and unground WGE samples were placed in ziplock bags, sealed, and stored at

-20°C until analysis.

4.3.3. Speciﬁc Mechanical Energy

The speciﬁc mechanical energy (SME) in Watt hours/kg was calculated
according to the equation of Brent et a1 (1997):
SME = [actual screw speed rpm / rated screw rpm] x [% torque / 100] x [motor power in
W / feed rate in kg/hr]

Rated screw speed = 500 rpm; motor power = 2000 W

81

4.3.4. Optimization of Ultrasonic Extraction Time for Ginsenosides from Wheat-
Ginseng Extrudates

Four mL of 70% (v/v) aq. methanol were added to GP (0.] g). Fourteen mL
of 70% (v/v) aq. methanol were added to WP (0.9 g), WF-GP (0.9 g WF + 0.1 g GP), or
WGE samples (1.0 g) extruded under different conditions. Each mixture was
ultrasonically extracted for 60, 90, 120, or 150 min using an Ultrasonicator (Model FS
14H, Fisher Scientiﬁc, Pittsburgh, PA, USA, Power: 155W) at room temperature, and
then centrifuged at 11,200 x g for 20 min using a centrifuge (Model J2-21M, Beckman
Instruments Inc., Fullerton, CA, USA) at 25°C. The supernatant was evaporated using
a Micro Rotary Evaporator (Model 4214000, Labconco, Kansas City, MO, USA) at
50°C. Prior to RP-HPLC analysis, the residue was dissolved in 2 mL of 70% (v/v) aq.
methanol and ﬁltered through a 0.45 um nylon ﬁlter membrane (Millipore, Ireland).
Each sample was extracted in duplicate and two chromatographic runs were performed

for each extract.

4.3.5. Fractionation and Identification of Ginsenosides by RP-HPLC

RP-HPLC was carried out on a Millenium 2010 HPLC Workstation,
consisting of a Waters 600E multi-solvent delivery system (Waters, Milford, MA, USA),
a temperature control module, and a 996 photodiode array detector. Separation was
performed at 30°C on a Phenomenex 300 RP, Jupiter C-l8 column (4.6 x 250 mm, 5 pm
particle diameter; Phenomenex, Torrance, CA, USA). A Phenomenex security guard
with the same packing material served as the guard column.

The solvents were A: water (distilled water ﬁltered through Milli-Q-Plus,

82

Millipore), and B: HPLC grade acetonitrile. Separation was achieved using the
following gradient: 0-5 min, 20-30% B; 5-25 min, 30-40% B; 25-30 min, 40-20% B.
The solvent ﬂow rate was 1 mL/min and the injection volume was 20 uL. The solvents
were purged with helium at the rate of 20 mL/min. The column was equilibrated for 10
min with 20% acetonitrile prior to injection. The UV detection wavelength was set at
203 nm (Lau et al 2004), and the detector output was transmitted simultaneously to the

computer for data storage and graphic representation.

4.3.6. Statistical Analysis

All statistical analyses were performed using SAS version 9.1 (SAS
Institute Inc., Cary, NC, USA). Analysis of variance (ANOVA) was performed using
the general linear models (GLM) procedure to determine signiﬁcant differences among
the samples. Means were compared by using Fisher’s least signiﬁcant difference
(LSD) procedure. Signiﬁcance was deﬁned at the 5% level. In addition, Pearson
correlation coefﬁcients were determined among the samples. The CORR procedure

was used to obtain correlation coefficients.

83

4.4. RESULTS AND DISCUSSION

4.4.]. Optimization of Ultrasonic Extraction Time for Ginsenosides from Wheat-
Ginseng Extrudates

Effects of ultrasonic extraction (UE) time of WF, GP, and WF-GP on
extractable G (Rbl , Re, and Rd) are presented in Table 4.3. No measurable G (Rbl, Re,
and Rd) in pure WF were found at any of the UE times studied. The quantities of G
Rbl, G Re, and G Rd extracted from both pure GP and WF-GP were not signiﬁcantly
different at all the ranges of UE time (60-150 min) studied. However, the quantities of
G extracted from WGE samples extruded at 30% feed moisture, 200 rpm screw speed,
and 120°C zone 5 barrel temperature increased with increasing UE time, and ﬁnally
reached a plateau after UE time of 120 min (Figure 4.1). All the studied WGE samples
showed the same trend (Appendix 3). The ﬁndings established that the maximum
extraction of extractable G from WGE samples produced under each of the different

extrusion process conditions could be achieved by ultrasonication for 120 min or longer.

4.4.2. Changes in Ginsenosides during Extrusion Cooking

The quantities of G (Rbl, Re, and Rd) ultrasonically extracted for 150 min
from WF-GP and WGE samples extruded at different conditions are listed in Table 4.4.
The quantities of G Rb] and G Rc extracted from WGE samples extruded at 110°C zone
5 barrel temperature were not changed upon extrusion cooking. However, the
quantities of G Rd extracted ﬁom the same WGE samples were signiﬁcantly increased
upon extrusion cooking. The quantities of each of G Rbl, G Re, and G Rd extracted
from WGE samples extruded at a zone 5 barrel temperature of more than 120°C were

signiﬁcantly higher than those of their counterpart G extracted from the control WF-GP.

84

This ﬁnding was consistent with Ha et a1 (2005) who showed signiﬁcant increases in the
quantities of G Rbl, G Rb2, G Re, and G Rd after an extrusion process of raw Panax
ginseng.

Some studies have been conducted to compare G compositions of White and
red ginseng (Ha et a1 2005; Kim et al 2000; Kwon et al 2001; Lau et a1 2003; Lau et al
2004). The G composition of red ginseng was obviously different from that of white
ginseng, because of the loss of malonyl G during the steaming process used to produce
red ginseng (Fuzzati 2004). Shibata (2001) noted that white ginseng includes the
malonyl esters of G Rbl, G Re, and G Rd, and that the malonyl group, which is
originally attached at the 6"-position of the glycosyl moieties of G Rbl, G Re, and G Rd,
is easily lost during the steaming process when making red ginseng. Ren and Chen
(1999) reported that malonyl G are thermally very unstable; therefore, they are readily
demalonylated by heating. According to Fuzzati (2004), the acidic malonyl G
(malonyl G Rb] , malonyl G Re, and malonyl G Rd) degraded or converted into the
corresponding neutral G (G Rbl, G Re, and G Rd, respectively) during the steaming
process. Therefore, it is postulated in the present study that extrusion cooking under a
zone 5 barrel temperature of 2 120°C could stimulate the loss of malonyl acid from
glycosyl moieties of malonyl G Rbl, malonyl G Re, and malonyl G Rd, subsequently
inducing an increase in the quantities of G Rbl, G Re, and G Rd, respectively, in WGE
samples.

RP-HPLC chromatograms of G ultrasonically extracted for 150 min ﬁom
non-extruded WF-GP and WGE samples extruded at 25% feed moisture, 300 rpm screw
speed, and 140°C zone 5 barrel temperature are shown in Figure 4.2. The RP-HPLC

chromatograms of the WF-GP and the WGE samples were found to be distinctively

85

different, indicating that when WF-GP was extruded, the HPLC proﬁle of G was
changed. In particular, G Rg2 (peak 3) and G Rg3 (peak 5), which were not present in
WF-GP, were found in the WGE sample. Furthermore, the quantities of G Rg2 and G
Rg3 were dependent on the extrusion process conditions studied (Table 4.4); increases in
the quantities of G Rg2 and G Rg3 were observed when feed moisture was decreased,
and screw speed and barrel temperature each increased.

Wang et a1 (2006) studied the changes in G in berries of Panax quinquefolius
after steaming treatment (loo-120°C for 1 hr). They found that the amounts of G (Rg2,
Rg3, Rh], and Rh2) in the steamed samples were increased as compared to those in raw
samples. They reported that these G can be used as marker compounds to distinguish
red ginseng from white ginseng. Kim et al (2000) evaluated the effect of steaming
Panax ginseng, at 100, 110, and 120°C for 2 hr using an autoclave, on its chemical
components. They observed that G (F4, Rg3, and Rg5) were not found in the raw
ginseng, but in the steamed ginseng samples. According to Lau et al (2003), when
Panax notoginseng was steamed at 120°C for 1, 2, 3, and 9 hr, G (Rbl, Rd, and Re)
signiﬁcantly decreased and four new major peaks appeared in the RP-HPLC
fractionation which had not been present in chromatograms of the raw ginseng sample.
They reported that the new peaks were associated with the unique G which are present
only in red ginseng products, and that the quantities of the new G in the steamed samples
increased with increasing duration of the steaming process. According to Ha et a1
(2007), less polar G, including G (Rg3, Rg5, Rkl, and F4), are typically considered as G
unique to red ginseng products.

Ha et al (2005) employed an extrusion process to produce red ginseng

products using raw Panax ginseng. The extrusion conditions used were feed moisture

86

of 15 or 22% (wb), screw speed of 200 rpm, and barrel temperatures of 110/110/60/40°C
(the barrel zones of 1/2/3/4, respectively). They found an increase in the G Rg group
(G Rgl, G Rg2, and G Rg3) in the ﬁnal extruded samples. Shibata (2001) reported that
the glycosyl moieties at the C-20 position of the protopanaxadiol type G, including G
Rbl, G Rb2, G Re, and G Rd, are partly degraded to produce G Rg3 during the steaming
process to make red ginseng. Popovich and Kitts (2004) showed that G Rg3 are
naturally absent in Panax quinquefolius, but are produced by a thermal process. They
also reported that the formation of G Rg3 is associated with the breakdown of the more
abundant G Rb] and G Rc. On the other hand, G Rg2 can be produced by the
protopanaxatriol type G, such as G Re, in the same way (Kim et al 2000). Thus, it is
suggested in the present study that the presence of G Rg3 and G Rg2 in WGE samples is
attributed to the chemical degradation and/or conversion of the thermolabile
protopanaxadiol type G and protopanaxatriol type G, respectively, during high
temperature extrusion cooking. Furthermore, it is suggested that, utilizing raw ginseng
as one of the ingredients, extruded nutraceutical WF snack products can be produced by
extrusion cooking and yield red ginseng components in the extrudates.

To investigate the effects of extrusion process variables on changes in the
quantities of G Rg2 and G Rg3 in WGE samples, WF-GP were extruded at 24 different
extrusion process conditions. The quantities of G Rg2 ultrasonically extracted for 150
min from non-extruded WF-GP samples and from WGE samples extruded under
different conditions are listed in Table 4.4. The SME values obtained from different
extrusion process conditions studied are presented in Table 4.5. G Rg2 were found in
all the WGE samples studied, and the quantities of G Rg2 were signiﬁcantly affected by

extrusion process variables studied.

87

First of all, increasing feed moisture from 25 to 35%, at constant zone 5
barrel temperature and screw speed (130°C and 300 rpm, respectively), resulted in a
signiﬁcant decrease in the quantities of G Rg2 in WGE samples, from 0.15 to 0.08 mg
per 100 mg GP, and signiﬁcantly reduced SME values from 268.5 to 243.9 Wh/kg.
The ﬁnding (a decrease in SME values with increasing feed moisture) was consistent
with observations by Ryu and Ng (2001), who reported that lower feed moisture induces
higher viscosity of the melt during extrusion cooking and this can lead to an increase in
the degree of mechanical energy input on the extruded products. Therefore, it is
suggested that in the present study, the increased mechanical energy input caused by the
lower feed moisture extrusion cooking condition could facilitate the loss of the glycosyl
moieties at the C-20 position of the protopanaxatriol type G, subsequently causing an
increase in the quantities of G Rg2 in WGE samples.

Secondly, elevating screw speed from 200 to 300 rpm, at constant zone 5
barrel temperature and feed moisture (140°C and 25%, respectively), signiﬁcantly
increased the quantities of G Rg2 in WGE samples ﬁ'om 0.09 to 0.20 mg per 100 mg GP,
and led to a signiﬁcant increase in SME values from 131.8 to 262.8 Wh/kg. According
to Frame (1994), the degree of ﬁll in the barrel and the viscosity of the dough can be
diminished with increasing the screw speed during extrusion cooking. However, in the
present study, the increase in screw speed was large enough to overcome a slight
reduction in the % torque from the motor, resulting in a higher SME. Thus, it appears
in the present study that more protopanaxatriol type G could be readily converted to G
Rg2 at the higher screw speed extrusion process conditions, due to higher mechanical
energy input occurring during high screw speed extrusion cooking conditions.

Finally, increasing zone 5 barrel temperature from 110 to 140°C, at constant

88

feed moisture and screw speed (25% and 300 rpm, respectively), caused a signiﬁcant
increase in the quantities of G Rg2 in WGE samples, from 0.09 to 0.20 mg per 100 mg
GP. As already mentioned, the traditional processing to make red ginseng includes
steaming raw ginseng at 90-100’C for 2-3 hr. Therefore, it was hypothesized in the
present study that temperature higher than these could improve the production of red
ginseng components and shorten the processing time. The ﬁnding indicated that higher
barrel temperature during extrusion cooking is indeed one of the important factors that
increases the quantities of G Rg2 in WGE samples.

As reported earlier, increasing SME values (i.e., the lower feed moisture or
higher screw speed extrusion cooking conditions) were associated with production of
greater quantities of G Rg2 in WGE samples. However, the increase in barrel
temperature from 110 to 140°C at these extrusion process conditions caused an increase
in the quantities of G Rg2 in WGE samples, despite signiﬁcantly decreased SME values
(from.282.9 to 262.8 Wh/kg). The trend of decreasing SME values with increasing
barrel temperature is generally consistent with other published results (Altan et a1 2008;
Guha et al 1997; 110 et a1 1999; Ryu and Ng 2001; Tanhehco and Ng 2005). Lower
SME values are a result of the lower viscosity of the melt at higher temperatures,
thereby leading to lower mechanical energy input on the extruded products (Altan et a1
2008; Ryu and Ng 2001).

Based on all of the results (on effects of extrusion process variables on the
increase in G Rg2 in WGE samples) obtained in the present study, it was suggested that
increasing barrel temperature played the more dominant role in the conversion of
protOpanaxatriol type G into G Rg2 than did mechanical energy input during extrusion

cooking of WF-GP.

89

The quantities of G Rg3 ultrasonically extracted for 150 min from WF-GP
and WGE samples extruded under different conditions are listed in Table 4.4. In
particular, G Rg3 are valued for their pharmaceutical activities. Numerous studies have
focused on G Rg3 to evaluate the effects of heating processes on producing red ginseng
products (Ha et a1 2005; Kim et al 2000; Popovich and Kitts 2004; Wang et al 2005).
Kim et al (2000) observed that during a steaming process of raw Panax ginseng,
increasing steaming temperature from 100 to 120°C resulted in an increase in the
quantities of G Rg3. In work done by Wang et a1 (2006), when berries of Panax
quinquefolius were steamed at 120°C, the quantities of G Rg3 were signiﬁcantly higher
than for those steamed at 100°C.

In the present study, unlike G Rg2 which were found in all the WGE samples
produced, G Rg3 were present in WGE samples produced at only the following three
extrusion process conditions: (1) 25% feed moisture, 300 rpm screw speed, and 130°C
zone 5 barrel temperature, (2) 25% feed moisture, 300 rpm screw speed, and 140°C zone
5 barrel temperature, (3) 30% feed moisture, 300 rpm screw speed, and 140°C zone 5
bzurel temperature. Of these, extrusion process condition (2) produced the highest
quantities of G Rg3, and is the extrusion condition that also produced the highest
quantities of G Rg2. In particular, the extrusion condition (2) had the highest zone 5
barrel temperature (140°C), the highest screw speed (300 rpm), and the lowest feed
moisture (25%), indicating that an increase in severity of the extrusion conditions could

improve the conversion of protopanaxadiol type G to G Rg3 in the WGE samples.

4.4.3. Correlations among Ginsenosides and Product Temperature

90

Pearson’s coefﬁcients of correlation (r) among G (Rbl, Re, Rd, and Rg2) and
product temperature are reported in Table 4.6. The quantities of G Rbl were found to
be positively associated with the amounts of G Rc (r = 0.97, p 5 0.001), G Rd (r = 0.90,
p 5 0.001), and G Rg2 (r = 0.47, p S 0.05). Product temperature represents the actual
temperature of the dough at the exit die during extrusion cooking. The quantities of G
Rbl, Re, Rd, and Rg2 were positively correlated (r = 0.90, p S 0.00]; r = 0.86, p S
0.00]; r = 0.76, p _<_ 0.001; r = 0.61, p S 0.01, respectively) with the product temperature,
indicating that increasing product temperature not only could increase the chemical loss
of malonyl acid from glycosyl moieties of the malonyl G (Rbl, Re, and Rd), but also

could improve the conversion efficiency of protopanaxatriol type G to G Rg2.

91

4.5. SUMMARY

The present study focused on changes in chemical compositions of G after
extrusion cooking of a blend of WF-GP. Extrusion cooking led not only to an increase
in the quantities of G Rbl, G Re, and G Rd in WGE samples, but also to the production
of new G Rg2 and G Rg3 in certain WGE samples, indicating that changes in chemical
composition of G can take place during high temperature extrusion cooking. The
quantities of G Rg2 and G Rg3 in WGE samples were signiﬁcantly affected by the
extrusion process variables (feed moisture, screw speed, and barrel temperature) studied.
The optimal extrusion process condition in the present study, producing the maximum
quantities of G Rg2 and G Rg3 in WGE samples, was 25% feed moisture, 300 rpm
screw speed, and 140°C zone 5 barrel temperature. Moreover, the presence of G Rg2
and G Rg3 in WGE samples offered the possibility of the production of extruded

nutraceutical WF snack foods containing red ginseng components.

92

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Kim, W. Y., Kim, J. M., Han, S. B., Lee, S. K, Kim, N. D., Park, M. K, Kim, C. K,
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Li, T. S. C., Mazza, G., Cottrell, A. C., and Gao, L. 1996. Ginsenosides in roots and
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94

cooking of cereal product. Cereal Chem. 52:283-297.

Popovich, D. G., and Kitts, D. D. 2004. Generation of ginsenosides Rg3 and Rh2 from
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Rinaldi, V. E. A., Ng, P. K. W., and Bennink, M. R. 2000. Effects of extrusion on
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Zazueta-Morales, J ., Martinez-Bustos, F., Jacobo-Valenzuela, N., Ordorica-Falomir, C.,

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95

Table 4.1.

Extrusion Conditions for a Wheat Flour-Ginseng Powder (GP) Blend

 

 

 

 

 

(10% GP, w/w)
Feed Moisture Screw Speed Barrel Temperature Zones 1-5
(%, wb) (rpm) (’0'

25 200 40/70/90/110/110
30 200 40/70/90/110/110
35 200 40/70/90/110/110

' ' ' ' is" ' ' ' " ' ' ' 300 """"" 40707901107110 """
30 300 40/70/90/110/110
35 300 40/70/90/110/110
25 200 40/70/90/110/120
30 200 40/70/90/110/120
35 200 40/70/90/110/120

' ' ' ' 55' ' ' ' - ' ' ' "3’00 """"" 4'0/70790/1'107130 """
30 300 40/70/90/110/120
35 300 40/70/90/110/120
25 200 40/70/90/110/130
30 200 40/70/90/110/130
35 200 40/70/90/110/130

' " " " 55' " ' " " " ' ' 300 """""" 40707901107130 """
30 300 40/70/90/110/130
35 300 40/70/90/110/130
25 200 40/70/90/110/140
30 200 40/70/90/110/140
35 200 40/70/90/110/140

' ' ' ' 55' ' " ' " ' ' " 300 """"" iii/707901107140 """
30 300 40/70/90/110/140
35 300 40/70/90/110/140

 

' Zone 5 - nearest to exit die.
Feed rate - 2.0 kg/hr, db

96

Table 4.2.

Screw Conﬁguration for Twin-Screw Extruder

 

 

Zone Type‘II Flight Angle (°)
I 5D Twin __
II 3D Twin _

7 FP 30

III SD Twin _

3D Twin _
N 3 FP 60
3 RP 30
2D Single _
4 FP 60

V 3 RP 30
2D Single _
Die

 

" D: screw diameter (19 mm); P: kneading paddle (0.25 D); FP: forwarding paddle; RP: reversing
paddle; Twin: twin lead screw; Single: single lead screw; Die: 3mm single round type exit die.

97

Table 4.3.
Effects of Ultrasonic Extraction (UE) Time of Wheat Flour (WF, 0.9 g), Ginseng
Powder (GP, 0.] g), and a WF-GP (0.9 g WF + 0.1 g GP) Blend on Extractable

Ginsenosides (Rbl, Rc, and Rd)

 

 

 

Sample UE time Ginsenosides (mg/100 mg GP)
(min) Rbl Rc Rd
GP 60 0.86a 0.51a 0.24a
90 0.85a 0.50a 0.25a
120 0.86a 0.52a 0.25a
150 0.86a 0.51a 0.24a
WF 60 ND ND ND
90 ND ND ND
120 ND ND ND
150 ND ND ND
WF-GP 60 0.86a 0.52a 0.25a
90 0.85a 0.51a 0.24a
120 0.86a 0.51a 0.25a
150 0.86a 0.51a 0.25a

 

Values with different letters within the same column differ significantly (P<0.05).

ND: not detectable.

98

Table 4.4.
Quantities of Ginsenosides (Rbl, Rc, Rd, Rg2, and Rg3) Ultrasonically Extracted
for 150 min from a Wheat Flour (WE-Ginseng Powder (GP) Blend (10% GP, w/w)

and Wheat-Ginseng Extrudates

 

Extrusion Conditions Ginsenosides (mg/100 mg GP)

 

 

 

 

 

 

M/S/T' Rb] Rc Rd Rg2 Rg3
WF-GP (non-extruded) 0.86d 0.51d 0.25e ND ND
25/200/110 0.86d 0.51d 0.28cd 0.06h ND
30/200/110 0.87d 0.51d 0.29c 0.06h ND
35/200/110 0.87d 0.51d 0.26de 0.05h ND
' " " 3573007110 """ 07800" ' ' 0.500 ' " 3725.5 " ' 0.090 ' ' 'N'n' '
30/300/110 0.85d 0.50d 0.27cd 0.06h ND
35/300/110 0.85d 0.5]d 0.27cd 0.06h ND
25/200/120 1.03ab 0.61a 0.32ab 0.08f ND
30/200/120 1.00b 0.56c 0.32ab 0.08f ND
35/200/120 0.96c 0.55c 0.32ab 0.07g ND
' " ' 2573007120 """ 17000” ' ' 0.02; ' ' "0733.10 " ' 0.010 ' ' ”Nb" "
30/300/120 0.96c 0.58b 0.32ab 0.08f ND
35/300/120 0.96c 0.58b 0.3lb 0.08f ND
25/200/130 1.04a 0.61a 0.32ab 0.08f ND
30/200/130 1.04a 0.62a 0.34a 0.07g ND
35/200/130 1.04a 0.63a 0.33ab 0.07g ND
' " " 2573007130 """ 1704; ' ' 0.023 ' ' "03:00 ' ' 0.1'50 " ' 0 050 "
30/300/130 1.05a 0.61a 0.32ab 0.09e ND
35/300/130 1.05a 0.62a 0.32ab 0.08f ND
25/200/140 1.06a 0.61a 0.32ab 0.09e ND
30/200/140 1.04a 0.61a 0.3lab 0.09e ND
35/200/140 1.05a 0.62a 0.32ab 0.07g ND
' ' " 2573007140 """ 17.05.? ' ' 0.013 " " “073300 " ' 0.20; ' ' 0.07; '
30/300/140 1.05a 0.61a 0.33ab 0.12c 0.05h
35/300/140 1.04a 0.62a 0.32ab 0.09ef ND

 

Values with different letters within the same column differ significantly (P<0.05).
'M: Feed moisture (%, wb); S: screw speed (rpm); T: zone 5 barrel temperature (°C).
ND: not detectable.

99

Table 4.5.

System Variables Related to Different Extrusion Process Variables of Wheat-

 

 

 

 

 

 

Ginseng Extrudates
Extrusion Process Variables System Variables
Feed Moisture Screw Speed Temperature' . SMEb P'I‘c
(% wb) (rpm) (°C) (W 110(8) (°C)
25 200 110 157.8h ll7jkl
30 200 110 154.0hi 116k]
35 200 110 146.6hi 1151
' ' ' 25 """" 3 00 """ 1'10 """ 2 820; """ 119} ' ' '
30 300 110 259.8cde ll8jk
35 300 110 252.0defg ll7jkl
25 200 120 155.0hi l30h
30 200 120 153.2hi 1271
35 200 120 133 8] 1261
' ' ' 25 """" 3 00 """ 1'20 """ 2 747850 """ 1320 ' ' '
30 300 120 253.5def 13lh
35 300 120 249.3efg l30h
25 200 130 146.01 145fg
30 200 130 148.0hi 145efg
35 200 130 129 2] 144f
' ' "2'5 """" 3 00 """ 1'30 """ 2 08510 """ 1272 ' ' '
30 300 130 261.9cd 147ef
35 300 130 243.9fg 146efg
25 200 140 131.8j 156bc
30 200 140 130.6j 154cd
35 200 140 117 2k 153d
' ' ' '2'5 """" 3 00 """ 1'40 """ 2 02.001 """ 103; ' ' '
30 300 140 256.8de 157b
35 300 140 241.8g 152d

 

Values with different letters within the same column differ significantly (P<0.05).
”Temperature: zone 5 barrel temperature.

l’SME: specific mechanical energy.

‘PT: product temperature.

100

Table 4.6.

Pearson’s Coefﬁcients of Correlation (r) among Ginsenosides (Rbl, Rc, Rd, and

 

 

Rg2) and PT'
Rb] Rc Rd Rg2 PT
Rbl 097*** 0.90*** 047* 0.90***
Re 090*** 0.46* 0.86***
Rd 049* 0.76***
Rgl 0.61"
PT

 

*, **, ***, Significantly different at the 5%, 1%, and 0.1% levels, respectively.

‘PT: product temperature.

101

 

 

 

 

-0-Rc
+Rd
—O—Rg2

 

 

 

 

142
E: -
CD 11)
E”
c 0.8 "
c
y-r
is
g 0.6 -
8
IE
8 (L4 '
0
'5 0.2 -
04)
30

60

90 120 150 180
IﬂEthne(nﬁn)

Fig. 4.1. Effects of ultrasonic extraction (UE) time of wheat-ginseng extrudates

extruded at 30% feed moisture, 200 rpm screw speed, and 120°C zone 5 barrel

temperature on extractable ginsenosides (Rbl, Rc, Rd, and Rg2). GP: ginseng

powder.

102

Absorbance (AU)

Absorbance (AU)

Fig. 4.2. RP-HPLC chromatograms of ginsenosides ultrasonically extracted for 150
min from a non-extruded blend of wheat ﬂour (WE-ginseng powder (GP) and a
wheat-ginseng extrudate (WGE) extruded at 25% feed moisture, 300 rpm screw

speed, and 140°C zone 5 barrel temperature.

0.15

0.12

0.09

0.06

0.03

0.00

0.15

0.12

0.09

0.06

0.03

0.00

 

 

 

 

 

 

 

 

 

 

 

10 15 20 25 30
Time (min)
1
WGE
4
3
10 15 20 25 30

Time (min)

103

l: Rh]; 2: Re; 3: Rg2; 4: Rd; S: Rg3.

CHAPTER 5
Effects of Extrusion Process Variables on Quality

Properties of Wheat-Ginseng Extrudates

104

5.]. ABSTRACT

The present study was conducted to investigate the effects of extrusion
process variables (feed moisture, screw speed, and barrel temperature) on the physical
[expansion ratio, water absorption index (WAI), and water solubility index (WSI)],
pasting, and thermal properties of wheat- ginseng extrudates (WGE). A wheat ﬂour-
ginseng powder (GP) blend (10% GP, w/W) was extruded in a twin—screw extruder (L/D
ratio of 25:1) with full factorial combinations of feed moisture (25, 30, and 35%), screw
speed (200 and 300 rpm), and zone 5 barrel temperature (110, 120, 130, and 140°C).
The expansion ratios of WGE samples were signiﬁcantly increased with decreasing feed
moisture, decreasing screw speed, and increasing barrel temperature. Increasing feed
moisture signiﬁcantly increased WAI values of WGE samples and signiﬁcantly
decreased WSI values of WGE samples. However, an increase in either of screw speed
or barrel temperature caused a signiﬁcant decrease in WAI values of WGE samples and
a signiﬁcant increase in WSI values of WGE samples. Rapid Visco Analyzer peak
viscosity values of WGE samples were signiﬁcantly affected by changes in extrusion
process variables studied, indicating that the degree of starch degradation and/or
gelatinization in WGE samples is a very important factor associated with their peak
viscosity. WAI values of WGE samples were positively correlated (r = 0.88, p S
0.001) with peak viscosity values of WGE samples, whereas WSI values of WGE
samples were negatively correlated (r = -0.82, p 5 0.001). Increasing feed moisture
resulted in an increase in values of transition peak temperature (Tp) of WGE samples,
whereas increasing screw speed and barrel temperature each led to a decrease in Tp

values of WGE samples, determined by differential scanning calorimetry.

105

5.2. INTRODUCTION

The quality properties of ﬁnal extruded products are considerably affected by
extrusion process variables, such as feed rate, feed moisture, screw conﬁguration, screw
speed, diameter of exit die, barrel temperature, and the addition of other materials
(Anderson and Ng 2001, 2003; Ding et al 2006; Hagenimana et a1 2006; Ryu and Ng
2001; Singh and Singh 2004). These variables can be optimized during the extrusion
process to improve the quality properties of ﬁnal extruded products, including physical
[expansion ratio, color, bulk density, water absorption index (WAI), and water solubility
index (W 81)], pasting, and thermal characteristics (Anderson and Ng 2001; Ding et al
2005; Ding et al 2006; Guha et a1 1998; Kaletunc and Breslauer 1993; McPherson et a1
2000; Ozcan and Jackson 2005; Ryu and Ng 2001).

It has been reported that texture is one of the most important factors in the
quality properties of extruded snack products, and that texture is associated with an
extrudate’s expansion ratio (Ali et al 1996). Ryu and Ng (2001) observed that the
expansion ratio of wheat ﬂour (WF) extrudates or cornmeal extrudates signiﬁcantly
increased with decreasing feed moisture, and Hagenimana et al (2006) reported that rice
ﬂour extrudates extruded at higher feed moisture levels are harder after cooling than
those with lower feed moistures.

Pasting properties of extrudates have been studied using a Rapid Visco
Analyzer (RVA). Ozcan and Jackson (2005) reported that extruded corn starch shows
lower RVA viscosity proﬁles than raw corn starch because of starch degradation during
extrusion cooking. Bhattacharya et a1 (1999) evaluated the pasting properties of a
potato and WF blend extruded in a twin-screw extruder at different moisture contents

(l6-21%) and screw speeds (200-400 rpm). They noted that the degree of starch

106

gelatinization that occurred during extrusion cooking affected the viscosity of the
extrudates.

Differential scanning calorimetry (DSC) has been applied to investigate the
thermal properties of extruded corn starch (Blanche and Sun 2004; McPherson et al
2000; Ozcan and Jackson 2005), corn ﬂour (Kaletunc and Breslauer 1993), and WP
(Anderson and Ng 2001). Anderson and Ng (2001) investigated the transition peak
temperature (Tp), and transition enthalpy (AI-I) of non-extruded and extruded WF
samples and they reported that the value of Tp is particularly associated with the heat
stability of the non-extruded and extruded samples.

Ginseng is one of the most widely used herbal medicines in the world. It
has been known that ginseng exhibits various pharmaceutical effects, including
antioxidation, antistress, immunostimulation, and anticancer effects (Bae et a1 2002; Kang
et a1 2006; Shibata 2001; Yun et al 2001).

In recent years, many consumers have become interested in nutraceutical
foods. Ginseng can be a health supplement for consumers. Ginseng is usually
consumed in the forms of raw plant materials, dried plant materials, extracts, and
commercial products such as capsules, tablets, drinks, and jellies in the world. The
addition of ginseng to common cereal ﬂours (wheat, corn, and rice) for producing
nutraceutical extruded products (snacks or instant powder) could be of interest to
consumers. The extrusion process is fundamentally extremely complicated due to its
high degree of process variability, thus the product quality of any ﬁnal extrudates can be
considerably inﬂuenced by even small changes in extrusion processing conditions.
However, no studies have been found evaluating the effects of extrusion process

variables on the product quality of ﬁnal extruded nutraceutical WF products containing

107

ginseng root powder. Therefore, the present study was designed to investigate the
quality properties [physical (expansion ratio, WAI, and WSI), pasting, and thermal
characteristics] of wheat-ginseng extrudates (WGE) produced with a twin-screw

extruder at different extrusion process conditions.

108

5.3. MATERIALS AND METHODS
5.3.]. Materials

Soft WF was obtained from the Mennel Milling Co. in Fostoria, OH, USA.
The raw ginseng root (Panax ginseng C. A. Meyer), harvested aﬁer four years growth
and milled into powder (particle size of S 167 um), was cultivated in Geumsan, Korea,
and purchased at a ginseng market. WP and GP were analyzed for their moisture
contents using AACC] Approved Method 44-15A (AACCI, 2000), and the moisture

contents of WP and GP were 11.2% (wb) and 8.3% (wb), respectively.

5.3.2. Extrusion Cooking

Extrusion cooking was described in Chapter 4 of this dissertation.

5.3.3. Speciﬁc Mechanical Energy
Speciﬁc mechanical energy (SME) was described in Chapter 4 of this

dissertation.

5.3.4. Expansion Ratio

After drying WGE samples extruded under different conditions in an air oven
(45°C) overnight, 8 digital caliper (Model CD-6"CS, Mitutoyo Corp., Japan) was used to
measure their diameters. To calculate the expansion ratio, the average measurement of

10 WGE diameters was divided by the die diameter of 3 mm.

5.3.5. Water Absorption Index (WAI) and Water Solubility Index (W SI)

109

WA] and WSI of WGE samples extruded under different conditions were
measured according to the procedure of Jin et al (1995) with some modiﬁcations. Two
g of ground WGE samples were combined with 20 mL of distilled water in 30 mL round
bottom centrifuge tubes. The tubes were allowed to sit for 10 min and were inverted 3
times each at 5 and 10 min. After 10 min, the suspensions were centrifuged (Model J2-
21M, Beckman Instruments Inc., Fullerton, CA, USA) at 1000 x g for 25 min. The
supernatant was collected and WA] was calculated as:

WAI = weight of pellet/sample weight (db)
WSI was measured by drying the supernatant in an air oven overnight at 60°C. WSI
was calculated as:

WSI = weight of dried supernatant/sample weight (db)

5.3.6. Pasting Properties

Pasting properties (values of peak, trough, breakdown, ﬁnal viscosity, and
setback) of a non-extruded WF-ginseng powder (GP) blend (3.15 g WF + 0.35 g GP)
and ground WGE samples extruded under different conditions were analyzed. Pasting
properties were determined using a RVA (Model 4, Newport Scientiﬁc Inc.,
Warriewood, Australia). For each analysis, 3.5 g of a sample (14% wb) were mixed
with 25 mL of distilled water. The proﬁle for analysis was Standard Method 1
according to AACCI Approved Method 76-21 (AACCI, 2000) and data ﬁ'om the RVA
were processed by Thermocline version 1.2 software (Newport Scientiﬁc Inc.,

Warriewood, Australia).

5.3.7. Thermal Properties

110

Thermal properties of a non-extruded WF-GP blend (9 mg WF + 1 mg GP)
and ground WGE samples extruded under different conditions were analyzed using a
DSC (Model TA 2910, TA Instruments, Newcastle, DE, USA). Ten mg of a sample
were weighed into a DSC aluminum pan (Model 0319-1525, PerkinElmer Inc., Shelton,
CT, USA) and 20 uL of distilled water were added using a micro-syringe. The sample
pan was hermetically sealed and allowed to equilibrate overnight at room temperature.
Samples were heated from 30 to 200°C at the heating rate of 10°C/min. A sealed
empty pan was used as a reference. Tp and AH were recorded and analyzed using TA

Universal Analysis Software (version 3.6, TA Instruments, Newcastle, DE, USA).

5.3.8. Statistical Analysis

All statistical analyses were performed using SAS version 9.1 (SAS
Institute Inc., Cary, NC, USA). Analysis of variance (ANOVA) was performed using
the general linear models (GLM) procedure to determine signiﬁcant differences among
the samples. Means were compared by using Fisher’s least signiﬁcant difference
(LSD) procedure. Signiﬁcance was deﬁned at the 5% level. In addition, Pearson
correlation coefﬁcients were determined among the samples. The CORR procedure

was used to obtain correlation coefﬁcients.

1]]

5.4. RESULTS AND DISCUSSION

5.4.1. Expansion Ratio

The effects of extrusion process variables (feed moisture, screw speed, barrel
temperature) on the expansion ratio of WGE samples are reported in Table 5.].
Increasing feed moisture from 25 to 35%, at constant screw speed and zone 5 barrel
temperature (200 rpm and 140°C, respectively), caused a signiﬁcant decrease in the
expansion ratios of WGE samples from 2.81 to 2.13. The trend of decreasing
expansion ratio of extrudates with increasing feed moisture has also been noted by Ding
et al (2005), who observed that increasing feed moisture in the extrusion cooking of rice
ﬂour induces a signiﬁcant decrease in the expansion ratio of the rice-based extrudates.
Ryu and Ng (2001) noted that higher feed moisture for the extrusion cooking of WF or
whole cornmeal decreases the expansion ratios of those extrudates. They reported that
not only bubble growth but also bubble shrinkage during extrusion cooking can be
inﬂuenced by the viscosity of the melt in a barrel (Ryu and Ng 2001). More
speciﬁcally, the lower viscosity of a melt at higher feed moisture extrusion process
conditions can cause an increase in the shrinkage and collapse of bubbles at the exit die,
resulting in a reduction in the expansion ratio of the extrudates. In work conducted by
Ding et a1 (2006), it was shown that feed moisture was found to be the main factor
affecting the expansion ratio of WF extrudates. They reported that the expansion ratio
of extrudates is attributed to the elasticity properties of the melt. They also reported
that the elasticity of the melt at higher feed moisture extrusion conditions decreases due
to plasticization of the melt, thus the SME is reduced and starch gelatinization is also
considerably reduced, subsequently causing a decrease in the expansion ratio of the

extrudates.

112

An increase in screw speed from 200 to 300 rpm, at constant feed moisture
and zone 5 barrel temperature (25% and 140°C, respectively), led to the signiﬁcant
reduction in the expansion ratios of WGE samples ﬁom 2.81 to 1.99. A change in
screw speed can affect residence time, the network of starch and proteins, and the melt
viscosity, thereby affecting the expansion ratio of extrudates (Colonna et a1 1998).
Blanche and Sun (2004) reported that melt viscosity decreased with increasing screw
speed from 300 to 400 rpm during extrusion cooking of corn starch, causing a decrease
in pressure, subsequently lessening expansion ratio of the extrudates. In addition,
according to Anderson and Ng (2003), the trend of decreasing expansion ratio at higher
screw speed extrusion conditions is a result of breakdown of the feed materials into
smaller molecules, which may prevent bubble formation.

The expansion ratios of WGE samples signiﬁcantly increased from 2.23 to
2.81 with increasing zone 5 barrel temperature from 110 to 140°C, at constant feed
moisture and screw speed (25% and 200 rpm, respectively). This ﬁnding was
consistent with Ding et a1 (2005), who reported that higher barrel temperatures led to
increased expansion ratios of rice flour extrudates. In the present study, the
increasingly higher barrel temperature extrusion cooking could have caused increasingly
greater ﬂashing-off of moisture upon die exit, inducing an increase in the expansion
ratio of WGE samples. This occurrence is mainly due to a great difference between the
exit die temperature and room temperature, causing an instant expansion of the

extrudates upon exiting the die due to the very high water vaporization rate.

5.4.2. Water Absorption Index (WAI) and Water Solubility Index (W SI)

113

The effects of extrusion process variables on WAI and WSI of WGE samples
are listed in Table 5.1. WAI values of WGE samples produced under different
extrusion process conditions ranged from 6.53 to 8.73 g/g, and WSI values of WGE
samples ranged ﬁ'om 0.06 to 0.26 g/g.

The WAI of extrudates measures the amount of water held by the starch after
dispersion of starch in excess water and may be related to the degree of starch damage
due to gelatinization and fragmentation of starch during high temperature and shear
extrusion cooking (Anderson and Ng 2003). The WSI measures the amount of soluble
components released from starch upon extrusion and is considered an indicator of
degradation of molecular components (Kirby et al 1988).

In the present study, increasing feed moisture from 25 to 35%, at constant
screw speed and zone 5 barrel temperature (200 rpm and 140°C, respectively),
signiﬁcantly increased WAI values of WGE samples ﬁom 6.70 to 7.68 g/g, whereas it
signiﬁcantly decreased WSI values of WGE samples ﬁ'om 0.12 to 0.07 g/g. Higher
WAI values and lower WSI values of WGE samples with increasing feed moisture may
be associated with a decrease in the degradation of starch granules during the higher feed
moisture extrusion cooking. During extrusion cooking at higher feed moisture levels,
more water acts as a plasticizer, decreasing the degree of degradation of starch granules,
leading to an increase in WAI values of WGE samples and a decrease in WSI values of
WGE samples (Hagenimana et al 2006).

Secondly, an increase in screw speed from 200 to 300 rpm, at constant feed
moisture and zone 5 barrel temperature (25% and 110°C, respectively), resulted in a
signiﬁcant decrease in WA] values of WGE samples ﬁom 8.04 to 7.28 g/g, but caused a

signiﬁcant increase in WSI values of WGE samples from 0.09 to 0.13 g/g. Anderson

114

and Ng (2003) noted that WAI values of WF extrudates are signiﬁcantly decreased with
increasing screw speed, because increasing screw speed leads to more damaged polymer
chains, therefore decreasing the ability of starch molecules to bind more water molecules.
This is also in agreement with Jin et al (1995) who reported that higher screw speeds
result in lower WAI values of corn meal extrudates using a twin screw extruder.

Finally, WAI values of WGE samples were signiﬁcantly reduced from 8.04
to 6.70 g/ g, and WSI values of WGE samples were signiﬁcantly increased from 0.09 to
0.12 gg when zone 5 barrel temperature was increased ﬁom 110 to 140°C, at a constant
feed moisture and screw speed (25% and 200 rpm, respectively). Ding et al (2006) also
observed the same result in the extrusion cooking of WF extrudates. They reported that
lower WAI values and higher WSI values of the extrudates are the result of higher
degree of starch gelatinization occurring during higher temperature extrusion cooking.
It is suggested in the present study that extrusion cooking under higher zone 5 barrel
temperature conditions could increase the degree of wheat starch gelatinization, which
causes lower WAI values and higher WSI values of WGE samples. Furthermore, it is
plausible that at higher barrel temperatures, the combination of thermal and mechanical
energies could have fully cooked the starch, subsequently increasing the degree of starch

gelatinization and/or degradation.

5.4.3. Pasting Properties

RVA pasting curves of a non-extruded blend of WF-GP and a ground WGE
sample, extruded at 30% feed moisture, 200 rpm screw speed, and 110°C zone 5 barrel
temperature, are illustrated in Figure 5.1. During the initial 4 min, RVA viscosity of

the WGE sample was higher than that of WF-GP, indicating that the gelatinized starch

115

can hydrate faster than unmodiﬁed starch in the non-extruded blend. However, RVA
viscosity of WGE samples remained low throughout the heating cycle (SO-95°C), in
contrast to that of WF-GP which had substantial increases. The result can be related to
the degradation and/or gelatinization of starch granules occurring during extrusion
cooking. This reduced the swelling capacity of amylose and amylopectin in extruded
starches.

RVA peak viscosity values of ground WGE samples produced under
different extrusion process conditions ranged from 36.3 to 206.1 RVU (Table 5.2).
Increasing feed moisture ﬁ'om 25 to 35%, at a constant screw speed and zone 5 barrel
temperature (200 rpm and 120°C, respectively), signiﬁcantly increased the peak
viscosity values of WGE samples from 113.9 to 176.5 RVU. However, peak viscosity
values of WGE samples were signiﬁcantly decreased from 78.2 to 36.3 RVU with
increasing screw speed from 200 to 300 rpm, at a constant feed moisture and zone 5
barrel temperature (25% and 140°C, respectively). Increasing zone 5 barrel
temperature from 110 to 140°C, at constant feed moisture and screw speed (25% and
200 rpm, respectively), signiﬁcantly decreased peak viscosity values of WGE samples
from 118.9 to 78.2 RVU. The effects of extrusion process variables on peak viscosity
values of WGE samples were generally consistent with other previous published results
(Blanche and Sun 2004; Guha et al 1998).

In the present study, it is speculated that a decrease in the peak viscosity
values of WGE samples resulted from an increase in severity of extrusion process
conditions. It is possible that extrusion cooking at the higher screw speed led to more
starch degradation, subsequently producing WGE samples with much lower pasting

peak viscosity values. With increasing barrel temperature, the starches were likely

116

gelatinized to a greater extent, resulting in reduced peak viscosity values of the ground
WGE samples. Blanche and Sun (2004) observed that higher feed moisture and lower
screw speed each cause an increase in the peak viscosity values of corn starch extrudates.
They reported that the increased peak viscosity of the corn starch extrudates is due to the
presence of starch granules remaining ungelatinized upon extrusion cooking.
According to Guha et a1 (1998), on the other hand, who showed that the peak viscosity
values of rice ﬂour extrudates decrease with increasing barrel temperature and screw
speed, the degree of breakdown of a starch granule is dependent on the type of starch,
mechanical shear, temperature and chemical agents present during the gelatinization of

starch.

5.4.4. Thermal Properties

DSC thermograms of a non-extruded blend of WF-GP and a WGE sample,
extruded under 30% feed moisture, 300 rpm screw speed, and 140°C zone 5 barrel
temperature, are shown in Figure 5.2. Tp and AH values of WGE samples extruded
under different conditions are listed in Table 5.2. A gelatinization peak was observed
on the DSC thermogram obtained for WF-GP, and Tp and AH values of WF-GP were
694°C and 3.36 J/g, respectively. However, WGE samples did not exhibit a
gelatinization peak on the DSC thermogram. Tp and AH values of WGE samples
extruded under different conditions were observed in the range from 153.7 to 172.3°C
and in the range from 1.75 to 6.60 J/g, respectively.

Ozcan and Jackson (2005) suggested that a gelatinization peak was not found
on the DSC thermogram obtained for corn starch extrudates, since the starch granules

were already melted and gelatinized during extrusion cooking. Blanche and Sun

117

(2004) noted that high shear and pressure extrusion cooking can produce damaged and
depolyrnerized starch molecules, so that no gelatinization peak is found on the DSC
thermograms of these corn starch extrudates. Therefore, it seems consistent in the
present study that no gelatinization peak was found on the DSC thermograms for the
ground WGE samples due to the gelatinization and/or breakdown of starch granules
during extrusion cooking.

In the present study, increasing feed moisture ﬁ'om 25 to 35%, at constant
screw speed and zone 5 barrel temperature (300 rpm and 140°C, respectively), resulted
in a signiﬁcant increase in Tp values of WGE samples from 153.7 to 159.7°C. On the
other hand, Tp values of WGE samples were signiﬁcantly decreased from 161.5 to
153.7°C with increasing screw speed from 200 to 300 rpm, at constant feed moisture and
zone 5 barrel temperature (25% and 140°C, respectively). Increasing zone 5 barrel
temperature ﬁ'om 110 to 140°C, at constant feed moisture and screw speed (30% and
200 rpm, respectively), signiﬁcantly reduced Tp values of WGE samples from 170.3 to
162.6’C. Anderson and Ng (200]) evaluated the effects of twin-screw extrusion
processing on the thermal properties of WF extrudates. They observed that increasing
screw speed ﬁ'om 240 to 320 rpm and die temperature from 120 to 160°C each decreased
Tp values of WF extrudates, noting that extrusion cooking under lower screw speed or
lower die temperature improved heat stability of the extrudates. In general, heat
stability of an extrudate describes the resistance to breakdown of molecular aggregate
structures formed during extrusion cooking by gelatinized starch and/or denatured
proteins. It was hypothesized in the present study that higher Tp values of WGE
samples would be associated with a higher melting temperature of WGE samples,

subsequently increasing the heat stability of WGE samples. Based on the results

118

obtained in the present study, it is suggested that lower feed moisture, higher screw
speed, and higher barrel temperature extrusion conditions each could diminish the heat

stability of WGE samples.

5.4.5. Correlations among Extrudate Quality Properties and System Variables

Pearson’s coefﬁcients of correlation (r) among quality properties and system
variables (SME and product temperature) are reported in Table 5.3. Product
temperature represents the actual temperature of the dough at the exit die during
extrusion cooking. For the WGE samples studied, product temperature of WGE
samples was found to be negatively associated (r = -0.71, p 5 0.001) with WAI values
and positively associated (r = -0.45, p S 0.05) with WSI values. SME values were
negatively related (r = -0.45, p S 0.05) to WAI values and negatively related (r = -0.79, p
5 0.001) to peak viscosity values of WGE samples. In contrast, SME values were
positively related (r = 0.69, p s 0.01) to WSI values of WGE samples. Tanhehco and
Ng (2005) also found a negative relationship between SME values and WAI values of
ground WF extrudates as well as a positive correlation between SME values and WSI
values of ground WF extrudates. They attributed their ﬁndings to the fact that SME
values reﬂect degree of starch ﬁagmentation.

As expected, WAI values of WGE samples were negatively correlated (r =
-O.76, p 5 0.001) with the respective WSI values. WAI values were positively
correlated (r = 0.88, p 5 0.001) with peak viscosity values, whereas WSI values were
negatively correlated (r = -0.82, p 5 0.001) with peak viscosity values for the WGE
samples in the present study. The positive relationship between WAI and peak

viscosity of the extrudates can be probably explained by the fact that both WAI and peak

119

viscosity values describe the degree of starch degradation and/or gelatinization occurring
during extrusion cooking (Anderson and Ng 2003; Guha et a1 1998). For example,
increasing barrel temperature increased the starch gelatinization during extrusion
cooking, subsequently leading to a reduction in both WAI and peak viscosity values of
WGE samples (Ding et a1 2006; Guha et a1 1998).

Tp values of WGE samples were positively correlated (r = 0.88, p S. 0.001)
with their respective WAI values as well as positively correlated (r = 0.86, p 5 0.001)
with peak viscosity values. Based on these results, it is suggested that heat stability of
WGE samples could be increased if the degree of starch gelatinization and/or

degradation were decreased during extrusion cooking.

120

5.5. SUMMARY

Effects of extrusion process variables (feed moisture, screw speed, and barrel
temperature) on the quality properties (physical, pasting, and thermal) of WGE samples
produced under different extrusion conditions were investigated. During extrusion
cooking, the melt viscosity, bubble growth or shrinkage, and the degree of gelatinization
and/or degradation could inﬂuence the ﬁnal quality of WGE samples. The data on
quality properties obtained in the present study also suggested that the expansion ratio,
WAI, WSI, peak viscosity, and Tp of WGE samples were affected by each of the
extrusion process variables tested in the present study. Data obtained from the present
extrusion study may be helpful in predicting the expected performance of extruded
materials in investigations into the potential use of WF mixed with ginseng, or other
nutraceutical materials, for the improvement of nutritional quality of ﬁnal extruded

products.

121

5.6. LITERATURE CITED

AACC International. 2000. Approved Methods of the American Association of Cereal
Chemists. 10th Ed. Method 44-15A and 76-2]. The Association: St. Paul, MN.

Ali, Y., Hanna, M. A., Chinnaswamy, R. 1996. Expansion characteristics of extruded
corn grits. LWT. 29:702-707.

Anderson, A. K, and Ng, P. K W. 2001. Thermal and hydrophobic properties of
extruded wheat ﬂour components as measured by differential scanning calorimetry
and ﬂuorescence spectroscopy. Food Sci. Biotechnol. 10: 156-160.

Anderson, A. K, and Ng, P. K. W. 2003. Physical and microstructural properties of
wheat ﬂour extrudates as affected by vital gluten addition and process conditions.
Food Sci. Biotechnol. 12:23-28.

Bae, E. A., Han, M. J., Choc, M. K, Park, S. Y., and Kim, D. H. 2002. Metabolism of
20(8) and 20(R) ginsenoside Rg3 by human intestinal bacteria and its relation to in
Vitro biological activities. Biol. Pharm. Bull. 25:58-63.

Bhattacharya, S., Sudha, M. L., and Rahim, A. 1999. Pasting characteristics of an
extruded blend of potato and wheat ﬂours. J. Food Eng. 40:107-111.

Blanche, S., and Sun, X. 2004. Physical characterization of starch extrudates as a
function of melting transitions and extrusion conditions. Adv. Polym. Technol. 23:
277-290.

Colonna, P., Tayeb, J., and Mercier, C. 1998. Extrusion cooking of starch and starchy
products. Pages 247-319 in: Extrusion cooking. C. Mercier, P. Linko, and J. M.
Harper, eds. AACC International: St. Paul, MN.

Ding, Q. B., Ainsworth, P., Plunkett, A., Tucker, G., and Marson, H. 2005. The effect of
extrusion conditions on the physicochemical properties and sensory characteristics of
rice-based expanded snacks. J. Food Eng. 66:283-289.

Ding, Q. B., Ainsworth, P., Plunkett, A., Tucker, G., and Marson, H. 2006. The effect of
extrusion conditions on the ﬁrnctional and physical properties of wheat-based
expanded snacks. J. Food Eng. 73:142-148.

Guha, M., Ali, S. 2., and Bhattacharya, S. 1998. Effect of barrel temperature and screw
speed on rapid viscoanalyser pasting behaviour of rice extrudate. Int. J. Food Sci.
Technol. 33:259-266.

Hagenimana, A., Ding, X., and Fang, T. 2006. Evaluation of rice ﬂour modiﬁed by
extrusion cooking. J. Cereal Sci. 43:38-46.

Jin, J., Hsieh, F., and Huff, H. E. 1995. Effects of soy ﬁber, salt, sugar, and screw speed
on physical properties and microstructure of corn meal extrudate. J. Cereal Sci.

122

22: 185-194.

Kaletunc, G., and Breslauer, K J. 1993. Glass transitions of extrudates: relationship with
processing-induced fragmentation and end-product attributes. Cereal Chem. 70:548-
552.

Kang, K. S., Kim, H. Y., Yamabe, N., and Yokozawa, T. 2006. Stereospeciﬁcity in
hydroxyl radical scavenging activities of four ginsenosides produced by heat
processing. Bioorg. Med. Chem. Lett. 16:5028-5031.

Kirby, A. R., Ollett, A. L., Parker, R., and Smith, A. C. 1988. An experimental study of
screw conﬁguration effects in the twin-screw extrusion-cooking of maize grits. J.
Food Eng. 8:247-272.

McPherson, A. E., Bailey, T. B., and Jane, J. 2000. Extrusion of cross-linked
hydroxypropylated corn starches. 1. Pasting properties. Cereal Chem. 77:320-325.

Ozcan, S., and Jackson, D. S. 2005. Functionality behavior of raw and extruded corn
starch mixtures. Cereal Chem. 82:223-227.

Ryu, G. H., and Ng, P. K. W. 2001. Effects of selected process parameters on expansion
and mechanical properties of wheat ﬂour and whole cornmeal extrudates. Starch-
Starke 53:147-154.

Shibata, S. 2001. Chemistry and cancer activities of ginseng saponins and some related
triterpenoid compounds. J. Korean Med. Sci. 16:828-37.

Singh, J., and Singh, N. 2004. Effect of process variables and sodium alginate on
extrusion behavior of nixtamalized corn grit. Int. J. Food Prop. 7:329-340.

Tanhehco, E. J ., and Ng, P. K W. 2005. The effects of extrusion cooking and milling on
the instant properties of wheat powders. Food Sci. Biotechnol. 14:758-765.

Yun, T. K, Lee, Y. S., Lee, Y. H., Kim, S. 1., and Yun, H. J. 2001. Anticarcinogenic

effect of Panax ginseng C. A. Meyer and identiﬁcation of active compounds. J.
Korean Med. Sci. 16:86-18.

123

Table 5.1.
Effects of Extrusion Process Variables on the Expansion Ratio (ER), Water

Absorption Index (WAI), and Water Solubility Index (W SI) of Wheat-Ginseng

 

 

 

 

 

 

Extrudates
Extrusion Process Variables ER WAI WSI
Feed Moisture Screw Speed TemperatureII (g/g) (g/g)
(%, Wb) (rpm) (°C)
25 200 110 2.230 8.04011 0.091jk
30 . 200 110 2.1 If 8.471) 0.08”
35 200 110 1.99g 8 73a 0 06m
_ - - 2'5- - - - - - 300 ------ 1 I0 ----- 132-if - .7-28-gli- - “-0.13; -
30 300 110 1.89] 7 67f 0 12g
35 300 110 1 85k 7 86de 0 09ljk
25 200 120 2.45c 7.31gh 0 llgh
30 200 120 2.25e 7 92d 0 091jk
35 200 120 1.97gh 8 21¢ 0 08k]
- - - 25. - - - - - 500 ------ l 2.0 ----- 1799}- - .6-91-1- - -0130. '-
30 300 120 1.99g 7 171] 0 l3ef
35 300 120 1.94111 7.38g 0 10111
25 200 130 2.511) 6 701k 0 12g
30 200 130 2.24e 7.31gh 0 0911
35 200 130 1.98g 7 92d 0 071m
- - - 25. - - - - - 300 ------ l 50 ----- 138-g- - .6531:- - -5230- -
30 300 130 1.97gh 7 22gb 0 18d
35 300 130 1.97gh 7.21gh 0.11g
25 200 140 2.81a 6 703k 0 12fg
30 200 140 230d 7 28gb 0 0911‘]
35 200 140 2.13f 7 68ef 0 071m
- - - '55- - - - - - 300 ------ 1 30 ----- 1:99}- - .656:- - —0-26a- '-
30 300 140 1.99g 6 8lij 0 19011
35 300 140 1.96gh 7.1411 0.13ef

 

Values with different letters within the same column differ significantly (P<0.05).
”Temperature: zone 5 (nearest exit die) barrel temperature.

124

Effects of Extrusion Process Variables on Peak Viscosity (PV),

Table 5.2.

Transition Peak Temperature (Tp), and Transition Enthalpy (AH) of Wheat-

 

 

 

 

 

 

Ginseng Extrudates
Extrusion Process Variables PV Tp AH
Feed Moisture Screw Speed Temperature'I (RVUb) (°C) (J/ g)
(%, wb) (rpm) (°C)
25 200 110 118.9ef 167.8cd 2.65ijk
30 200 110 138.0d 170.3b 3.70fgh
35 200 110 206.1a 172.3a 1.75jk
' " ' 55' ' ' ' ' ' 306 """ 1 i0 """ 575.3111" ' '1'5311' " "6706.6 '
30 300 110 62.8k 163.4ef 5.04cde
35 300 110 111.8fgh 164.0e 2.72ij
25 200 120 113.9efg 166.1d 3.94fgh
30 200 120 121.3e 167.9cd 4.56def
35 200 120 176.5b 168.7bc 3 43ghi]
' ' ' 55' ' ' ' " ' 301') """ 1 50 """ 471.7ni'n' '1'1'soTsiij' 379E161-
30 300 120 55.6kl 161.8fgh 5.89abc
35 300 120 97.1i 162.3efg 3.90fgh
25 200 130 112.5fg 162.4efg 3.76fgh
30 200 130 110.1gh 162.9efg 3.78fgh
35 200 130 159.4c 163.9e 3.81fgh
' " ' 55' " ' ' ' ' 306 """" 1 50 """ 3-85no ' 3567231" "2757le '
30 300 130 55.9kl 159.9j 5.39bcd
35 300 130 83.7j 160.2ij 3.53ghi
25 200 140 78.2j 161.5ghi 5.4lbcd
30 200 140 103.9hi 162.6efg 3.49ghij
35 200 140 138.4d 163.6ef 4.37efg
' " ' 35' " ' " " ' 306 """ 1 30 """ 375.50" ' "153773 ' '3T4z'hii "
30 300 140 49.0lm 156.2lm 6.60a
35 300 140 62.8k 159.7jk 4.55def

 

Values with different letters within the same column differ significantly (P<0.05).
”Temperature: zone 5 (nearest exit die) barrel temperature.
“RVU: Rapid Visco Units.

125

Table 5.3.

Pearson’s Coefﬁcients of Correlation (r) among Quality Properties [Expansion
Ratio (ER), Water Absorption Index (WAI), Water Solubility Index (W SI), Peak
Viscosity (PV), Transition Peak Temperature (Tp), and Transition Enthalpy (AH)]
of Wheat-Ginseng Extrudates and System Variables [Speciﬁc Mechanical Energy

(SME) and Product Temperature (PT)]

 

 

ER WAI WSI PV Tp AH SME PT

ER - 0.22 - 0.18 0.13 0.17 0.05 - 0.63*** 0.27
WAI - 0.76*** 0.82*** 0.88*** - 0.38 - 0.45* - 0.7l***
WSI - 0.82*** - 0.81*** 0.23 0.69"”r 0.47*

PV 0.86“W - 0.52“ - 0.79*** - 0.39

Tp - 0.42* - 0.67" - 0.67"

AH 0.31 0.24
SME . - 0.02

PT

 

*, **, ***, Significantly different at the 5%, 1%, and 0.1% levels, respectively.

126

 

 

 

 

 

 

300 100
Temperature profile
250 -
(A) - 80
S 200 " E:
> - 60 v
a 2
g; 150 *3
g o
.21 - 40 3'
> o
100 - i“
- 20
50 (B)
.'
0 I l I J l 1 l l 1 l l l 0
0 2 4 6 8 10 12

Time (min)

Fig. 5.1. Rapid Visco Analyzer pasting curves of a non-extruded wheat ﬂour (WF)-
ginseng powder (GP) blend (10% GP, w/w), and a wheat-ginseng extrudate (W GE)
extruded at 30% feed moisture, 200 rpm screw speed, and 110°C zone 5 barrel

temperature. Viscosity reported in Rapid Visco Units (RVU).

127

 

 

 

 

 

 

-1 '
-1.2 -
E»
E -1.4 - WGE
B
a
u:
‘5 -1.6 -
:
WF-GP
-1.s -
'2 I I I
o 50 100 150 200

Temperature (°C)

Fig. 5.2. Differential scanning thermograms of a wheat-ginseng extrudate (WGE)
extruded at 30% feed moisture, 300 rpm screw speed, and 140°C zone 5 barrel
temperature, and a non-extruded wheat flour (WF) and ginseng powder (GP)

blend (10% GP, w/w).

128

CHAPTER 6
Physicochemical Properties of Bread and Cookies
Baked from Wheat Flour Blended with Ginseng

Powder

129

6.]. ABSTRACT

This study investigated (1) the effects of adding ginseng powder (GP) on
Farinogrgh, thermal, and pasting properties of hard wheat ﬂour (HWF) and soft wheat
ﬂour (SWF), (2) the quality properties of hard wheat-ginseng bread (10% GP, w/w) and
soft wheat-ginseng cookies (10% GP, w/w), and (3) the changes in ginsenosides (G,
ginseng saponins) in hard wheat-ginseng bread and soft wheat-ginseng cookies upon
baking. Farinograph data showed that water absorption values of HWF-GP and SWF-
GP were signiﬁcantly higher than those of the respective HWF and SWF. The dough
made from HWF-GP was weaker and less stable than that made from HWF alone.
Differential scanning calorimetry data showed that HWF-GP and SWF-GP have higher
transition onset temperature values than do their respective HWF and SWF, whereas
HWF and SWF show larger transition enthalpy values for gelatinization than do their
respective blends. Rapid Visco Analyzer data showed that the addition of GP to either
HWF or SWF signiﬁcantly decreased the peak viscosity of the samples. The bread loaf
weight increased and the bread loaf volume decreased when GP was added to HWF
bread formulations. The width of the cookie sample baked from SWF-GP was
signiﬁcantly smaller than that of the cookie sample baked from SWF. Individual G
(Rbl, Rc, Rd, Rg2, and Rg3) were fractionated and identiﬁed by RP-HPLC from GP,
ground bread crust and ground bread crumb of samples baked from HWF and HWF-GP,
and ground cookie samples baked from SWF and SWF-GP. The quantities of G (Rbl,
Rc, and Rd) extracted from the bread crust and bread crumb samples baked from HWF-
GP, and the cookie samples baked from SWF-GP were signiﬁcantly higher than those of
the G extracted from pure GP, weight per original weight GP, indicating the conversion

of malonyl G (Rbl, Re, and Rd) into G (Rbl, Re, and Rd, respectively) during bread-

130

and cookie-baking. G Rg2 were found in the bread crust and bread crumb of samples
baked from HWF-GP, whereas G Rg3 were present only in the bread crust samples.

Ginsenosides Rg2 and Rg3 were not present in cookie samples baked from SWF-GP.

131

6.2. INTRODUCTION

Common wheat is divided into soft wheat and hard wheat varieties,
depending on its kernel texture (Hoseney 1998). In particular, soft wheat has lower
protein content and weaker protein strength as compared to hard wheat (Atwell 2001;
Posner 2000). Hard wheat ﬂour (HWF) is used mainly for the production of bread
products, while soft wheat ﬂour (SWF) is usually used to produce chemically-leavened
products, such as cookies, cakes, pastries, crackers, pie dough, and pretzels.

Bread, a critical staple food, is consumed in some form or another all over
the world. In general, the major ingredient for making bread is HWF, with other
ingredients being yeast, salt, and water. Over the years, many different food
ingredients, such as barley and soy flours (Basman et a1 2003), ﬁber (Dalgetty and Baik
2006; Filipovic et a1 2007; Mohamed et al 2008; Park et a1 1997; Pomeranz et al 1977),
folic acid (Osseyi et a1 2001), immature wheat meal (Mujoo and Ng 2003),
polysaccharides (Fan et al 2006), vitamin E (Ranhotra et al 2000), and whole waxy
wheat ﬂour (Hung et a1 2007), have been included in bread formulations to improve the
nutritional value.

Cookies are one of the most popular bakery products because of their
convenience, ready-to-eat nature, and long shelf-life (Singh and Mohamed 2007).
Recently, the addition of ﬁber to cookie formulations has been studied to make
nutraceutical cookie products (Bilgicli et al 2007; Larrea et a1 2005; Uysal et al 2007).

Ginseng has long been used in Asian countries as a traditional medicine
(F uzzati 2004). Ginsenosides (G, ginseng saponins) are the main active components of
ginseng, one of the most popular herbal medicines worldwide (Ren and Chen 1999).

Over 30 G have been identiﬁed; among these, Rbl, Rb2, Rc, Rd, and Re are the most

132

abundant (Corbit et al 2005; Kim et al 2005). On the other hand, G compound Rg3 is
only present in ginseng after special treatment of the ginseng root, which yields so-
called red ginseng (Bae et al 2002; Kim et al 2000; Popovich and Kitts 2004; Shibata
2001; Wang et al 2006). In terms of health beneﬁts, this red ginseng is considered
vastly superior to untreated ginseng, or so-called white ginseng (Ha et al 2005; Ha et a1
2007; Kim et al 2000; Kwon et al 2001; Lau et a1 2004; Popovich and Kitts 2004; Wang
et al 2006).

So—called healthy foods, especially those with nutraceutical properties, are in
great demand in our health conscientious society. Bread and cookies are good
candidates in this respect, for a portion of their wheat ﬂour to be replaced with
nutraceutical ingredients, such as ginseng. However, no studies have reported
evaluating the product quality of and ginsenosides content in baked HWF bread and
SWF cookies that incorporate ginseng root powder (GP). Thus, the objectives of this
study were to investigate (l) the effects of adding GP on the Farinograph, thermal, and
pasting properties of HWF and SWF, (2) the quality properties of hard wheat-ginseng
bread and soft wheat-ginseng cookies, and (3) the changes in G in hard wheat-ginseng
bread and soft wheat-ginseng cookies upon baking, relative to their respective unbaked

doughs.

133

6.3. MATERIALS AND METHODS

6.3.1. Materials

HWF and SWF were obtained from the Mennel Milling Co. in Fostoria, OH,
USA. The raw ginseng root (Panax ginseng C. A. Meyer), harvested after four years
growth and milled into powder (particle size of S 167 um), was cultivated in Geumsan,
Korea, and purchased at a ginseng market. Standards for ginsenosides Rbl, Rc, Rd,

Rg2, and Rg3 were provided by ILHWA Co. (Kuri, Korea).

6.3.2. Chemical Analysis

HWF, SWF, and GP samples were analyzed for their moisture and protein
contents using AACCI Approved Methods 44-15A and 46-13 (AACCI, 2000),
respectively (Table 6.1). Protein contents were calculated by multiplying the nitrogen

content by the conversion factor 5.7, and reported on a 14% moisture basis.

6.3.3. Farinograph Properties

Farinograph properties of HWF (50 g), SWF (50 g), a blend of HWF-GP (45
g HWF + 5 g GP), and a blend of SWF-GP (45 g SWF + 5 g GP) were determined using
AACCI Approved Method 54-21 (AACCI 2000). Optimal water absorption, dough

development time, stability, and mixing tolerance index were determined.

6.3.4. Thermal Properties
A differential scanning calorimeter (DSC Model TA 2910, TA Instruments,
Newcastle, DE, USA) was used to measure the thermal properties of HWF (10 mg),

SWF (10 mg), GP (10 mg), HWF-GP (9 mg HWF + 1 mg GP), and SWF-GP (9 mg

134

SWF + 1 mg GP). Each sample was weighed into a DSC aluminum pan (Model 0319-
1525, PerkinElmer lnc., Shelton, CT, USA), and 20 uL of distilled water were added
using a micro-syringe. The sample pan was hermetically sealed and allowed to
equilibrate overnight at room temperature. Samples were heated from 30 to 200°C at
the heating 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 (AH)
were recorded and analyzed using the TA Universal Analysis Software (version 3.6, TA

Instruments, Newcastle, DE, USA).

6.3.5. Pasting Properties

The pasting properties of HWF (3.5 g), SWF (3.5 g), GP (3.5 g), HWF-GP
(3.15 g HWF + 0.35 g GP), and SWF-GP (3.15 g SWF + 0.35 g GP) were analyzed
using a Rapid Visco Analyzer (RVA Model 4, Newport Scientiﬁc Inc., Warriewood,
Australia). The pasting properties included the following parameters: peak viscosity —
highest viscosity during heating; trough viscosity — lowest viscosity following peak
viscosity; breakdown viscosity - peak viscosity minus trough; ﬁnal viscosity — the
viscosity at the completion of the heating cycle; setback — ﬁnal viscosity minus peak
viscosity. Each sample (14%, wb) was mixed with 25 mL of distilled water. The
proﬁle of the analysis was Standard Method 1 according to AACCI Approved Method
76-21 (AACCI, 2000) and data obtained from the RVA were processed by Thermocline

version 1.2 software (Newport Scientiﬁc Inc., Warriewood, Australia).

6.3.6. Breadmaking

Breads were prepared 1‘10me (100 g) and from HWF-GP (90 gHWF+10 g

135

GP) according to the optimized straight-dough breadmaking method (AACCI Approved
Method lO—lOB; AACCI 2000) with some modiﬁcations. The formulation of bread dough
also included sugar (6.0 g), salt (1.5 g), shortening (3.0 g), yeast (5.3 g, Safrnex, Milwaukee,
WI, USA), ascorbic acid (5.0 mg), and water based on optimum water absorption ﬁ'om the
Farinograph. Breads were baked at 215°C for 24 min in a rotary oven (National MFG Co.,
Lincoln, NE, USA), removed ﬁom their pans, and then cooled at room temperature for 2 hr
prior to quality analysis. The bread-baking procedures were performed in duplicate for each

sample.

6.3.7. Bread Quality

The cooled loaves were weighed and loaf volumes were determined by the
rapeseed displacement procedure in a volume-measuring apparatus. After loaf volumes were
measured, loaves were placed into individual ziplock bags, sealed, and stored for 24 hr at
room temperature prior to bread ﬁrmness analysis. All subsequent bread ﬁrmness
determinations were performed on the central bread crumb. Two slices (each 12.5 mm thick)
of bread were cut from the center of loaves using a cutter (Model FF-3199, Hobart MFG. Co.,
Troy, OH, USA). 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 2 slices was determined

with an acryl probe (38 mm diameter) at a compression rate of 1.7 mm/s.

6.3.8. Cookiemaking
Wire-cut cookies were prepared according to AACCI Approved Method 10-54

(AACCI 2000). Cookies were prepared ﬁom SWF (40 g) and ﬁ'om SWF-GP (36 g SWF + 4

136

g GP). In addition, the formulation of cookie dough also included sucrose (12.8 g),
granulated brown sugar (4 g), non-fat dry milk (0.4 g), salt (0.5 g), sodium bicarbonate (0.4 g),
shortening (16 g), high fructose corn syrup (0.6 g), ammonium bicarbonate (0.2 g), and water
(8.7 g). Cookies were baked at 205°C for 11 min in a rotary oven (National MFG Co.,
Lincoln, NE, USA) and then put on racks to cool to room temperature for 30 min prior to

quality analysis. The cookie-baking procedures were performed in duplicate for each sample.

6.3.9. Cookie Quality

Weight, width, and thickness of the cookies were evaluated after the cookies were
cooled. Two cookies which were made in the same batch were placed together in a ziplock
bag, sealed, and stored for 24 hr at room temperature prior to measuring breaking strength
(hardness). Hardness of cookies was determined from the peak of the displacement plot of
a cookie during shearing in 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.

6.3.10. Sample Preparation for RP-HPLC

Crust portions of bread samples were obtained from the top 1 cm of a bread
slice (12.5 mm thickness) which had been cut ﬁ'om the central section of a bread loaf.
Central crumb portions (2cm x 2 cm) of breads were cut from the center of the same
bread slice (12.5 mm thickness). Samples of bread crust, bread crumb, and cookies
were frozen and dried in a freeze dryer for 24 hr. After drying, portions (about 10 g) of

each sample were ground to a particle size of S 330 um (54 mesh screen) using a coffee

137

grinder (Braun Inc., Wobum, MA, USA). After grinding, moisture contents of ground
samples were determined according to AACCI Approved Method 44-15A (AACCI,
2000). Ground samples were placed in ziplock bags, sealed, and stored at -20°C until

analysis.

6.3.11. Extraction of Ginsenosides

Four mL of 70% (v/v) aq. methanol were added to GP (0.1 g). Fourteen mL
of 70% (v/v) aq. methanol were added to ground bread crust and to ground bread crumb
samples ( 1.11 g, based on 0.1 g GP) baked from HWF and HWF-GP, and to ground
cookie samples (1.72 g, based on 0.1 g GP) baked from SWF and SWF-GP. Each
mixture was ultrasonically extracted for 150 min using an Ultrasonicator (Model FS 14H,
Fisher Scientiﬁc, Pittsburgh, PA, USA, Power: 155W) at room temperature and then
centrifuged at 11,200 x g for 20 min using a centrifuge (Model J2-21M, Beckman
Instruments Inc., Fullerton, CA, USA) at 25°C. The supernatant was evaporated using
a Micro Rotary Evaporator (Model 421-4000, Labconco, Kansas City, MO, USA) at
50°C. Prior to RP-HPLC analysis, the residue was dissolved in 2 mL of 70% (v/v) aq.
methanol and ﬁltered through a 0.45 pm nylon ﬁlter membrane (Millipore, Ireland).
Each sample was extracted in duplicate and two chromatographic runs were performed

for each extract.

6.3.12. Fractionation and Identiﬁcation of Ginsenosides by RP-HPLC
RP-HPLC was carried out on a Millenium 2010 HPLC Workstation,
consisting of a Waters 600E multi-solvent delivery system (Waters, Milford, MA, USA),

a temperature control module, and a 996 photodiode array detector. Separation was

138

performed at 30°C on a Phenomenex 300 RP, Jupiter C-l8 column (4.6 x 250 mm, 5 pm
particle diameter; Phenomenex, Torrance, CA, USA). A Phenomenex security guard
with the same packing material served as the guard column.

The solvents were A: water (distilled water ﬁltered through Milli-Q-Plus,
Millipore), and B: HPLC grade acetonitrile. Separation was achieved using the
following gradient: 0-5 min, 20-30% B; 5-25 min, 30-40% B; 25-30 min, 40-20% B.
The solvent ﬂow rate was 1 mL/min and the injection volume was 20 uL. The solvents
were purged with helium at the rate of 20 mL/min. The column was equilibrated for 10
min with 20% acetonitrile prior to injection. The UV detection wavelength was set at
203 nm (Lau et a1 2004), and the detector output was transmitted simultaneously to the

computer for data storage and graphic representations.

6.3.13. Statistical Analysis

All statistical analyses were performed using SAS version 9.1 (SAS
Institute Inc., Cary, NC, USA). Analysis of variance (ANOVA) was performed using
the general linear models (GLM) procedure to determine signiﬁcant differences among
the samples. Means were compared by using Fisher’s least signiﬁcant difference

(LSD) procedure. Signiﬁcance was deﬁned at the 5% level.

139

6.4. RESULTS AND DISCUSSION

6.4.1. Farinograph Properties

Farinograph properties of HWF, HWF-GP, SWF, and SWF-GP are shown in
Table 6.2. The addition of GP to either HWF or SWF signiﬁcantly increased water
absorption values, indicating that water absorption capacity of GP is higher than that of
either HWF or SWF alone. The dough development time and stability value of HWF-
GP were signiﬁcantly lower than those of the HWF, whereas the mixing tolerance index
value was signiﬁcantly higher when GP was added to HWF. In the present study, the
ﬁndings indicated that the dough made from HWF-GP is weaker and less stable during
mixing than that made from HWF alone. The deterioration in each of the Farinograph
properties could be related to the lower amounts of gluten proteins in HWF-GP, due to a
diluting effect with the addition of GP, as compared to HWF. On the other hand, the
dough development time, stability, and mixing tolerance index values of SWF-GP were
not signiﬁcantly different from those of the SWF. Since the SWF already has very
weak dough properties, the addition of GP did not signiﬁcantly further weaken the SWF
dough samples, though there was a trend of slight decreases in the measured

Farinograph properties.

6.4.2. Thermal Properties

Thermal properties of HWF, SWF, GP, HWF-GP, and SWF-GP are
presented in Table 6.3. To and Tp values of both HWF-GP and SWF-GP were
signiﬁcantly higher than those of HWF and SWF, respectively. On the other hand, AH
values of the blends were signiﬁcantly lower than those of the respective HWF and SWF

samples. Pure GP did not exhibit an endothermic peak.

140

Nashed et a1 (2003) observed that values of To and Tp of wheat starch
increased with the addition of glycerol, while increasing the water content resulted in
decreases in the values of To and Tp. They reported that adding glycerol delays the
gelatinization process of wheat starch rather than assisting it. However, water behaves
as a plasticizer, leading to a decrease in the values of To and Tp. Adding gluten protein
(Mohamed and Rayas-Duarte 2003), soy protein concentrate (Li et a1 2007), and sugars
(Baek et al 2004) to wheat starch, corn starch, and corn starch, respectively, has been
shown to increase values of To and Tp of the starches. In the present study, because
GP demonstrated no endothermic peak, it can be assumed that wheat starch plays a
major role in the thermal behavior of the blends. The observation (an increase in
values of To with the addition of GP to either HWF or SWF) indicated that adding GP to
either HWF or SWF can make wheat starch gelatinization more difﬁcult due to the
competition for water between wheat starch and GP components. This absorption
competition limits the water available for gelatinization of wheat starch, resulting in
increased To of the blends. This speculation is conﬁrmed by the Farinograph water
absorption data (Table 6.2). On the other hand, a decrease in AH with the addition of
GP to either HWF or SWF could be due to the proportionate reduction in the amounts of
HWF or SWF in the blends. In other words, the quantity of wheat starch (g starch/g
sample) in pure HWF or in SWF is greater than in their respective blends, increasing
energy requirements to gelatinize these amounts of starch, resulting in increased values

of AH.

6.4.3. Pasting Properties

141

Effects of addition of GP on the pasting properties of HWF and SWF were
studied via the RVA. The pasting properties (peak, trough, breakdown, ﬁnal viscosity,
and setback) of HWF, SWF, GP, HWF-GP, and SWF-GP are reported in Table 6.4.
SWF had higher values for peak, trough, breakdown, ﬁnal viscosity, and setback than
did HWF. This ﬁnding was good agreement with Ragaee and Abdel-Aal (2006), who
reported that higher RVA viscosity values (peak, trough, breakdown, ﬁnal viscosity, and
setback) of SWF, as compared to HWF, are attributed to the presence of more starch
granules within SWF and this results in increased swelling capacity of the sample. All
RVA values for HWF and SWF were signiﬁcantly higher than those of pure GP and of
their respective blends, except for the trough viscosity for HWF and its blend, which
were similar. Pure GP had very low viscosity values for all of the RVA parameters
studied, indicating that pure GP has very low swelling capacity. Therefore, it is
suggested in the present study that a decrease in peak viscosity of each blend relative to
its parent ﬂour was probably a result of the lower wheat starch content within the blends

(see end of section 6.4.2) as well as a low swelling capacity of the GP.

6.4.4. Bread Quality Properties

Quality properties of bread samples baked from HWF and HWF-GP are
listed in Table 6.5. A small but signiﬁcant increase in the loaf weight of the baked
bread sample was noted when GP was added to HWF. As was noted earlier, GP had a
higher water absorption capacity than did HWF. Thus, it is plausible that HWF-GP
could absorb more water than did HWF during bread-baking, subsequently increasing

the bread loaf weight.

142

The addition of GP to the HWF bread formulation led to a signiﬁcant
decrease in the loaf volume of the bread sample. The result in the present study was
consistent with the studies on bread baked from HWF and ﬁber (Dalgetty and Baik
2006; Hung et al 2007; Mujoo and Ng 2003; Pomeranz et a1 1977). Pomeranz et al
(1977) reported that the addition of different ﬁbers (cellulose, oat bulls, and wheat bran)
to the HWF bread formulation decreased the loaf volume of the bread samples mainly
due to decreased gas retention. Dalgetty and Baik (2006) reported that the loaf weight
of the bread sample increases and loaf volume of the bread sample decreases when bulls
and soluble ﬁbers (isolated from peas, lentils, and chickpeas) are added to HWF.
Filipovic et al (2007) observed that the loaf volume of a bread sample made ﬁ'om a
blend of HWF and sugar-beet ﬁber is lower than that of a bread sarnpie made ﬁ'om HWF
alone. Mujoo and Ng (2003) studied the physicochemical properties of bread baked
from HWF blended with immature wheat meal. They reported that the loaf volume of
the bread sample baked from a blend of HWF-immature wheat meal is lower than that of
the bread sample baked from HWF, and suggested that the reduced loaf volume of the
bread sample results from reduced gas retention. In the present study, a decrease in the
loaf volume of the bread sample baked from the HWF-GP blend could be explained by
lower amounts of wheat gluten in the bread formulation, resulting in a weaker gluten
matrix with the reduced ability to retain gases created during fermentation, and hence a
decrease in the loaf volume of the bread sample baked from the blend.

No effect on the crumb ﬁrmness of the bread sample was observed when GP
was added to HWF. It had been expected that the ﬁrmness of the bread samples baked
from HWF-GP would be increased due to the lower loaf volume that compacts the

crumb structure. In the present study, however, lack of change in the ﬁrmness of the

143

bread samples baked from the blend could be associated with not only increased water

content in the formula, but also higher moisture retention capacity during baking.

6.4.5. Cookie Quality Properties

Quality properties of cookie samples baked ﬁom SWF and SWF-GP are
shown in Table 6.6. Weight, thickness, and hardness of the cookie samples baked from
the blend were not signiﬁcantly different from those of the cookie samples baked from
SWF. However, a signiﬁcant decrease in width of the cookie samples was noted when
GP was added to SWF.

Uysal et a1 (2007) studied the effect of incorporation of dietary ﬁber (from
apples, lemons, wheat, and wheat bran) on the spread ratio of wire-cut cookies. They
reported that adding ﬁber to the SWF cookie formulation reduced the spread ratio of
cookie samples. Miller et al (1997) reported that cookie spread is dependent on the
water absorption properties of the ﬂour sample. They suggested that ﬂours with higher
water absorption capacity decrease the cookie spread since this higher absorption
decreases the amount of water available to dissolve the sugar in the cookie formulation.
According to Yamazaki (1962), ﬂours which have a low water retention capacity are
considered good quality ﬂours in terms of cookie production, producing large spread
cookies. In the present study, it was conﬁrmed that the higher water absorption

capacity of the blend is associated with a decrease in the width of the cookie samples.

6.4.6. Changes in Ginsenosides during Bread-Baking and Cookie-Baking
The quantities of G (Rbl, Rc, Rd, Rg2, and Rg3) ultrasonically extracted for

150 min from GP, bread crusts and bread crumbs from HWF and HWF-GP bread

144

samples, and cookies baked ﬁom SWF and SWF-GP are shown in Table 6.7. The
quantities of G Rbl, G Rc, and G Rd extracted from the bread crust and crumb samples
baked from the blend and the cookie samples baked from the blend were signiﬁcantly
higher than their respective quantities extracted from GP. In addition, G Rg2 were
found in the bread crust and crumb samples baked from HWF-GP. Interestingly, the
amounts of G Rg2 extracted from the bread crust sample baked from HWF-GP were
signiﬁcantly higher than the amounts extracted from the bread crumb of the same
sample. G Rg3 were only present in the bread crust sample baked from the blend.
However, no G (Rbl, Rc, Rd, Rg2, and Rg3) in the bread crust and bread crumb of
samples baked from HWF and the cookie samples baked ﬁom SWF were found.

The types of G extracted from ginseng are quite different depending on the
processing methods used, such as in the production of white and red ginseng (Ha et a1
2005; K0 et al 2005; Shibata 2001). Ha et al (2005) showed a signiﬁcant increase in
the quantities of G Rbl, G Rb2, G Re, and G Rd after extrusion processing of raw Panax
ginseng. Shibata (2001) noted that white ginseng includes the malonyl esters of G Rbl,
G Rb2, G Re, and G Rd, while the malonyl group, which is originally attached at the 6"-
position of the glycosyl moieties of G Rbl, G Rb2, G Rc, and G Rd, is easily lost during
the steaming process used to make red ginseng. Fuzzati (2004) reported that the acidic
malonyl G (malonyl G Rbl, malonyl G Rb2, malonyl G Rc, and malonyl G Rd) are
thermally very unstable; they degraded or converted into the corresponding neutral G (G
Rbl, G Rb2, G Re, and G Rd, respectively) during the steaming process in red ginseng
production.

Based on results in the present study, it is proposed that the high temperature

bread- and cookie-baking processes could lead to the loss of malonyl acid from the

145

glycosyl moieties of malonyl G Rbl, malonyl G Re, and malonyl G Rd, thereby leading
to increases in the amounts of G Rbl, G Rc, and G Rd in the bread crust and bread
crumb of samples baked from HWF-GP and in the cookie samples baked from SWF-GP.
Furthermore, the amounts of G Rbl, G Rc, and G Rd extracted from the bread crust and
bread crumb samples baked from HWF-GP were higher than those extracted from the
cookie samples baked from SWF-GP. It is speculated that not only the higher baking
temperature (215°C) but also the longer baking time (24 min) used for bread-baking
could cause an increase in the amounts of extracted G, as compared to cookie-baking
conditions (205°C and 11 min).
Wang et al (2006) noted that the amounts of G Rg2, G Rg3, G Rhl, and G
Rh2 increase after steaming process of raw Panax quinquefolius and they reported that
these G can be used as marker compounds to distinguish red ginseng from white ginseng.
Ha et a] (2005) also found an increase in the G Rg group (G Rgl, G Rg2, and G Rg3) in
the ﬁnal extruded Panax ginseng samples. Shibata (2001) reported that the glycosyl
moiety at the C-20 position of the protopanaxadiol type G, including G Rbl, G Rb2, G
Re, and G Rd, is partly degraded to produce G Rg3 during the steaming process to make
red ginseng. Popovich and Kitts (2004) showed that G Rg3 are naturally absent in
Panax quinquefolius, but are produced by a thermal process. On the other hand, G Rg2
can be produced by the protopanaxatriol type G, such as G Re, in the same way (Kim et
a1 2000). In the present study, results indicated that the bread-baking conditions used
could lead to the chemical degradation and/or conversion of the thermolabile
protopanaxadiol type G and protopanaxatriol type G into G Rg3 and G Rg2, respectively.
Kim et al (2000) observed that during steaming processing of raw Panax

ginseng, increasing the steaming temperature from 100 to 120°C resulted in an increase

146

in the quantities of G Rg2 and G Rg3. In work conducted by Wang et al (2006), when
berries of Panax quinquefolius were steamed at 120°C, the quantities of G Rg2 and G
Rg3 increased signiﬁcantly compared with those steamed at 100°C. In the present
study, it was hypothesized that higher temperatures could chemically stimulate an
increase in the conversions of protopanaxatriol type G and protopanaxadiol type G to G
Rg2 and G Rg3, respectively. It is suggested in the present study that the greater
amounts of G Rg2, as well as the presence of G Rg3, in the crusts of bread samples
baked from the HWF-GP blend, as compared to the bread crumb sample baked from the
same blend, could be due to a difference in the actual temperature between the outside

crust and the inside crumb during bread-baking.

147

6.5. SUMMARY

The addition of GP to either HWF or SWF induced an increase in water
absorption values, increased To and Tp values, and decreased the RVA viscosity values
during the heating process. It can be concluded that the higher water absorption
capacity of GP interferes with the moisture uptake of wheat starch in the blends, and
hence diminishes the effective moisture contents of the starch available to aid
gelatinization. The addition of GP to the HWF bread formulations and to the SWF
cookie formulations led to decreases in the loaf volumes of the bread and in the widths
of the cookies. It is presumed that the temperatures used for bread- and cookie-baking
caused increases in the amounts of G (Rbl, Re, and Rd) in the samples. New G
compounds (Rg2 and Rg3), and highly desirable ones pharmaceutically, were found
only in bread samples baked from the HWF-GP blend, with G Rg3 appearing in the
crust but not the crumb of bread samples. Although the quality properties of the bread
samples baked from HWF-GP and the cookie samples baked from SWF-GP were
slightly lower than those of their counterpart samples without GP, the slight decreases in
quality seem outweighed by the presence of nutraceutical components, G, in the
products baked ﬁ'om the blends. Therefore, the production of bread and cookies which
incorporate ginseng powder can broaden the utilization of ginseng, and the products can

be regarded as possible health-promoting nutraceutical foods.

148

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151

Table 6.1.
Moisture and Protein Contents of Hard Wheat Flour (HWF), Soft Wheat Flour

(SWF), and Ginseng Powder (GP) Samples

 

 

Sample Moisture Content (%, wb) Protein Content (%)'I
HWF 13.2 12.7
SWF 11.2 8.3
GP 8.3 11.2

 

'Protein percentages are on a 14% moisture basis.

152

Table 6.2.
Farinograph Properties of Hard Wheat Flour (HWF), a Blend of HWF-Ginseng

Powder (GP), Soft Wheat Flour (SWF), and SWF-GP‘

 

 

Sample Water Dough Stability Mixing
Absorption Development (min) Tolerance
(%) Time Index

(min) (BU)

HWF 63.0a 6.5a 4.5a 20c
HWF-GP 65.0b 4.5b 3.5b 60b
SWF 49.2d 1.5c 2.0c 95a
SWF-GP 51.6c 1.0c 1.0c 100a

 

Values with different letters within the same column differ signiﬁcantly (P<0.05).
'Flour-GP blends were a ratio of 9:1 w/w.
”BU: Brabender Units.

153

Table 6.3.
Thermal Properties'l of Hard Wheat Flour (HWF), Ginseng Powder (GP), a Blend

of HWF-GP, Soft Wheat Flour (SWF), and SWF-GP”

 

 

Sample To (°C) Tp (°C) AH (J/g)
HWF 60.0d 67.4c 4.27c
HWF-GP 61.7c 69.0a 3.36d
SWF 62.5b 68.2b 7.13a
SWF-GP 63.7 a 69.4a 6.73b
GP ND ND ND

 

Values with different letters within the same column differ signiﬁcantly (P<0.05).
ND: not detectable.
'To: transition onset temperature; Tp: transition peak temperature; AH: transition enthalpy.

l’l'r‘lour-GP blends were a ratio of 9:1 w/w.

154

Table 6.4.

Pasting Properties of Hard Wheat Flour (HWF), Ginseng Powder (GP), a Blend of

HWF-GP, Soft Wheat Flour (SWF), and SWF-GP‘I

 

 

Sample Peak Trough Breakdown Final Viscosity Setback
(RVU) (RVU) (RVU) (RVU) (RVU)
HWF 102.7c 29.8c 72.9c 79.4c 49.6c
HWF-GP 97.6d 28.8c 68.8d 71.9d 43.1d
SWF 287.3a 167.0a 120.2a 296.0a 128.6a
SWF-GP 242.8b 132.0b 110.7b 252.5b 120.4b
GP 25.2e 21.8d 3.4e 30.1e 8.3e

 

Values with different letters within the same column differ signiﬁcantly (P<0.05).

RVU: Rapid Visco Units.

'Flour-GP blends were a ratio of 9:1 w/w.

155

Table 6.5.
Quality Properties of Breads Baked from Hard Wheat Flour (HWF) and a Blend of

HWF-Ginseng Powder (GP)'l

 

 

Sample Weight (g) Volume (cm’) Firmness (N)
HWF 148.6b 725a 2.20a
HWF-GP 153.2a 650b 2.56a

 

Values with different letters within the same column differ signiﬁcantly (P<0.05).
'HWF-GP blend was a ratio of 9:1 w/w.

156

Table 6.6.
Quality Properties of Cookies Baked from Soft Wheat Flour (SWF) and a Blend of

SWF-Ginseng Powder (GP)"

 

Sample Weight (g) Width (cm) Thickness (cm) Hardness (N)

 

SWF 21.1a 16.1a 1.75a 42.5a

SWF-GP 20.5a 15.0b 1.68a 40.2a

 

Values with different letters within the same column differ signiﬁcantly (P<0.05).
'SWF-GP blend was a ratio of 9:] w/w.

157

Table 6.7.
Quantities of Ginsenosides (Rbl, Rc, Rd, Rg2, and Rg3) Ultrasonically Extracted
for 150 min from Ginseng Powder (GP), Bread Crusts and Bread Crumbs Baked
from Hard Wheat Flour (HWF) and a Blend of HWF-GP, and Cookies Baked from

Soft Wheat Flour (SWF) and a Blend of SWF-GP'I

 

 

 

Sample Ginsenosides (mg/100 mg GP)
Rbl Rc Rd Rg2 Rg3

GP 0.86c 0.51c 0.25b ND ND
Bread Crust

HWF ND ND ND ND ND

HWF-GP 0.96a 0.59a 0.33a 0.17a 0.07
Bread Crumb

HW F ND ND ND ND ND

HWF-GP 0.98a 0.59a 0.31a 0.06h ND
Cookie

SWF ND ND ND ND ND

SWF-GP 0.89b 0.56b 0.33a ND ND

 

Values with different letters within the same column differ signiﬁcantly (P<0.05).
ND: not detectable.
'Flour-GP blends were a ratio of 9:1 w/w.

158

CHAPTER 7

GENERAL CONCLUSIONS

159

It has been known that ginseng exhibits various pharmaceutical effects,
including antioxidation, antistress, immunostimulation, and anticancer effects. Though
ginseng has been consumed in the form of raw plant materials, dried plant materials,
extracts, and commercial products such as capsules, tablets, drinks, and jellies in the
world, the production of nutraceutical wheat ﬂour (WF) snack foods, bread, and cookies
that incorporate ginseng has not been reported. The present study investigated (1) the
optimum extraction conditions of ginsenosides (G, ginseng saponins) from a blend of
WF-ginseng powder (GP), (2) the effects of extrusion process variables on quantities
and types of extractable G in wheat-ginseng extrudates (WGE), (3) the quality properties
of WGE extruded under different conditions, and (4) physicochemical properties of
bread and cookies baked from WF blended with GP, and quantities and types of
extractable G in these products.

While studying the optimum extraction conditions of G ﬁom WF-GP,
interactions between WF components and G were observed in WF-GP heated at 90°C.
The interactions were found to occur between the wheat starch fraction (SF) and G as
well as between the gluten fraction (GF) and G. Furthermore, the degree of
interactions between the SF and G was greater than that between the GF and G in the
heated WF-GP. The interactions between WF components and G signiﬁcantly
decreased the amounts of G extractable from the heated WF-GP samples. These
interactions were disruptable, and the degree of disruption increased with increasing
ultrasonic extraction time (i.e., total energy input). Ultrasonic extraction may have
facilitated the access of the solvent [70% (v/v) aq. methanol] to the G, resulting in an
increase in the extractable quantities of individual G. Based on the results obtained in

the present study, the optimum extraction conditions to achieve the maximum extraction

160

of G from WF-GP were a sample-to-solvent ratio of 0.071 :1 (mg/pl.) and an
ultrasonication time of 90 min.

Effects of extrusion process variables (feed moisture, screw speed, and barrel
temperature) on extractable G in WGE were investigated. It was revealed that an
increase in the quantities of ginsenosides Rbl, Re, and Rd in WGE samples is a result of
the degradation and/or conversion of malonyl ginsenosides Rbl, Re, and Rd into
ginsenosides Rbl, Rc, and Rd, respectively, and that the presence of the new
ginsenosides Rg2 and Rg3 is due to the chemical modiﬁcation of thermolabile
protopanaxatriol type G and protopanaxadiol type G into G Rg2 and G Rg3, respectively.
It was observed that the presence and quantities of G Rg2 and G Rg3 are signiﬁcantly
affected by extrusion process variables studied. In particular, increasing zone 5 barrel
temperature played the more dominant role in the conversion of protopanaxatriol type G
into G Rg2 than did other processing variables (e.g., screw speed and feed moisture).
The optimal extrusion process condition in the present study, producing the maximum
quantities of G Rg2 and G Rg3 in WGE samples, was 25% feed moisture, 300 rpm
screw speed, and 140°C zone 5 barrel temperature.

The physical [expansion ratio, water absorption index (WAI), and water
solubility index (WSI)], pasting, and thermal properties of WGE samples extruded under
different conditions were determined. It was found that expansion ratio, WAI, WSI,
peak viscosity and transition peak temperature of WGE samples were dependent on
extrusion process variables studied, indicating that the ﬁnal quality properties of WGE
samples can be controlled by the combination of extrusion process variables tested in the
present study.

Farinograph, thermal, and pasting properties of hard wheat ﬂour (HWF), soft

161

wheat ﬂour (SWF), and blends of HWF-GP and SWF-GP were analyzed. Farinograph
data revealed that the dough made from HWF-GP was weaker and less stable than that
made from HWF alone. Differential scanning calorimetry data indicated that adding
GP to either HWF or SWF can inhibit wheat starch gelatinization due to the competition
for water between wheat starch and GP components. Rapid Visco Analyzer data
showed that a decrease in peak viscosity of HWF-GP or SWF-GP relative to HWF or
SWF, respectively, is probably a result of the lower wheat starch content in the blends as
well as a low swelling capacity of the GP.

Bread samples were prepared from HWF and HWF-GP, and cookie samples
were prepared ﬁ'om SWF and SWF-GP. A signiﬁcant decrease in the loaf volume of
the bread and in the width of the cookie was observed when GP was added to the bread
formulation and the cookie formulation. It was found that bread- and cookie-baking
can cause an increase in the amounts of G Rbl, G Re, and G Rd in bread samples baked
from HWF-GP and cookie samples baked from SWF-GP. Although new G Rg2 were
found in both bread crumb and bread crust samples baked from HWF-GP, new G Rg3
were present only in the bread crust sample.

Based on the overall ﬁndings obtained in the above studies, interactions
between WF components and G, including interactions between the SF and G and
between the GF and G, did occur in a blend of WF-GP heated at 90°C. G had a higher
degree of and/or stronger interactions with the SF than with the GP in WF-GP. These
interactions could be disrupted, given the proper extraction conditions. An increase in
the amounts of G (Rbl, Re, and Rd) in WGE, HWF-GP bread, and SWF-GP cookie
samples explained the chemical degradation of acidic malonyl G (Rbl, Re, and Rd) to

neutral G (Rbl, Re, and Rd, respectively) during the extrusion and baking processes.

162

The presence of new ginsenosides (Rg2 and Rg3) in WGE and HWF-GP bread samples
indicated that both extrusion and bread-baking processes can produce the more desirable
ginsenosides. A higher zone 5 barrel temperature (2 130°C) during extrusion cooking
and higher actual bread-baking temperature (i.e., bread crust) improved the conversion
efﬁciency of thermolabile protopanaxatriol type G and protopanaxadiol type G into G
Rg2 and G Rg3, respectively. A healthy extruded snack product with nutraceutical
properties and acceptable quality properties can be optimized by the combinations of
extrusion process variables. Furthermore, the production of WF snacks, bread, and
cookies containing ginseng can broaden the utilization of ginseng and the products can

be considered as possible nutraceutical foods.

163

CHAPTER 8

FUTURE RECOMMENDATIONS

164

The following are recommendations for further research:

(1) To better understand the types of interactions (i.e., physical and chemical
interactions) between wheat ﬂour (WF) components and ginsenosides (G,
ginseng saponins) in a heated blend of WF-ginseng powder (GP), other
analytical tools, such as confocal laser microscopy, X-ray diffractometry, and
nuclear magnetic resonance spectroscopy, should be used.

(2) To improve not only the chemical degradation of malonyl G Rbl , malonyl G Rc,
and malonyl G Rd into G Rbl, G Re, and G Rd, respectively, but also the
conversion efﬁciency of protopanaxadiol type G and protopanaxatriol type G
into G Rg3 and G Rg2, respectively, in wheat-ginseng extrudates, more severe
extrusion process conditions should be considered: e.g., high shear screw
conﬁgurations, zone 5 barrel temperature 2 150°C, screw speed 2 400 rpm, and
feed moisture S 20%.

(3) To improve the loaf volume of the bread sample baked from a blend of hard
WF-GP, effects of adding dough enhancers such as vital gluten and ascorbic acid

to the bread formulation should be determined.

165

APPENDIX 1.
Effects of Sample Heating of Wheat Flour (WF, 0.9 g), Ginseng Powder (GP, 0.1 g),
and a Blend of WF-GP (0.9 g WF + 0.1 g GP) on Subsequent Extractable

Ginsenosides (Rbl, Re, and Rd)

 

 

 

Sample Temperature Ginsenosides (mg/100 mg GP)
(°C) Rbl Rc Rd
GP 25 0.87 0.54 0.26
50 0.89 0.55 0.26
70 0.87 0.53 0.26
90 0.89 0.54 0.27
WF 25 ND ND ND
50 ND ND ND
70 ND ND ND
90 ND ND ND
WF-GP 25 0.87 0.54 0.26
50 0.86 0.53 0.26
70 0.79 0.51 0.23
90 0.72 0.47 0.21

 

ND: not detectable.

166

APPENDIX 2.

Moisture Contents of Wheat-Ginseng Extrudates Extruded at Different Conditions

 

Extrusion Process Variables

 

 

 

 

 

Feed Moisture Screw Speed Temperature' Moisture Content”
(%, wb) (rpm) (°C) (%, wb)

25 200 110 8.2
30 200 110 8.3
35 200 110 8 2

""25- """" 3710""--'i'15""""-72' -----
30 300 110 8 l
35 300 110 7 l
25 200 120 8 0
30 200 120 9.3
35 200 120 8 2

""25- ''''' 330'"'--123""""-72' -----
30 300 120 6 l
35 300 120 5 0
25 200 130 7 8
30 200 130 6 6
35 200 130 5 9

""25- """" shinn'isﬁuunn'Es' """
30 300 130 5 5
35 300 130 5 4
25 200 140 6 6
30 200 140 6 5
35 200 140 5 9

"'"25' """" 3710"""14Tr"""-'-55 """"
30 300 140 5 4
35 300 140 5.6

 

'Temperature: zone 5 (nearest exit die) barrel temperature.
hMoisture content was determined immediately after drying at 45°C overnight.

167

APPENDIX 3.
Effects of Ultrasonic Extraction (UE) Time on Extractability of Ginsenosides (Rbl,

Rc, Rd, Rg2, and Rg3) from Wheat-Ginseng Extrudates Extruded at Different

 

 

 

Conditions
Extrusion UE Time Ginsenosides (mg/100 mg Ginseng Powder)
Conditions (min) Rbl RC Rd R32 Rg3
_MS/T'
25/200/110 60 0.64 0.43 0.21 0.05 ND
90 0.78 0.48 0.26 0.05 ND
120 0.87 0.52 0.28 0.06 ND
150 0.87 0.51 0.28 0.06 ND
30/200/110 60 0.64 0.42 0.20 0.05 ND
90 0.80 0.46 0.25 0.05 ND
120 0.86 0.52 0.28 0.06 ND
150 0.87 0.51 0.29 0.06 ND
35/200/110 60 0.63 0.41 0.20 0.04 ND
90 0.78 0.45 0.22 0.04 ND
120 0.86 0.52 0.26 0.05 ND
150 0.87 0.51 0.26 0.05 ND
25/300/110 60 0.60 0.41 0.19 0.05 ND
90 0.80 0.47 0.23 0.07 ND
120 0.85 0.51 0.28 0.09 ND
150 0.86 0.50 0.28 0.09 ND
30/300/110 60 0.56 0.36 0.19 0.05 ND
90 0.76 0.46 0.23 0.05 ND
120 0.85 0.50 0.27 0.06 ND
150 0.85 0.50 0.27 0.06 ND
35/300/110 60 0.62 0.41 0.21 0.05 ND
90 0.82 0.47 0.23 0.05 ND
120 0.85 0.51 0.27 0.06 ND
150 0.85 0.51 0.27 0.06 ND

 

'M: feed moisture (%, wb); S: screw speed (rpm); T: zone 5 barrel temperature (°C).
ND: not detectable.

168

APPENDIX 3.

 

 

 

Continued
Extrusion UE Time Ginsenosides (mg/100 mg Ginseng Powder)
Conditions min
M/S/T‘ ( ) Rbl Rc Rd Rg2 Rg3
25/200/120 60 0.69 0.44 0.21 0.05 ND
90 0.79 0.48 0.25 0.05 ND
120 1.02 0.61 0.31 0.08 ND
150 1.03 0.61 0.32 0.08 ND
30/200/120 60 0.53 0.37 0.19 0.05 ND
90 0.79 0.49 0.23 0.06 ND
120 0.99 0.57 0.31 0.07 ND
150 1.00 0.56 0.32 0.07 ND
35/200/120 60 0.60 0.41 0.19 0.05 ND
90 0.79 0.46 0.23 0.06 ND
120 0.96 0.55 0.32 0.07 ND
150 0.96 0.55 0.32 0.07 ND
25/300/120 60 0.66 0.43 0.21 0.06 ND
90 0.80 0.48 0.23 0.08 ND
120 1.01 0.62 0.33 0.11 ND
150 1.01 0.62 0.33 0.11 ND
30/300/120 60 0.55 0.38 0.19 0.05 ND
90 0.79 0.49 0.24 0.06 ND
120 0.96 0.58 0.32 0.08 ND
150 0.96 0.58 0.32 0.08 ND
35/300/120 60 0.64 0.41 0.19 0.06 ND
90 0.84 0.49 0.24 0.07 ND
120 0.96 0.58 0.31 0.08 ND
150 0.96 0.58 0.31 0.08 ND

 

'M: feed moisture (%, wb); S: screw speed (rpm); T: zone 5 barrel temperature (°C).

ND: not detectable.

169

APPENDIX 3.

 

 

 

Continued
Extrusion UE Time Ginsenosides (mg/100 mg Ginseng Powder)
Conditions min
M/S/T' ( ) Rbl Rc Rd Rg2 Rg3
25/200/130 60 0.71 0.45 0.22 0.06 ND
90 0.81 0.48 0.24 0.07 ND
120 1.04 0.62 0.33 0.08 ND
150 1.04 0.61 0.32 0.08 ND
30/200/130 60 0.71 0.44 0.22 0.05 ND
90 0.82 0.48 0.24 0.06 ND
120 1.05 0.62 0.33 0.07 ND
150 1.04 0.62 0.33 0.07 ND
35/200/130 60 0.68 0.44 0.22 0.05 ND
90 0.81 0.49 0.23 0.05 ND
120 1.04 0.62 0.32 0.07 ND
150 1.04 0.63 0.33 0.07 ND
25/300/130 60 0.69 0.45 0.22 0.08 0.04
90 0.80 0.49 0.24 0.11 0.05
120 1.04 0.62 0.32 0.14 0.06
150 1.04 0.62 0.32 0.15 0.06
30/300/130 60 0.71 0.43 0.22 0.06 ND
90 0.80 0.49 0.24 0.07 ND
120 1.04 0.61 0.32 0.09 ND
150 1.05 0.61 0.32 0.09 ND
35/300/130 60 0.68 0.45 0.23 0.05 ND
90 0.78 0.48 0.22 0.07 ND
120 1.04 0.62 0.32 0.08 ND
150 1.05 0.62 0.32 0.08 ND

 

'M: feed moisture (%, wb); S: screw speed (rpm); T: zone 5 barrel temperature (°C).

ND: not detectable.

170

APPENDIX 3.

 

 

 

Continued
Extrusion UE Time Ginsenosides (mg/100 mg Ginseng Powder)
Conditions min
M/S/T‘ ( ) Rbl Rc Rd Rg2 Rg3
25/200/140 60 0.70 0.43 0.21 0.07 ND
90 0.78 0.48 0.22 0.08 ND
120 1.06 0.61 0.32 0.09 ND
150 1.06 0.61 0.32 0.09 ND
30/200/140 60 0.66 0.44 0.21 0.06 ND
90 0.80 0.48 0.23 0.08 ND
120 1.04 0.62 0.31 0.09 ND
150 1.04 0.61 0.31 0.09 ND
35/200/140 60 0.65 0.43 0.21 0.05 ND
90 0.79 0.49 0.23 0.06 ND
120 1.06 0.62 0.32 0.07 ND
150 1.05 0.62 0.32 0.07 ND
25/300/140 60 0.66 0.44 0.21 0.14 0.06
90 0.79 0.48 0.23 0.16 0.06
120 1.06 0.61 0.32 0.19 0.07
150 1.05 0.61 0.33 0.30 0.07
30/300/140 60 0.64 0.43 0.21 0.09 0.04
90 0.79 0.49 0.24 0.10 0.05
120 1.04 0.61 0.32 0.11 0.05
150 1.05 0.61 0.32 0.12 0.05
35/300/140 60 0.63 0.43 0.21 0.06 ND
90 0.78 0.49 0.23 0.08 ND
120 1.03 0.62 0.32 0.09 ND
150 1.04 0.62 0.32 0.09 ND

 

'M: feed moisture (%, wb); S: screw speed (rpm); T: zone 5 barrel temperature (°C).

ND: not detectable.

171

APPENDIX 4.
Die Pressure Related to Different Extrusion Process Variables of Wheat-Ginseng

Extrudates

 

Extrusion Process Variables

 

 

 

 

 

Feed Moisture Screw Speed Temperature‘ Die Pressure

1%.wb) (rpm) (°C) (Psi)

25 200 110 705

30 200 110 585

35 200 110 325
""2'5""'"30'6 """" rio """" 4'15""

30 300 110 375

35 300 110 230

25 200 120 600

30 200 120 500

35 200 120 275
""2"5"""'307r """" 1'2'0 """" 2'95""

30 300 120 315

35 300 120 190

25 200 130 510

30 200 130 220

35 200 130 100
""2'5'-""'30'6 """" 130 """"" 1'36""

30 300 130 70

35 300 130 40

25 200 140 350

30 200 140 220

35 200 140 120
""2'5""""3071 """" 1'50 """" 1'06""

30 300 140 90

35 300 140 40

 

”Temperature: zone 5 (nearest exit die) barrel temperature.

172

APPENDIX 5.
Residence Time Related to Different Extrusion Screw Speeds of Wheat-Ginseng

Extrudates

 

Extrusion Process Variables

 

 

Feed Moisture Temperature‘ Screw Speed Residence Time
(%, Wb) (°C) (rpm) (sec)
25 120 200 67
25 120 300 63

 

'Temperature: zone 5 (nearest exit die) barrel temperature.

173

APPENDIX 6.

Pasting Properties of Wheat-Ginseng Extrudates Extruded at Different Conditions

 

 

 

 

 

Extrusion Conditions Trough Breakdown Final Setback
M/S/T'| (RVU) (RVU) Viscosity (RVU)
(RVU)

25/200/110 64.3 54.7 106.0 41.8
30/200/110 66.3 71.7 104.3 38.0
35/200/110 27.5 178.6 49.9 22.4

' ' '2'5/306/1'16 ' " ' ' 1'63 ' ' ' ' 5977 """ 3'15 """ 1 15' ' '
30/300/110 14.2 48.6 25.1 10.9
35/300/110 23.2 88.6 41.5 18.3
25/200/120 64.8 49.1 105.9 41.1
30/200/120 47.9 73.4 83.4 35.5
35/200/120 42.0 134.5 80.7 38.7

' " '2'5/306/126 " " ' ' 1'23 ' ' " ' 5'23 '''' 271.5 """ 1 2'1" ' "
30/300/120 17.0 38.6 32.0 15.0
35/300/120 20.5 76.7 35.6 16.1
25/200/130 61.7 50.3 100.1 37.9
30/200/130 28.6 81.5 54.2 25.6
35/200/130 17.6 141.8 36.7 19.1

" ' '2'5/30'6/1'36' """ s .'1' ' " ‘ ' 30.3 """ {7.2 """ 5.4" " '
30/300/130 15.5 40.4 30.3 14.8
35/300/130 10.5 73.3 20.5 10.0
25/200/140 33.4 44.8 57.6 24.3
30/200/140 28.3 75.6 53.4 25.1
35/200/140 19.4 119.0 41.8 22.4

' ' '2'5/305/1'45 """ 7 .3 ' ' ' ' 28:5 """ 2'25 """ 1 3.7" ' '
30/300/140 10.7 38.3 21.0 10.0
35/300/140 10.1 52.8 19.6 9.5

 

'M: feed moisture (%, wb); S: screw speed (rpm); T: zone 5 barrel temperature (°C).
RVU: Rapid Visco Units.

174

APPENDIX 7.
Thermal Properties' of Hard Wheat Flour (HWF), Ginseng Powder (GP), Blends
of HWF-GP (10, 20, and 30% GP, w/w), Soft Wheat Flour (SWF), and Blends of

SWF-GP (10, 20, and 30% GP, w/w)

 

 

Sample To (°C) Tp (°C) AH (J/g)

HWF 60.0 67.4 4.27

GP ND ND ND

90% HWF-10% GP 61.7 68.9 3.36
80% HWF-20% GP 62.4 69.4 2.89
70% HWF-30% GP 63.3 69.8 2.29
SWF 62.5 68.2 7.13

90% SWF-10% GP 63.7 69.4 6.73
80% SWF-20% GP 64.7 69.9 4.67
70% SWF-30% GP 65.6 70.7 3.81

 

‘To: transition onset temperature; Tp: transition peak temperature; AH: transition enthalpy.
ND: not detectable.

175

APPENDIX 8.
Pasting Properties of Hard Wheat Flour (HWF), Ginseng Powder (GP), Blends of
HWF-GP (10, 20, and 30% GP, w/w), Soft Wheat Flour (SWF), and Blends of

SWF-GP (10, 20, and 30% GP, w/w)

 

 

Sample Peak Trough Breakdown Final Setback
(RVU) (RVU) (RVU) Viscosity (RVU)
(RVU)
HWF 102.7 29.8 72.9 79.4 49.6
GP 25.2 21.8 3.4 30.1 8.3
90% HWF-10% GP 97.5 28.8 68.8 71.8 43.0
80% HWF-20% GP 86.1 26.3 59.8 59.3 33.0
70% HWF-30% GP 79.7 25.6 54.1 56.1 30.0
SWF 287.3 167.0 120.2 296.0 128.6
90% SWF-10% GP 242.8 132.0 110.7 252.5 120.4
80% SWF-20% GP 199.6 103.8 95.8 199.2 95.4
70% SWF-30% GP 163.1 81.6 81.7 153.0 71.3

 

RVU: Rapid Visco Units.

176

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