Ruyi Wu Thesis

AN ABSTRACT OF THE THESIS OF
Ruyi Wu for the degree of Master of Science in Food Science and Technology
presented on December 12, 2008.
Title: Effects of Refrigeration Storage and Processing Technologies on the Bioactive
Compounds and Antioxidant Capacities of Blackberries (‘Marion’ and ‘Evergreen’).
Abstract approved:
Yanyun Zhao
The objective of this project was to investigate the effects of refrigeration
storage and processing technologies on the quality and nutraceutical benefit of
blackberries. ‘Marion’ and ‘Evergreen’, the two major blackberry varieties in Oregon,
were evaluated in this study. For refrigeration storage, fresh fruit were packed in clam-
shell containers right after harvest and stored at 2.0 ± 0.2 oC and 95 ± 2% relative
humidity for 9 or 7 days for ‘Marion’ and ‘Evergreen’, respectively. For evaluating
processing effect, fresh fruit were individually quick frozen at -23 oC and then subjected
to different processing treatments including freeze drying, hot-air drying, canning-in-
water, canning-in-20° Brix sucrose syrup, and making into jam. Processed berry
products were stored at room temperature at 25 ± 2 oC and 30 ± 2% relative humidity
for up to 6 months. Physicochemical properties including pH, titratable acidity (TA),
total soluble solids (TSS) and moisture content (MC), bioactive compounds including
total phenolics (TPC) and total monomeric anthocyanins (ACY), and antioxidant
capacities evaluated as radical scavenging activity (RSA), oxygen radical absorbance
capacity (ORAC) and ferric reducing ability of plasma (FRAP) were monitored in fresh
blackberries and processed blackberry products right after processing and during
storage. Frozen fruit were used as a control for evaluating processing effects. TA
reduced by 36.8% and 46.2% in ‘Marion’ and ‘Evergreen’, respectively (P<0.05) at the
end of refrigeration storage compared to that at day 0. TPC and ACY decreased
significantly in both varieties. ORAC declined by 20% and FRAP increased by 18.75%,
respectively in ‘Evergreen’ (P<0.05) while remained stable in ‘Marion’ during
refrigeration storage. Right after processing, TA in all the processed ‘Marion’ products
lost significantly, while TSS increased in dried blackberries and blackberry jams of both
varieties. Freeze drying increased TPC by 27% and 21% in ‘Marion’ and ‘Evergreen’,
respectively (P<0.05), whereas hot-air drying reduced ACY by 56% in ‘Marion’ and
84% in ‘Evergreen’ (P<0.05). The content of measured bioactive compounds lost 31%
to 70% (P<0.05) in canned products except TPC in ‘Marion’ canned-in-water. ORAC
and FRAP lost 21%-61% in canned ‘Marion’ (P<0.05). RSA increased in canning-in-
water fruit while decreased in canning-in-sucrose syrup for both varieties. Making
blackberries to jams resulted in extensive losses of bioactive compounds and
antioxidant capacity in both varieties. Over 6–month storage, TA deceased 9% and 12%
in ‘Marion’ and ‘Evergreen’ jams, respectively (P<0.05). The significant losses of ACY
in hot-air dried, canned blackberries and blackberry jams, and RSA in frozen, dried and
canned blackberries, ORAC in hot-air dried and canned-in-water blackberries of both
varieties were also observed. Two varieties of blackberries varied in physicochemical
properties, content of bioactive compounds and antioxidant capacities, with higher

values of TA, MC, TPC, ACY, RSA and FRAP observed in ‘Marion’ than those in
‘Evergreen’.
©Copyright by Ruyi Wu
December 12, 2008
All Rights Reserved
Effects of Refrigeration Storage and Processing Technologies on the Bioactive
Compounds and Antioxidant Capacities of Blackberries (‘Marion’ and ‘Evergreen’)
by
Ruyi Wu
A THESIS
submitted to
Oregon State University
in partial fulfillment of
the requirements for the
degree of
Master of Science
Presented December 12, 2008
Commencement June 2009

Master of Science thesis of Ruyi Wu presented on December 12, 2008.
APPROVED:
Major Professor, representing Food Science and Technology
Head of the Department of Food Science and Technology
Dean of the Graduate School
I understand that my thesis will become part of the permanent collection of Oregon
State University libraries. My Signature below authorizes release of my thesis to any
reader upon request.
Ruyi Wu, Author

ACKNOWLEDGEMENTS
This comprehensive work can not be completed successfully by me alone. Here
is my sincere acknowledgement to all the individuals generously contributing to my
research and giving support in my everyday life in Corvallis.
I would like to express my foremost appreciation to my advisor, Professor
Yanyun Zhao for offering me the opportunity to study and work in the Department of
Food Science and Technology at Oregon State University. The academic knowledge I
learned and research experiences I obtained will be invaluable for my career. I am also
grateful to her patient guidance, insightful advices, and constant encouragement
throughout my study at OSU.
I greatly appreciate my graduate committee members, Drs. James Kennedy,
Andrew Ross and Kenneth Johnson for dedicating your precious times and efforts to
review my thesis project, give critical and constructive suggestions and serve on my
committee.
I would like to recognize the great assistance I received from Mr. Brian Yorgey
for helping me process canned blackberries and blackberry jams and the lab activities in
FST 210 class, Mr. Jeff Clawson for operating hot-air drier and freeze drier in FST pilot
plant, and Mr. James Batdorff for solving equipment problems. Without your help, I
would not have the blackberry products for my thesis research.

Many thanks to Dr. James Kennedy and Mr. Seth Cohen for providing me the
facilities to run HPLC and advices in analyzing HPLC data, to Dr. Mark Daeschel for
kindly giving me the permission to conduct preliminary study on the HPLC instrument
in his lab, to Mr. Bob Durst for his professional suggestions on the sample extraction
procedures, and to Dr. Balz Frei and Ms. Deborah Hobbs at the Linus Pauling Institute
for their considerable work on conducting the ORAC and FRAP assays of my samples.
I am also very grateful to my colleagues in Dr. Zhao’s research group, Dr.
Daniel Lin for transferring extraction skills to me and Dr. Jingyun Duan for her multiple
helps during my experiments.
My gratitude also goes to all the other faculty and staff in this department and
especially the main office ladies, Linda Hoyser, Linda Dunn, Iowa Williams, Debby
Yacas, Christina Hull and Tamara Critchfield. And particular thanks to all my friends in
Corvallis for their continuous care and friendship.
Finally, I would like to deliver my deepest appreciation to my parents and
relatives in China and the United States for their endless encouragement, understanding,
patience, support and love which help me overcome the difficulties throughout my life.
TABLE OF CONTENTS
Page
1. CHAPTER 1 Introduction...................................................................................... 1
2. CHAPTER 2 Literature Review............................................................................. 6
2.1 Introduction....................................................................................................... 6
2.2 Overview of blackberries……………………………………………............... 7
2.2.1 History and botanical classification of blackberries………………….…... 7

2.2.2 Cultivars of blackberries………………………………………….............. 7
2.2.3 Production of blackberries………………………………………............... 9
2.2.4 Physiological ripening of blackberries………………………………….... 9
2.2.5 Chemical composition of blackberries………………………………..….. 10
2.2.5.1 Carbohydrates……………………………………………………....... 11
2.2.5.2 Organic acids……………………………………………………….... 11
2.2.5.3 Phenolic compounds of blackberries………………………………..... 12
2.2.5.4 Enzymes in blackberries…………………………………………........ 15
2.2.5.5 Vitamins and minerals……………………………………................... 16
2.2.6 Health benefit of blackberries…………………………………………….. 16
2.2.7 Quality of fresh blackberries……………………………………............... 17
2.2.7.1 Fruit size………………………………………………………........... 17
2.2.7.2 Color……………………………………………………………......... 18
2.2.7.3 Firmness………………………………………………………............ 19
2.2.7.4 Fruit decay………………………………………………………........ 19
2.2.7.5 Weight loss………………………………………………................... 20
2.2.7.6 Total soluble solids, Brix, and acidity……………………………...... 20
2.2.7.7 Nutritional values……………………………………………….......... 21
2.2.8 Pre-harvest factors affecting quality of fresh blackberries………….......... 21
2.2.8.1 Genotype and cultivars……………………………………………...... 21
2.2.8.2 Maturity……………………………………………………………..... 22
2.2.8.3 Climate, location, season and year…………………………................ 23
2.2.8.4 Cultivation technique……………………………………………….... 24
2.2.8.5 Pests and diseases……………………………………………….......... 24
2.3 Post-harvest handling technologies………………………………………....... 25
2.3.1 Introduction………………………………………………………………. 25
2.3.2 Refrigeration storage…………………………………………………....... 25
2.3.3 Controlled atmosphere storage (CAS) and modified atmosphere
package (MAP)………………………………………………………................. 26
TABLE OF CONTENTS (Continued)
Page
2.3.4 Edible coatings…………………………………………………............... 27

2.3.5 Other possible technologies……………………………………................ 28
2.4 Processing of blackberries…………………………………………................ 29
2.4.1 Introduction……………………………………………………............... 29
2.4.2 Freezing………………………………………………………………….. 30
2.4.3 Canning…………………………………………………………………... 31
2.4.4 Dehydration………………………………………………………….…... 33
2.4.5 Jam and jelly processing………………………………………................. 34
2.4.6 Juice and concentrate…………………………………………………...... 35
2.4.7 Berry wine……………………………………………………………....... 37
2.4.8 Other application of blackberries……………………………………….... 37
2.5 Conclusion………………………………………………………………..….. 38
2.6 Literature cited................................................................................................. 39
3. CHAPTER 3 Effects of Refrigeration Storage and Processing Technologies

on the Bioactive Compounds and Antioxidant Capacities of Blackberries
(‘Marion’ and ‘Evergreen’)…………………………………………………...……. 51
ABSTRACT……………………………………………………………………... 52
INTRODUCTION………………………………………………………………. 55
MATERIALS AND METHODS………………………………………………... 57
RESULTS AND DISCUSSION……………………………………………….... 65
ACKNOWLEDGEMENTS…………………………………………………...… 99
LITERATURE CITED………………………………………………………….. 100
4. CHAPTER 4 General Conclusion……………………………………................. 105
BIBLIOGRAPHY……………………………………………………………….... 108
TABLE OF CONTENTS (Continued)
Page
APPENDIX…………………………………………………………………..… 122
Effect of Edible Coatings on the Quality of Fresh Blueberries (Duke and Elliott)
under Commercial Storage Conditions…………………………........................ 123
LIST OF FIGURES
Figure Page
2.1 Diagrammatic scheme of juice processing…………………………………… 36
3.1 Flow diagram of experimental design………………………………............... 58
3.2 Changes of selected physicochemical properties of two blackberry

varieties (‘Marion’ and ‘Evergreen’) during refrigeration storage
(2.0 ± 0.2 oC, 95 ± 2% RH)………….... ……………………………………...…. 68
3.3 Changes of bioactive compounds in two blackberry varieties

(‘Marion’ and ‘Evergreen’) during refrigeration storage
(2.0 ± 0.2 oC, 95 ± 2% RH)…………………………………...... ………........…... 71
3.4 Changes of antioxidant capacities in two blackberry varieties

(‘Marion’ and ‘Evergreen’) during refrigeration storage
(2.0 ± 0.2 oC, 95 ± 2% RH)…………… ……………………..............…………... 72
3.5 Physicochemical properties of two blackberry varieties (‘Marion’

and ‘Evergreen’) after processing and during room temperature storage
(25 ± 2 oC, 30 ± 2% RH) ….…..........................................................................….. 76
3.6 Bioactive compounds of two blackberry varieties (‘Marion’ and

‘Evergreen’) after processing and during room temperature storage
(25 ± 2 oC, 30 ± 2% RH) ……...................................................................….……. 80
3.7 Antioxidant capacities of two blackberry varieties (‘Marion’ and

‘Evergreen’) after processing and during room temperature storage
(25 ± 2 oC, 30 ± 2% RH) ….…….......................................................................…. 82
LIST OF TABLES
Table Page
2.1 Types and cultivars of blackberries…………………………………………… 9
2.2 Carbohydrate composition of fresh blackberry fruit (100g)………...…...….... 11
2.3 Vitamin composition of fresh blackberry fruit (100g)……………......………. 16
2.4 Mineral composition of fresh blackberry fruit (100g)………………..……….. 16
2.5 Average fruit weight of fresh blackberries……………………...…………….. 18
2.6 Range of colorimetric parameters of thornless blackberries………………….. 18
3.1 ANOVA results of two blackberry varieties (‘Marion’ and ‘Evergreen’)

during refrigeration storage (2.0 ± 0.2 oC, 95 ± 2% RH)……………………...….. 66
3.2 ANOVA results of two blackberry varieties (‘Marion’ and ‘Evergreen’)

after processing and during room temperature storage (25 ± 2 oC, 30 ± 2% RH)... 74
3.3 Physicochemical properties and contents of bioactive compounds in two

blackberry varieties (‘Marion’ and ‘Evergreen’) at harvest…………………..…. 87
3.4 Mean comparison of physicochemical parameters, bioactive compounds

and antioxidant capacities two blackberry varieties (‘Marion’ and ‘Evergreen’)
throughout refrigeration storage (2.0 ± 0.2 oC, 95 ± 2% RH) …...…………..….. 92
3.5 Mean comparison of physicochemical parameters, bioactive compounds

and antioxidant capacities of processed blackberry (‘Marion’ and ‘Evergreen’)
during room temperature storage (25 ± 2 oC, 30 ± 2% RH) …………………..... 94
3.6 Correlation coefficients for physicochemical parameters and bioactive

compounds measured in two blackberry varieties ('Marion’ and ‘Evergreen’)
during refrigeration storage (2.0 ± 0.2 oC, 95 ± 2% RH) ……………………..... 96
3.7 Correlation coefficients for physicochemical parameters and bioactive

compounds measured in processed blackberry (‘Marion’ and ‘Evergreen’)
during room temperature storage (25 ± 2 oC, 30 ± 2% RH) ……………….…... 97
LIST OF APPENDIX FIGURES
Figure Page
A.1 Physicochemical properties of ‘Duke’ fruit during room temperature

storage……………………………………………………………………………. 138
A.2 Physicochemical properties of ‘Elliott’ fruit during room temperature

storage……………………………………………………………………………. 141
A.3 Antioxidant content and total phenolic content of ‘Duke’ fruit during
room temperature storage………………………………………………………... 147
A.4 Antioxidant content and total phenolic content of ‘Elliott’ fruit during
room temperature storage………………………………………………………… 149
LIST OF APPENDIX TABLES
Table Page
A.1 ANOVA results of ‘Duke’ and ‘Elliott’ fruit during cold storage……………. 133
A.2 ANOVA results of ‘Duke’ and ‘Elliott’ fruit during room temperature
storage……………………………………………………………………………... 134
A.3 Overall means of main effects on pH, TA, TSS and FN of ‘Duke’
and ‘Elliott’ fruit during 1-week cold storage…………………………………....... 135
A.4 Physicochemical and nutritional properties of ‘Duke’ and ‘Elliott’

fruit during 1-week cold storage……………………………………………….….. 136
Compounds and Antioxidant Capacities of Blackberries
(‘Marion’ and ‘Evergreen’)
CHAPTER 1
Introduction
Berry fruit are pronounced in vitamins, minerals, folic acid, dietary fiber and
phytochemicals, particularly phenolic compounds including flavonoids, tannins and
phenolic acids. Epidemiological evidences have promoted the consumption of berry
fruit in regular diet against chronic diseases via bioavailable phytochemicals
participating in chemical reactions and biological metabolism in human body (1-5).
As one of the most popular berry crops, blackberry (Rubus spp.) is high in
anthocyanins, ellagic acid, and antioxidant capacity with variation of phytochemical
concentrations by genotype, climate, harvest season, location and cultivation conditions
(6-9). Thorny ‘Marion’ (Rubus ursinus) and thornless ‘Evergreen’ (Rubus laciniatus)
are the two widespread cultivars of trailing blackberries grown in Oregon and
dominantly utilized for processing (10, 11). Recently, more interests have arisen in
analyzing the composition of polyphenolics, anthocyanins and ellagintannins in fresh
blackberries by High Performance Liquid Chromatography (HPLC) (12-14).
Fresh blackberries are easily bruised by rough handling after harvest because of
their soft tissue. In addition, blackberries are extremely perishable with only 2-3 days of
2
shelf life at -0.6-0°C and 90-95% relative humidity (15). Shrinkage, bleeding, changes
of physicochemical and nutritional properties, and mold growth commonly occur during
post-harvest refrigeration storage as a result of moisture loss, mechanical injury, fruit
respiration, and microbiological contamination (16-20).
Accordingly, multiple technologies have been employed to process fresh
blackberries into various value-added products for long-term preservation. For instance,
freeze processing aims to prolong the shelf-life of blackberries by inactivating bacteria
and enzyme activity and suppressing respiration of fruit at low temperature (21).
Canning and jam processing thermally destroy pathogenic and spoilage
microorganisms, inactivate enzymes, and eventually eliminate oxygen in the final
products (22). Drying removes water from fresh fruit and minimizes water activity
essential for microorganism growth (23). Besides extension of the shelf- life,
applications of post-harvest processing technologies also diversify the types of
blackberry products. For example, juices blended with blackberries and other fruit offer
delicious and nutritional liquid products to the market (24). Moreover, blackberry
incorporated candies, cereals, yogurt, ice creams, smoothies, and pies have become
more popular.
Therefore, it is worthwhile investigating the impacts of post-harvest storage and
processing on the significant bioactive compounds and antioxidant capacities of
blackberries, thus establishing the scientific references for consumers when making
purchase decision and for manufacturers to advance the technologies satisfying both
safety and nutrition requirements. Accordingly, the objective of this study was to
3
evaluate the effects of post-harvest refrigeration storage, processing technologies
including freezing, hot-air drying, freeze drying, canning-in-syrup and canning-in-
water, and jam manufacturing, as well as post-processing storage in room conditions on
the basic physicochemical properties, important bioactive compounds, and antioxidant
capacities of ‘Marion’ and ‘Evergreen’ blackberries.
Literature cited
(1) Beattie, J.; Crozier, A.; Duthie, G.G. Potential health benefits of berries. Curr. Nutr.
Food Sci. 2005, 1, 71-86.
(2) Shukitt-Hale, B.; Lau, F.C.; Joseph, J.A. Berry fruit supplementation and the aging
brain. J. Agric. Food Chem. 2008, 56, 636-641.
(3) Tsuda, T. Regulation of adipocyte function by anthocyanins; possibility of

preventing the metabolic syndrome. J. Agric. Food Chem. 2008, 56, 642-646.
(4) Prior, R.L.; Wu, X.L.; Gu, L.W.; Hager, T.J.; Hager, A.; Howard, L.R. Whole
berries versus berry anthocyanins: interactions with dietary fat levels in the
C57BL/6J mouse of obesity. J. Agric. Food Chem. 2008, 56, 647-653.
(5) Seeram, N.P. Berry fruits for cancer prevention: current status and future prospects.
J. Agric. Food Chem. 2008, 56, 630-635.
(6) Zadernowski, R.; Naczk, M.; Nesterowicz, J. Phenolic acid profiles in some small
berries. J. Agric. Food Chem. 2005, 53, 2118-2124.
(7) Siriwoharn, T.; Wrolstad, R.E.; Finn, C.E.; Pereira, C.B. Influence of cultivar,
maturity, and sampling on blackberry (Rubus L. Hybrids) anthocyanins,
polyphenolics, and antioxidant properties. J. Agric. Food Chem. 2004, 52, 8021-
8030.
(8) Thomas, R.; Woods, F.M.; Dozier, W.A; Ebel, R.C.; Nesbitt, M.; Wilkins, B.;
Himelrick, D. Cultivar variation in physiochemical and antioxidant activity of
Alabama-grown blackberries. Small Fruits Rev. 2005, 4, 57-71.
4
(9) Reyes-Carmona, J.; Yousef, G.G.; Martinez-Peniche, R.A.; Lila, M.A. Antioxidant
capacity of fruit extracts of blackberry (Rubus sp.) produced in different climatic
regions. J. Food Sci. 2005, 70, S497-S503.
(10) Strik, B.C.; Clark, J.R.; Finn, C.E.; Bañados, M.P. Worldwide blackberry
production. HortTechnology 2007, 17, 205-213.
(11) Finn, C.E.; Strik B.C. Blackberry cultivars for Oregon. Oregon State University
Extension Publications 2008, EC 1617-E, 1-12.
(12) Siriwoharn, T.; Wrolstad, R.E. Polyphenolics composition of Marion and Evergreen
blackberries. J. Food Sci. 2004, 69, FCT233-FCT240.
(13) Fan-Chiang, H.; Wrolstad, R.E. Anthocyanin pigment composition of blackberries.

J. Food Sci. 2005, 70, C198-C202.
(14) Hager, T.J.; Howard, L.R.; Liyanage, R.; Lay, J.O.; Prior, R.L. Ellagitannin
composition of blackberry as determined by HPLC-ESI-MS and MALDI-TOF-MS.
J. Agric. Food Chem. 2008, 56, 661-668.
(15) Salunkhe, D.K.; Desai, B.B. Small fruits-berries. In Postharvest Biotechnology of

Fruits; Salunkhe, D.K.; Desai, B.B., Eds.; CRC press: Boca Raton, FL, 1984; Vol. 1
pp 111-112.
(16) Bower, C. Postharvest handling, storage, and treatment of fresh market berries. In
Berry fruit: value-added products for health promotion; Zhao, Y., Eds.; CRC press:
New York, 2007; Chapter 9, pp 262-284.
(17) Perkins-Veazie, P.; Collins, J.K.; Clark, J.R. Shelf-life and quality of ‘Navaho’ and
‘Shawnee’ blackberry fruit stored under retail storage conditions. Journal of food
quality 1999, 22, 535-544.
(18) Perkins-Veazie, P.; Kalt, W. Postharvest storage of blackberry fruit does not
increase antioxidant levels. Acta Horticulture 2002, 585, 521-524.
(19) Antunes, L.E.C.; Duarte, J.; De Souza, C.M. Postharvest conservation of blackberry
fruits. Pesquisa Agropecuaria Brasileira 2003, 38, 413-419.
(20) Kalt, W.; Forney, C.F.; Martin, A.; Prior, R.L. Antioxidant capacity, vitamin C,
phenolics, and anthocyanins after fresh storage of small fruits. J. Agric. Food Chem.
1999, 47, 4638-4644.
5
(21) Hui, Y.H.; Lim, M.; Nip, W.; Smith, J.S.; Yu, P.H.F. Principles of food processing.
In Food processing: principles and applications; Smith, J.S.; Hui, Y.H., Eds.;
Blackwell: Ames, Iowa, 2004; Chapter 1, pp17-19.
(22) Körmendy, I. Fruit processing: principles of heat treatment. In Handbook of fruits

and fruit processing; Hui, Y.H.; Barta, J.; Cano, M.P.; Gesek, T.; Sidhu, J.S.; Sinha,
N.K., Eds.; Blackwell: Ames, Iowa, 2006; Chapter 3, pp 45-57.
(23) Barta, J. Fruit drying principles. In Handbook of fruits and fruit processing; Hui,
Y.H.; Barta, J.; Cano, M.P.; Gesek, T.; Sidhu, J.S.; Sinha, N.K., Eds.; Blackwell:
Ames, Iowa, 2006; Chapter 5, pp 81-94.
(24) Rutledge, P. Production of non-fermented fruit products. In Fruit processing;

Arthey, D.; Ashurst, P.R., Eds.; Blackie Academic & Professional: London, U.K.,
1996; Chapter 4, pp 70-93.
6
CHAPTER 2
Literature Review
2.1 Introduction
With benefits from climatic and soil conditions suitable for the growth of blackberry
plants, Pacific Northwest especially Oregon in the United States is one of the largest
production regions and leading suppliers of blackberry fruit in the world (1).
Blackberries are well-known as an excellent nutritional resouce for a healthy diet in
humans and have been nourishing people as a type of small fruit for centuries (2, 3).
This chapter will introduce the history, botanical classification, cultivars, production
and physiological changes during ripening of blackberries, and summarize the chemical
composition, health benefits and quality characteristics of fresh blackberries. Pre-
harvest practices related to the quality of blackberries at harvest and post-harvest
processing technologies applied for preserving fresh blackberries are covered as well.
7
2.2 Overview of blackberries
2.2.1 History and botanical classification of blackberries
Blackberries are in Plantae Kingdom, Magnoliophyta Division, Magnoliopsida
Class, Rosaceae Family, and Rubus Genus. In the United States, species
R.allegheniensis, R. argutus, R. cuneifolius, and R. Canadensis grow in north while R.
trivialis is southeastern species. The Pacific Northwest region of the US has native
specise R. ursinus and non-native species R. lacinatus introduced from Europe in 1860
(4).
Blackberry plants with trailing, erect or semi-erect canes are generally cultivated in
either sandy or clay soil with pH from 5 to 7. They can be propagated by cutting root,
suckering, layering tip and cutting leaf stem. Pinky white flowers of blackberries are
generated in the second year of planting. Blackberry (Rubus spp.) fruit are formed on a
delicate cluster of fragile drupelets with attractive color (5).
2.2.2 Cultivars of blackberries
Trailing, erect, and semi-erect are recognized as the three major types of
blackberries grown in the United States classified by cane architecture. Erect
blackberries with firm texture but weak flavor were mainly bred in Arkansas for the
fresh market. The cultivars of erect blackberries harvested from July to August include
‘Cherokee’, ‘Arapaho’, ‘Natchez’, ‘Shawnee’, ‘Kiowa’, ‘Chickasaw’, ‘Quachita’,
‘Apache’, and ‘Navaho’ (Table 2.1). Semi-erect blackberries with high yield were
mainly breed in Maryland and Illinois for the fresh market as well. Their cultivars
8
harvested from July to October include ‘Loch Tay’, ‘Loch Ness’, ‘Hull Thornless’,
‘Triple Crown’, ‘Doyle’s’, ‘Black Satin’, ‘Smoothstem’, ‘Dirksen Thornless’,
‘Thornfree’, and ‘Chester Thornless’ (Table 2.1). Trailing blackberries predominately
developed in Oregon for both fresh market and processing have two species, native
Rubus ursinus and Europe origin Rubus laciniatus. Many cultivars belong to trailing
blackberry such as ‘Obsidian’, ‘Metolius’, ‘Silvan’, ‘Siskiyou’, ‘Black Butte’,
‘Cascade’, ‘Pacific’, ‘Santiam’, ‘Olallie’, ‘Black Diamond’, ‘Kotala’, ‘Marion’, ‘Black
Pearl’, ‘Wild Treasure’, ‘Nightfall’, ‘Waldo’ and ‘Evergreen’ (Table 2.1).
‘Marion’ blackberries with dark red color, medium size, medium seeds and
excellent flavor grown in very vigorous and thorny plant and ‘Evergreen’ blackberries
with dark black color, small size, large seeds and good flavor grown in vigorous and
thornless plant stand for the native Rubus ursinus species and the imported Rubus
laciniatus species of trailing blackberries, respectively. Both ‘Marion’ and ‘Evergreen’
are in high yielding and favorable to be commercially processed into different
blackberry products for year-round availability (6).

9
Table 2.1 Types and cultivars of blackberries

Blackberries
Types Erect blackberries Semi-erect blackberries Trailing blackberries
Cherokee Loch Tay Marion
Arapaho Loch Ness Evergreen
Natchez Hull Thornless Black diamond
Shawnee Triple Crown Waldo
Kiowa Doyle’s Black Pearl
Cultivars
Chickasaw Black Satin Obsidian
Quachita Smoothstem Metolius
Apache Dirksen Thornless Siskiyou
Navaho Thornfree Silvan
Chester Thornless Nightfall
Source: Adapted from Finn et al. (6).
2.2.3 Production of blackberries
The planted area of blackberries increased 45% in ten years from 1995, reaching
49,508 acres with 154,643 tons of worldwide production in 2005. Mainly through
alternative year production system and machine harvest system, Oregon supplied
25,185 tons of blackberries in 7,754 acres in 2005, which topped in cultivation area and
total production of blackberries in the United States and accounted for approximate
16% of worldwide production of blackberries. ‘Marion’ and Thornless ‘Evergreen’
were reported as two important cultivars composing of 61% and 11% of blackberries in
Oregon, respectively (7).
2.2.4 Physiological ripening of blackberries
Fruit ripening refers to the process of fruit developing to edible food. There are a
series of physical and chemical changes involved in fruit ripening. Non-climacteric
pattern of ripening is recognized in blackberries (8).

10
First of all, softening occurs in blackberries as the maturity increases, which has
been attributed to the breakdown of cellular substances such as pectin, cellulose,
hemicellulose and other polysacchrides through hydration (9).
Secondly, glucose, fructose and sucrose are three main carbohydrates partly from
the hydrolysis of starch within fruit. These sugars accumulate during fruit ripening,
resulting in increased soluble solids content or brix (10). In contrast, organic acids are
consumed as one of the substrates in respiration course of fruit and decomposed under
light and ambient temperature.
Thirdly, chlorophyll degrades concurrently with the synthesis of anthocyanins,
representing the color changes of blackberries from green, red, to black as maturity
progresses (11).
Fourthly, polymerization leads to a reduction in phenolics, flavor compounds
formed and antioxidant capacity enhanced as detected in blackberries from previous
studies (12, 13). Finally, the production of ethylene, a traditional physiological index in
fruit ripening, is not consistent in all the blackberry cultivars (14).
2.2.5 Chemical composition of blackberries
Nutrition fact of fresh blackberries (Rubus spp.) used as reference is updated and
accessible in national nutrient database released by USDA (15). But chemical
compositions of blackberries are subjected to variability in cultivars, harvest seasons,
locations, maturity stage and other pre-harvest factors. The principal chemical
compositions in fresh blackberries are elaborated as follows.

11
2.2.5.1 Carbohydrates
Following water, carbohydrates are the second most abundant components in fresh
blackberries. They are the essential element which provides consumers with the
fundamental sensory attribute of sweetness in most fruit. Fruit ripening actually is the
process of accumulation of sugars and reduction of acids. The contents of subgroup of
carbohydrates in 100g of fresh blackbeerries are listed in Table 2.2 (15), indicating
glucose and fructose are the two major sugars in ripe blackberries in general.
Table 2.2 Carbohydrate composition of fresh blackberry fruit (100g)

Carbohydrate Sugar Sucrose Glucose Fructose Maltose Galactose
(g) (g) (g) (g) (g) (g) (g)
9.61±0.00 4.88±0.69 0.07±0.00 2.31±0.30 2.40±0.37 0.07±0.00 0.03±0.03
Values are means ± standard deviation (n=2-5).
Source: Adapted from USDA national nutrient database for standard reference (15).
2.2.5.2 Organic acids
Similar to carbohydrates, organic acids are another indicator to evaluate the basic
taste of sourness in most fruit. Moreover, it is a factor affecting the stabilization of
pigments in blackberries. According to the study by Kafkas et al. in investigating five
cultivars of blackberries, ‘Navaho’, ‘Chester Thornless’, ‘Jumbo’, ‘Bursa 2’, and ‘Loch
Ness’ grown in Turkey, malic acid accounts for 0.6% to 1.1% on weight, thus
statistically a major organic acid detected in fresh blackberries by HPLC compared to
ascorbic acid and citric acid (16).

12
2.2.5.3 Phenolic compounds of blackberries
As product from plant secondary metabolism, phenolics can be classified into
phenolic acids, flavonoids, hydrolyzable tannins and other phytochemicals such as
lignans, sterols, and stilbenes (17). Phenolics in fruit are biologically synthesized as
plants grow and fruit ripen generally through hydroxylation, methylation, esterification
and glycosylations involving Phenylalanine Ammonialyse (PAL), Cinnamate 4-
Hydroxylase (C4H) and Hydroxycinnamate CoA Ligase (CoAL), which are affected by
environmental conditions such as light, temperature and nutrients in the plants and pre-
harvest practices as well (17). From the standpoint of sensory properties, phenolics
provide consumers with the perception of bitterness, astringency and the color of fruit
(17).
According to Macheix et al., hydroxybenzoic acids (HBA) and hydroxycinnamic
acids (HCA) are the two dominant types of phenolic acids in blackberries (17).
Zadernowski et al. reported that free, ester bonded and glycoside bonded phenolic acids
constitute 3.3%, 53.1% and 43.6% of total phenolic acids respectively in Rubus
plicatus. There are significantly more HCA existing in blackberries than HBA.
Specifically, salicylic is in the highest fraction (524.1 mg/Kg of dry matter of fresh
berries) of hydroxybenzoic acids (HBA) (18). Hydroxycaffeic (627.6 mg/Kg of dry
matter of fresh berries), m-coumaric (596.6 mg/ Kg of dry matter of fresh berries) and
3, 4-dimethoxycinnamic (501.9 mg/Kg of dry matter of fresh berries) topped as
hydroxycinnamic acids (HCA) in phenolic acids profile (18).

13
As the water-soluble pigments belong to the category of flavonoids in fruit and
vegetables, anthocyanins are responsible for the appealing color of blackberries. They
are in glycosidic form of anthocyanidins (17). Depending on the positions and numbers
of hydrogen, hydroxyl, and methoxyl groups in the basic chemical structure, six
anthocyanidins including Pelargonidin (Pg), Cyanidin (Cy), Delphinidin (Dp), Peonidin
(Pn), Petunidin (Pt), and Malvidin (Mv) are naturally occurring and widely recognized
in berry fruit (19). Cho et al. reported that cyanidin 3-glucoside, cyanidin 3-rutinoside,
cyanidin 3-xyloside, cyanidin 3-malonylglucoside, and cyanidin 3-dioxalylglucoside are
the five common anthocyanins present in blackberries. Among them, cyanidin 3-
glucoside was identified as primary composition (20). Wu et al. indicated that
nonacylated anthocyanins contributed 94% and 97% to the total level of anthocyanins in
the overall of four blackberry varieties studied and ‘Marion’ blackberry, respectively,
and 90% and 76% of anthocyanins in overall blackberries and ‘Marion’ blackberry,
respectively are monoglycosides (21).
Total anthocyanin contents in five genotypes of blackberries harvested in Oregon in
2000 were reported as 164 mg/100g fresh weight (2). Significantly higher contents of
anthocyanins (245 mg/100g fresh weight) were identified in blackberries without
specific information about harvest time and location in another study (21). Results in
these two studies were both roughly in the range of 114.39 to 241.54 mg/100g of fresh
blackberries with six cultivars, ‘Apache’, ‘APF-12’, ‘Arapaho’, ‘Chickasaw’, ‘Kiowa’,
and ‘Navaho’ (20). Significantly lower contents of anthocyanins (88.7 mg/100g fresh
weight) were in ‘Black Diamond’, ‘Smoothstem’, ‘Thornless Boy Sembes’, ‘Darrow’,

14
‘Chester’, ‘Hull Thornless’, and ‘Black Satin’ blackberries (22). These numbers also
fell in the range of anthocyanin content from 70.3 to 201 mg/100g fresh weight of 18
varieties of blackberries harvested in 51 locations (23).
Flavonol is considered to be another largest group of flavonoids mainly in the flesh
of blackberries. There were 102 to 160 mg of flavonol composed of quercetin 3-
galactoside, quercetin 3-glucoside, quercetin 3-rutinoside, quercetin 3-xyloside,
quercetin 3-xylosylglucuronide, and quercetin 3-glucosylxyloside in every Kg of fresh
weight of ‘Apache’, ‘APF-12’, ‘Arapaho’, ‘Chickasaw’, ‘Kiowa’, and ‘Navaho’
blackberries (20). ‘Evergreen’ blackberries showed a significantly higher content of
flavonol than that of ‘Marion’ blackberries harvested in Oregon in 1999 (24).
Tannins are polyphenolics polymerized with proteins, carbohydrates and other
molecules, providing astringency in fruit and vegetables (25). They are traditionally
divided into hydrolysable tannins and condensed tannins based on structural variations.
Hydrolysable tannins are created by esterification of hydroxyl groups in carbohydrates
usually glucoside and carboxyl groups in phenolic acids such as gallic acid,
hexahydroxydiphenic acid, and ellagic acid (17).
Like other berries, blackberries are abundant in hydrolysable tannins (19). More
studies have focused on the analysis of ellagitannins in blackberries. It has been
confirmed that ellagitannins are mostly distributed in seeds other than flesh (24, 26). Six
ellagitannins including pedunculagin, casuarictin/potentillin, castalagin/ vescalagin,
lambertianin A/sanguiin H-6, lambertianin C, and labertianin D have been identified by
virtue of HPLC-ESI-MS (High Performance Liquid Chromatography-ElectroSpray

15
Ionization-Mass Spectrometry) instrument (27). Besides ellagitannins, free, acylated
and glycosylated ellagic acids are also available in blackberry fruit. Ellagic acid
derivatives were significantly higher in the whole ‘Evergreen’ blackberries (3.62
mg/100g FW) than in ‘Marion’ blackberries (1.64 mg/100g FW) (24).
Condensed tannins or proanthocyanidins are another division of tannins rich in the
kingdom of plants. Structurally, they are complex of flavonoids linked through carbon-
carbon bonds which are not hydrolysable (28). Contents of proathocyanidins composed
of catechin and epicatechin were only detected in blackberry seeds ranging from 48.8 to
58.5 and 9.26 to 11.3 mg/100g FW, respectively (24).
In addition, blackberries contain secoisolariciresinol, which is a kind of lignans
resulted from the interactions between two cinnamic acids and approved to have
anticancer effect (29).
2.2.5.4 Enzymes in blackberries
Enzymes play an important role in boosting chemical reactions which consequently
result in quality deterioration of berry fruit in color, texture, flavor, and nutritional value
during ripening, post-harvest handling, and storage. Peroxidase and polyphenol oxidase
actively participate in enzymatic browning by catalyzing the oxidation of phenols to
quinones which further interact with proteins to form complex with large molecular
weight (30, 31), resulting in the consumption of phenolics and the brown color on fruit.
Tissue softening partly caused by the decomposition of cell wall materials involves
enzymes as well, such as cellulase and hemicellulase. Polygalacturonase and pectate

16
lyase break down pectin, while glucanase, xyloglucan, transglycosidase, xylanase, and
β-xylosidase decompose hemicellulose (32).
2.2.5.5 Vitamins and minerals
Blackberries are excellent source of numerous essential vitamins including vitamin
C, vitamin B groups, vitamin A, vitamin K, and folate, and minerals including
potassium, magnesium, calcium, phosphorus, and iron. The distribution of nutrients in
100g of fresh blackberries released by USDA is summarized in Table 2.3 and 2.4.
Table 2.3 Vitamin composition of fresh blackberry fruit (100g)

β
Vitamin A Vitamin Vitamin E Vitamin K Folate Choline
Carotene
(IU) C (mg) (mg) (mcg) (mcg) (mg)
(mcg)
214±39.42 21±0.00 1.17± 0.05 19.8±2.08 25±2.01 8.5±0.00 128±23.65
Table 2.4 Mineral composition of fresh blackberry fruit (100g)

Ca Fe Mg P K Na Zn Cu Se Mn
(mg) (mg) (mg) (mg) (mg) (mg) (mg) (mg) (mcg) (mg)
29 0.62 20 22 162 1.0 0.53 0.165 0.4 0.646
±6.74 ±0.08 ±1.26 ±0.79 ±0.00 ±0.00 ±0.04 ±0.03 ±0.00 ±0.23
2.2.6 Health benefit of blackberries
Several potential health benefits associated with the intake of phytochemicals in
blackberries have been confirmed by many scientific evidences. For example,
polyphenolics demonstrated an anti-cancer effect through quenching inflammatory and

17
inhibition of carcinogenic proliferation (33). It was also found that phytochemicals
reduced the risk of coronary heart disease in many pathways, such as intervening the
LDL oxidation and biosynthesis of cholesterol (34). Furthermore, Barbara et al.
declared that polyphenolics relieved the stress in nervous system against brain aging
(35). Finally, two recent biological studies claimed that anthocyanins were effective in
the prevention of obesity (36, 37).
2.2.7 Quality of fresh blackberries
The objective of cultivation practice, pre-harvest, and post-harvest handling is to
provide high quality of blackberries in fresh market and for subsequent processing. The
quality of blackberries is generally evaluated by the following quality characteristics.
2.2.7.1 Fruit size
Fruit size has direct effect on the marketability and acceptance of blackberries in
both fresh market and processing plant. Traditionally, large fruits are preferred by
consumers (38). Moyer et al. reported that a 100 g of blackberries contained 27 to 108
fruits based on an investigation on five genotypes (2). Eyduran et al. recorded that fresh
blackberries weighed 2.01 g (cv. ‘Black Satin’) to 9.25 g (cv. ‘Black butte’) per fruit
(Table 2.5) (39).

18
Table 2.5 Average fruit weight of fresh blackberries

Erect blackberries
Cultivars Cherokee Navaho Arapaho
Fruit weight
2.30 3.03 3.09
(g/berry)
Semi-erect blackberries
Cultivars Black Satin Ness Chester Thornless
Fruit weight
2.01 2.82 5.19
(g/berry)
Trailing blackberries
Cultivars Marion Nightfall Black Butte
Fruit weight
5.10 6.20 9.25
(g/berry)
Source: Adapted from Eyduran et al. (39).
2.2.7.2 Color
The shiny dark red or black color of blackberries is very attractive to consumers.
Fading of the color and degradation of the pigments are associated with quality
deterioration of blackberry fruit from the consumer standpoint. Colorimeters are widely
used to measure the surface color of fruit by using the Hunter L*a*b* system (40, 12,
41). Color parameters such as L*, a*, and b* stand for lightness, red to green, and
yellow to blue, respectively. L* values of four dull-black thorny erect blackberries were
20.4 to 25.3 (40). L*, a*, and b* values of nine wild Turkey blackberries harvest in
2005 were 17.35, 10.41 and 3.62, respectively (12). The complete colorimetric profile
of nine thornless blackberries (‘Bursa-1’, ‘Bursa-2’, ‘Bursa-3’, ‘C. Thornless’, ‘Bartin’,
‘Loch Ness’, ‘Navaho’, ‘Jumbo’, and ‘D. Thornless’) is summarized in Table 2.6 (41).
Table 2.6 Range of colorimetric parameters of thornless blackberries

Colorimetric measurements L* a* b* Chroma Hue Angle
Data Range 10.5-14.5 3.7-7.4 1.3-3.4 4.0-8.1 16.7-24.8
Source: Adapted from Turemis et al. (41).
19
2.2.7.3 Firmness
Fruit firmness is considered to be one of the critical texture properties because
blackberries are very susceptible and fruit softening occurrs as a result of degradation of
pectin existed in the cell wall of fruit (42). Firmness of blackberries could be measured
by shear, extrusion, compression, and penetration force using a Texture Analyzer (43,
11). The means of shear, extrusion, compression and penetration force applied on
frozen and thawed ‘Thornfree’ blackberries harvested in Spain at the commercial
maturity stage were reported as 16.02N, 20.11N, 20.63N and 8.85N, respectively (43).
For fresh ‘Navaho’ harvested in the US in 1996, the compression and penetration force
decreased during ripening approximately from 42 N to 2 N and from 0.9 N to 0.2 N,
respectively (11).
2.2.7.4 Fruit decay
Spoilage microorganisms mostly fungi grow and propagate rapidly by sourcing
nutrients from fruit tissues under warm weather and refrigeration storage condition with
high humidity, which consequently causes fruit decay (44). The physical damages of
fruit skin during and after harvest facilitate the overspread of microbial infection, thus
accelerating the decay of blackberries (44). Specifically, blackberries are prone to grow
grey molds (Botrytis cinerea) in the high moisture environment (45, 46). The
occurrence of fruit decay is detrimental for marketing fresh blackberries. Hence decay
rate is a necessary criterion to assess the quality of the fruit. The percentage of decay
depends on the cultivar and maturity stage, ranging from 0 to 40.6% after 7-d
20
refrigeration storage in ‘Choctaw’, ‘Cheyenne’, ‘Navaho’ and ‘Shawnee’ blackberries
(47).
2.2.7.5 Weight loss
Weight loss is a basic parameter to evaluate quality of blackberries during post-
harvest storage (48, 49). Loss of moisture is the major reason for weight loss of berries
as a result of post-harvest respiration and transmission (50). Besides, evaporation of
aroma compounds also partly contributes to weight loss in the fruit. A 0.8% to 3.3% of
weight loss was found in four blackberry cultivars (‘Choctaw’, ‘Cheyenne’, ‘Navaho’
and ‘Shawnee’) with three ripen stages (mottled, shiny black and dull black) during 7-d
refrigeration storage (47).
2.2.7.6 Total soluble solids, Brix, and acidity
Total soluble solids (TSS) reflect seeds and sugar content in the blackberries. The
seediness affects the taste and chewability of the fruit and the processing of fresh
blackberries (38).
Brix degree is defined as the percentage (w/w) of sugar (sucrose) present in food
samples. Fructose, glucose and sucrose are the three primary types of sugar and malic
acid is the dominant organic acid in blackberries (51).
Sugar content gradually elevates but the acidity drops as fruit ripen. The ratio of
sugar and acidity varies in different varieties of blackberries at harvest and is a good
indicator of balance between sweetness and sourness of fruit. Moreover, the sugar
21
content of fruit determines the amount of sugar addition to make shelf-stable blackberry
products, such as jams, jellies, and juices (38). The stability of some bioactive
compounds is heavily influenced by pH and acidity of the fruit.
Reyes-Carmona et al. reported a range of 2.33 (wild) to 4.28 (‘Evergreen’) for pH,
7.5% (wild) to 16.1% (‘Evergreen’) for TSS, and 1.02% (‘Evergreen’) to 4.22% (wild)
citric acid equivalent for titratable acidity (TA) in 11 cultivars of blackberries harvested
in different locations (52). Perkins-Veazie et al. also found TSS as low as 4.7% in
mottled ‘Cheyenne’ and TA 0.8% in dull black ‘Navaho’ and TA 0.84% in ‘Chester’ at
harvest (47).
2.2.7.7 Nutritional values
Blackberries are excellent sources of dietary fibers, vitamins and minerals (Table
2.3 and 2.4). Blackberries are well-known for their high content of antioxidants, total
phenolics, anthocyanins, ellagic acids, and other phytochemicals (13, 24). The exact
composition of nutraceuticals in blackberries is highly dependent on numerous pre-
harvest and post-harvest factors (1, 48).
2.2.8 Pre-harvest factors affecting quality of fresh blackberries
2.2.8.1 Genotype and cultivars
Perkins-Veazie et al. found that erect blackberry cultivars of ‘Arapaho’ and
‘Navaho’ had higher ratio of TSS and acidity, firmer texture at 5 °C or 10 °C, and
longer shelf-life (7-10d) at 2 °C than ‘Choctaw’ and ‘Shawnee’ blackberries (49).

22
Connor et al. reported that total phenolic and total anthocyanin content and
antioxidant capacity evaluated by FRAP significantly differed in 12 shiny blackberry
cultivars (‘Shawnee’, ‘Tayberry’, ‘Hull Thornless’, ‘Logan’, ‘Black Butte’, ‘Ranui’,
‘Navaho’, ‘Silvan’, ‘Siskiyou’, ‘Kotata’, ‘Marion’, and ‘Boysen’) (53, 54). Among
them, ‘Boysen’ contained the highest total phenolics, ‘Marion’ showed the highest
antioxidant capacity, and ‘Kotata’ possessed the highest total anthocyanins.
Siriwoharn et al. found that physicochemical parameters including TSS, TA and
Brix/TA, total phenolics, total anthocyanins, quantitative compositional distribution of
anthocyanins and polyphenolics, and antioxidant capacity evaluated by ORAC and
FRAP were considerably varied among blackberry cultivars (‘Marion’, ‘Waldo’,
‘Evergreen’, ‘Chester’, ‘Silvan’, ‘NZ’, and ‘ORUS’). ‘Evergreen’ demonstrated the
highest antioxidant capacity in both ORAC (75.5 µmol TE/g FW) and FRAP values
(91.5 µmol TE/g FW) (13).
Reyes-Carmona et al. indicated that genotypes, other than climate and region, was a
major factor affecting acidity, soluble solid content, polyphenols, and antioxidant
capacity in ripen ‘Brazos’, ‘Tupi’, ‘Comanche’, and wild blackberries harvested in
Mexico, and ‘Marion’, ‘Evergreen’, and ‘Siskiyou’ harvested in Oregon (52).
2.2.8.2 Maturity
Some studies have addressed the effect of maturity stage on the quality of
blackberry fruit. Perkins-Veazie et al. found that mottled (50% black) erect blackberry
cultivars, ‘Cheyenne’, ‘Navaho’, ‘Choctaw’, and ‘Shawnee’, were firmer, more acidic,
23
and less sweet than shiny black and dull black fruit of same blackberry cultivars. More
anthocyanins were accumulated in dull black blackberry fruit but deteriorated more
rapidly (47). Woods et al. studied thornless blackberries including ‘Apache’, ‘Arapaho’,
‘Chester’, ‘Loch Ness’, ‘Navaho’, and ‘Triple Crown’, and reported that pH increased,
TA declined, TSS/TA ratio increased, and antioxidant capacity increased along with
increased fruit maturity at harvest from red, mottled, shiny black to dull black (55).
Siriwoharn et al. also observed a significant increase of total anthocyanins and TSS
and loss of acids along with the ripening in ‘Marion’ and ‘Evergreen’ blackberries
harvested in Oregon in 2002 (13).
Wang and Lin investigated three cultivars of thornless blackberries, ‘Chester’,
‘Hull’, and ‘Triple Crown’ in Maryland and found that the anthocyanins were
synthesized while the level of total phenolics declined with the development of
blackberry fruit. Interestingly, ORAC values consistently decreased for fruit from green
to pink stage then increased from pink to ripe stage (56).
Tosun et al. confirmed the decrease in total phenolics and increase in anthocyanins
along with maturity in a study on blackberries harvested in Turkey in 2005 (12).
2.2.8.3 Climate, location, season and year
The qualities of berry crops and berry fruit are strongly dependent on macroclimate
such as temperature, light, carbon dioxide, and water, as well as microclimate such as
spacing between plants, pruning, and training during pre-harvest practice (48).
24
Connor et al. reported that antioxidant and total phenolic content of blackberries
harvested from both New Zealand and Oregon turned out significantly different in 12
shiny-black blackberries across the years which may result from the slightly changed
climate (53). Qian and Wang found that volatile acids, alcohols, aldehydes, ketones,
terpenes, terpenoids, and esters varied in ‘Marion’ and ‘Evergreen’ blackberries grown
in 1999, 2001, and 2002 in Oregon (57).
2.2.8.4 Cultivation technique
The optimal amount of water and nutrients including N, P, K.Ca, and Mg influences
crop growth, leaf production, and fruit ripening. The lack of water may result in soft and
less sweet fruit, but the excess of water may promote fruit bruise and decay. Application
of sufficient and balanced nutrients is critical for the quality of berry fruit at harvest
(48).
In addition, the organic and sustainable cultivation practices produced significantly
higher fresh-weight based total phenolics in frozen, freeze-dried, and air-dried ‘Marion’
blackberries harvested in Oregon than conventional ones (58).
2.2.8.5 Pests and diseases
Occurrence of pests and diseases, such as yellow vein disease (YVD), phragmidium
violaceum, and arthropod pests jeopardized the production of blackberry crops and the
safety, quality, and processing properties of blackberry fruit (59-61). On the other side,
25
the residual of herbicides, pesticides, and any other chemicals used against disease and
pest infection of berry crop could cause some safety concerns as well (62).
2.3 Post-harvest handling technologies
2.3.1 Introduction
Post-harvest handling practices basically involve four steps: harvesting, pre-cooling,
packaging, transportation and storage (63). Fresh blackberry fruit are extremely
perishable with a very short shelf-life. Hence, retaining quality and extending shelf-life
of fresh fruit throughout post-harvest handling are essential. A diverse array of storage
conditions, package technologies, and post-harvest handling technologies has been
investigated and introduced to the blackberry industry.
2.3.2 Refrigeration storage
The principle of refrigeration storage is to slow down the metabolism of fruit by
reducing temperature (64). Several previous studies have addressed the effect of
refrigeration storage on the quality of blackberries (65, 66). Cultivars mostly affect the
preservation of quality of fresh blackberries during refrigeration storage. Perkins-Veazie
et al. observed that shiny black erect blackberry cultivars, ‘Navaho’, ‘Cheyenne’, and
‘Shawnee’ were successfully stored in plexiglass boxes under 2 °C and 95% relative
humidity for 7 days without significant changes in firmness, pH, soluble solids, and
titratable acidity, but up to 3.4% of initial weight was lost (65). Antunes et al. found that
26
‘Brazos’ and ‘Comanche’ blackberries had 9 days of shelf-life at 2 °C. Fruit lost mass,
titratable acid and soluble solids but gained pH along with the storage time (66).
Harvest maturity is also important to the performance of fresh blackberries after
harvest. Perkins-Veazie et al. reported that ‘Navaho’ blackberries harvested at 60%
ripeness was marketable after 21 days while ‘Shawnee’ blackberries deteriorated after
14 days of storage at 2 °C and 95% relative humidity in polyethylene clamshell boxes
with paper absorbents. However, decay rate surged in both varieties of blackberries
during refrigeration storage (49).
Again, Perkins-Veazie et al. found that erect blackberries ‘Navaho’ and ‘Shawnee’
deteriorated more rapidly and lost more weight at 5 °C than at 2 °C storage (49, 67). As
for antioxidant capacity, Perkins-Veazie et al. reported no significant changes in ORAC
values of erect blackberries, ‘Navaho’, ‘Kiowa’, ‘Shawnee’, ‘Arapaho’, and ‘Choctaw’
harvested in Oklahoma in 1998 after 7 days of storage at 2 °C and 95% relative
humidity (68).
2.3.3 Controlled atmosphere storage (CAS) and modified atmosphere packaging
(MAP)
CAS is to modify the composition of gas in the environment where fruit are stored
for prolonging the shelf-life of fresh fruit (44). The quality of fruit can be better
preserved in atmosphere composed of a high concentration of carbon dioxide and low
concentration of oxygen, leading to delayed respiration and transpiration of fruit,
controlled growth of aerobic microorganism and insects, and lowered enzymatic

27
activity. However, the high percentage of carbon dioxide may potentially retard the
production of fruit aroma compounds and result in off-flavor (44). CAS generally
enriches carbon dioxide up to 10-20% and reduces oxygen to 5-10% in blackberry (44).
Perkins-Veazie and Collins found that CAS effectively reduce the incidence of decay
and retain titratable acid in both ‘Navaho’ and ‘Arapaho’ blackberries during the
storage at 2 °C (69). However, Agar et al. reported CAS not effective in retaining
ascorbic acid in thornfree blackberries (70).
On the same basis of CAS principles, modified atmosphere packaging (MAP) aims
to change the rate of respiration within the food and transpiration between food and air
outside the package by adjusting the regular composition of the atmosphere inside the
food package. Gas composition, storage temperature, and package material with
different permeabilities need to be carefully selected according to the characteristics of
food, the cultivar or type of raw food materials, maturity of fresh produce, and some
mechanical injuries for the well-signed MAP (71, 72). Few studies investigated the
effectiveness of MAP technology to extend the shelf-life of blackberries, but the grape
berries (73), apples (74), blueberries (75) and cherries (76).
2.3.4 Edible coatings
As a novel “invisible packaging” technology, edible coatings have been developed
with the purpose of extending the shelf-life of fresh fruits and vegetables. Edible
coatings work as gas and water barriers to minimize the respiration rate and moisture
28
loss. Antimicrobial agents may be incorporated into edible coatings to control
microorganism infection in fresh produce (77-79).
Polysaccharides, proteins and lipids may be used for forming coatings applied on
berry fruit. Several studies showed chitosan (1, 4-linked 2-amin -2-deoxy–β–D-glucan)
coated fresh strawberries with improved qualities including reduced weight loss and
decay and elevated firmness (78-82). Meneghel et al. reported that sodium alginate
coating on ‘Comanche’ blackberries provided ready-to-eat fruit with comparable
physical, chemical and sensory properties to fresh ones (83).
Protein-based coatings, such as caseinate and whey protein not only controlled gas
transfer during post-harvest ripening, but also provided additional proteins (84, 85). The
effectiveness of protein based coatings could be advanced by addition of lipids and
combination with other physical technologies such as irradiation (86). Lipids with
moisture resistant properties are traditionally used to retain fruit quality by holding
sufficient water in fruit tissues (85).
Previous studies have shown that kiwifruits, pineapple, and cherries have extended
shelf-life via coating of SemperfreshTM which is a sucrose-fatty acid ester (87-89).
However, to our best knowledge, no published study has evaluated the application of
protein and lipid based coating on fresh blackberries.
2.3.5 Other possible technologies
Exposure to ultraviolet (UV) light gave rise to an increase in antioxidant capacity
and a reduction in strawberry, blueberry and mango decay (90-92), and helped maintain
29
physicochemical properties, antioxidant capacity, and firmness in boysenberry fruit
(93). Gamma ray specifically inhibited mold growth under refrigerated storage (94, 95).
However, the carcinogenic radioactivity perception of consumers and the possible high
price both limited the wide application of irradiated food (96). Refrigeration storage at
2°C with 0.3 ppm ozone treatment destroyed fungi and preserved the color and
anthocyanins in thornless blackberries for 12 days (97).
Treatments of fruit using methyl jasmonate (MJ), melaleuca alternifolia (tea-tree oil
or TTO) and ethanol (EtOH) successfully prevented the decay, improved antioxidant
capacity and radical scavenging activity of ‘Triple Crown’ blackberries (98). Besides,
recent study further reported that methyl jasmonate (MJ) treatment on thornless
‘Chester’, ‘Hull’, and ‘Triple Crown’ blackberries remarkably increased flavonoid
content, antioxidant and anti-cancer capacities (99). Thus volatile application may be a
promising technology to stimulate the increase of nutritional contents and health benefit
of blackberries (99).
2.4 Processing of blackberries
2.4.1 Introduction
Due to their short production season and limited shelf-life, fresh blackberries are
commonly processed into frozen and dried products which are then applied to
confectionary, baking, cereal, dairy, and formulated food industry, or made into canned
fruit, juices, jams and jellies for providing diverse blackberry products in the market.
30
The quality of raw fruit and the specific processing technology directly impact the
quality of the final processed products.
The principles and research achievements of the major processing technologies
applied to fresh blackberries are summarized in the following sections. The principles
of these processing technologies are to destroy microorganisms, reduce water content,
and inhibit enzyme activity, thus resulting in safe, high quality, and longer shelf-life
products. On the other hand, these processing technologies may decrease the nutritional
contents, advance the structural transformation, and diminish biological activities of
some nutrients (100).
2.4.2 Freezing
During the freezing process, temperature and water activity of fresh food drop down
to decelerate chemical, microbiological, and biochemical reactions. The quality of
frozen food is challenged by moisture migration, drip loss, large crystals, flavor
sublimation, and solute precipitation which in turn is determined by freezing rate and
subsequent storage conditions (101).
Freezing rate is influenced by the characteristics of food samples such as size,
surface area, heat conductivity, processing parameters such as temperature and type of
freezing methods, and package material (101). The typical freezing methods used in
berry processing include air-blast freezing, spiral belt freezing, fluidized bed freezing,
and cryogenic freezing (102).

31
Liquid nitrogen application with high freezing rate had least impact on the structure
of frozen wild blackberries compared with static and plate freezing methods (103).
González et al. reported that Spanish wild blackberries harvested at commercial
ripening stage in 1996 showed significant drop in total anthocyanins, total phenolics,
and ellagic acid from 306.26 to 258.84 mg cyanin-3-glu/100 g FW, 977 to 903.61 mg
GAE/100 g FW, 25.93 to 19.34 mg/100 g FW, respectively after cryogenic freezing by
liquid nitrogen at -80°C, while total anthocyanins and phenolics stayed stable against
ellagic acid conversely rallied during the storage of frozen blackberries packed in
polyethylene bags for up to 6 months at -24°C (104). Schmidt et al. also pointed out that
individual quick freezing (IQF) had good retention of antioxidant activity and total
phenolics in North America wild blueberries and cultivated blueberries grown in New
Jersy and Michigan (105). In contrast, Ngo et al. found that air-blast freezing removed
the glossiness on the surface of six genotypes of strawberries, ‘Ovation’, ‘Totem’,
‘Puget Reliance’, ‘2273-1’, ‘1843-1’,and ‘1723-1’ obtained in Oregon in 2005 (106).
Vacuum impregnation of corn syrup, pectin and minerals has shown to be effective
pretreatments prior to freezing for maintaining the firmness and preventing drip loss of
frozen blackberries (107).
2.4.3 Canning
Canning is a thermal process for preserving food by destroying pathogenic and
spoilage microorganisms during processing and preventing their growth during post-
processing storage (108). To perform a successful canning process, numerous factors

32
have to be considered such as pH of the food, heat resistance of microorganisms, types
of container, and storage conditions (108). Canned blackberries are produced by going
through several procedures including filling fruit into metal cans, hot-syrup infusion,
exhausting, sealing, and applying high heat (109).
A very few studies have addressed the quality of canned blackberries although
canned fruit are the traditional shelf-stable products on the market. Hager et al. reported
that 22% and 27% of ORAC and 10.5% and 17.8% of total monomeric anthocyanins
accompanied with darkening of polymeric color were lost in 40° Brix Sweetose syrup
and water canned 2005 shiny black ‘Apache’ blackberries harvested in Arkansas,
respectively. Afterwards, a 6 months post-processing storage at 25 °C led to 60.6% and
65.8% losses in total monomeric anthocyanins, respectively, but no significant changes
in ORAC were observed (110). Consistent results were found in a similar study on fully
ripen ‘Bluecrop’ blueberries harvested in Arkansas in 2005. Brownmiller et al. found
that 46% and 42% of ORAC and 28% and 34% of total monomeric anthocyanins
accompanied with darkening of polymeric color were lost in blueberries canned in 40°
Brix Sweetose syrup and water, respectively right after canning process. The following
room temperature storage didn’t change ORAC of the canned blueberries, but huge
decline in total monomeric anthocyanins (71% and 62% for canned-in-syrup and
canned-in-water blueberries, respectively) was observed (111). Ngo et al. also showed
that 68.8% of total anthocaynins and 23.5% of total phenolics in the whole canned
strawberries in 20° Brix sucrose syrup were lost after canning and 60 days of room
temperature storage (106).

33
However, Chaovanalikit and Wrolstad indicated that in canned Bing cherries with
19° Brix sucrose syrup, no apparent loss of anthocyanins until storage for 5 months was
observed, while total phenolics remarkably increased after processing and during 5
month of storage at both 2°C and 22 °C (112).
2.4.4 Dehydration
Dehydration serves as a food preservation method by releasing bounded water and
removing water from tissues of food. The efficiency of dehydration varies with
processing conditions such as temperature, air-flow velocity, time, dehydration method,
and physical properties of food samples such as shape, dimension, size, surface area,
density, and original moisture content (113). Apparent changes include shrinkage,
bleeding and solute diffusion, and nutritional loss, which are related to the specific
dehydration process. Water and gas resistance, as well as light protection of the package
play important roles in the quality of dehydrated food during storage. Commonly used
dehydration methods are sun drying, hot-air drying, spray drying, vacuum drying, and
freeze drying (114).
Asami et al. reported that freeze drying was comparable to individually quick
freezing under air-blast tunnel at -32°C in respect to holding total phenolic in ‘Marion’
blackberry cultivated by conventional, organic, and sustainable practices and grown in
Oregon, whereas air-drying significantly lowered total phenolic content from 15.6% to
21.1% compared to frozen controls (58). Yurdugul claimed that freeze-dry was also an
34
excellent technology for preserving physical, physicochemical, and nutritional qualities
of dried ‘alpine’ strawberries (115).
Combinations of multiple dehydration technologies such as microwave-vacuum
drying, microwave drying with osmotic dehydration pretreatment, and innovation of
new technologies such as sequential infrared radiation and freeze-drying (SIPFD) have
become the trend in the research field to improve drying efficiency, reduce drying cost,
and assure quality of dried berries (116-118).
2.4.5 Jam and jelly processing
Jam and jelly processes use combination of heat, sugar, and acid to kill
microorganisms, inactivate enzymes, and lower water activity with the gel formation.
Uniform dissolution of ingredients and building up polysaccharides cross-linked
network holding water molecules by interaction between pectin and sugars under acidic
condition are critical for successful manufacture of jams and jellies. A few operation
units such as homogenizing, boiling, filling, and gel-setting are involved in jam and
jelly processing (119).
Amakura et al. showed stable total phenolic content (76.23-76.91 mg/100 g fresh
fruit) and DPPH radical scavenging activity during blackberry jam processing (120).
Häkkinen et al. also found that making strawberry jams exerted a minor effect on
quercetin and kaempferol which are two specific flavonoids (121). Nevertheless, a
study by Ngo et al. concluded that long-time heat treatment during jam manufacture
35
was detrimental to the bioactive compounds, leading to an approximate 70% and 20%
reduction of total anthocyanin and total phenolic content in ‘Totem’ strawberries (106).
2.4.6 Juice and concentrate
Berry juices supply the market as a liquid form of product. The traditional fruit juice
production is charted in Figure2.1 (122). Addition of appropriate enzymes catalyzes
cellular collapse and juice release in berry juice production (123). But endogenous
polyphenol oxidase is responsible for the loss of polyphenolics throughout all the steps
of juice processing (124).
Hager et al. reported that an approximate 67% loss of total monomeric
anthocyanins, 55% loss of ORAC value as well as a small increase in polymeric color
occurred in juice processing of shiny black ‘Apache’ blackberries (110). And over the 6
months of room temperature storage, ORAC value of blackberry juice showed no
significant changes while total monomeric anthocyanins fell by 69-75% (110).
Brownmiller et al. reported consistent results were demonstrated in a similar study
on ‘Bluecrop’ blueberries, in which more total monomeric anthocyanins were removed
and less polymeric color were formed in blueberry juice clarified by centrifuge
compared to non-clarified juice (111).
Skrede et al. revealed that most anthocyanins in frozen ‘Bluecrop’ blueberries were
not completely extracted into juice but were enriched in its press-cake. Because of the
heat treatment involved in production and enzymatic degradation in juice portion, the
leftover anthocyanins were limited in the final juice product (124).

36
Fruits reception Grading, washing
Juice extraction Extractor, squeezer
Clarification Centrifuge,
filtration
Blending
Pasteurization Evaporation Essence
Frozen storage Concentrate

Aseptic storage blending
Not-from-concentrate Cool down

Juice
Frozen storage
Concentrate
Figure 2.1 Diagrammatic scheme of juice processing (122).

37
2.4.7 Berry wine
Blackberries have been widely used for fruit wine manufacturing. In the production
of blackberry wine, juice is extracted from washed fresh or frozen fruit by milling,
crushing, maceration, or pressing with the aid of enzyme and heating treatments,
followed by the fermentation of juice with yeast, sugar and acid at 20-25 °C, maturation
at 7-15 °C, pasteurization or hot-filling (125). Addition of urea and SO2 during
fermentation is conducted for elevating the fermentation rate and inhibiting the growth
of wild yeast. (125). Blending with other sweeteners, flavor compounds, and stabilizers
are optional prior to pasteurization and filling for improving the organoleptic characters
of blackberry wine (125). Unfortunately, the loss of anthocyanins during blackberry
wine production was detected (126).
2.4.8 Other applications of blackberries
On the demand of functional food in the worldwide market and thanks to the
advancement of food technology, blackberries have been extensively incorporated into
conventional foods based on its recognized health benefits, appealing color, and
fantastic flavor, consequently enhanced the nutritional aspects of conventional foods
and expanded the diversity of food for consumers.
Dairy products such as yogurts, ice-creams, smoothies, bakery products such as
pies, energy bars, muffins, cakes, and cereals are the major food matrix to carry
blackberries. Furthermore, blackberries are significant contributors to blended fruit
juice popular in the market. Nonetheless, the bioavailability of food products fortified
38
with blackberries other than the individual blackberry fruit is unknown. A synergistic
effect with nutrients has been proposed in the fundamental research (34), but the
interactions among different bioactive compounds and between basic food matrix and
nutraceticals, as well as their impact on health benefit and bioavailability in blackberry
products are likely trends for future research.
2.5 Conclusion
Post-harvest handling and processing technologies are critical for delivering
blackberry fruit with high quality and safety from field through processing plant to
dining table at home. Although abundant blackberry products are available for
satisfying consumers’ demands for tasty and nutritious products, limited information is
available about the impacts of post-harvest handling and processing technologies on the
quality and nutritional value of blackberries.
Therefore, it is necessary to explore how the basic quality, nutritional contents, and
bioactive compounds of blackberries change over refrigeration storage and during
common processing procedures and subsequent storage in order to provide consumers
with various forms of blackberry products at a maximum nutritional level.

39
2.6 Literature cited
(1) Ravai, M. Quality attributes of berry crops. Cereal Foods World 1996, 41, 773-775.
(2) Moyer, R.A.; Hummer, K.E.; Finn, C.E.; Frei, B.; Wrolstad, R.E. Anthocyanins,
phenolics, and antioxidant capacity in diverse small fruits: Vaccinium, Rubus, and
Ribes. J. Agric. Food Chem. 2002, 50, 519-525.
(3) Seeram, N.P. Berry fruits: compositional elements, biochemical activities, and the
impact of their intake on human health, performance, and disease. J. Agric. Food
Chem. 2008, 56, 627-629.
(4) Rieger, M. Blackberries and raspberries (Rubus spp.). In Introduction to fruit crops;
Rieger, M., Eds.; Food Products Press: New York, 2006; Chapter 6, pp 89-104.
(5) Huxley, A.; Griffiths, M.; Levy, M. Rubus. In Dictionary of Gardening; Huxley, A.;
Griffiths, M.; Levy, M., Eds.; Stockton Press: New York, 1992; Vol. 4, pp 140-145.
(6) Finn, C.E.; Strik, B.C. Blackberry cultivars for Oregon. Oregon State University
(7) Strik, B.C.; Clark, J.R.; Finn, C.E.; Bañados, M.P. Worldwide blackberry
production. HortTechnology 2007, 17, 205-213.
(8) Thompson, A.K. Fruit ripening conditions. In Fruit and vegetables: harvesting,
handling and storage, 2nd ed.; Thompson A.K., Eds.; Blackwell: Oxford, U.K.,
2003; Chapter 10, pp 86-86.
(9) Sharma, R.M.; Singh, R.R. Harvesting, postharvest handling and physiology of
fruits and vegetables. In Postharvest technology of fruits and vegetables; Verma,
L.R.; Joshi, V.K., Eds.; Indus Pub. Co.: New Delhi, India, 2000; Vol. 1, pp 127-134.
handling and storage, 2nd ed.; Thompson, A.K., Eds.; Blackwell: Oxford, U.K.,
2003; Chapter 10, pp 89-90.
(11) Perkins-Veazie, P.; Clark, J.R.; Huber, D.J.; Baldwin, E.A. Ripening physiology in
‘Navaho’ thornless blackberries: color, respiration, ethylene production, softening,
and compositional changes. Journal of the American society for Horticultural
Science 2000, 125, 357-363.
(12) Tosun, I.; Ustun, N.S.; Tekguler, B. Physical and chemical changes during ripening
of blackberry fruits. Sci.Agric. 2008, 65, 87-90.
40
8030.
(14) Burdon, J.N.; Sexton, R. Fruit abscission and ethylene production of 4 blackberry
cultivars (Rubus spp.). Annals of applied biology 1993, 123, 121-132.
(15) United States Department of Agriculture (USDA) National Nutrient Database for
standard reference. http://www.nal.usda.gov/fnic/foodcomp/search/index.html
(accessed November 2008).
(16) Kafkas, E.; Kosar, M.; Türemis, N.; Baser, K.H.C. Analysis of sugars, organic acids
and vitamin C contents of blackberry genotypes from Turkey. Food Chem. 2006,
97, 732-736.
(17) Macheix, J.; Fleuriet, A.; Billot, J. The main phenolics of fruit. In Fruit phenolics;
Macheix, J.; Fleuriet, A.; Billot, J., Eds.; CRC Press: Boca Raton, Florida, 1990;
Chapter 1, pp 1-98.
(18) Zadernowski, R.; Naczk, M.; Nesterowicz, J. Phenolic acid profiles in some small
(19) Howard, L.R.; Hager, T.J. Berry fruit phytochemicals. In Berry fruit: value-added
products for health promotion; Zhao, Y., Eds.; CRC press: New York, 2007;
Chapter 3, pp 74-98.
(20) Cho, M.J.; Howard, L.R.; Prior, R.L.; Clark, J.R. Flavonoid glycosides and
antioxidant capacity of various blackberry, blueberry and red grape genotypes
determined by high-performance liquid chromatography/mass spectrometry. J. Sci.
Food Agric. 2004, 84, 1771-1782.
(21) Wu, X.; Beecher, G.R.; Holden, J.M.; Haytowitz, D.B.; Gebhardt, S.E.; Prior, R.L.
Concentrations of anthocyanins in common foods in the United States and
estimation of normal consumption. J. Agric. Food Chem. 2006, 54, 4069-4075.
(22) Benvenuti, S.; Pellati, F.; Melegari, M.; Bertelli, D. Polyphenols, anthocyanins,
ascorbic acid, and radical scavenging activity of Rubus, Ribes, and Aronia. J. Food
Sci. 2004, 69, FCT164- FCT169.

J. Food Sci. 2005, 70, C198-C202.
41
(25) Edwin, H. Polyphenol complexation. In Plant polyphenols; Edwin, H., Eds.;

Cambridge: New York, 1989; Chapter 4, pp 154-214.
(26) Daniel, E.M.; Krupnick, A.S.; Heur, Y.; Blinzler, J.A.; Nims, R.W.; Stoner, G.D.
Extraction, stability, and quantitation of ellagic acid in various fruits and nuts. J.
Food Composit. Annal. 1989, 2, 338-349.
J. Agric. Food Chem. 2008, 56, 661-668.
(28) Lewis, N.G.; Yamamoto, E. Tannins - their place in plant metabolism. In Chemistry
and significance of condensed tannins; Hemingway, R.W.; Karchesy, J.J.; Branham,
S.J., Eds.; Plenum Press: New York, 1989; pp 23-43.
(29) Thompson, L.U. Role of lignans in carcinogenesis. In Phytochemicals in human

health protection, nutrition, and plant defense; Romeo, J.T., Eds.; Kluwer
Academic/Plenum Publishers: New York, 1999; Chapter 3, pp 51-61.
(30) Talcott, S.T. Chemical components of berry fruits. In Berry fruit: value-added
(31) González, E.M.; Ancos, B.D.; Cano, M.P. Partial characterization of peroxidase and
polyphenol oxidase activities in blackberry fruits. J. Agric. Food Chem. 2000, 48,
5459-5464.
(32) Martínez, G.A.; Civello, P.M. Effect of heat treatments on gene expression and
enzyme activities associated to cell all degradation in strawberry fruit. Postharv.
Biol. Tech. 2008, 49, 38-45.
J. Agric. Food Chem. 2008, 56, 630-635.
(34) Liu, R.H. The potential health benefits of phytochemicals in berries for protecting
against cancer and coronary heart disease. In Berry fruit: value-added products for
health promotion; Zhao, Y., Eds.; CRC press: New York, 2007; Chapter 6, pp 187-
199.
42
(36) Tsuda, T. Regulation of adipocyte function by anthocyanins; possibility of

preventing the metabolic syndrome. J. Agric. Food Chem. 2008, 56, 642-646.
(38) Cahn, H.; DeFrancesco, J.; Nelson, E.; Martin, L. Fruit quality evaluation of
raspberries and blackberries at North Willamette Research and Extension Center.
Special report / Agricultural Experiment Station, Oregon State University 1992,
892, 2-9.
(39) Eyduran, S.P. Eyduran, E.; Agaoglu, Y.S. Estimation of fruit weight by cane traits
for eight American blackberries (Rubus fructicosus L.) cultivars. Journal of
Biotechnology 2008, 7, 3031-3038.
(40) Perkins-Veazie, P.; Collins, J.K.; Clark, J.R. Fruit characteristics of some erect
blackberry cultivars. HortSci. 1993, 28, 853-853.
(41) Turemis, N.; Kafkas, S.; Kafkas, E.; Onur, C. Fruit characeristics of nine thornless
blackberry genotypes. Journal American Pomological Society 2003, 57, 161-165.
(42) Thompson, A.K. Fruit ripening conditions. In Postharvest Technology of Fruit and
Vegetables; Thompson, A.K., Eds.; Blackwell Science: Harlow, Oxford, 1996;
(43) Sousa, M.B.; Canet, W.; Alvarez, M.D.; Fernandez, C. Effect of processing on the
texture and sensory attributes of raspberry (cv. Heritage) and blacbkerry (cv.
Thornfree). Journal of Food Engineering 2007, 78, 9-21.
(45) Archbold, D.D.; HamiltonKemp, T.R.; Barth, M.M.; Langlois, B.E. Identifying
natural volatile compounds that control gray mold (Botrytis cinerea) during
postharvest storage of strawberry, blackberry, and grape. J. Agric. Food Chem.
1997, 45, 4032-4037.
(46) Nordskog, B.; Stensvand, A.; Heiberg, N. Fungi occuring on aerial constituents of
cultivated blackberry (Rubus fruticosus L.) in Norway. Acta Agriculturae
Scandinavica Section B-Soil and Plant Sci. 2003, 53, 21-28.
43
(47) Perkins-Veazie, P.; Collins, J.K.; Clark, J.R. Cultivar and maturity affect
postharvest quality of fruit from erect blackberries. HortSci. 1996, 31, 258-261.
(48) Prange, R.K.; DeEll, J.R. Preharvest factors affecting postharvest quality of berry
crops. HortSci. 1997, 32, 824-829.
‘Shawnee’ blackberry fruit stored under retail storage conditions. Journal of Food
Quality 1999, 22, 535-544.
(50) Lin, D.; Zhao, Y. Innovations in the development and application of edible coatings
for fresh and minimally processed fruits and vegetables. Comprehensive Reviews in
Food Science and Food Safety 2007, 6, 1-13.
(51) Talcott, S.T. Chemical components of berry fruits. In Berry fruit: value-added
(53) Connor, A.M.; Finn, C.E.; Alspach, P.A. Genotypic and environmental variation in
antioxidant activity and total phenolic content among blackberry and hybridberry
cultivars. Journal of the American Society for Horticultural Sci. 2005, 130, 527-533.
(54) Connor, A.M.; Finn, C.E.; Alspach, P.A. Genetic and enviromental variation in
anthocyanins and their relationship to antioxidant activity in blackberry and
hybridberry cultivars. Journal of the American Society for Horticultural Sci. 2005,
130, 680-687.
(55) Woods, F.M.; Dozier, W.A.; Ebel, R.C.; Thomas, R.; Nesbitt, M.; Wilkins, B.S.;
Himelrick, D.G. Cultivar and maturity effects on fruit quality and antioxidant
properties in blackberry. HortSci. 2006, 41, 1043-1043.
(56) Wang, S.Y.; Lin, H. Antioxidant activity in fruits and leaves of blackberry,
raspberry, and strawberry varies with cultivar and developmental stage. J. Agric.
Food Chem. 2000, 48, 140-146.
(57) Qian, M.C.; Wang, Y. Seasonal variation of volatile composition and odor activity
value of ‘Marion’ (Rubus spp. hyb) and ‘Thornless Evergreen’ (R.laciniatus L.)
blackberries. J. Food Sci. 2005, 70, C13-C19.
44
(58) Asami, D.K.; Hong, Y.; Barrett, D.M.; Mitchell, A.E. Comparison of the total
phenolic and ascorbic acid content of freeze-dried and air-dried Marionberry,
strawberry, and corn grown using conventional, organic, and sustainable agricultural
practices. J. Agric. Food Chem. 2003, 51, 1237-1241.
(59) Susaimuthu, J.; Tzanetakis, I.E.; Gergerich, R.C.; Kim, K.S.; Martin, R.R. Viral
interactions lead to decline of blackberry plants. Plant Disease 2008, 92, 1288-
1292.
(60) Gomez, D.R.; Evans, K.J.; Baker, J.; Harvey, P.R.; Scott, E.S. Dynamics of
introduced populations of Phragmidium violaceum and implications for biological
control of European blackberry in Australia. Applied and Environmental
Microbiology 2008, 74, 5504-5510.
(61) Gordon, S.C.; Woodford, J.A.T.; Birch, A.N.E. Arthropod pests of Rubus in
Europe: pest status, current and future control strategies. Journal of Horticultural
Sci. 1997, 72, 831-862.
(62) Amano, S.; Yano, H. Evaluation of the practical performance of a commercially

available ELISA kit in the analysis of residual imidacloprid in agricultural products.
Journal of Pesticide Sci. 2005, 30, 249-253.
2003; pp 19-112.
2003; Chapter 7, pp 59-60.
(65) Perkins-Veazie, P.; Collins, J.K.; Clark, J.R. Changes in blackberry fruit quality
during storage. Acta Horticulturae 1993, 352, 87-90.
(66) Antunes, L.E.C.; Duarte, J.; de Souza, C.M. Postharvest conservation of blackberry
(67) Perkins-Veazie, P.; Collins, J.K.; Clark, J.R. Cultivar and storage temperature
effects on the shelflife of blackberry fruit. Fruit Varieties Journal 1999, 53, 201-
208.
increase antioxidant levels. Acta Horticulturae 2002, 585, 521-524.
45
(69) Perkins-Veazie, P.; Collins, J.K. Quality of erect-type blackberry fruit after short
intervals of controlled atmosphere storage. Postharv. Biol. Tech. 2002, 25, 235-239.
(70) Agar, I.T.; Streif, J.; Bangerth, F. Effect of high CO2 and controlled atmosphere
(CA) on the ascorbic and dehydroascorbic acid content of some berry fruits.
Postharv. Biol. Tech. 1997, 11, 47-55.
2003; Chapter 5, pp 38-46.
(72) Fonseca, S.C.; Oliveira, F.A.R.; Brecht, J.K. Modeling respiration rate of fresh
fruits and vegetables for modified atmosphere packages: a review. Journal of Food
Engineering 2002, 52, 99-119.
(73) Martinez-Romero, D.; Guillen, F.; Castillo, S.; Valero, D.; Serrano, M. Modified
atmosphere packaging maintains quality of table grapes. J. Food Sci. 2003, 68,
1838, 1843.
(74) Rocha, A.M.C.N.; Barreiro, M.G.; Morais, A.M.M.B. Modified atmosphere

package for apple ‘Bravo de Esmolfe’. Food Control 2004, 15, 61-64.
(75) Song, Y.; Vorsa, N.; Yam, K.L. Modeling respiration-transpiration in a modified
atmosphere packaging system containing blueberry. Journal of Food Engineering
2002, 53, 103-109.
(76) Petracek, P.D.; Joles, D.W.; Shirazi, A.; Cameron, A.C. Modified atmosphere
packaging of sweet cherry (Prunus avium L., ev. ‘Sams’) fruit: metabolic responses
to oxygen, carbon dioxide, and temperature. Postharv. Biol. Tech. 2002, 24, 259-
270.
(77) Park, H.J. Development of advanced edible coatings for fruits. Trends Food Sci.
Tech. 1999, 10, 254-260.
(78) Han, C.; Zhao Y.; Leonard, S.W.; Traber, M.G. Edible coatings to improve
storability and enhance nutritional value of fresh and frozen strawberries (Fragaria ×
ananassa) and raspberries (Rubus ideaus). Postharv. Biol. Tech. 2004, 33, 67-68.
(79) Park, S.I.; Stan, S.D.; Daeschel, M.A.; Zhao Y. Antifungal coatings on fresh
strawberries (Fragaria x ananassa) to control mold growth during cold storage. J.
Food Sci. 2005, 70, 202-207.
46
(80) Vargas, M.; Albors, A.; Chiralt, A.; Gonzalez-Martınez, C. Quality of cold-stored
strawberries as affected by chitosan–oleic acid edible coatings. Postharv. Biol. Tech.
2006, 41, 164–171.
(81) Ribeiro, C.; Vicente, A.A.; Teixeira, A.J.; Miranda, C. Optimization of edible
coating composition to retard strawberry fruit senescence. Postharv. Biol. Tech.
2007, 44, 63–70.
(82) Hernandez-Munoz, P.; Almenar, E.; Del Valle, V.; Velez, D.; Gavara, R. Effect of
chitosan coating combined with postharvest calcium treatment on strawberry
(Fragaria × ananassa) quality during refrigeration storage. Food Chem. 2008, 110,
428-435.
(83) Meneghel, R.F.D.; Benassi, M.D.; Yamashita, F. Sodium alginate coating for
blackberry (Rubus ulmifolius) fruits. Semina-Cienclas Agrarias 2008, 29, 609-617.
(84) Mei, Y.; Zhao, Y. Using edible coating to enhance nutritional and sensory qualities
of babay carrots. J. Food Sci. 2002, 65, 1964-1968.
(85) Khwaldia, K.; Perez, C.; Banon, S.; Desobry, S.; Hardy, J. Milk proteins for edible
films and coatings. Crit. Rev. Food Sci. Nutr. 2004, 44, 239–251.
(86) Vachon, C.; D’Aprano, G.; Lacroix, M.; Letendre, M. Effect of edible coating
process and irradiation treatment of strawberry Fragaria spp. on storage-keeping
quality. J. Food Sci. 2003, 68, 608-612.
(87) Fisk, L.C.; Silver, M.A.; Strik, B.C.; Zhao, Y. Postharvest quality of hardy kiwifruit
(Actinidia arguta ‘Ananasnaya’) associated with packaging and storage conditions.
(88) Nimitkeatkai, H.; Srilaong, V.; Kanlayanarat, S. Effect of edible coating on

Pineapple fruit quality during cold storage. Acta Hort. 2006, 712, 643-647.
(89) Yaman, Ö.; Bayoindirli, L. Effect of an edible coating and cold storage on shelf-life
and quality of cherries. Lebensmittel-Wissenschaft Und-Technologie-Food Sci
Technol. 2002, 35, 146-150.
(90) Erkan, M.; Wang, S.Y.; Wang, C.Y. Effect of UV treatment on antioxidant capacity,
antioxidant enzyme activity and decay in strawberry fruit. Postharv. Biol. Tech.
2008, 48, 163-171.
(91) Gonzalez-Aguilar, G.A.; Villegas-Ochoa, M.A.; Martinez-Tellez, M.A.; Gardea,

A.A.; Ayala-Zavala, J.F. Improving antioxidant capacity of fresh-cut mangoes
treated with UV-C. J. Food Sci. 2007, 72, S197-S202.
47
(92) Perkins-Veazie, P.; Collins, J.K.; Howard, L. Blueberry fruit response to postharvest
application of ultraviolet radiation. Postharv. Biol. Tech. 2008, 47, 280-285.
(93) Vicente, A.R.; Repice, B.; Martinez, G.A.; Chaves, A.R.; Civello, P.M.; Sozzi, G.O.
Maintenance of fresh boysenberry fruit quality with UV-C light and heat treatments
combined with low storage temperature. Biotech. 2004, 79, 246-251.
(94) Hussain, P.R.; Meena, R.S.; Dar, M.A.; Mir, M.A.; Shafi, F.; Wani, A.M. Effect of
gamma-irradiation and refrigerated storage on mold growth and keeping quality of
strawberry (Fragaria sp) cv ‘Confitura’. Journal of Food Sci. & Tech. 2007, 44, 513-
516.
(95) Cheour, F.; Mahjoub, A. Delayed ripening and senescence of strawberry (Fragaria ×
ananassa Duch.) by gamma irradiation. Sciences Des Aliments 2003, 23, 355-366.
(96) Frenzen, P.D.; Majchrowicz, A.; Buzby, J.C.; Imhoff, B. Consumer acceptance of
irradiated meat and poultry products. Agriculture information bulletin from United
States Department of Agriculture/Economic Research Service 2000 (Aug), 757, 1-8.
(97) Barth, M.M.; Zhou, C.; Mercier, J.; Payne, F.A. Ozone storage effects on
anthocyanin content and fungal growth in blackberries. J. Food Sci. 1995, 60, 1286-
1288.
(98) Chanjirakul, K.; Wang, S.Y.; Wang, C.Y. Siriphanich, J. Natural volatile treatments
increase free-radical scavenging capacity of strawberries and blackberries. J. Sci.
Food Agric. 2007, 87, 1463-1472.
(99) Wang, S.Y.; Bowman, L.; Ding, M. Methyl jasmonate enhances antioxidant activtiy
and flavonoid content in blackberries (Rubus sp.) and promotes antiproliferation of
human cancer cells. Food Chem. 2008, 107, 1261-1269.
(100) Hui, Y.H.; Lim, M.; Nip, W.; Smit,h J.S.; Yu, P.H.F. Principles of food processing.
Blackwell: Ames, Iowa, 2004; Chapter 1, pp 15-28.
(101) Hui, Y.H.; Lim, M.; Nip, W.; Smith, J.S.; Yu, P.H.F. Principles of food processing.
Blackwell: Ames, Iowa, 2004; Chapter 1, pp17-19.
(102) Zhao, Y. Freezing process of berries. In Berry fruit: value-added products for health
promotion; Zhao, Y., Eds.; CRC press: New York, 2007; Chapter 10, pp 291-310.
48
(103) Marti, J.; Aguilera, J.M. Effects of freezing rate on the mechanical characteristics
and microstructure of blueberries and wild blackberries. Revista De Agroquimica Y
Tecnologia De Alimentos 1991, 31, 493-504.
(104) González, E.M.; de Ancos, B.; Cano, M.P. Relation between bioactive compounds
and free radical-scavenging capacity in berry fruits during frozen storage. J. Sci.
Food Agric. 2003, 83, 722-726.
(105) Schmidt, B.M.; Erdman, J.W.; Lila, M.A. Effects of food processing on blueberry
antiproliferation and antioxidant activity. J. Food Sci. 2005, 70, S389-S394.
(106) Ngo, T.; Wrolstad, R.E.; Zhao, Y. Color quality of Oregon strawberries-impact of
genotype, composition, and processing. J. Food Sci. 2007, 72, C25-C32.
(107) Xie, J.; Zhao, Y. Improvement of physicochemical and nutritional qualities of

frozen Marionberry by vacuum impregnation pretreatment with cryoproctectants
and minerals. Journal of Horticultural Science & Biotechnology 2003, 78, 248-253.
(108) Ramaswamy, H.S.; Chen, C.R. Canning Principles. In Handbook of vegetables

preservation and processing; Hui, Y.H.; Ghazala, S.; Graham, D.M.; Murrell, K.D.;
Nip, W., Eds.; Marcel Dekker: New York, 2004; Chapter 4, pp 273- 282.
(109) Burrows, G. Production of thermally processed and frozen fruit. In Fruit processing;
Arthey, D.; Ashurst, P.R., Eds.; Blackie Academic & Professional: London, UK,
1996; Chapter 6, pp 135-164.
(110) Hager, T.J.; Howard, L.R.; Prior, R.L. Processing and storage effects on monomeric
anthocyanins, percent polymeric color, and antioxidant capacity of processed
blackberry products. J. Agric. Food Chem. 2008, 56, 689-695.
(111) Brownmiller, C.; Howard, L.R.; Prior, R.L. Processing and storage effects on
monomeric anthocyanins, percent of polymeric color, and antioxidant capacity of
processed blueberry products. J. Food Sci. 2008, 73, H72-H78.
(112) Chaovanalikit, A.; Wrolstad, R.E. Anthocyanin and polyphenolic composition of

fresh and processed cherries. J. Food Sci. 2004, 69, FCT73-FCT83.
(113) Barta, J. Fruit dyring principles. In Handbook of fruits and fruit processing; Hui,
Y.H.; Barta, J.; Cano, M.P.; Gusek, T.; Sidhu, J.S.; Sinha, N.K., Eds.; Blackwell:
(114) Figuerola, F.E. Dehydration of berries. In Berry fruit: value-added products for
333.
49
(115) Yurdugul, S. An evaluation of the retention of quality characteristics in fresh and

freeze-dried alpine strawberries. International Journal of Food Science and
Technology 2008, 43, 865-870.
(116) Bohm, V.; Kuhnert, S.; Rohm, K.; Scholze, G. Improving the nutritional quality of
microwave-vacuum dried strawberries: A preliminary study. Food Science and
Technology International 2006, 12, 67-75.
(117) Shih, C.; Pan, Z.; McHugh, T.; Wood, D.; Hirschberg, E. Sequential infrared
radiation and freeze-drying method for producing crispy strawberries. Transactions
of the Asabe 2008, 51, 205-216.
(118) Venkatachalapathy, K.; Raghavan, G.S.V. Microwave drying of osmotically

dehydrated blueberries. Journal of Microwave Power and electromagnetic Energy
1998, 33, 95-102.
(119) Vibhakara, H.S.; Bawa, A.S. Manufacturing jams and jellies. In Handbook of fruits
and fruit processing; Hui, Y.H.; Barta, J.; Cano, M.P.; Gusek, T.; Sidhu, J.S.; Sinha,
N.K., Eds.; Blackwell: Ames, Iowa, 2006; Chapter 11, pp 8189-204.
(120) Amakura, Y.; Umino, Y.; Tsuji, S.; Tonogai, Y. Influence of jam processing on the
radical scavenging activity and phenolic content in berries. J. Agric. Food Chem.
2000, 48, 6292-6297.
(121) Häkkinen, S.H.; Kärenlampi, S.O.; Mykkänen, H.M.; Törrönen, A.R. Influence of
domestic processing and storage on flavonol contents inberries. J. Agric. Food
Chem. 2000, 48, 2960-2965.
(122) Hui, Y.H. Fruits: Orange juice processing. In Food processing: principles and
applications; Smith, J.S.; Hui, Y.H., Eds.; Blackwell: Ames, Iowa, 2004; Chapter
21, pp 361-390.
(123) Rutledge, P. Production of non-fermented fruit products. In Fruit processing;

Arthey, D.; Ashurst, P.R., Eds.; Blackie Academic & Professional: London, UK,
1996; Chapter 4, pp 70-95.
(124) Skrede, G.; Wrolstad, R.E.; Durst, R.W. Changes in anthocyanins and polyphenolics
during juice processing of highbush blueberries (vaccinium corymbosum L.). J.
Food Sci. 2000, 65, 357-364.
(125) Jarvis, B. Cider, perry, fruit wines and other alcoholic fruit beverages. In Fruit
processing; Arthey, D.; Ashurst, P.R., Eds.; Blackie Academic & Professional:
London, UK, 1996; Chapter 5, pp 121-127.
50
(126) Rommel, A.; Wrolstad, R.E.; Heatherbell, D.A. Blackberry juice and wine-
processing and storage effects on anthocyanin composition, color and apprearance.
J. Food Sci. 1992, 57, 385-391.
51
CHAPTER 3

Compounds and Antioxidant Capacities of Blackberries
Ruyi Wu1, Balz Frei2, James A. Kennedy1, and Yanyun Zhao1,3
1
Department of Food Science & Technology, Oregon Sate University, Corvallis, OR
97331, USA.
2
Linus Pauling Institute, Oregon Sate University, Corvallis, OR 97331, USA.
3
Corresponding author: E-mail: yanyun.zhao@oregonstate.edu
To be submitted to Journal of Agricultural Food and Chemistry

American Chemical Society, 1155 Sixteenth Street N.W., Washington, DC 20036
52
ABSTRACT
Blackberry fruits have attractive color, unique flavor, and contain bioactive
compounds associated with a number of health benefits that have been confirmed by
scientific evidences. However, their perishable nature and short harvest season make it
challenging to preserve their quality. Multiple post-harvest processing technologies
have been applied to extend the shelf-life of fresh blackberries. This study investigated
the effects of refrigeration storage and different processing technologies including
freezing, freeze drying, hot-air drying, canning and jam processing, as well as 6-month
subsequent storage at room temperature on the physicochemical properties (pH,
titratable acidity (TA), total soluble solids (TSS) and moisture content (MC)), bioactive
compounds (total phenolics (TPC) and total monomeric anthocyanins (ACY)), and
antioxidant capacities evaluated as radical scavenging activity against DPPH (RSA),
oxygen radical absorbance capacity (ORAC) and ferric reducing ability of plasma
(FRAP) of two blackberry varieties, ‘Marion’ and ‘Evergreen’.
Refrigeration storage at 2.0 ± 0.2 oC and 95 ± 2% relative humidity (RH) decreased
TA by 36.8% in ‘Marion’ at 9 days and 46.2% in ‘Evergreen’ at 7 days. TPC and ACY
fluctuanted in ‘Marion’ while continuously declined in ‘Evergreen’ along with
refrigeration storage time. Similar to TPC and ACY, RSA fluctuated in ‘Marion’, but
was stable in ‘Evergreen’. ORAC decreased by 20% and FRAP increased by 18.75% in
‘Evergreen’ at the end of refrigeration storage, while no significant changes in ‘Marion’
were observed.
53
TA declined in all processed ‘Marion’. The increases in TSS of dried fruit and jams
were observed in both blackberry varieties. Freeze drying increased TPC (21%), ACY
(5.5%), and RSA (14%) in ‘Evergreen’ and TPC in ‘Marion’ (27%) compared to frozen
control. Hot-air drying resulted in significant (P<0.05) loss of ACY (56%), ORAC
(37%), and FRAP (27%) in ‘Marion’, and TPC (37%), ACY (84%), and RSA (13%) in
‘Evergreen’. Canning significantly reduced the content of bioactive compounds by 31%
to 70% in both varieties except TPC in canned-in-water ‘Marion’. ORAC and FRAP
reduced in canned ‘Marion’ by 21% to 61%. Canning-in-water increased RSA (22% in
‘Marion’ and 8% in ‘Evergreen’) while canning-in-20° Brix sucrose syrup decreased
RSA by 31% in both varieties. Jam processing resulted in 67% to 84% loss of TPC and
ACY, 78% to 80% loss of RSA, and 65% to 77% loss of ORAC and FRAP in both
blackberry varieties.
The 6-month post-process storage at 25 ± 2 oC and 30 ± 2% RH decreased TA of
jams by 9% in ‘Marion’ and 12% in ‘Evergreen’, but no significant (P>0.05) changes in
TA in canned and dried samples. ACY declined in hot-air dried and canned blackberries
from 21% to 57%, and jams from 56% to 66% in both varieties, RSA consistently fell in
all frozen, dried, and canned samples from 24% to 86%, ORAC reduced in hot-air dried
(13% for ‘Marion’ and 45% for ‘Evergreen’) and canned-in-water samples (68% for
‘Marion’ and 38% for ‘Evergreen’) during room storage.
‘Marion’ had significantly (P<0.05) higher TA, MC, TPC, ACY, RSA and FRAP
values than those of ‘Evergreen’, but lower pH, TSS, and ORAC values. TA, TPC,
54
RSA, and FRAP were positively correlated to each other in both fresh and processed
blackberry products.
55
INTRODUCTION
Berry fruits have been widely recognized as an excellent source of bioactive
phenolic compounds including flavonoids, phenolic acid, and tannins (1), providing
positive impacts both individually and synergistically against cardiovascular disease,
cancer, inflammatory, aging, obesity, diabetes, and other chronic diseases (2-7).
Blackberries have attracted substantial attention nowadays due to their high
anthocyanins and ellagitannin content, high antioxidant activities, and favorable flavor
demonstrated in numerous studies (8-10). The production of blackberries in the United
States was 35,099 tons in 2005, which accounted for the highest portion of the
worldwide production of blackberries in 2005 (11). Unfortunately, the fragile surface
and high post-harvest respiration rate of blackberries contributes significantly to their
quality, nutritional, and microbiological deterioration, resulting in limited shelf-life and
diminished heath benefits for fresh market (12). Refrigeration storage is the most
common practice used for fresh blackberry preservation (13-15).
Due to the short production season and perishable characteristics of blackberries,
most fresh fruit is processed into frozen, dried, and canned forms, or processed into
jams, jellies, and juices for longer storage to satisfy various markets and consumer
demands (16). Freeze processing is generally considered as the least destructive
preservation technology for phenolic compounds in berries and is recommended as a
pretreatment for manufacturing other berry products, although the physical and
nutritional quality of frozen berries can be affected by freezing methods, package
materials, storage conditions, and blackberry varieties and maturity stage (17).
56
Drying is a traditional method used to preserve fresh berries by reducing their water
activity. Dried fruit have many different applications in snacks, breakfast, and
formulated foods (18). The quality of dried berries is determined by the type of drying
method applied, water activity of the final product, and package and storage conditions.
For ‘Marion’ blackberry, freeze drying has been shown to retain higher phenolics than
that of hot-air drying (19). Freeze drying has also been shown to help preserve
anthocyanins and antioxidant capacity of Saskatoon berries (20).
Canning process uses high temperature to destroy microorganisms and creates
hermetical sealing (21). Our previous study reported that the canning of Oregon
strawberries in syrup increased their total phenolics and anthocyanins substantially (22).
A recent study also concluded that canning increased polymeric color, but decreased
anthocyanin content and antioxidant capacity of blackberries, while storage at 25 oC did
not change antioxidant capacity significantly, but led to the continuous gain of
polymeric color and loss of anthocyanins (23).
Berry jams are one of the most important dietary forms of berry fruit consumption
(24). Previous studies have indicated that a small portion of flavonols was lost when
making strawberry jams via cooking berries with sugar for 30 min (25) while total
phenolics were well retained during blackberry jam processing (26).
The extent to which nutraceuticals preservation in blackberry products heavily
depends on the specific processing technology, the variety of the blackberries,
production location, maturity and time when harvested, and storage conditions (16).
Therefore, more comprehensive studies are necessary to understand the impact of

57
different processing technologies on the nutraceutical benefits of various forms of
blackberry products. Such information is essential for processors to better design the
processing procedures and for consumers to understand the health benefit of product
when purchasing.
The objective of this study was to investigate the changes of total phenolics, total
monomeric anthocyanins, and antioxidant capacities in two blackberry varieties
(‘Marion’ and ‘Evergreen’) during refrigeration storage of fresh fruit and when the fruit
were subjected to different processing treatments including freezing, drying, canning,
jam-making, and subsequent 6-month storage in room condition. Basic
physicochemical properties of fresh and processed berries were also investigated.
MATERIALS AND METHODS
Fruit. Two blackberry varieties, ‘Marion’ and ‘Evergreen’, were obtained from
Scenic Fruit Co. (Gresham, OR, USA) and RainSweet, Inc. (Salem, OR, USA) in early
July and early September, 2007, respectively. Fresh fruit were shipped under
refrigeration to the Department of Food Science & Technology (FST) at Oregon State
University the day after harvest. For refrigeration storage, fresh fruit were packed in 10
oz (300 g) clam-shell containers and stored at 2.0 ± 0.5 oC and 95 ± 2% RH at FST pilot
plant under dark. For preparing frozen samples, fresh fruit were washed with 20 ppm
chlorinated water for 80 sec prior to freezing process. ‘Marion’ fruit were individually
frozen on stainless steel trays in an air blast freezer at -23 oC in the FST pilot plant,
while fresh ‘Evergreen’ fruit were individually frozen at the RainSweet facility under
58
similar conditions. Frozen fruit were packed into 2 L wide-mouth glass jars (Ball
Corp., Muncie, IN, USA) with lids and stored at -23 ± 2 oC for up to 6 months (Figure
3.1).
Fresh Blackberries
Wash
Freeze Refrigeration storage

(2.0±0.2oC, 95±2% RH)
Con FD HD Can Jam

0d
1d
0m 0m 0m 0m 0m 3d
Room storage 3m 3m 3m 3m 3m 5d
(25±2oC, 30±2%) 6m 6m 6m 6m 6m 7d
9d
Figure 3.1 Flow diagram of experimental design. Con: Control (frozen); FD: Freeze
drying; HD: Hot-air drying; Can: Canning; Jam: Jam processing.
Dry processing. Both freeze and hot-air dryings were applied in this study. For
freeze drying, whole fruits were laid on aluminum trays in a freeze drier (Hull Corp.,
Hatboro, PA, USA) and frozen under a condenser temperature of -45 oC for about 1 h.
After the chamber temperature of the drier reached to -45 oC, vacuum was applied for
achieving a maximum vacuum pressure of 30 Pa. It took approximately 72 h for the
fruit to be fully dried at the shelf temperature (the temperature on the shelf of the
59
chamber when drying) of 35 oC and 50% RH. For hot-air drying, whole fruit were dried
in the MP-2000 Enviro-Pak conventional drier (Division of Tech-Mark, Inc.,
Clackamas, OR, USA) under a shelf temperature of 50 oC and 50% RH with 50% air
speed for 48 h. Dried blackberries were packaged in moisture-resistant plastic Ziploc
bags (Johnson & Son, Inc., Racine, WI, USA) and stored in desiccators (Space Saver
Vacuum desiccators, Scienceware, Pequannock, NJ, USA) with drierite (anhydrous
calcium sulfate, W.A. Hammond drierite company Ltd., Xenia, OH, USA) under dark at
25 ± 2 oC and 30 ± 2% RH for up to 6 months.
Jam processing. To make blackberry jams composed of 45% fruit and 55% sugar,
frozen blackberries were thawed overnight at room temperature and then crushed in a
stainless steel kettle (DN-30 Groen MFG. Co., Chicago, IL, USA). Crushed fruit were
weighed and the Brix of the fruit was measured by using a refractometer (RFM81 Multi
Scale Automatic, Bellingham+Stanley Inc., Atlanta, Georgia). The pH of ‘Evergreen’
blackberries was titrated (Brinkmann Digital Buret, BRAND, W-Germany) to 3.2 by
1% W/V citric acid (Integra Chemical Company, Renton, Washington, USA). The fruit
puree was transferred into a jacketed steam kettle (D-20 Groen MFG. Co., Chicago, IL,
USA) heated up to ~49 oC, subsequently mixed with pre-measured amounts of cane
sugar (G&H, distributed by Domino Foods Inc., Yonkers, NY, USA) and 150 grade
rapid-set pectin (Pacific pectin Inc., Oakhurst, CA, USA) by stirring to avoid clumping.
Fruit mixtures were heated to boiling, and the Brix was checked until reaching
approximately 68 oBrix. The finished product was hot-filled into pasteurized 400 mL
Mason jars with caps (Ball Corp., Muncie, IN, USA). The filled jars were inverted to
60
pasteurize the lids for 3 min and then allowed to cool and set up in the upright position
at room temperature. Blackberry jams were stored in the dark cardboard boxes at 25 ±
2 oC and 30 ± 2% RH for up to 6 months.
Canning processing. Approximate 210 g of frozen blackberries were filled into
No. 300 plain can (300 X 407, Ball Corp., Muncie, IN, USA) with C-enameled ends. A
180 g of boiling 20 oBrix sucrose syrup or distilled (DI) water were added into the can,
leaving ½ inch headspace. The cans with the lids on went through a steam tunnel (Dixie
Canner, Athens, Georgia, USA) at around 95 oC and heated up. After coming out of the
steam tunnel, the hot cans were immediately sealed by using the Automatic Master-
Sealer (Canning devices, Inc., Manitowoc, WI, USA). The sealed cans were placed in
boiling water for 15 min and then cooled in cold running water. The temperature of the
first can coming out of the steam tunnel and the vacuum of the finished cans were
checked by a thermometer (Weston, Model 2261, CSI International Inc., USA) and
vacuum gauge (Marshalltown, Model 89011, USA), respectively during the processing.
After cooling down, the cans were stored in the dark cardboard boxes at 25 ± 2 oC and
30 ± 2% RH for up to 6 months.
Measurement of pH, total soluble solids and titratable acidity. A 20 g of fresh,
frozen and canned blackberries and blackberry jams, or 5 g of freeze and hot-air dried
blackberries were crushed and homogenized with 180 mL of DI water separately for 1
min by using a 12-speed blender (Osterizer, Jarden Corp., Mexico). Total soluble solids
(TSS) of the homogenized solution were measured by a refractometer (RA-250, kem,
Kyoto Electronics Manufacturing Co. Ltd, Japan) right after blending. The solution was
61
then filtered through a filter paper (Qualitative 15 cm Whatman International Ltd.
Maidstone, England) and 30 mL of filtered solution was collected for testing pH using a
pH meter (IQ240, IQ Scientific Instruments, Inc., San Diego, CA, USA) and titratable
acidity (TA) by titrating (Brinkmann Digital Buret, BRAND, W-Germany) with 0.1 N
aqueous NaOH (J.T.Baker, Mallinckrodt Baker, Inc., Phillipsburg, NJ, USA) until a
final pH of 8.2 was reached. TA results were expressed as mass percentage of malic
acid based on fresh or dried weight of blackberry samples.
Measurement of moisture content. Approximate 10 g of blackberry samples (Mi)
(TL-204 Analytical Balance, Denver Instrument Company, city, State, USA) were
placed into a pre-weighed glass dish (Md). Samples were dried in a convection oven
(STM 40, Precision Scientific Inc., Chicago, IL, USA) at 70 oC for 6 h and then
removed to a vacuum oven (Forma Scientific Inc., Marietta, OH, USA) set at 30 Pa
vacuum and 70 oC until fully dry. The final weight (Mf) of dried sample with the dish
was measured. Moisture content (MC) on a fresh weight basis was calculated as:
(M i + M d ) − M f
MC = × 100%
Mi
Sample extraction. The extraction of blackberry samples for analyzing bioactive
compounds and antioxidant capacity was conducted according to a modified method by
Rodriguez -Saona and Wrolstad (27). A 10 g of fresh, frozen, canned blackberries, or 2
g of hot-air and freeze dried sample was powdered under liquid nitrogen in a stainless
steel Waring blender (Model 1001, Waring Products Co., Winsted, CT, USA). Fresh,
frozen and canned blackberries were extracted by 100% aqueous acetone with 0.01%
(v/v) HCl (EMD Chemicals Inc., Gibbstown, NJ, USA) once followed by 70% (v/v)
62
aqueous acetone with 0.01% (v/v) HCl twice with 0.5 min of ultrasound in an ultrasonic
cleaner at 50/60 Hz, 117 V and 185 W (Branson B-220H 19, 6"W X 10”L X 4”D,
SmithKline Co., Shelton, CT, USA). The mixtures were centrifuged at 4000g for 5 min
(CL International Clinical Centrifuge, International Equipment Co., Boston, MA, USA),
the supernatants were gathered in plastic bottles, and 50 mL chloroform (Mallinckrodt
Baker Inc., Phillipsburg, NJ, USA) was added into the bottles to partition the filtrate.
After the solvent in bottles was centrifuged at 3000g for 30 min under refrigerated
conditions, the chloroform was discarded through the separation funnels (Pyrex,
Corning Inc., Lowell, MA, USA) and the aqueous portion was transferred to the boiling
flask (Kontes, Kimble Chase Life Science and Research Products, Vineland, NJ, USA).
After removing the remaining organic solvents in the extract by rotary evaporator
(Brinkmann Instruments, Westbury, NY) under vacuum pressure of 30 Pa and with
immersion of the flask with samples in water bath (CMS, Model No: 392-142, Lab-Line
Instruments Inc., Melrose park, IL, USA) at 40 oC for 1 min, the extract was transferred
to a 50 mL volumetric flask (Kontes, Kimble Chase Life Science and Research
Products, Vineland, NJ, USA) and then transferred into 2.0 mL microtubes (Axygen
Scientific, Union city, CA, USA). Triplicate extractions were performed for each
blackberry sample throughout all the experiments.
Analysis of total phenolic content (TPC). TPC was determined by the Folin-
Ciocalteau colorimetric method (28). A 0.5 mL of diluted sample extract and 0.5 mL of
50, 100, 150, or 200 ppm gallic acid (Sigma Chemical Co. MO, USA) solutions were
vortexed with 7.5 mL DI water and 0.5 mL Folin-Ciocalteau reagent (Sigma Chemical
63
Co. MO, USA) in a series of test tubes, respectively, and a 0.5 mL DI water was used as
control. After setting up at room temperature for 10 min, the solutions were mixed with
3 mL of 20% (w/v) Na2CO3 (Sigma Chemical Co. MO, USA) and placed in water bath
at 40 oC for 20 min. Samples taken out of the water bath were immediately cooled to
room temperature in an ice bath for 3 min. The absorbance of the samples and standards
were measured spectrophotometrically (UV160U Shimadzu spectrophotometer, Kyoto,
Japan) at 765 nm. TPC was calculated as mg of gallic acid equivalents (GAE) per gram
fresh weight (FW) or dried weight (DW) of sample.
Analysis of total monomeric anthocyanins. Total monomeric anthocyanins were
quantified by the pH-differential method (29). The sample extract was diluted in both
0.025 M potassium chloride (EMD Chemicals Inc., Gibbstown, NJ, USA) buffer
(pH=1.0) and 0.4 M sodium acetate (EMD Chemicals Inc., Gibbstown, NJ, USA) buffer
(pH=4.5), respectively. After equilibration at room temperature for 15 min, the
absorbance of two dilutions was analyzed by a UV160U spectrophotometer (Shimadzu
Corp., Kyoto, Japan) at both 510 nm and 700 nm. Total monomeric anthocyanins (mg
cyanidin-3-glu equivalent/g FW or DW) were calculated according to the equation:
[( A
510 ]
− A700 ) pH 1.0 − ( A510 − A700 ) pH 4.5 × 449.2 × DF × 1000 / 26900
Analysis of radical scavenging activity. The DPPH (1, 1-diphenyl-2-
picryhydrazyl) assay was carried out to measure the radical scavenging activity of
blackberry samples (30). A 0.5 mL of diluted sample extract and 0.004, 0.01, 0.02,
0.03, or 0.04 mg/mL ascorbic acid (Mallinckrodt Baker Inc., Phillipsburg, NJ, USA)
were mixed with 1 mL of DPPH solvent (Kasel Kogyo Co. Ltd, Tokyo, Japan) in a
64
series of 2.5 mL disposal cuvettes (PLASTIBRAND, Postfach, Germany). The solution
was set at room temperature for 5 min and the absorbance was determined by a
spectrophtometer at 517 nm. The absorbance of ascorbic acid was used as standard, and
radical scavenging activity of samples was reported as mg of ascorbic acid equivalents
(AAE) per gram FW or DW of sample.
Analysis of antioxidant capacity. Oxygen radical absorbance capacity (ORAC)
assay (31) and ferric reducing ability of plasma (FRAP) assay (32) were conducted to
evaluate antioxidant capacity of blackberry samples. To measure ORAC, 30 µl sample
(diluted as necessary) and 0, 10, 20, 40 µM Trolox were placed in pre-warmed plates,
and then 200 µl pre-warmed PE reagent (solution of β-Phycoerythrin) was added into
each plate. After the filled plates were incubated for 1 h at 37 oC, 70 µl AAPH reagent
(2,2’-azobis(2-amidino-propane) dihydrochloride, MW = 271.17) was added into each
plate to trigger the chemical reaction. The changes in PE fluorescent value of samples
were read kinetically and recorded every 2 min and lasted for 2 h. The area of positive
changes in the curve was measured as antioxidant capacity. The ORAC values were
finally expressed as micromoles of Trolox equivalent (TE) per gram of FW or DW
sample. To measure FRAP, 40 µl sample (diluted as necessary) and 0, 62.5, 125, 250,
500 µM Trolox were placed in plates, and a 300 µl of pre-warmed FRAP reagent (30
mL of 300 mM acetate buffer with 3 mL of 10 mM 2,4,6-tri(2-pyridil)-s-triazine
solution and 3 mL of 20 mM FeCl3 solution ) was added into each plate. After the filled
plates were incubated for 15 min at 37 oC, the absorbances of solutions were analyzed at
65
550 nm. The final FRAP values were also reported as micromoles of Trolox equivalent
(TE) per gram of FW or DW sample.
Statistical analysis. The measurements of all the parameters were completed at
every storage time for both blackberry varieties with three replicates. Data analyses
were performed by analysis of variance (ANOVA) and general linear model (GLM)
using SAS statistical software 9.01 (SAS institute, Cary, NC). Multiple comparisons
among the treatments with significant differences tested in ANOVA were conducted by
using LSD (least significant difference) at P<0.05.
RESULTS AND DISCUSSION
Effect of refrigeration storage on physicochemical parameters, bioactive
compounds and antioxidant capacities of blackberries. Fresh ‘Marion’ and
‘Evergreen’ blackberries were able to be stored at 2.0 ± 0.2 oC and 96 ± 2% RH for 9
and 7 days, respectively before significant quality deterioration and decay occurred
(<10% decay). According to the ANOVA results, refrigeration storage significantly
(P<0.05) affected pH, TA, TPC, ACY, and RSA in ‘Marion’, and TSS, TA, TPC, ACY,
ORAC, and FRAP in ‘Evergreen’ (Table 3.1).

66
Table 3.1 ANOVA results of two blackberry varieties (‘Marion’ and ‘Evergreen’) during refrigeration storage (2.0 ± 0.2 oC,
95 ± 2% RH)
Marion
* ns **
Factor pH TSS TA MCns TPC*** ACY*** RSA*** ORACns FRAPns
df 5 5 5 5 5 5 5 5 5
SS 0.10 4.09 1.51 0.0005 694.59 87645.18 948.66 23917.26 180165.32
Storage
MS 0.02 0.82 0.30 0.0001 138.92 17529.04 189.73 4783.45 36033.06
Time
F 3.53 0.69 9.69 6.13 63.21 39.14 17.64 2.63 2.77
Pr>F 0.03 0.64 0.001 0.05 <0.0001 <0.0001 <0.0001 0.08 0.07
Evergreen
ns * ***
Factor pH TSS TA MCns TPC*** ACY*** RSAns ORAC* FRAP***
df 4 4 4 4 4 4 4 4 4
SS 0.32 6.65 0.13 0 308.15 74209.51 7.12 7688.91 2495.93
Storage
MS 0.08 1.66 0.03 0 77.04 18552.38 1.78 1922.23 623.98
Time
F 2.04 3.84 19.69 3.56 52.79 18.46 0.84 4.28 14.84
Pr>F 0.16 0.04 <0.0001 0.05 <0.0001 0.0001 0.53 0.03 <0.001
* =p< 0.05; ** =p< 0.01; *** =p< 0.001; ns= no significance.
TSS: total soluble solids; TA: titratable acidity; MC: moisture content; TPC: total phenolics content; ACY: total monomeric
anthocyanins; RSA: radical scavenging activity; ORAC: oxygen radical absorbance capacity; FRAP: ferric reducing ability
of plasma.
df: degrees of freedom; SS: sum of squares; MS: mean square; F: calculated F-value; Pr: probability.
67
The pH value of ‘Marion’ increased from initial 3.20 to 3.33 while TA declined
from initial 2.50% to 1.58% after 9 days of storage (Figure 3.2). Meanwhile, TA of
‘Evergreen’ decreased 46.15% during the 7 days of storage (P<0.05), from initial 0.52%
to 0.28% (Figure 3.2). These results can be explained that organic acids were
consumed due to fruit respiration and exposure to light during post-harvest storage. TSS
of ‘Marion’ didn’t change (P>0.05) throughout the refrigeration storage, same as the
‘Evergreen’ after 3 days (Figure 3.2), indicating that the sugar gaining during fruit
ripening may compensate for the lost sugar, used as a substrate for fruit respiration (33-
35).
68
5.5
Marion
20
Evergreen Marion Evergreen
5.0
Total Soluble Solids (%)

18
4.5 16
pH
4.0 14
3.5 12
3.0 10
2.5 8
0 1 3 5 7 9 0 1 3 5 7 9
Storage time (Day) Storage time (Day)
3.0 90
Marion Evergreen Marion Evergreen
Titratable acidity (Malic acid,
2.5
Moisture content (%)

85
2.0
g/100g FW)
1.5 80
1.0
75
0.5
0.0 70
0 1 3 5 7 9 0 1 3 5 7 9
Figure 3.2 Changes of selected physicochemical properties of two blackberry varieties (‘Marion’ and ‘Evergreen’) during
refrigeration storage (2.0 ± 0.2 oC, 95 ± 2% RH).
69
Distinctive trends on TPC were observed in two blackberry varieties during
refrigeration storage (Figure 3.3). In ‘Marion’, TPC didn’t change at the 1st day of
storage (57.57-55.27 mg GAE/g DW), rose significantly by the 3rd day (66.38 mg
GAE/g DW), but decreased to 49.38 mg GAE/g DW at the 9th day. In ‘Evergreen’,
TPC was reduced 27.72% at the 3rd day of storage (P<0.05), but no significant change
during the rest of storage, was 22.49 mg GAE/g DW at day 7. These different patterns
may attribute to the diverse constitutions or proportions of phenolic compounds in two
varieties, as well as different polyphenoloxidases activities that are essential to the
phenolics degradation in fruit (33, 35).
ACY showed similar changes as TPC during refrigeration storage (Figure 3.3). In
‘Marion’, ACY decreased from 11.02 to 9.39 mg cyanidin-3-glu equivalent/g DW at the
1st day, but increased at the 2nd day, and then fluctuated afterwards, reached the lowest
number of 9.17 mg cyanidin-3-glu equivalent/g DW at day 9 (P<0.05). ACY in
‘Evergreen’ increased 13.73% after the 1st day of storage (P<0.05), but decreased
continuously during the rest of storage, had a 25% reduction from day 1 to day 7
(P<0.05). According to Kalt et al. (36), ambient temperature at some extent promoted
the formation of anthocyanins, and the stability of anthocyanins was impacted by pH,
light, enzymes and even fruit braisins (37). The ACY in both blackberry varieties
declined at the end of refrigeration storage probably because the elevated pH during
storage accelerated the degradation of anthocyanins and the refrigeration temperature
was too low to accumulate anthocyanins (36).

70
With respect to antioxidant capacity, RSA in ‘Marion’ showed a similar trend as
ACY (Figure 3.4), suggesting that anthocyanins probably had great contribution to
radical scavenging activity against DPPH radicals during refrigeration storage. No
change in RSA for ‘Evergreen’ was observed. No changes in ORAC for ‘Evergreen’
during the first 5 days of storage (278.61-302.26 umol TE/g DW), but a 20.08%
decrease during the last 2 days (day 5 to 7) (P<0.05), while the FRAP values of
‘Evergreen’ increased 13.72% during the 1st day of storage, and continuously increased
during the rest of storage, reached to 241.86 umol TE/g DW at day 7, 18.75% higher
than the value at day 0 (P<0.05) (Figure 3.4). For ‘Marion’, no significant changes
(P>0.05) in ORAC and FRAP were observed during refrigeration storage with a mean
value of 174.59 umol TE/g DW and 757.09 umol TE/g DW, respectively (Figure 3.4).
In previous studies, no significant change in ORAC was reported for blackberries
stored at 2 oC (13), neither for blueberries stored above 10 oC for 8 days (36), but
significant increases in ORAC and anthocyanins were found in strawberries and
raspberries stored at room temperature (36). Therefore, increasing the post-harvest
storage temperature may potentially promote the synthesis of anthocyanins and other
bioactive compounds.
71
90 15
Total monomeric anthocyanins (mg

cyanidin-3-glu equavalent/g DW)
Total phenolics (mg GAE/g DW)
80 Marion Evergreen Marion Evergreen
70 13
60 11
50
40 9
30
7
20
10 5
0 1 3 5 7 9 0 1 3 5 7 9
Figure 3.3 Changes of bioactive compounds in two blackberry varieties (‘Marion’ and ‘Evergreen’) during refrigeration
storage (2.0 ± 0.2 oC, 95 ± 2% RH).
72
120
Radical Scavenging Activity (mg 400
Marion Evergreen Marion Evergreen
100 350
ORAC (umol TE/g DW)

300
80
AAE/g DW)
250
60 200
40 150
100
20
50
0 0
0 1 3 5 7 9 0 1 3 5 7 9
1200
Marion Evergreen
1000
FRAP (umol TE/g DW)
800
600
400
200
0
0 1 3 5 7 9
Storage time (Day)
Figure 3.4 Changes of antioxidant capacities in two blackberry varieties (‘Marion’ and ‘Evergreen’) during refrigeration
storage (2.0 ± 0.2 oC, 95 ± 2% RH).
73
Effect of processing treatments on physicochemical parameters, bioactive
compounds and antioxidant capacities of blackberries. Overall, processing
technologies and post-processing storage at room temperature significantly (P<0.05)
affected the measured physicochemical parameters, bioactive compounds and
antioxidant capacities of both blackberry varieties (Table 3.2).

74
Table 3.2 ANOVA results of two blackberry varieties (‘Marion’ and ‘Evergreen’) after processing and during room
temperature storage (25 ± 2 oC, 30 ± 2% RH)
Marion
Factors pH TSS TA MC TPC ACY RSA ORAC FRAP
df 5 5 5 5 5 5 5 5 5
SS 0.25 28062.13 405.72 6.55 5008.04 3364765.75 4695.68 703547.41 2224544.3
Processing Technologies MS 0.05 5612.43 81.14 1.31 1001.61 672953.15 939.14 140709.48 444908.86
F 16.68 2097.17 137.74 64354 432.77 855.46 220.83 188.83 236.99
Pr＞F <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001
df 2 2 2 2 2 2 2 2 2
SS 0.002 14.59 5.32 0.0001 64.06 50233.8 1973.46 40475.79 21145.41
Storage Time MS 0.0008 7.30 2.66 0.00007 32.03 25116.9 986.73 20237.9 10572.71
F 0.26 2.73 4.51 3.55 13.84 31.93 232.02 27.16 5.63
Pr＞F 0.77 0.08 0.02 0.04 <0.0001 <0.0001 <0.0001 <0.0001 0.007
Evergreen
Factors pH TSS TA MC TPC ACY RSA ORAC FRAP
df 5 5 5 5 5 5 5 5 5
SS 8.43 29620.33 24.57 7.04 3402.16 4547562.25 2791.44 441475.56 665227.35
Processing Technologies MS 1.69 5924.07 4.91 1.41 680.43 909512.45 558.29 88295.11 133045.47
F 36.57 1332.39 26.05 30416.3 210.39 1427.04 75.49 98.9 58.92
Pr＞F <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001
df 2 2 2 2 2 2 2 2 2
SS 0.22 27.98 0.58 0.0001 38.93 44679.92 2126.38 16338.07 22009.96
Storage Time MS 0.11 13.99 0.29 0.00005 19.47 22339.96 1063.19 8169.03 11004.98
F 2.43 3.15 1.53 1.12 6.02 35.05 143.75 9.15 4.87
Pr＞F 0.10 0.06 0.23 0.34 0.006 <0.0001 <0.0001 0.0006 0.013
of plasma. df: degrees of freedom; SS: sum of squares; MS: mean square; F: calculated F-value; Pr: probability.
75
Comparing with frozen samples, pH increased in hot-air dried (8%) and canned
‘Marion’ (4% and 9% in canned-in-water and canned-in-syrup, respectively), and TA
decreased in processed ‘Marion’, with the greatest loss (75%) in jam, followed by
canned-in-syrup (69%), canned-in-water (42%), hot-air dried (29%), and freeze dried
(15%) (Figure 3.5). Whereas no significant changes in TA and pH of hot-air dried and
canned ‘Evergreen’, a 53% increase in TA of freeze dried ‘Evergreen’ was observed.
Besides, jam processing of ‘Evergreen’ resulted in a 32% decrease and 60% increase in
pH and TA, respectively, probably due to the addition of citric acid during jam
processing.
Blackberry jams had 460% and 400% higher TSS than frozen ‘Marion’ and
‘Evergreen’ respectively (Figure 3.5), right after processing due to the addition of sugar
and water evaporation during cooking. Drying removed most of the water from fresh
berries, resulting in 340% and 424% increase in TSS of freeze-dried ‘Marion’ and
‘Evergreen’, and 306% and 189% increase in hot-air dried ‘Marion’ and ‘Evergreen’,
respectively (Figure 3.5). Canned-in-water ‘Marion’ and ‘Evergreen’ had 38% and
53% lower TSS than frozen samples, respectively, possibly due to the diffusion of water
from canning solution to the fruit and leaching of the solutes from fruit to the solution,
which was confirmed by the higher moisture content of canned-in-water blackberries
compared to frozen samples (Figure 3.5). The combined effects of water diffusion and
sugar addition in canned-in-syrup products led to a 39% increase in TSS for ‘Marion’,
but no significant change in TSS for ‘Evergreen’ was observed (P>0.05).

76
Marion Evergreen
5.0
0 month 3 month 6 month 5.0 0 month 3 month 6 month
4.5
4.5
4.0
4.0
pH
3.5
pH
3.5
3.0 3.0
2.5 2.5
2.0 2.0
FC FD HD CW CS JAM FC FD HD CW CS JAM
Marion Evergreen

15 15
0 month 3 month 6 month 0 month 3 month 6 month
12 12
g/100g DW)
g/100g DW)
9 9
6 6
3 3
0
0
FC FD HD CW CS JAM
FC FD HD CW CS JAM
Figure 3.5 Physicochemical properties of two blackberry varieties (‘Marion’ and ‘Evergreen’) after processing and during
room temperature storage (25 ± 2 oC, 30 ± 2% RH) (FC: Frozen control; FD: freeze dried blackberries; HD: Hot-air dried
blackberries; CW: Canned-in-water blackberries; CS: Canned-in-sucrose syrup blackberries; Jam: Blackberry jam).
77
Marion Evergreen
0 month 3 month 6 month

Total soluble solids (%)
80 0 month 3 month 6 month
Total soluble solids (%)

80
60 60
40 40
20 20
0 0
Marion Evergreen
100 0 month 3 month 6 month 100 0 month 3 month 6 month

80 80
60 60
40 40
20 20
0 0
Figure 3.5 Physicochemical properties of two blackberry varieties (‘Marion’ and ‘Evergreen’) after processing and during
blackberries; CW: Canned-in-water blackberries; CS: Canned-in-sucrose syrup blackberries; Jam: Blackberry jam)
(Continued).
78
Processing technologies showed different effects on the bioactive compounds of the
two blackberry varieties (Figure 3.6). Freeze drying, recognized as the best technology
to make high-quality dried products (38), increased TPC of ‘Marion’ and ‘Evergreen’
27% and 21% on a dry weight basis, respectively. Freeze drying also increased ACY
5.5% in ‘Evergreen’, but decreased ACY 25% in ‘Marion’. Although the enzymes in
blackberries were inactivated by freezing at low temperature, they may recover during
drying at 35 oC shelf temperature, thus accelerating the degradation of monomeric
anythocyanins and the polymerization with tannins, polysaccharides and other
flavonoids (39, 40), which may explain the observed losses of ACY in ‘Marion’. The
destruction of anthocyanins in freeze dried ‘Marion’ was not as high as that in hot-air
dried fruit because the low temperature and vacuum in freeze drying protected the fruit
phenolics from oxidation. On the other hand, fruit cell destruction during freezing and
ice sublimation contributed to the extraction of anthocyanins (19), which possibly led to
the observed increase (5.5%) in anthocyanins in ‘Evergreen’ and explained the higher
TPC in freeze dried blackberries than that in frozen controls. Moreover, different
compositions of anthocyanins in the two blackberry varieties could be another plausible
reason for the varying contents of anthocyanins observed on freeze dried samples (41).
In contrast, hot-air drying promoted the oxidation and condensation of phenolic
compounds (19), resulting in 56% and 84% reduction in ACY of ‘Marion’ and
‘Evergreen’, respectively (Figure 3.6). The higher loss of TPC (37%) was found in hot-
air dried ‘Evergreen’ than in ‘Marion’, probably due to the smaller size and larger
surface area of ‘Evergreen’ than those of ‘Marion’.

79
Canned-in-water fruit had 47% and 35% reduction in ACY in ‘Marion’ and
‘Evergreen’, respectively, and 31% decrease in TPC in ‘Evergreen’ compared with
frozen controls, while canned-in-syrup had 36% and 54% TPC reduction in ‘Marion’
and ‘Evergreen’, respectively, and 70% ACY reduction in both varieties (Figure 3.6).
These results may be attributed to the thermal degradation of ACY and preferential
dissolution of ACY into canning solution (23). Anthocyanins are the predominant
phenolic compounds in blackberries, it is expected that TPC had a similar trend as
anthocaynins. However, less deduction of TPC than that of ACY in both varieties was
observed, probably due to the undergoing polymerization of monomeric anthocyanins
which produced polymers detected as phenolics and consumed large amount of
monomeric anthocyanins during canning process (23).
Blackberry jams showed the lowest TPC and ACY among all processed products,
only 33% of TPC and 20% of ACY retained comparing with frozen controls (Figure
3.6). This may be explained as condensation and polymerization of phenolic
compounds were accelerated after cellular crush and long-time cooking of the fruit
under the exposure to oxygen and heat during jam processing (25).
80
Marion Evergreen

40 40
30 30
20 20
10 10
0 0
Marion Evergreen
Anthocyanins (mg cyanidin-3-glu
Anthocyanins (mg cyanidin-3-glu

10
0 month 3 month 6 month 10 0 month 3 month 6 month
8
equivalent/g DW)
equavalent/g DW)
8
6 6
4 4
2 2
0 0
Figure 3.6 Bioactive compounds of two blackberry varieties (‘Marion’ and ‘Evergreen’) after processing and during room
temperature storage (25 ± 2 oC, 30 ± 2% RH) (FC: Frozen control; FD: freeze dried blackberries; HD: Hot-air dried
81
Different processing technologies showed different impacts on the antioxidant
capacities of the blackberries (Figure 3.7). Freeze drying didn’t change RSA, ORAC
and FRAP of the fruit except 14% increase in RSA of ‘Evergreen’. Hot-air dry resulted
in 37% and 27% decrease in ORAC and FRAP of ‘Marion’, respectively, and 13%
decrease in RSA of ‘Evergreen’. Antioxidant capacity was also remarkably reduced in
canned-in-syrup fruit with 31% loss in RSA and 50-60% loss in ORAC of both varieties
and 58% loss in FRAP of ‘Marion’. Canning-in-water increased RSA of ‘Marion’ 22%,
decreased ORAC and FRAP of ‘Marion’ 34% and 21%, respectively, whereas increased
RSA and FRAP of ‘Evergreen’ 8% and 110%, respectively. Although the degradation
of phenolics theoretically diminishes antioxidant capacity of heat-treated blackberries,
some polymers formed through Maillard reaction during thermal process may possibly
compensate or even increase the ability against certain type of radicals (42). Similar to
TPC and ACY, the greatest loss of antioxidant capacity was detected in jams with about
20% RSA, 20%-30% ORAC, and 30-35% FRAP remained in finished jams compared
to frozen controls.
82
Marion Evergreen
Radical scavenging activity
Radical scavenging activity

60 0 month 3 month 6 month 0 month 3 month 6 month
60
(mg AAE/g DW) 50
(mg AAE/g DW)

50
40 40
30 30
20 20
10 10
0 0
Marion
Evergreen

ORAC (umol TE/g DW)
ORAC (umol TE/g DW)

400 400
300 300
200 200
100 100
0 0
Figure 3.7 Antioxidant capacities of two blackberry varieties (‘Marion’ and ‘Evergreen’) after processing and during
83
Marion Evergreen
FRAP (umol TE/g DW)

FRAP (umol TE/g DW)
800 800
600 600
400 400
200 200
0 0
Figure 3.7 Antioxidant capacities of two blackberry varieties (‘Marion’ and ‘Evergreen’) after processing and during room
temperature storage (25 ± 2 oC, 30 ± 2% RH) (FC: Frozen control; FD: freeze dried blackberries; HD: Hot-air dried
blackberries; CW: Canned-in-water blackberries; CS: Canned-in-sucrose syrup blackberries; Jam: Blackberry jam)
(Continued).
84
Effect of post-processing storage on physicochemical parameters, bioactive
compounds and antioxidant capacities of blackberries. During post-process room
storage at 25 ± 2 oC and xx RH, significant changes in pH were only observed in jams
and canned-in-syrup fruit in both blackberry varieties (Figure 3.5). The pH of the jams
increased by 6% and 10% in ‘Marion’ and ‘Evergreen’, respectively, while the pH of
canned-in-syrup blackberries rose by 9% in ‘Evergreen’, fell by 3.5% in ‘Marion’ at the
end of 6 month storage. A 16% decrease in TA and 20% increase in TSS of ‘Marion’
over 6 months of frozen storage at -23 oC were observed (Figure 3.5), possibly caused
by the temperature fluctuation in the freezer and oxidation of the fruit. After 6 months
of room storage, TA of ‘Marion’ and ‘Evergreen’ jams lost 9% and 12%, respectively,
TSS in canned-in-water and canned-in-sugar ‘Marion’ decreased by 5% and increased
by 4%, respectively (Figure 3.5), as a result of water and sugar penetrated into the fruit,
and TSS of hot-air dried ‘Marion’ was down 17%, while up 20% in freeze dried
‘Marion’. No significant changes in TA of canned and dried fruits (both varieties) were
observed during room storage (Figure 3.5). Moisture content of canned blackberries
slightly increased over 6-month of room storage, by 1.1% for canned-in-water samples
for both varieties and 1.3% for canned-in-syrup ‘Marion’ (P<0.05) attributing from
gradual water penetration to canned fruit immersed in the syrup. Moisture content in
dried samples was continuously lost by 13% in hot-air dried ‘Marion’ and 35% in hot-
air dried ‘Evergreen’, and 15% in freeze-dried ‘Evergreen’ at the end of 6-month
storage (Figure 3.5), suggesting the water loss of samples through package.
85
Canned fruit (both varieties) showed increased TPC during room storage, where
TPC of canned-in-water ‘Evergreen’ increased 40% and that of canned-in-syrup
‘Marion’ increased 22% at the end of 6-month storage (Figure 3.6). This is partly
because of the destroyed fruit cellular structure after long-term immersing in canning
solution, making the phenolics extraction easier (25, 43). At the end of 6-month room
storage, ACY was significantly destroyed in all thermally processed ‘Marion’ and
‘Evergreen’, including jams (56%, 66%), canned-in-water (21%, 57%), canned-in-syrup
(42%, 53%), and hot-air dried (48%, 24%) (Figure 3.6), as a result of ACY’s
condensation and complexion with other chemical compounds accelerated by heat-
stable peroxidase in shelf-stable products under room temperature (23).
RSA fell down significantly in all blackberry samples except jams, 39% and 82%
reduction in frozen, 29% and 53% in freeze dried, 47% and 86% in hot-air dried, 62%
and 24% in canned-in-water, and 49% and 32% in canned-in-syrup ‘Marion’ and
‘Evergreen’ after 6-month room storage, respectively (Figure 3.7). These results could
be explained by the bioactive compounds against DPPH free radicals were possibly
sensitive to oxygen and gradually leached into the canning solution during room
storage. ORAC decreased 13% and 45% in hot-air dried, and 68% and 38% in canned-
in-water ‘Marion’ and ‘Evergreen’, respectively (Figure 3.7). Finally, there was 18%
decrease in FRAP of frozen ‘Marion’, 57% loss in ‘Marion’ jams, 34% decrease in
canned-in-syrup ‘Evergreen’, whereas 28% increase in canned-in-water ‘Marion’ and
17% increase in freeze dried ‘Evergreen’ (Figure 3.7).

86
Comparison of our results about fresh blackberries with previously reported
data. Table 3.3 reported the mean values of physicochemical parameters and bioactive
compounds of two blackberry varieties prior to cold storage.

87
Table 3.3 Physicochemical properties and contents of bioactive compounds in two blackberry varieties (‘Marion’ and
‘Evergreen’) at harvest
Variety pH TSS TA MC TPC ACY RSA ORAC FRAP
3.16 12.85 2.45 0.82 57.57 11.02 90.33 231.57 819.46
Marion
±0.05 ±1.07 ±0.22 ±0.00 ±1.64 ±0.22 ±2.37 ±32.93 ±7.61
4.20 17.85 0.52 0.75 31.46 7.58 17.24 297.87 203.66
Evergreen
±0.01 ±0.65 ±0.05 ±0.00 ±0.67 ±0.18 ±2.99 ±4.81 ±6.05
Data are expressed as means ± standard deviation (n=3).
TSS: total soluble solids, %; TA: titratable acidity, g malic acid/100g FW; MC: moisture content, %;
TPC: total phenolics content, mg GAE(Gallic acid equivalent/g DW ); ACY: total monomeric anthocyanins, mg cyanidin-3-
glu equivalent/g DW; RSA: radical scavenging activity, mg AAE(Ascorbic acid equivalent)/g DW; ORAC: oxygen radical
absorbance capacity, umol TE (Trolox equivalent)/g DW; FRAP: ferric reducing ability of plasma, umol TE (Trolox
equivalent)/g DW.
88
The pH and TSS of ‘Marion’ (3.16, 12.85 %) and ‘Evergreen’ (4.40, 17.85%)
(Table 3.3) were in good agreement with those of ripe ‘Marion’ and ‘Evergreen’
reported by Reyes-Carmona et al. (44) and Siriwoharn et al. (8) (3.13 and 13.5%, and
4.28 and 15.7 %, respectively), while our TSS of ‘Evergreen’ was much higher than that
in Greece (11.5 %) (45). TA of ‘Marion’ (2.45%) reported as percent of malic acid
equivalent in this study (Table 3.3) was significantly higher than 1.28% for ripe
‘Marion’, similar to 2.32% for under-ripe ‘Marion’ unveiled as percent of citric acid
equivalent reported by Siriwoharn et al. (8), and our TA of ‘Evergreen’ (0.52% malic
acid equivalent) was close to 0.47% citric acid equivalent for overripe fruit reported by
the same author (8).
TPC in ‘Marion’ at harvest, 10.35 mg GAE/g FW or 57.57 mg GAE/g DW (Table
3.3), was consistent with the values of 10.05 mg GAE/g FW for blackberries harvested
in 2002 (8), but higher than 8.44 mg GAE/g FW for the same variety of fruit harvested
in 1999 (9). However, the mean TPC of ‘Evergreen’, 7.97 mg GAE/g FW or 31.46 mg
GAE/g DW was nearly 20% lower than that reported by Siriwoharn et al. (8), 9.60 mg
GAE/g FW for ripe fruit, but 50% higher than that measured in Greece in ‘Evergreen’
(2007) at 20.61 mg GAE/g DW (45).
ACY of ‘Marion’ at the harvest from this study (1.98 mg cyd-3-glu equivalent/g FW
or 11.02 mg cyd-3-glu equivalent/g DW) (Table 3.3) was slightly lower than that of
‘Marion’ reported by Siriwoharn et al. (8, 9), 2.21 and 2.25 mg cyd-3-glu equivalent/g
FW, while ‘Evergreen’ (1.92 mg cyd-3-glu equivalent/g FW or 7.58 mg cyd-3-glu

89
equivalent/g DW) had higher value than that of the same variety reported by Siriwoharn
et al. (8, 9), 1.31 and 1.54 mg cyd-3-glu equivalent/g FW.
ORAC in ‘Marion’ at the harvest, 41.67 umol TE/g FW or 231.57 umol TE/g DW
(Table 3.3), was about two third of that reported by Siriwoharn et al. (8), 60.9 umol
TE/g FW for ripe fruits, but still significantly higher than 20.3-24.6 umol TE/g FW of
fruit juices from other varieties (‘Chester thornless’, ‘Hull thornless’, ‘Triple Crown’)
(46). ORAC in ‘Evergreen’ at harvest, 75.43 umol TE/g FW or 297.87 umol TE/g DW
(Table 3.3), was significantly higher than that in ‘Marion’ in this study, and coincided
with the number reported by Siriwoharn for the same variety (8), 75.5 umol TE/g FW.
As opposite to ORAC values, FRAP of ‘Evergreen’ (51.57 umol TE/g FW or
203.66 umol TE/g DW) (Table 3.3) was significantly lower than that of ‘Marion’
(147.38 umol TE/g FW, 819.46 umol TE/g DW) (Table 3.3), and both of them ran out
of the range of 63.5-97.3 umol TE/g FW in several selected varieties of blackberries
reported by Siriwoharn et al. (8).
The physicochemical properties and bioactive compounds of fresh blackberries are
influenced by the fruit variety, genotype, maturity stage, harvest location and time,
environmental condition and practice technology of cultivation, extraction procedures
and solvents, and even the testing assays based on distinctive mechanisms. For
example, ORACPE is considered to underestimate antioxidant capacity compared to
ORACFL because of the binding effect between B-PE and polyphenolics (47, 48). This
may explain the ORACPE values of ‘Marion’ and ‘Evergreen’ from this study (231.57
umol TE/g DW and 297.87 umol TE/g DW, respectively) were significantly lower than
90
the ORACFL values of the same varieties harvested in Oregon (423.0 umol TE/g DW
and 366.8 umol TE/g DW) reported by Reyes-Carmona et al. (44).
Comparison of our results about processed fruit with previously reported data.
Total monomeric ACY of frozen (2.48 mg/g FW), canned-in-syrup (2.21 mg/g FW) and
canned-in-water ‘Apache’ blackberries (2.04 mg/g FW) right after processing reported
by Hager et al. (23) were significantly higher than those of ‘Marion’ (1.42, 0.54, 0.52
mg/g FW) and ‘Evergreen’ blackberries (1.32, 0.47, 0.61 mg/g FW) from this study
(Figure 3.6). Canning process resulted in about 60% loss of ACY in ‘Marion’ and
‘Evergreen’ while minor loss in ‘Apache’ blackberries. Throughout 6-month storage,
total monomeric ACY was stable in both frozen ‘Marion’ (1.42-1.42 mg/g FW) and
‘Evergreen’ (1.30-1.33 mg/g FW) (Figure 3.6), whereas degraded 25%-44% in canned
‘Marion’ and 52%-62% in ‘Evergreen’ (Figure 3.6), which was consistent with data
reported by Hager et al. (23), in which were 65.8% and 60.6% losses in canned-in-syrup
and canned-in-water ‘Apache’ blackberries, respectively. Similar to ACY, ORAC of
frozen (97.2 umol TE/g FW), canned-in-syrup (75.8 umol TE/g FW), and canned-in-
water ‘Apache’ blackberries (71.1 umol TE/g FW) right after processing (23) were
significantly higher than those of ‘Marion’ (70.17, 34.65, 32.42 umol TE/g FW) and
‘Evergreen’ blackberries (54.88, 31.95, 43.67 umol TE/g FW) from this study (Figure
3.7). Significant losses of ORAC were detected in canned-in-water ‘Marion’ (68%) and
‘Evergreen’ (38%) (Figure 3.7), but not in ‘Apache’ blackberries (23) at the end of
room temperature storage.

91
Our results revealed that the initial TPC of frozen ‘Marion’ blackberries was about
5.00 mg GAE/g FW and showed comparable amount to hot-air dried samples (Figure
3.6). However, TPC of frozen ‘Marion’ (4.00 mg GAE/g FW) reported by Asami et al.
(19) was lower than our results and the value decreased significantly after hot-air
drying.
Comparison between ‘Marion’ and ‘Evergreen’ blackberries. Two blackberry
varieties evaluated in this study showed statistical differences in all measured
physicochemical and nutraceutical parameters (Table 3.4).

92
Table 3.4 Mean comparison of physicochemical parameters, bioactive compounds and antioxidant capacities of two
blackberry varieties (‘Marion’ and ‘Evergreen’) throughout refrigeration storage (2.0 ± 0.2 oC, 95 ± 2% RH)
3.19 13.18 2.16 0.82 59.67 10.06 79.58 174.59 757.09
Marion
±0.10 b ±1.04 b ±0.33 a ±0.01 a ±6.51 a ±0.74 a ±7.96 a ±51.90 b ±140.61 a
4.42 16.91 0.39 0.76 27.19 7.61 16.65 283.73 227.90
Evergreen
±0.23 a ±0.89 a ±0.10 b ±0.01 b ±4.80 b ±0.78 b ±1.42 b ±29.50 a ±14.43 b
Data are expressed as means ± standard deviation (n=3);
Values with the same letters are not significant different (LSD, p< 0.05).
TSS: total soluble solids, %; TA: titratable acidity, g malic acid/100g FW; MC: moisture content, %;
TPC: total phenolics content, mg GAE (Gallic acid equivalent/g DW); ACY: total monomeric anthocyanins, mg cyanidin-3-
glu equivalent/g DW; RSA: radical scavenging activity, mg AAE (Ascorbic acid equivalent)/g DW; ORAC: oxygen radical
equivalent)/g DW.
93
‘Marion’ had significantly (P<0.05) higher TA, MC, TPC, ACY, RSA and FRAP
values than those of ‘Evergreen’, but lower pH, TSS, and ORAC values. The pH and
TSS of ‘Evergreen’ was about 130% higher than those of ‘Marion’, while TA of
‘Marion’ was about 500% higher than that of ‘Evergreen’ (Table 3.4). These
differences can lead to the different flavors of the two varieties: ‘Marion’ tasted sour,
while ‘Evergreen’ was sweet. More seeds in ‘Evergreen’ would be another possible
reason for its high TSS value (9). According to Mitcham (49), Mazza and Miniati (50),
the lower pH that is favorable to form and stabilize ACY is probably responsible for the
higher ACY content in ‘Marion’ than that in ‘Evergreen’. The higher ACY content of
‘Marion’ was corresponded to its higher TPC than those of ‘Evergreen’, which
confirmed the results by Siriwoharn and Wrolstad (9). Scavenging activity against
DPPH radicals and FRAP in plasma in ‘Marion’ were about 500% and 300% higher
than those in ‘Evergreen’, respectively, possibly because ‘Marion’ contains more
antioxidants, such as ascorbic acid that is capable of reducing DPPH radicals and ferric
at low pH (51). Whereas, the smaller fruit size and larger surface area of ‘Evergreen’
probably resulted in 160% higher ORAC than those of ‘Marion’ (52). Last but not least,
slightly higher moisture content was observed in ‘Marion’ (82%) than that in
‘Evergreen’ (76%), making ‘Marion’ juicier (Table 3.4).
The means of all parameters measured in all the processed blackberry products were
compared statistically (Table 3.5).

94
Table 3.5 Mean comparison of physicochemical parameters, bioactive compounds, and antioxidant capacities of processed
blackberry (‘Marion’ and ‘Evergreen’) during room temperature storage (25 ± 2 oC, 30 ± 2% RH)
3.23 33.49 6.54 0.49 27.33 4.01 27.52 240.30 513.86
Marion
±0.10 b ±23.21 a ±2.89 a ±0.35 a ±9.99 a ±2.56 a ±12.08 a ±123.64 a ±217.44 a
4.12 31.97 2.00 0.48 23.23 3.52 18.20 209.51 301.91
Evergreen
±0.45 a ±23.74 a ±0.79 b ±0.36 a ±8.67 b ±3.00 a ±10.85 b ±101.59 a ±131.03 b
Data are expressed as means ± standard deviation (n=3);
Values with the same letters are not significant different (LSD, p< 0.05).
TSS: total soluble solids, %; TA: titratable acidity, g malic acid/100g DW; MC: moisture content, %;
TPC: total phenolics content, mg GAE (Gallic acid equivalent/g DW); ACY: total monomeric anthocyanins, mg cyanidin-3-
glu equivalent/g DW; RSA: radical scavenging activity, mg AAE (Ascorbic acid equivalent)/g DW; ORAC: oxygen radical
equivalent)/g DW.
95
‘Marion’ had significantly lower pH and higher TA than ‘Evergreen’ after
processing and during room temperature storage (P<0.05) (Table 3.5). TSS and MC of
processed two blackberry variety products didn’t show differences as seen in fresh fruit
during refrigeration storage (Table 3.4). Means of TPC, RSA, and FRAP in ‘Marion’
products over room storage were nearly 46%, 35%, and 68% of those in fresh ‘Marion’
over refrigeration storage, but still showed significantly (P<0.05) higher values than that
of processed ‘Evergreen’ products (Table 3.5). There were no significant differences
between two varieties in means of ACY and ORAC of processed blackberry products
observed over room storage, but about 40% and 50% of ACY in fresh ‘Marion’ and
‘Evergreen’ were remained in processed ‘Marion’ and ‘Evergreen’ products,
respectively over room storage (Table 3.5).
Correlation among measured physicochemical parameters, bioactive
compounds and antioxidant capacities. Correlation coefficients among the measured
physicochemical parameters, bioactive compounds, and antioxidant capacities in two
blackberry varieties during refrigeration and room temperature storage are presented in
Tables 3.6 and 3.7, respectively.

96
Table 3.6 Correlation coefficients for physicochemical parameters and bioactive compounds measured in two blackberry
varieties (‘Marion’ and ‘Evergreen’) during refrigeration storage (2.0 ± 0.2 oC, 95 ± 2% RH)
pH TSS TA MC TPC ACY RSA ORAC FRAP
pH 1 0.85*** -0.97*** -0.94*** -0.95*** -0.87*** -0.96*** 0.75*** -0.91***
TSS - 1 -0.91*** -0.87*** -0.84*** -0.79*** -0.89*** 0.71*** -0.9***
TA - - 1 0.95*** 0.95*** 0.89*** 0.97*** -0.74*** 0.94***
MC - - - 1 0.95*** 0.91*** 0.97*** -0.80*** 0.9***
TPC - - - - 1 0.91*** 0.94*** -0.76*** 0.9***
ACY - - - - - 1 0.9*** -0.63*** 0.82***
RSA - - - - - - 1 -0.77*** 0.92***
ORAC - - - - - - - 1 -0.65***
FRAP - - - - - - - - 1
of plasma.
97
Table 3.7 Correlation coefficients for physicochemical parameters and bioactive compounds measured in processed
blackberry (‘Marion’ and ‘Evergreen’) during room temperature storage (25 ± 2 oC, 30 ± 2% RH)
pH TSS TA MC TPC ACY RSA ORAC FRAP
pH 1 -0.27** -0.71*** 0.12 0.0003 0.04 -0.19 0.01 -0.31**
TSS - 1 0.03 -0.86*** -0.23* -0.12 -0.22* -0.07 -0.36***
TA - - 1 -0.06 0.53*** 0.48*** 0.58*** 0.55*** 0.77***
MC - - - 1 0.007 0.14 0.12 -0.05 0.17
TPC - - - - 1 0.68*** 0.62*** 0.70*** 0.73***
ACY - - - - - 1 0.49*** 0.86*** 0.49***
RSA - - - - - - 1 0.66*** 0.71***
ORAC - - - - - - - 1 0.63***
FRAP - - - - - - - - 1
of plasma.
98
Results showed that TSS had positive correlation to pH (r=0.85), but negative to
TA (r= -0.91) (Table 3.6), demonstrating the typical physicochemical changes with acid
converting to sugar in fruit during post-harvest storage (33). TA was positively, while
pH was negatively correlated to ACY, TPC, RSA and FRAP of fresh blackberries
during refrigeration storage (Table 3.6), and TA also correlated to TPC (r=0.53), RSA
(r=0.58), ORAC (0.55) and FRAP (0.77) of processed blackberry products during room
temperature storage (Table 3.7), indicating the low pH and high acidic environment
stabilizing ACY. Moisture content of processed blackberry products was negatively
related to TSS during room temperature storage (Table 3.7), which could be explained
by the loss of water corresponded with increase of soluble solids in the fruits.
FRAP, RSA and TPC were positively correlated in both fresh and processed
blackberries because all these 3 assays were based on the electron transfer reaction to
quantify the reduction capacity of samples against Ferric (Fe Ш), DPPH radical and F-C
reagent, respectively. The strong correlation between TPC and RSA, TPC and FRAP of
both fresh and processed blackberries also confirmed that phenolic compounds
including phenolic acid, flavonoids, and tannins are the major sources of antioxidants in
blackberries (48). Moreover, the linear correlations between ACY and TPC (r=0.91),
RSA (r=0.90) and FRAP (r=0.82) in fresh blackberries (Table 3.6), ACY and ORAC
(r=0.86) in processed blackberries were found (Table 3.7), which can be explained as
ACY is the major contribution to the total phenolics, providing antioxidant capacity in
fresh blackberries (48).

99
Finally, moderately positive correlation between ORAC and FRAP (r=0.63) was
observed in processed blackberry products over room temperature storage (Table 3.7),
but not for fresh blackberry during refrigeration storage. Although both ORAC and
FRAP are widely acceptable methods to measure antioxidant capacity of berries, ORAC
assay basically relies on hydrogen atom transfer reaction to evaluate the capacity of
donating hydrogen against peroxyl radicals other than reducing power which FRAP
assay relies on (47). The sensitivity of antioxidants for specific assay, various
compositions or proportions of antioxidants in two varieties of blackberries and
interactions among several types of antioxidants may partly induce different ORAC and
FRAP values used for evaluating antioxidant capacity (53). ORAC values showed no
significant changes in ‘Marion’ during refrigeration storage, which rendered the narrow
ORAC data range for ‘Marion’ (Figure 3.4). Therefore, it is difficult to identify the
relationship between ORAC and FRAP in this study.
ACKNOWLEDGEMENTS
The authors acknowledge Scenic Fruit Co. (Gresham, OR, USA) for donating ‘Marion’
blackberries, Mr. Brian Yorgey for helping make canned blackberries and blackberry
jams, Mr. Jeff Clawson for running the driers, and Ms. Deborah Hobbs for the analysis
of ORAC and FRAP values of the samples.

100
LITERATURE CITED
(1) Seeram, N.P. Berry fruits: compositional elements, biochemical activities, and the
impact of their intake on human health, performance, and disease. J. Agric. Food
Chem. 2008, 56, 627-629.
(2) Liu, S.; Manson, J.E.; Lee, I.M; Cole, S.R.; Hennekens, C.H.; Willett, W.C.;
Buring, J.E. Fruit and vegetable intake and risk of cardiovascular disease: the
women’s health study. Am. J. Clin. Nutr. 2000, 72, 922-928.
J. Agric. Food Chem. 2008, 56, 630-635.
(6) Kraft, T.F.B.; Dey, M.; Rogers, R.B.; Ribnicky, D.M.; Gipp, D.M.; Cefalu, W.T.;
Raskin, I.; Lila, M.A. Phytochemical composition and metabolic performance-
enhancing activity of dietary berries traditionally used by native North Americans.
J. Agric. Food Chem. 2008, 56, 654-660.
(7) Liu, R.H. The potential health benefits of phytochemicals in berries for protecting
199.
8030.
J. Agric. Food Chem. 2008, 56, 661-668.
101
(11) Strik, B.C. Berry crops: Worldwide area and production systems. In Berry fruit:
value-added products for health promotion; Zhao, Y., Eds.; CRC press: New York,
2007; Chapter 1, pp 9-11.
increase antioxidant levels. Acta Horticulture 2002, 585, 521-524.
quality 1999, 22, 535-544.
(15) Antunes, L.E.C.; Duarte, J.; De Souza, C.M. Postharvest conservation of blackberry
(16) Rickman, J.C.; Barrett, D.M.; Bruhn, C.M. Review: Nutritional comparison of fresh,
frozen and canned fruits and vegetables. Part 1. Vitamins C and B and phenolic
compounds. J. Sci. Food Agric. 2007, 87, 930-944.
(17) Zhao, Y. Freezing process of berries. In Berry fruit: value-added products for health
(18) Figuerola, F.E. Dehydration of berries. In Berry fruit: value-added products for
333.
(19) Asami, D.K.; Hong, Y.; Barrett, D.M.; Mitchell, A.E. Comparison of the total
phenolic and ascorbic acid content of freeze-dried and air-dried Marionberry,
strawberry, and corn grown using conventional, organic, and sustainable agricultural
practices. J. Agric. Food Chem. 2003, 51, 1237-1241.
(20) Kwok, B.H.L.; Hu, C.; Durance, T.; Kitts, D.D. Dehydration techniques affect
phytochemical contents and free radical scavenging activities of Saskatoon berries
(Amelanchier alnifolia Nutt.). Food Sci. 2004, 69, S122-S126.
(21) Ramaswamy, H.S.; Meng, Y. Commercial canning of berries. In Berry fruit: value-
added products for health promotion; Zhao, Y., Eds.; CRC press: New York, 2007;
Chapter 12, pp 336-361.
(22) Ngo, T.; Wrolstad, R.E.; Zhao, Y. Color quality of Oregon strawberries-Impact of
102
(23) Hager, T.J.; Howard, L.R.; Prior, R.L. Processing and storage effects on monomeric
(24) Figuerola, F.E. Berry jams and jellies. In Berry fruit: value-added products for
385.
(25) Häkkinen, S.H.; Kärenlampi, S.O.; Mykkänen, H.M.; Tőrrőnen, A.R. Influence of
domestic processing and storage on flavonol contents in berries. J. Agric. Food
Chem. 2000, 48, 2960-2965.
(26) Amakura, Y.; Umino, Y.; Tsuji, S.; Tonogai, Y. Influence of jam processing on the
radical scavenging activity and phenolic content in berries. J. Agric. Food Chem.
2000, 48, 6292-6297.
(27) Rodriguez-Saona, L.E.; Wrolstad, R.E. Extraction, isolation and purification of

anthocyanins. In Current Protocols in Food Analytical Chemistry; John Wiley &
Sons: New Jersy, 2001; F1.1.1-F1.1.11.
(28) Singleton, V.L.; Rossi, J.A. Colorimetry of total phenolics with phosphomolybdic -
phosphotungstic acid reagents. Ameri. J Enol. Viticult. 1965, 16, 144-158.
(29) Giusti, M.M.; Wrolstad, R.E. Characterization and measurement of anthocyanins by

UV-Visible spectroscopy. In Current Protocols in Food Analytical Chemistry; John
Wiley & Sons, Inc.: New Jersy, 2001; F1.2.1-F1.2.13.
(30) Brand-Williams, W.; Cuvelier, M.E.; Berset, C. Use of a free radical method to
evaluate antioxidant activity. Lebensmittel-Wissenschaft Technol. 1995, 28, 25-30.
(31) Cao, G.; Alessio, H.M.; Cutler, R.G. Oxygen-radical absorbance capacity assay for
antioxidants. Free Radic Biol Med. 1993, 14, 303-311.
(32) Benzie, I. F.F.; Strain, J.J. The ferric reducing ability of plasma (FRAP) as a
measure of “Antioxidant Power”: The FRAP Assay. Analytical Biochem. 1996, 239,
70-76.
(33) Thompson, A.K. Fruit ripening conditions. In Postharvest Technology of Fruit and
Vegetables; Thompson, A.K. Eds.; Blackwell Science Ltd.: Oxford, 1996; pp 170-
178.
103
(34) Sharma, R.M.; Singh, R.R. Harvest, postharvest handling and physiology of fruits
and vegetables. In Postharvest technology of fruits and vegetables: handling,
processing, fermentation, and waste management; Verma, L.R.; Joshi, V.K., Eds.;
Indus Pub. Co., New Delhi, 2000; Vol. 1, pp 95-147.
(35) Tucker, G.A. Flavor changes. In Biochemistry of fruit ripening; Seymour, G.B.;
Taylor, J.E.; Tucker, G.A., Eds.; Chapman & Hall: London, 1993; pp 9-14.
(36) Kalt, W.; Forney, C.F.; Martin, A.; Prior, R.L. Antioxidant capacity, vitamin C,
1999, 47, 4638-4644.
(37) Tucker, G.A. Colour changes. In Biochemistry of fruit ripening; Seymour, G.B.;
Taylor, J.E.; Tucker, G.A., Eds.; Chapman & Hall: London, 1993; pp 14-17.
(38) Shishehgarha, F.; Makhlouf, J.; Ratti, C. Freeze-drying characteristics of

strawberries. Drying technology 2002, 20, 131-145.
(39) Ancos, B.D.; Sánchez-Moreno, C.; Pascual-Teresa, S.D.; Cano, M.P. Fruit freezing
principles. In Handbook of fruits and fruit processing, Hui, Y.H.; Barta, J.; Cano,
M.P.; Gesek, T.; Sidhu, J.S.; Sinha, N.K., Eds.; Blackwell: Ames, Iowa, 2006; Part
І, pp 59-80.
(40) Shahidi, F.; Naczk, M. Contribution of phenolic compounds to flavor and color
characteristics of foods. In Phenolics in food and nutraceuticals; Shahidi, F.; Naczk,
M., Eds.; CRC press: New York, 2004; Chapter 9, pp 443-469.

J. Food Sci. 2005, 70, C198-C202.
(42) Brownmiller, C.; Howard, L.R.; Prior, R.L. Processing and storage effects on
(43) Boziotis, C.A. Diffusion of sugar, acids and pigments during processing and storage
of canned berries. M.S. Thesis, Oregon State University, Corvallis, OR, 1963.
104
(45) Pantelidis, G.E.; Vasilakakis, M.; Manganaris, G.A.; Diamantidis, G. Antioxidant

capacity, phenol, anthocyanin and ascorbic acid contents in raspberries,
blackberries, red currants, gooseberries and Cornelian cherries. Food Chem. 2007,
102, 777-783.
(46) Wang, S.Y.; Lin, H. Antioxidant activity in fruits and leaves of blackberry,
raspberry, and strawberry varies with cultivar and developmental stage. J. Agri.
Food Chem. 2000, 48, 140-146.
(47) Huang, D.; Ou, B.; Prior, R.L. The chemistry behind antioxidant capacity assays. J.
Agri. Food Chem. 2005, 53, 1841-1856.
(48) Howard, L.R.; Hager, T.J. Berry fruit phytochemicals. In Berry fruit: value-added
(49) Mitcham, E. Quality of berries associated with preharvest and postharvest

conditions. In Berry fruit: value-added products for health promotion; Zhao, Y.,
Eds.; CRC press: New York, 2007; Chapter 7, pp 212-220.
(50) Mazza, G.; Miniati, E. Color stabilization and intensification. In Anthocyanins in

fruits, vegetables and grains; Mazza, G.; Miniati, E., Eds.; CRC press: Boca Raton,
FL, 1993; pp10-20.
(51) Scalzo, J.; Mezzetti, B.; Battino, M. Total antioxidant capacity evalution: Critical
steps for assaying berry antioxidant features. BioFactors 2005, 23, 221-227.
(52) Prior, R.L.; Cao, G.; Martin, A.; Sofic, E.; McEwen, J.; O’ Brien, C.; Lischner, N.;
Ehlenfeldt, M.; Kalt, W.; Krewer, G.; Mainland, C.M. Antioxidant capacity as
influenced by total phenolic and anthocyanin content, maturity, and variety of
Vaccinium species. J. Agri. Food Chem. 1998, 46, 2686-2693.
(53) Aguilar-Garcia, C.; Gavino, G.; Baragano-Mosqueda, M.; Hevia, P.; Gavino, V.C.
Correlation of tocopherol, tocotrienol, γ-oryzanol and total polyphenol content in
rice bran with different antioxidant capacity assays. Food Chem. 2007, 102, 1228-
1232.
105
CHAPTER 4
General Conclusion
Refrigeration storage, processing technologies and subsequent room storage
affected physicochemical parameters, contents of bioactive compounds, and antioxidant
capacities of ‘Marion’ and ‘Evergreen’ blackberries. Fruit ripening and decay
underwent in blackberries during post-harvest refrigeration storage, leading to only 9
days of shelf-life of ‘Marion’ and 7 days of shelf-life of ‘Evergreen’ at 2 ± 0.2 oC and
96 ± 2% RH in this study. Generally, refrigeration storage didn’t well retain the level of
bioactive compounds investigated in this study, and had inconsistent influences on
antioxidant capacity of two varieties. These results confirmed previous findings and
suggested that other treatments along with refrigeration storage are necessary for
prolonging shelf-life and retaining nutraceutical benefits of blackberries in fresh market.
Freeze drying increased total soluble solids and total phenolic content but didn’t
significantly change ORAC or FRAP in both blackberry varieties. Hot-air drying
enhanced total soluble solids while decreased total monomeric anthocyanin content in
both varieties and reduced antioxidant capacities of ‘Marion’. Canning diminished total
monomeric anthocyanins heavily in all the blackberry samples. Two types of canning
106
solutions, water and 20° Brix sucrose syrup exerted different effects on canned
products. Canning-in-sucrose syrup doubled the loss of total monomeric anthocyanins
compared with canning-in-water. Canning-in-water significantly increased, but
canning-in-sucrose syrup decreased radical scavenging activities. Besides, the losses of
TA and ORAC were observed in canned blackberries except canned-in-water
‘Evergreen’. Total phenolics, total monomeric anthocyanins, radical scavenging
activities against DPPH, ORAC and FRAP consistently dropped in blackberry jams of
both varieties.
Post-processing room storage didn’t change TA of canned and dried blackberries
but decreased that of blackberry jams. Total monomeric anthocyanins in canned, hot-air
dried blackberries and blackberry jams decreased significantly and only freeze dried
blackberries well preserved total phenolics and monomeric anthocyanins. Antioxidant
capacities of blackberry samples reduced or remained stable during room storage except
minor increase of radical scavenging activities in ‘Marion’ jams, FRAP in canned-in-
water ‘Marion’ and freeze-dried ‘Evergreen’. Our results showed that among all tested
processing technologies in this study, freeze drying best retained bioactive compounds
and antioxidant capacity in processed blackberries. In contrast, jam manufacturing was
the most detrimental mainly due to the application of high temperature for a long time.
Moreover, cultivar variations were detected in original nutraceutical contents in
fresh blackberries and their changes over post-harvest storage, processing and
subsequent storage in this study. Therefore, more blackberry cultivars should be
included in the future investigations.

107
Finally, the correlations among physicochemical properties, bioactive compounds
and antioxidant activities were observed and discussed. Such correlations would need
more data from other cultivars of blackberries to confirm.

108
BIBLIOGRAPHY
Agar, I.T.; Streif, J.; Bangerth, F. Effect of high CO2 and controlled atmosphere (CA)
on the ascorbic and dehydroascorbic acid content of some berry fruits. Postharv. Biol.
Tech. 1997, 11, 47-55.
Aguilar-Garcia, C.; Gavino, G.; Baragano-Mosqueda, M.; Hevia, P.; Gavino, V.C.
Correlation of tocopherol, tocotrienol, γ-oryzanol and total polyphenol content in rice
bran with different antioxidant capacity assays. Food Chem. 2007, 102, 1228-1232.
Amakura, Y.; Umino, Y.; Tsuji, S.; Tonogai, Y. Influence of jam processing on the
radical scavenging activity and phenolic content in berries. J. Agric. Food Chem. 2000,
48, 6292-6297.
Amano, S.; Yano, H. Evaluation of the practical performance of a commercially

available ELISA kit in the analysis of residual imidacloprid in agricultural products.
Journal of Pesticide Sci. 2005, 30, 249-253.
Ancos, B.D.; Sánchez-Moreno, C.; Pascual-Teresa, S.D.; Cano, M.P. Fruit freezing
principles. In Handbook of fruits and fruit processing; Hui, Y.H.; Barta, J.; Cano, M.P.;
Gesek, T.; Sidhu, J.S.; Sinha, N.K., Eds.; Blackwell: Ames, Iowa, 2006; Part І, pp 59-
80.
Antunes, L.E.C.; Duarte, J.; De Souza, C.M. Postharvest conservation of blackberry

Archbold, D.D.; HamiltonKemp, T.R.; Barth, M.M.; Langlois, B.E. Identifying natural
volatile compounds that control gray mold (Botrytis cinerea) during postharvest storage
of strawberry, blackberry, and grape. J. Agric. Food Chem. 1997, 45, 4032-4037.
Asami, D.K.; Hong, Y.; Barrett, D.M.; Mitchell, A.E. Comparison of the total phenolic
and ascorbic acid content of freeze-dried and air-dried Marionberry, strawberry, and
corn grown using conventional, organic, and sustainable agricultural practices. J. Agric.
Food Chem. 2003, 51, 1237-1241.
Barta, J. Fruit drying principles. In Handbook of fruits and fruit processing; Hui, Y.H.;
Barta, J.; Cano, M.P.; Gesek, T.; Sidhu, J.S.; Sinha, N.K., Eds.; Blackwell: Ames, Iowa,
2006; Chapter 5, pp 81-94.
Barth, M.M.; Zhou, C.; Mercier, J.; Payne, F.A. Ozone storage effects on anthocyanin
content and fungal growth in blackberries. J. Food Sci. 1995, 60, 1286-1288.
Beattie, J.; Crozier, A.; Duthie, G.G. Potential health benefits of berries. Curr. Nutr.
Food Sci. 2005, 1, 71-86.
109
Benvenuti, S.; Pellati, F.; Melegari, M.; Bertelli, D. Polyphenols, anthocyanins, ascorbic
acid, and radical scavenging activity of Rubus, Ribes, and Aronia. J. Food Sci. 2004,
69, FCT164- FCT169.
Benzie, I.F.F.; Strain, J.J. The ferric reducing ability of plasma (FRAP) as a measure of
“Antioxidant Power”: The FRAP Assay. Analytical Biochem. 1996, 239, 70-76.
Bohm, V.; Kuhnert, S.; Rohm, K.; Scholze, G. Improving the nutritional quality of
microwave-vacuum dried strawberries: A preliminary study. Food Science and
Technology International 2006, 12, 67-75.
Bower, C. Postharvest handling, storage, and treatment of fresh market berries. In Berry
fruit: value-added products for health promotion; Zhao, Y., Eds.; CRC press: New
York, 2007; Chapter 9, pp 262-284.
Boziotis, C.A. Diffusion of sugar, acids and pigments during processing and storage of
canned berries. M.S. Thesis, Oregon State University, Corvallis, OR, 1963.
Brand-Williams, W.; Cuvelier, M.E.; Berset, C. Use of a free radical method to evaluate
antioxidant activity. Lebensmittel-Wissenschaft Technol. 1995, 28, 25-30.
Brownmiller, C.; Howard, L.R.; Prior, R.L. Processing and storage effects on
Burdon, J.N.; Sexton, R. Fruit abscission and ethylene production of 4 blackberry

cultivars (Rubus spp.). Annals of applied biology 1993, 123, 121-132.
Burrows, G. Production of thermally processed and frozen fruit. In Fruit processing;

Arthey, D.; Ashurst, P.R., Eds.; Blackie Academic & Professional: London, UK, 1996;
Cahn, H.; DeFrancesco, J.; Nelson, E.; Martin, L. Fruit quality evaluation of raspberries
and blackberries at North Willamette Research and Extension Center. Special report /
Agricultural Experiment Station, Oregon State University 1992, 892, 2-9.
Cao, G.; Alessio, H.M.; Cutler, R.G. Oxygen-radical absorbance capacity assay for
antioxidants. Free Radic Biol Med. 1993, 14, 303-311.
Chanjirakul, K.; Wang, S.Y.; Wang, C.Y. Siriphanich, J. Natural volatile treatments
increase free-radical scavenging capacity of strawberries and blackberries. J. Sci. Food
Agric. 2007, 87, 1463-1472.
110
Chaovanalikit, A.; Wrolstad, R.E. Anthocyanin and polyphenolic composition of fresh

and processed cherries. J. Food Sci. 2004, 69, FCT73-FCT83.
Cheour, F.; Mahjoub, A. Delayed ripening and senescence of strawberry (Fragaria ×

ananassa Duch.) by gamma irradiation. Sciences Des Aliments 2003, 23, 355-366.
Cho, M.J.; Howard, L.R.; Prior, R.L.; Clark, J.R. Flavonoid glycosides and antioxidant
capacity of various blackberry, blueberry and red grape genotypes determined by high-
performance liquid chromatography/mass spectrometry. J. Sci. Food Agric. 2004, 84,
1771-1782.
Connor, A.M.; Finn, C.E.; Alspach, P.A. Genotypic and environmental variation in
antioxidant activity and total phenolic content among blackberry and hybridberry
Connor, A.M.; Finn, C.E.; Alspach, P.A. Genetic and enviromental variation in
anthocyanins and their relationship to antioxidant activity in blackberry and hybridberry
Daniel, E.M.; Krupnick, A.S.; Heur, Y.; Blinzler, J.A.; Nims, R.W.; Stoner, G.D.
Extraction, stability, and quantitation of ellagic acid in various fruits and nuts. J. Food
Composit. Annal. 1989, 2, 338-349.
Edwin, H. Polyphenol complexation. In Plant polyphenols; Edwin, H., Eds.;

Cambridge: New York, 1989; Chapter 4, pp 154-214.
Erkan, M.; Wang, S.Y.; Wang, C.Y. Effect of UV treatment on antioxidant capacity,
antioxidant enzyme activity and decay in strawberry fruit. Postharv. Biol. Tech. 2008,
48, 163-171.
Eyduran, S.P. Eyduran, E.; Agaoglu, Y.S. Estimation of fruit weight by cane traits for
eight American blackberries (Rubus fructicosus L.) cultivars. Journal of Biotechnology
2008, 7, 3031-3038.
Fan-Chiang, H.; Wrolstad, R.E. Anthocyanin pigment composition of blackberries. J.

Food Sci. 2005, 70, C198-C202.
Figuerola, F.E. Berry jams and jellies. In Berry fruit: value-added products for health
Figuerola, F.E. Dehydration of berries. In Berry fruit: value-added products for health
111
Finn, C.E.; Strik, B.C. Blackberry cultivars for Oregon. Oregon State University
Fisk, L.C.; Silver, M.A.; Strik, B.C.; Zhao, Y. Postharvest quality of hardy kiwifruit
(Actinidia arguta ‘Ananasnaya’) associated with packaging and storage conditions.
Fonseca, S.C.; Oliveira, F.A.R.; Brecht, J.K. Modeling respiration rate of fresh fruits
and vegetables for modified atmosphere packages: a review. Journal of Food
Engineering 2002, 52, 99-119.
Frenzen, P.D.; Majchrowicz, A.; Buzby, J.C.; Imhoff, B. Consumer acceptance of

irradiated meat and poultry products. Agriculture information bulletin from United
States Department of Agriculture/Economic Research Service 2000 (Aug), 757, 1-8.
Giusti, M.M.; Wrolstad, R.E. Characterization and measurement of anthocyanins by

UV-Visible spectroscopy. In Current Protocols in Food Analytical Chemistry; John
Wiley & Sons, Inc.: New Jersy, 2001; F1.2.1-F1.2.13.
Gomez, D.R.; Evans, K.J.; Baker, J.; Harvey, P.R.; Scott, E.S. Dynamics of introduced
populations of Phragmidium violaceum and implications for biological control of
European blackberry in Australia. Applied and Environmental Microbiology 2008, 74,
5504-5510.
Gonzalez-Aguilar, G.A.; Villegas-Ochoa, M.A.; Martinez-Tellez, M.A.; Gardea, A.A.;

Ayala-Zavala, J.F. Improving antioxidant capacity of fresh-cut mangoes treated with
UV-C. J. Food Sci. 2007, 72, S197-S202.
González, E.M.; Ancos, B.D.; Cano, M.P. Partial characterization of peroxidase and
polyphenol oxidase activities in blackberry fruits. J. Agric. Food Chem. 2000, 48, 5459-
5464.
González, E.M.; de Ancos, B.; Cano, M.P. Relation between bioactive compounds and
free radical-scavenging capacity in berry fruits during frozen storage. J. Sci. Food
Agric. 2003, 83, 722-726.
Gordon, S.C.; Woodford, J.A.T.; Birch, A.N.E. Arthropod pests of Rubus in Europe:
pest status, current and future control strategies. Journal of Horticultural Sci. 1997, 72,
831-862.
Hager, T.J.; Howard, L.R.; Liyanage, R.; Lay, J.O.; Prior, R.L. Ellagitannin
composition of blackberry as determined by HPLC-ESI-MS and MALDI-TOF-MS. J.
Agric. Food Chem. 2008, 56, 661-668.
112
Hager, T.J.; Howard, L.R.; Prior, R.L. Processing and storage effects on monomeric
Han, C.; Zhao, Y.; Leonard, S.W.; Traber, M.G. Edible coatings to improve storability
and enhance nutritional value of fresh and frozen strawberries (Fragaria × ananassa) and
raspberries (Rubus ideaus). Postharv. Biol. Tech. 2004, 33, 67-68.
Hernandez-Munoz, P.; Almenar, E.; Del Valle, V.; Velez, D.; Gavara, R. Effect of
chitosan coating combined with postharvest calcium treatment on strawberry (Fragaria
× ananassa) quality during refrigeration storage. Food Chem. 2008, 110, 428-435.
Howard, L.R.; Hager, T.J. Berry fruit phytochemicals. In Berry fruit: value-added
products for health promotion; Zhao, Y., Eds.; CRC press: New York, 2007; Chapter 3,
pp 74-98.
Huang, D.; Ou, B.; Prior, R.L. The chemistry behind antioxidant capacity assays. J.
Agri. Food Chem. 2005, 53, 1841-1856.
Hui, Y.H. Fruits: Orange juice processing. In Food processing: principles and
applications; Smith, J.S.; Hui, Y.H., Eds.; Blackwell: Ames, Iowa, 2004; Chapter 21,
pp 361-390.
Hui, Y.H.; Lim, M.; Nip, W.; Smith, J.S.; Yu, P.H.F. Principles of food processing. In
Food processing: principles and applications; Smith, J.S.; Hui, Y.H., Eds.; Blackwell:
Ames, Iowa, 2004; Chapter 1, pp17-19.
Hui, Y.H.; Lim, M.; Nip, W.; Smith, J.S.; Yu, P.H.F. Principles of food processing. In
Food processing: principles and applications; Smith, J.S.; Hui, Y.H., Eds.; Blackwell:
Hussain, P.R.; Meena, R.S.; Dar, M.A.; Mir, M.A.; Shafi, F.; Wani, A.M. Effect of
gamma-irradiation and refrigerated storage on mold growth and keeping quality of
strawberry (Fragaria sp) cv ‘Confitura’. Journal of Food Sci. & Tech. 2007, 44, 513-
516.
Huxley, A.; Griffiths, M.; Levy, M. Rubus. In Dictionary of Gardening; Huxley, A.;
Griffiths, M.; Levy, M., Eds.; Stockton Press: New York, 1992; Vol. 4, pp 140-145.
Häkkinen, S.H.; Kärenlampi, S.O.; Mykkänen, H.M.; Törrönen, A.R. Influence of

domestic processing and storage on flavonol contents in berries. J. Agric. Food Chem.
2000, 48, 2960-2965.
113
Jarvis, B. Cider, perry, fruit wines and other alcoholic fruit beverages. In Fruit
processing; Arthey, D.; Ashurst, P.R., Eds.; Blackie Academic & Professional: London,
UK, 1996; Chapter 5, pp 121-127.
Kafkas, E.; Kosar, M.; Türemis, N.; Baser, K.H.C. Analysis of sugars, organic acids and
vitamin C contents of blackberry genotypes from Turkey. Food Chem. 2006, 97, 732-
736.
Kalt, W.; Forney, C.F.; Martin, A.; Prior, R.L. Antioxidant capacity, vitamin C,
1999, 47, 4638-4644.
Khwaldia, K.; Perez, C.; Banon, S.; Desobry, S.; Hardy, J. Milk proteins for edible
films and coatings. Crit. Rev. Food Sci. Nutr. 2004, 44, 239–251.
Kraft, T.F.B.; Dey, M.; Rogers, R.B.; Ribnicky, D.M.; Gipp, D.M.; Cefalu, W.T.;
Raskin, I.; Lila, M.A. Phytochemical composition and metabolic performance-
enhancing activity of dietary berries traditionally used by native North Americans. J.
Agric. Food Chem. 2008, 56, 654-660.
Kwok, B.H.L.; Hu, C.; Durance, T.; Kitts, D.D. Dehydration techniques affect
phytochemical contents and free radical scavenging activities of Saskatoon berries
(Amelanchier alnifolia Nutt.). J. Food Sci. 2004, 69, S122-S126.
Körmendy, I. Fruit processing: principles of heat treatment. In Handbook of fruits and

fruit processing, Hui, Y.H.; Barta, J.; Cano, M.P.; Gesek, T.; Sidhu, J.S.; Sinha, N.K.,
Eds.; Blackwell: Ames, Iowa, 2006; Chapter 3, pp 45-57.
Lewis, N.G.; Yamamoto, E. Tannins - their place in plant metabolism. In Chemistry and
significance of condensed tannins; Hemingway, R.W.; Karchesy, J.J.; Branham, S.J.,
Eds.; Plenum Press: New York, 1989; pp 23-43.
Lin, D.; Zhao, Y. Innovations in the development and application of edible coatings for
fresh and minimally processed fruits and vegetables. Comprehensive Reviews in Food
Science and Food Safety 2007, 6, 1-13.
Liu, R.H. The potential health benefits of phytochemicals in berries for protecting
health promotion; Zhao, Y., Eds.; CRC press: New York, 2007; Chapter 6, pp 187-199.
Liu, S.; Manson, J.E.; Lee, I.M; Cole, S.R.; Hennekens, C.H.; Willett, W.C.; Buring,
J.E. Fruit and vegetable intake and risk of cardiovascular disease: the women’s health
study. Am. J. Clin. Nutr. 2000, 72, 922-928.
114
Macheix, J.; Fleuriet, A.; Billot, J. The main phenolics of fruit. In Fruit phenolics;
Macheix, J.; Fleuriet, A.; Billot, J., Eds.; CRC Press: Boca Raton, Florida, 1990;
Chapter 1, pp 1-98.
Marti, J.; Aguilera, J.M. Effects of freezing rate on the mechanical characteristics and
microstructure of blueberries and wild blackberries. Revista De Agroquimica Y
Tecnologia De Alimentos 1991, 31, 493-504.
Martinez-Romero, D.; Guillen, F.; Castillo, S.; Valero, D.; Serrano, M. Modified
atmosphere packaging maintains quality of table grapes. J. Food Sci. 2003, 68, 1838,
1843.
Martínez, G.A.; Civello, P.M. Effect of heat treatments on gene expression and enzyme
activities associated to cell all degradation in strawberry fruit. Postharv. Biol. Tech.
2008, 49, 38-45.
Mazza, G.; Miniati, E. Color stabilization and intensification. In Anthocyanins in fruits,

vegetables and grains; Mazza, G.; Miniati, E., Eds.; CRC press: Boca Raton, FL, 1993;
pp10-20.
Mei, Y.; Zhao, Y. Using edible coating to enhance nutritional and sensory qualities of
babay carrots. J. Food Sci. 2002, 65, 1964-1968.
Meneghel, R.F.D.; Benassi, M.D.; Yamashita, F. Sodium alginate coating for

blackberry (Rubus ulmifolius) fruits. Semina-Cienclas Agrarias 2008, 29, 609-617.
Mitcham, E. Quality of berries associated with preharvest and postharvest conditions. In

Moyer, R.A.; Hummer, K.E.; Finn, C.E.; Frei, B.; Wrolstad, R.E. Anthocyanins,
phenolics, and antioxidant capacity in diverse small fruits: Vaccinium, Rubus, and
Ribes. J. Agric. Food Chem. 2002, 50, 519-525.
Ngo, T.; Wrolstad, R.E.; Zhao, Y. Color quality of Oregon strawberries-impact of

Nimitkeatkai, H.; Srilaong, V.; Kanlayanarat, S. Effect of edible coating on Pineapple

fruit quality during cold storage. Acta Hort. 2006, 712, 643-647.
Nordskog, B.; Stensvand, A.; Heiberg, N. Fungi occuring on aerial constituents of

cultivated blackberry (Rubus fruticosus L.) in Norway. Acta Agriculturae Scandinavica
Section B-Soil and Plant Sci. 2003, 53, 21-28.
115
Pantelidis, G.E.; Vasilakakis, M.; Manganaris, G.A.; Diamantidis, Gr. Antioxidant

capacity, phenol, anthocyanin and ascorbic acid contents in raspberries, blackberries,
red currants, gooseberries and Cornelian cherries. Food Chem. 2007, 102, 777-783.
Park, H.J. Development of advanced edible coatings for fruits. Trends Food Sci. Tech.
1999, 10, 254-260.
Park, S.I.; Stan, S.D.; Daeschel, M.A.; Zhao, Y. Antifungal coatings on fresh
strawberries (Fragaria x ananassa) to control mold growth during cold storage. J. Food
Sci. 2005, 70, 202-207.
Perkins-Veazie, P.; Clark, J.R.; Huber, D.J.; Baldwin, E.A. Ripening physiology in
‘Navaho’ thornless blackberries: color, respiration, ethylene production, softening, and
compositional changes. Journal of the American society for Horticultural Science 2000,
125, 357-363.
Perkins-Veazie, P.; Collins, J.K.; Clark, J.R. Changes in blackberry fruit quality during
storage. Acta Horticulturae 1993, 352, 87-90.
Perkins-Veazie, P.; Collins, J.K.; Clark, J.R. Cultivar and maturity affect postharvest
quality of fruit from erect blackberries. HortSci. 1996, 31, 258-261.
Perkins-Veazie, P.; Collins, J.K.; Clark, J.R. Cultivar and storage temperature effects on
the shelflife of blackberry fruit. Fruit Varieties Journal 1999, 53, 201-208.
Perkins-Veazie, P.; Collins, J.K.; Clark, J.R. Fruit characteristics of some erect
blackberry cultivars. HortSci. 1993, 28, 853-853.
Perkins-Veazie, P.; Collins, J.K.; Clark, J.R. Shelf-life and quality of ‘Navaho’ and
quality 1999, 22, 535-544.
Perkins-Veazie, P.; Collins, J.K.; Howard, L. Blueberry fruit response to postharvest

application of ultraviolet radiation. Postharv. Biol. Tech. 2008, 47, 280-285.
Perkins-Veazie, P.; Collins, J.K. Quality of erect-type blackberry fruit after short
intervals of controlled atmosphere storage. Postharv. Biol. Tech. 2002, 25, 235-239.
Perkins-Veazie, P.; Kalt, W. Postharvest storage of blackberry fruit does not increase
antioxidant levels. Acta Horticulture 2002, 585, 521-524.
Petracek, P.D.; Joles, D.W.; Shirazi, A.; Cameron, A.C. Modified atmosphere
packaging of sweet cherry (Prunus avium L., ev. ‘Sams’) fruit: metabolic responses to
oxygen, carbon dioxide, and temperature. Postharv. Biol. Tech. 2002, 24, 259-270.
116
Prange, R.K.; DeEll, J.R. Preharvest factors affecting postharvest quality of berry crops.
HortSci. 1997, 32, 824-829.
Prior, R.L.; Cao, G.; Martin, A.; Sofic, E.; McEwen, J.; O’ Brien, C.; Lischner, N.;
Ehlenfeldt, M.; Kalt, W.; Krewer, G.; Mainland, C.M. Antioxidant capacity as
influenced by total phenolic and anthocyanin content, maturity, and variety of
Vaccinium species. J. Agri. Food Chem. 1998, 46, 2686-2693.
Prior, R.L.; Wu, X.L.; Gu, L.W.; Hager, T.J.; Hager, A.; Howard, L.R. Whole berries
versus berry anthocyanins: interactions with dietary fat levels in the C57BL/6J mouse of
obesity. J. Agric. Food Chem. 2008, 56, 647-653.
Qian, M.C.; Wang, Y. Seasonal variation of volatile composition and odor activity
value of ‘Marion’ (Rubus spp. hyb) and ‘Thornless Evergreen’ (R.laciniatus L.)
blackberries. J. Food Sci. 2005, 70, C13-C19.
Ramaswamy, H.S.; Chen, C.R. Canning Principles. In Handbook of vegetables

preservation and processing; Hui, Y.H.; Ghazala, S.; Graham, D.M.; Murrell, K.D.;
Nip, W., Eds.; Marcel Dekker: New York, 2004; Chapter 4, pp 273- 282.
Ramaswamy, H.S.; Meng, Y. Commercial canning of berries. In Berry fruit: value-

Chapter 12, pp 336-361.
Ravai, M. Quality attributes of berry crops. Cereal Foods World 1996, 41, 773-775.
Reyes-Carmona, J.; Yousef, G.G.; Martinez-Peniche, R.A.; Lila, M.A. Antioxidant

Ribeiro, C.; Vicente, A.A.; Teixeira, A.J.; Miranda, C. Optimization of edible coating
composition to retard strawberry fruit senescence. Postharv. Biol. Tech. 2007, 44, 63–
70.
Rickman, J.C.; Barrett, D.M.; Bruhn, C.M. Review: Nutritional comparison of fresh,
frozen and canned fruits and vegetables. Part 1. Vitamins C and B and phenolic
compounds. J. Sci. Food Agric. 2007, 87, 930-944.
Rieger, M. Blackberries and raspberries (Rubus spp.). In Introduction to fruit crops;

Rieger, M., Eds.; Food Products Press: New York, 2006; Chapter 6, pp 89-104.
Rocha, A.M.C.N.; Barreiro, M.G.; Morais, A.M.M.B. Modified atmosphere package for
apple ‘Bravo de Esmolfe’. Food Control 2004, 15, 61-64.
117
Rodriguez-Saona, L.E.; Wrolstad, R.E. Extraction, isolation and purification of

anthocyanins. In Current Protocols in Food Analytical Chemistry; John Wiley & Sons:
New Jersy, 2001; F1.1.1-F1.1.11.
Rommel, A. Wrolstad, R.E.; Heatherbell, D.A. Blackberry juice and wine-processing

and storage effects on anthocyanin composition, color and apprearance. J. Food Sci.
1992, 57, 385-391.
Rutledge, P. Production of non-fermented fruit products. In Fruit processing, Arthey,

D.; Ashurst, P.R., Eds.; Blackie Academic & Professional: London, U.K., 1996;
Salunkhe, D.K.; Desai, B.B. Small fruits-berries. In Postharvest Biotechnology of

Fruits; Salunkhe, D.K.; Desai, B.B., Eds.; CRC press: Boca Raton, FL, 1984; Vol. 1 pp
111-112.
Scalzo, J.; Mezzetti, B.; Battino, M. Total antioxidant capacity evalution: Critical steps
for assaying berry antioxidant features. BioFactors 2005, 23, 221-227.
Schmidt, B.M.; Erdman, J.W.; Lila, M.A. Effects of food processing on blueberry
antiproliferation and antioxidant activity. J. Food Sci. 2005, 70, S389-S394.
Seeram, N.P. Berry fruits: compositional elements, biochemical activities, and the
impact of their intake on human health, performance, and disease. J. Agric. Food Chem.
2008, 56, 627-629.
Seeram, N.P. Berry fruits for cancer prevention: current status and future prospects. J.
Agric. Food Chem. 2008, 56, 630-635.
Shahidi, F.; Naczk, M. Contribution of phenolic compounds to flavor and color

characteristics of foods. In Phenolics in food and nutraceuticals; Shahidi, F.; Naczk,
M., Eds.; CRC press: New York, 2004; Chapter 9, pp 443-469.
Sharma, R.M.; Singh, R.R. Harvest, postharvest handling and physiology of fruits and
vegetables. In Postharvest technology of fruits and vegetables: handling, processing,
fermentation, and waste management; Verma, L.R.; Joshi, V.K., Eds.; Indus Pub. Co.,
New Delhi, 2000; Vol. 1, pp 95-147.
Sharma, R.M.; Singh, R.R. Harvesting, postharvest handling and physiology of fruits
and vegetables. In Postharvest technology of fruits and vegetables; Verma, L.R.; Joshi,
V.K., Eds.; Indus Pub. Co.: New Delhi, India, 2000; Vol. 1, pp 127-134.
118
Shih, C.; Pan, Z.; McHugh, T.; Wood, D.; Hirschberg, E. Sequential infrared radiation
and freeze-drying method for producing crispy strawberries. Transactions of the Asabe
2008, 51, 205-216.
Shishehgarha, F.; Makhlouf, J.; Ratti, C. Freeze-drying characteristics of strawberries.

Drying technology 2002, 20, 131-145.
Shukitt-Hale, B.; Lau, F.C.; Joseph, J.A. Berry fruit supplementation and the aging
Singleton, V.L.; Rossi, J.A. Colorimetry of total phenolics with phosphomolybdic-

phosphotungstic acid reagents. Ameri. J Enol. Viticult. 1965, 16, 144-158.
Siriwoharn, T.; Wrolstad, R.E.; Finn, C.E.; Pereira, C.B. Influence of cultivar, maturity,
and sampling on blackberry (Rubus L. Hybrids) anthocyanins, polyphenolics, and
antioxidant properties. J. Agric. Food Chem. 2004, 52, 8021-8030.
Siriwoharn, T.; Wrolstad, R.E. Polyphenolics composition of Marion and Evergreen

Skrede, G.; Wrolstad, R.E.; Durst, R.W. Changes in anthocyanins and polyphenolics
during juice processing of highbush blueberries (vaccinium corymbosum L.). J. Food
Sci. 2000, 65, 357-364.
Strik, B.C. Berry crops: Worldwide area and production systems. In Berry fruit: value-
Chapter 1, pp 9-11.
Strik, B.C.; Clark, J.R.; Finn, C.E.; Bañados, M.P. Worldwide blackberry production.
HortTechnology 2007, 17, 205-213.
Song, Y.; Vorsa, N.; Yam, K.L. Modeling respiration-transpiration in a modified

atmosphere packaging system containing blueberry. Journal of Food Engineering 2002,
53, 103-109.
Sousa, M.B.; Canet, W.; Alvarez, M.D.; Fernandez, C. Effect of processing on the
texture and sensory attributes of raspberry (cv. Heritage) and blacbkerry (cv.
Thornfree). Journal of Food Engineering 2007, 78, 9-21.
Susaimuthu, J.; Tzanetakis, I.E.; Gergerich, R.C.; Kim, K.S.; Martin, R.R. Viral
interactions lead to decline of blackberry plants. Plant Disease 2008, 92, 1288-1292.
119
Talcott, S.T. Chemical components of berry fruits. In Berry fruit: value-added products
for health promotion; Zhao, Y., Eds.; CRC press: New York, 2007; Chapter 2, pp 55-
56.
Talcott, S.T. Chemical components of berry fruits. In Berry fruit: value-added products
for health promotion; Zhao, Y., Eds.; CRC press: New York, 2007; Chapter 2, pp 59-
62.
Thomas, R.; Woods, F.M.; Dozier, W.A; Ebel, R.C.; Nesbitt, M.; Wilkins, B.;
Himelrick, D. Cultivar variation in physiochemical and antioxidant activity of Alabama-
grown blackberries. Small Fruits Rev. 2005, 4, 57-71.
Thompson, A.K. Fruit ripening conditions. In Fruit and vegetables: harvesting,

handling and storage, 2nd ed.; Thompson, A.K., Eds.; Blackwell: Oxford, U.K., 2003;
pp 19-112.


Thompson, A.K. Fruit ripening conditions. In Postharvest Technology of Fruit and

Vegetables; Thompson, A.K., Eds.; Blackwell Science: Harlow, Oxford, 1996; Chapter
6, pp 170-188.


Thompson, L.U. Role of lignans in carcinogenesis. In Phytochemicals in human health

protection, nutrition, and plant defense; Romeo, J.T., Eds.; Kluwer Academic/Plenum
Publishers: New York, 1999; Chapter 3, pp 51-61.
Tsuda, T. Regulation of adipocyte function by anthocyanins; possibility of preventing

the metabolic syndrome. J. Agric. Food Chem. 2008, 56, 642-646.
120
Tosun, I.; Ustun, N.S.; Tekguler, B. Physical and chemical changes during ripening of
blackberry fruits. Sci.Agric. 2008, 65, 87-90.
Tucker, G.A. Colour changes. In Biochemistry of fruit ripening; Seymour, G.B.; Taylor,
J.E.; Tucker, G.A., Eds.; Chapman & Hall: London, 1993; pp 14-17.
Tucker, G.A. Flavor changes. In Biochemistry of fruit ripening; Seymour, G.B.; Taylor,
J.E.; Tucker, G.A., Eds.; Chapman & Hall: London, 1993; pp 9-14.
Turemis, N.; Kafkas, S.; Kafkas, E.; Onur, C. Fruit characeristics of nine thornless
blackberry genotypes. Journal American Pomological Society 2003, 57, 161-165.
United States Department of Agriculture (USDA) National Nutrient Database for

standard reference. http://www.nal.usda.gov/fnic/foodcomp/search/index.html (accessed
Nov 2008).
Vachon, C.; D’Aprano, G.; Lacroix, M.; Letendre, M. Effect of edible coating process
and irradiation treatment of strawberry Fragaria spp. on storage-keeping quality. J.
Food Sci. 2003, 68, 608-612.
Vargas, M.; Albors, A.; Chiralt, A.; Gonzalez-Martınez, C. Quality of cold-stored

strawberries as affected by chitosan–oleic acid edible coatings. Postharv. Biol. Tech.
2006, 41, 164–171.
Venkatachalapathy, K. Raghavan, G.S.V. Microwave drying of osmotically dehydrated

blueberries. Journal of Microwave Power and electromagnetic Energy 1998, 33, 95-
102.
Vibhakara, H.S.; Bawa, A.S. Manufacturing jams and jellies. In Handbook of fruits and
fruit processing; Hui, Y.H.; Barta, J.; Cano, M.P.; Gusek, T.; Sidhu, J.S.; Sinha, N.K.,
Eds.; Blackwell: Ames, Iowa, 2006; Chapter 11, pp 8189-204.
Vicente, A.R.; Repice, B.; Martinez, G.A.; Chaves, A.R.; Civello, P.M.; Sozzi, G.O.
Maintenance of fresh boysenberry fruit quality with UV-C light and heat treatments
combined with low storage temperature. Biotech. 2004, 79, 246-251.
Wang, S.Y.; Bowman, L.; Ding, M. Methyl jasmonate enhances antioxidant activtiy and
flavonoid content in blackberries (Rubus sp.) and promotes antiproliferation of human
cancer cells. Food Chem. 2008, 107, 1261-1269.
Wang, S.Y.; Lin, H. Antioxidant activity in fruits and leaves of blackberry, raspberry,
and strawberry varies with cultivar and developmental stage. J. Agric. Food Chem.
2000, 48, 140-146.
121
Woods, F.M.; Dozier, W.A.; Ebel, R.C.; Thomas, R.; Nesbitt, M. Wilkins, B.S.;
Himelrick, D.G. Cultivar and maturity effects on fruit quality and antioxidant properties
in blackberry. HortSci. 2006, 41, 1043-1043.
Wu, X.; Beecher, G.R.; Holden, J.M.; Haytowitz, D.B.; Gebhardt, S.E.; Prior, R.L.
Concentrations of anthocyanins in common foods in the United States and estimation of
normal consumption. J. Agric. Food Chem. 2006, 54, 4069-4075.
Xie, J.; Zhao, Y. Improvement of physicochemical and nutritional qualities of frozen

Marionberry by vacuum impregnation pretreatment with cryoproctectants and minerals.
Journal of Horticultural Science & Biotechnology 2003, 78, 248-253.
Yaman, Ö.; Bayoindirli, L. Effect of an edible coating and cold storage on shelf-life and
quality of cherries. Lebensmittel-Wissenschaft Und-Technologie-Food Sci Technol.
2002, 35, 146-150.
Yurdugul, S. An evaluation of the retention of quality characteristics in fresh and

freeze-dried alpine strawberries. International Journal of Food Science and Technology
2008, 43, 865-870.
Zadernowski, R.; Naczk, M.; Nesterowicz, J. Phenolic acid profiles in some small
Zhao, Y. Freezing process of berries. In Berry fruit: value-added products for health
122
APPENDIX
123
APPENDIX
under Commercial Storage Conditions
Ruyi Wu1, Daniel Lin1, Bernadine Strik2, and Yanyun Zhao1,3
1
Department of Food Science & Technology, Oregon Sate University, Corvallis, OR
97331, USA.
2
Department of Horticulture, Oregon Sate University, Corvallis, OR 97331, USA.
3
Corresponding author: E-mail: yanyun.zhao@oregonstate.edu.
To be submitted to HortTechnology
113 South West Street, Suite 200, Alexandria, VA 22314
124
under Commercial Storage Conditions
________________________
ADDITIONAL INDEX WORDS. Northern highbush blueberry, Vaccinium
corymbosum, cultivar, shelf-life, fruit quality, firmness, desiccation, antioxidants,
phenolics, ready-to-eat
_________________________
SUMMARY. The edible coatings, SemperfreshTM (SF), chitosan (CH), and calcium
caseinate (CC), were applied to ‘Duke’, and CC to ‘Elliott’ ripe fruit to evaluate effect
on quality during storage. Fruit were washed in 200 ppm chlorinated water before
applying coatings, then stored at 2 oC for 1 week, followed by storage at room
temperature (20±3 oC) for up to 12 days for quality evaluation. Storage conditions were
chosen to simulate the commercial situation of a brief cold storage period followed by
display/sale under un-refrigerated conditions. CH coating helped reduce the decay rate
of ‘Duke’ throughout the storage period. SF coating decreased weight loss of ‘Duke’
after 6 days of storage. CC coating significantly increased firmness of ‘Elliott’ after 6
days of storage. In general, ‘Duke’ fruit that were washed and coated had a higher
antioxidant content (AC) and total phenolic content (TPC) than non-washed fruit.
Washing and coating did not significantly affect AC and TPC of ‘Elliott’. Our results
suggest that edible coatings have potential for retaining quality of pre-washed, ready-to-
eat fresh blueberries under commercial storage conditions.

125
INTRODUCTION
Blueberries are well-known for having prominent antioxidant activities and a
high content of phenolic compounds and anthocyanins (Kalt et al., 2001; Zheng and
Wang, 2003). Consumption of blueberries has shown promise of health benefits,
including reduction of cholesterol level (Abidov et al., 2006; Kahlon and Smith, 2007),
anti-aging (Wilson et al., 2006), anti-inflammatory (Torri et al., 2007), anti-
carcinogenic (Seeram et al., 2006; Yi et al., 2006), prevention of urinary tract infection
and diabetes (Martineau et al., 2006), and improvement of eyesight and brain health
(Willis et al., 2005).
Fresh highbush blueberries have been reported to have a shelf-life of 1 to 8 weeks
depending on stage of fruit ripeness, method of harvest, presence of fruit disease, and
storage conditions (temperature, relative humidity, and atmosphere; Ballinger et al.,
1978; Beaudry et al., 1998; Galletta et al, 1971; Hancock et al., 2008). Post-harvest
respiration and transpiration cause quality deterioration of fresh fruit, limiting shelf-life.
In addition, bioactive compounds may degrade rapidly during post-harvest storage,
partly due to the oxidation of polyphenolics with exposure to light and oxygen
(Srivastava et al., 2007).
Several preservation technologies, including cold storage, UV irradiation, modified
atmosphere packaging and ozonation, have been used to reduce deterioration, prolong
shelf-life and retain the nutritional value of fresh blueberries (Chiabrando et al., 2006;
Connor et al., 2002; Trigo et al., 2006; Zheng et al., 2003). Since the first use of wax as
a coating material in the 12th to 13th century in China, edible coatings have been applied
126
for extending shelf-life of fresh produce (Baldwin et al., 1999; Han et al., 2004;
Nimitkeatkai et al., 2006; Park, 1999; Ribeiro et al., 2007; Vargas et al., 2006). Along
with increased interest in ready-to-eat and ‘invisible processed’ fruit with high quality
and safety, edible coatings may provide a means to provide pre-washed, ready-to-eat
blueberries. Edible coatings can possibly control the internal gas atmosphere of the
fruit, minimizing fruit respiration rate (Park, 1999) and may serve as a barrier to water
vapor, reducing moisture loss and delaying fruit dehydration (Baldwin et al., 1995). In
addition, some edible coating materials, such as chitosan, have shown antimicrobial
functions against the growth of certain microorganisms (Park et al., 2005; Zhang and
Quantick, 1998).
Polysaccharides, proteins, lipids and their combinations may be used as coating
materials for fresh produce (Baldwin et al., 1995). Chitosan (1, 4 - linked 2 - amino – 2
- deoxy – β – D - glucan), a derivative of chitin, has excellent film-forming and
antimicrobial functions and has been successfully used to control quality loss of fresh
strawberries and raspberries (Han et al., 2004; Park et al., 2005; Ribeiro et al., 2007;
Vargas et al., 2006), sliced mango fruits (Chien et al., 2007), citrus (Fornes et al., 2005),
fresh-cut water chestnut (Pen and Jiang, 2003), and many other fruits and vegetables
(Lin and Zhao, 2007). Caseinate, a milk-protein based material, has excellent oxygen
barrier properties and has been studied in carrots (Mei and Zhao, 2003), apples and
potatoes (Letien et al., 2001), celery (Avena-Bustillos et al., 1997), and strawberries
(Vachon et al., 2003) for controlling post-harvest respiration. SemperfreshTM is a
commercial coating product of sucrose-fatty acid ester, and was reported to effectively
127
decrease weight loss of hardy kiwifruit (Fisk et al., 2008), cherry (Yaman and
Bayoindirli, 2002), and summer squash (Kaynas and Ozelkok, 1999), and extend shelf-
life of pineapple for up to 5 weeks by preventing moisture loss (Nimitkeatkai et al.,
2006).
The objectives of this study were to investigate the effectiveness of chitosan,
calcium caseinate, and SemperfreshTM based coatings for enhancing the shelf-life and
retaining the antioxidant properties of pre-washed, ready-to-eat highbush blueberry
cultivars under commercial storage conditions.
MATERIALS & METHODS
FRUIT. Two cultivars of highbush blueberry, ‘Duke’ and ‘Elliott’, were hand
harvested by a commercial picking crew from a farm in Sheridan, Ore. in mid-July and
mid-Aug. 2006, respectively. Harvested fruit were immediately packed in a 6-fl oz
(170 g blueberry weight) vented, plastic “clam-shell” containers (industry standard) and
transported to the Food Science laboratory at Oregon State University, Corvallis, Ore.
COATINGS. Food-grade coating materials were used including: chitosan (CH:
Vanson Inc., Redmond, Wash.; 89.8% deacetyelation) extracted from shrimp shells;
calcium caseinate (CC: Alanate 385, NZMP, Santa Rosa, Cal.; 92.9% protein and 1.4%
calcium); and Semperfresh™ (SF: AgriCoat Industries Ltd., England; distributed by
Pace International, Seattle, Wash.) which is a mixture of sucrose esters of fatty acids,
sodium carboxymethlcellulose, and mono-diglycerides of fatty acids. Other materials

128
used in the coating formulation were glycerol (Fisher Scientific Inc., Fairawn, NJ) and
glacial acetic acid (Baker Adamson, Morristown, NJ).
A 2% (w/v) CH coating solution was prepared by dissolving CH in 1% aqueous
acetic acid with 50% glycerol (w/CH dry weight), adding 0.15% Tween 20 (w/w),
homogenizing (Polytron PT 10-35, Kinematica AG, Littau, Switzerland) for 90 sec at
3000g, and then storing overnight at room temperature. A 2% CC coating solution was
prepared by dissolved 2% CC in de-ionized water with addition of 50% glycerol (w/CC
dry weight) and 0.15% Tween 20 (w/w). The mixture was homogenized for 1 min at
3000g and shaken in a 60 ºC water bath for 30 min, followed by cooling to room
temperature. The SF coating solution was prepared by diluting 50% SF concentrate
with deionized water to 1%, and mixing with 50% glycerol (w/SF weight) and 0.15%
Tween 20 (w/w).
TREATMENT OF FRUIT. The major goal of applying coatings on fresh
blueberries is to develop ready-to-eat fresh fruit with a longer shelf-life. Hence, fruit
were first sanitized by washing in 200 ppm chlorinated water prior to coating
application. Washed ‘Duke’ fruit were randomly assigned to one of three coating
treatments: CH, CC or SF. A non-washed SF-coated treatment was included for
comparison. Washed ‘Elliott’ were coated with CC only, based on observations that
CC-coated ‘Duke’ fruit from the earlier harvest season had a more natural looking
“bloom” (natural waxy coating on un-washed fruit). A non-washed, CC-coated
treatment was included for comparison. In both cultivars, a non-washed (natural waxy
bloom present) and washed (in water, removing much of the bloom) control were used
129
as controls. All treatment coatings were applied twice, by dipping fruit in the coating
solution for 30 sec, draining on a stainless steel screen for 30 min, and then repeating
the same procedure to achieve a uniform surface coating.
To simulate commercial storage conditions, coated samples were re-packaged in the
vented plastic containers, stored in a cooler at 2 ± 1 ºC and 88% relative humidity in the
dark for 1 week, and then removed and placed at room temperature (retail display
condition) at 20 ± 3 ºC and 30% relative humidity under normal room light for 12 days
and changes in quality monitored.
FRUIT QUALITY PARAMETERS. All fruit quality parameters were measured
before cold storage and then at 0 (1 week after cold storage), 3, 6, 9 and 12 days at room
temperature. Blueberries taken out from one clamshell container were used for one
replication of quality measurement. Total soluble solid content (TSS), pH and titratable
acidity (TA) were measured following the procedures as described by Fisk et al. (2008),
where TA was reported as percent malic acid (mass/mass) on the basis of fresh weight
of fruit. Three replications were conducted for each treatment.
Firmness (FN) of fresh blueberries was determined by measuring compression force
using pre-calibrated BioWorks FirmTech2 Instrument (BioWorks, Inc., Wamego,
Kan.). A sub-sample of fruit (25 berries) were set on the indentures on the turntable,
and tested individually through the compression of the load cell with continuously
counterclockwise rotation of turntable after 25 fruits at a time. The force thresholds
were from 25 to 250 g. Percentage weight loss (WL) was calculated as weight change
of fruit at each sampling time divided by the initial weight of the fruit. Decay rate (DR)
130
of the fruit was defined as percentage of fruit with a visible lesion. Measurements were
conducted at storage times of 0, 3, 6, 9, and 12 days after room temperature with 3
replications per quality measurement.
EXTRACTION OF POLYPHENOLICS. The extraction of polyphenolics was
carried out using a modified method of Rodriguez -Saona and Wrolstad (2001).
Briefly, fruit were cryogenically powered with liquid nitrogen, a 5 g powered sample
was mixed with 100% (v/v) aqueous acetone (EMD Chemicals Inc., Gibbstown, NJ),
sonicated for 1 min and centrifuged for 5 min, and the remaining filtrate was re-
extracted twice using 70% (v/v) aqueous acetone. The aqueous phase at the top was
combined and transferred into a glass centrifuge bottle with 50 mL of chloroform
(Mallinckrodt Baker Inc., Phillipsburg, NJ) added and mixed. After centrifugation at
3000g for 30 min, the aqueous phase was collected to a rotary evaporator (Brinkmann
Instruments, Westbury, NY) to remove the residual acetone. The extract was diluted in
de-ionized water to desired concentration and stored at -70 oC until analysis.
TOTAL ANTIOXIDANT CONTENT ANT AND PHENOLICS. Total antioxidant
content (AC) was determined using DPPH assay (Brand-Williams et al., 1995). The
sample extract was mixed with 1.5 mL DPPH (1, 1-diphenyl-2-picryhydrazyl) (Kasel
Kogyo Co. Ltd, Tokyo, Japan) in a small screw-cap test tube, vortexed, and set at room
temperature for 5 min. The water soluble antioxidant content was determined
spectrophotometrically at 517 nm with the absorbance of ascorbic acid (Mallinckrodt
Baker Inc., Phillipsburg, NJ) used as a standard. Results with two replications were
131
expressed as mg of ascorbic acid equivalents (AAE) per gram fresh weight (FW) of
sample.
Total phenolic content (TPC) was measured by a modified method from Singleton
and Rossi (1965). A series of test tubes containing 7.5 mL de-ionized water and 0.5mL
Folin-Ciocalteu reagent (Sigma Chemical Co., MO) were prepared. A 0.5 mL diluted
extract was mixed with 0.5 mL of 50, 100, 150, 200 ppm gallic acid (Sigma Chemical
Co., MO) solutions, respectively, and 0.5 mL de-ionized water was used as control.
After the solutions were mixed by a vortexer and set at room temperature for 10 min,
they were mixed with 3 mL 20% (w/v) Na2CO3, then placed in water bath at 40 oC for
20 min. The samples were immediately cooled to room temperature in ice bath for 3
min. The absorbance of samples and a standard were analyzed by spectrophotometer at
765nm. TPC was calculated as mg of gallic acid equivalents (GAE) per gram FW of
sample. AC and TPC measurements were performed at storage times of 0, 6, 12 days
for both cultivars with two replicates.
STATISTICAL ANALYSES. Data analyses were performed by ANOVA (analysis
of variance) and GLM (general linear model) using SAS statistical software 9.01 (SAS
institute, Cary, NC). Multiple comparisons among the treatments with significant
differences tested in ANOVA were conducted by using LSD (least significant
difference) at P<0.05.
132
RESULTS & DISCUSSION
Based on ANOVA results, during 1-week cold storage, AC and TPC of ‘Duke’ and
pH and TA of ‘Elliott’ were affected (P<0.05) by coating treatments, while FN and TPC
of ‘Duke’ and TA and TPC of ‘Elliott’ were affected (P<0.05) by storage time (Table
1).
133
Table 1. ANOVA results of ‘Duke’ and ‘Elliott’ fruit during cold storage
Duke
Factors TSSa pH TAb FNc ACg * TPCh
df 5 5 5 5 5 5
SS 1.74 0.34 0.003 0.03 1.77 524.08
MS 0.35 0.01 0.0005 0.01 0.35 104.82
Coating Treatment F 1.15 0.27 0.09 2.64 11.23 6.17
Pr＞F 0.44 0.91 0.99 0.16 0.0003 0.005
df 1 1 1 1 1 1
SS 0.92 0.05 0.04 0.02 0.018 4144.72
Storage Time MS 0.92 0.05 0.04 0.02 0.018 4144.72
F 3.03 1.95 0.71 9.53 0.57 244
Pr＞F 0.14 0.22 0.44 0.03 0.47 <0.0001
Elliott
a
Factors TSS pH TAb FNc ACg TPCh *
df 3 3 3 3 3 3
SS 0.61 0.045 0.051 0.02 <0.0001 556.66
Coating Treatment MS 0.20 0.015 0.017 0.007 <0.0001 185.55
F 3.68 14.23 11.88 1.71 1.46 1.99
Pr＞F 0.16 0.028 0.036 0.34 0.30 0.19
df 1 1 1 1 1 1
SS 0.056 0.005 0.016 0.004 <0.0001 1119.32
Storage Time MS 0.056 0.005 0.016 0.004 <0.0001 1119.32
F 1.02 4.31 11.30 0.96 2.05 12.01
Pr＞F 0.39 0.13 0.044 0.4 0.19 0.0085
a
Total soluble solids; b Titratable acidity, reported as % malic acid (mass/mass) on the
basis of fresh weight of fruit; c Firmness, N; d Weight loss, %; e Decay rate, %;
f
Percentage of marketable fruits, %; g Antioxidant content, mg ACE/ g FW fruit;
h
Total phenolic content, mg GAC/g FW fruit; * Significant interaction between coating
treatment and storage time.
During room temperature storage, all the quality parameters measured in this study
were affected (P<0.05) by storage time for both cultivars, and coating treatments
impacted (P<0.05) WL and PMF in ‘Duke’ and all measured quality parameter except
TSS and TPC in ‘Elliott’ (Table 2).

134
Table 2. ANOVA results of ‘Duke’ and ‘Elliott’ fruit during room temperature
storage
Duke
Factors TSSa pH TAb FNc WLd DRe PMFf ACg TPCh
df 5 5 5 5 5 5 5 5 5
SS 2.24 0.93 0.009 0.044 16.71 179.44 155.84 0.42 181.76
Coating MS 0.45 0.019 0.001 0.009 3.34 35.89 31.17 0.08 36.35
Treatment F 1.41 1.33 1.33 1.67 2.99 2.45 3.45 1.83 0.97
Pr＞F 0.27 0.29 0.29 0.19 0.036 0.069 0.021 0.16 0.46
df 4 4 4 4 4 4 4 2 2
SS 5.71 0.42 0.11 0.12 383.76 2110.03 9649.26 2.89 2861.27
Storage MS 1.43 0.10 0.027 0.03 95.94 527.51 2412.31 1.45 1430.63
Time F 4.49 7.49 19.87 5.71 85.79 36.01 267.27 31.63 37.99
Pr＞F 0.01 <0.001 <0.0001 0.003 <.0001 <.0001 <.0001 <0.0001 <0.0001
Elliott
Factors TSSa pH TAb FNc WLd DRe PMFf ACg TPCh
df 3 3 3 3 3 3 3 3 3
SS 4.80 0.12 0.31 0.67 7.52 1725.16 1384.18 0.0004 454.26
Coating
MS 1.60 0.04 0.10 0.22 2.51 575.05 461.39 0.0001 151.42
Treatment
F 1.88 3.77 10.32 23.65 2.77 38.72 35.44 5.69 2.99
Pr＞F 0.15 0.02 <0.0001 <0.0001 0.06 <0.0001 <0.0001 0.012 0.07
df 4 4 4 3 3 4 4 2 2
SS 13.5 0.89 0.40 4.67 1171.80 3271.71 28703.30 0.004 3639.47
Storage
MS 3.38 0.22 0.10 1.56 390.60 817.93 7175.83 0.002 1819.74
Time
F 3.97 21.64 9.94 166.08 432.02 55.07 551.13 91.18 35.89
Pr＞F 0.008 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001
a
Total soluble solids; b Titratable acidity, reported as % malic acid (mass/mass) on the
basis of fresh weight of fruit; c Firmness, N; d Weight loss, %; e Decay rate, %;
f
Percentage of marketable fruits, %; g Antioxidant content, mg ACE/ g FW fruit;
h
Total phenolic content, mg GAC/g FW fruit.
pH, TOTAL ACIDITY AND TOTAL SOLUBLE SOLIDS. For ‘Duke’, pH, TA
and TSS were not significantly (P>0.05) affected by coating treatment or cold storage
time (Table 1), but pH tended to increase while TA and TSS decreased based on the
overall means of all the ‘Duke’ samples pre- and post- 1-week cold storage (Table 3).
Non-washed control (3.81, 13.67%) ranked the lowest mean in pH but highest in TSS
over cold storage compared to that in Washed (3.94, 13.17%) and Washed coated
‘Duke’ (3.92-3.96, 12.50%-12.83%) (Table 3 and 4). The overall means of TA in
‘Duke’ over cold storage fell in a narrow range of 0.62 to 0.65% (Table 3).
135
Table 3. Overall means of main effects on pH, TA, TSS and FN of ‘Duke’ and
‘Elliott’ fruit during 1-week cold storagea
Duke
pH TA TSS FN
Non-washedb 3.81 0.63 13.67 1.80
Washedc 3.94 0.61 13.17 1.74
Non-washed SFd 3.95 0.65 12.67 1.69
Coating e
Washed SF 3.94 0.63 12.83 1.68
Treatment f
Washed CH 3.96 0.64 12.50 1.67
Washed CCg 3.92 0.66 12.83 1.68
P value 0.91 0.99 0.44 0.16
Pre-CS 3.85 0.65 13.22 1.75
Storage
Post-CS 3.98 0.62 12.67 1.67
Time
P value 0.22 0.44 0.14 0.03
Elliott
b
Non-washed 3.25 1.10 14.17 1.94
Washedc 3.25 1.09 13.84 1.88
Coating h
Non-washed CC 3.23 1.19 14.50 1.80
Treatment
Washed CCg 3.07 1.29 13.84 1.85
P value 0.028 0.036 0.16 0.34
Pre-CS 3.18 1.12 14.00 1.84
Storage
Post-CS 3.22 1.21 14.17 1.89
Time
P value 0.13 0.044 0.39 0.4
a
Values are means (n=3); b No washing and no coating served as a control; c Washing
but no coating; d Coating of SemperfreshTM without washing; e Washing and coating of
SemperfreshTM; f Washing and coating of chitosan; g Washing and coating of calcium
caseinate; h Coating of calcium caseinate without washing.
136
Table 4. Physicochemical and nutritional properties of ‘Duke’ and ‘Elliott’ fruit during 1-week cold storagea
Duke
Treatments pH TA TSS FN AC TPC
b
Non-washed Pre-CS 3.66±0.40 0.61±0.09 14.0±1.00 1.80±0.03 3.38±0.13 4.16±0.10
Post -CS 3.95±0.11 0.65±0.05 13.3±0.58 1.81±0.01 2.64±0.02 2.12±0.03
c
Washed Pre-CS 3.77±0.08 0.67±0.06 13.7±0.58 1.79±0.03 2.42±0.04 3.49±0.08
Post -CS 4.11±0.15 0.55±0.10 12.7±0.58 1.70±0.07 2.55±0.08 2.11±0.07
d
Non-washed SF Pre-CS 3.83±0.14 0.72±0.13 12.3±1.15 1.70±0.05 2.57±0.22 3.59±0.29
Post -CS 4.08±0.12 0.58±0.09 13.0±0.00 1.68±0.01 2.49±0.11 2.45±0.22
Washed SFe Pre-CS 3.94±0.34 0.66±0.16 13.0±1.00 1.74±0.03 1.83±0.03 3.10±0.07
Post -CS 3.94±0.02 0.59±0.02 12.7±1.15 1.62±0.02 2.41±0.12 2.08±0.10
Washed CHf Pre-CS 4.09±0.21 0.56±0.11 13.3±0.58 1.75±0.08 2.81±0.08 4.02±0.15
Post -CS 3.83±0.12 0.71±0.09 11.7±0.58 1.59±0.01 2.69±0.49 2.60±0.30
g
Washed CC Pre-CS 3.84±0.03 0.70±0.04 13.0±1.00 1.73±0.09 2.24±0.11 3.32±0.43
Post -CS 3.99±0.23 0.61±0.13 12.7±0.58 1.62±0.05 2.79±0.16 2.42±0.19
Elliott
Non-washedb Pre-CS 3.20±0.03 1.08±0.05 14.0±1.00 1.87±0.08 7.13±0.14 4.99±0.96
Post -CS 3.30±0.08 1.11±0.06 14.3±0.58 2.00±0.03 6.65±0.10 5.53±0.15
Washedc Pre-CS 3.21±0.03 1.05±0.08 13.7±1.15 1.88±0.07 7.23±0.03 5.56±0.19
Post -CS 3.28±0.05 1.13±0.11 14.0±1.00 1.88±0.10 7.20±0.35 6.26±0.63
h
Non-washed CC Pre-CS 3.23±0.14 1.11±0.22 14.3±1.15 1.75±0.03 6.93±0.45 5.31±0.61
Post -CS 3.23±0.14 1.27±0.03 14.7±0.58 1.85±0.16 7.11±0.02 6.65±0.27
Washed CC Pre-CS 3.06±0.06 1.24±0.05 14.0±0.00 1.88±0.03 7.10±0.22 5.54±0.19
Post -CS 3.08±0.05 1.33±0.02 13.7±0.58 1.81±0.08 6.79±0.07 6.31±0.10
a
Values are means (n=3)
137
During room temperature storage of “Duke”, coating treatments didn’t show effect
on TA, pH and TSS (P>0.05), but the mean of TA declined significantly (P<0.05) over
time from the initial 0.62% to 0.43% at 9 days, mean of pH tended to increase during
the first 3 days, then declined over time reaching 3.84 at 9 days, significantly lower than
4.20 at 3 days. In contrast, TSS tended to decrease during the first 3 days, but increased
during the rest of storage, reached to 13.22% at 9 days which was significantly higher
than 12.00% at 3 days (Fig. 1).

138
16 5.0
Duke Duke
4.5
14
TSS (%)
pH
4.0
12
3.5
10 3.0
0 3 6 9 12 0 3 6 9 12
Room storage time (day) Room storage time (day)
0.8 2.2
Duke Duke
0.7 2.1
0.6 2.0
Firmness (N)
0.5 1.9
TA
0.4 1.8
0.3 1.7
0.2 1.6
0.1 1.5
0.0 1.4
0 3 6 9 12 0 3 6 9 12
Fig. 1. Physicochemical properties of ‘Duke’ fruit during room temperature storage. Since no significant differences
(P>0.05) in TSS, pH, TA and firmness among different treatments were identified, mean values of all treatments were
reported.
139
18 Duke
16
Non-washed Washed
14 Non-washed SF Washed SF
Washed CH Washed CC
12
Weight loss (%)
10
6
4
0
0 3 6 9 12
Room storage time (day)
50
Duke
45
40 Non-washed Washed
Non-washed SF Washed SF
35 Washed CH Washed CC
Decay rate (%)
30
25
20
15
10
5
0
0 3 6 9 12
Fig. 1. Physicochemical properties of ‘Duke’ fruit during room temperature

storage. Since no significant differences (P>0.05) in TSS, pH, TA and firmness
among different treatments were identified, mean values of all treatments were
reported. (Continued)
140
For ‘Elliott’, Washed CC had significantly (P<0.05) lower pH (3.06, 3.08), but
higher TA (1.24%, 1.33%) than Non-washed (3.20, 3.30; 1.08%, 1.11%) and Washed
(3.21, 3.28; 1.05%, 1.13%) both pre-and post- cold storage (Table 3). The means of pH
and TA increased from 3.18 to 3.22 and from 1.12% to 1.21%, respectively during cold
storage (Table 3). During room storage, CC coating showed significantly effect on pH
and TA, where Washed CC and Non-washed CC had significantly (P<0.05) lower pH
and higher TA (mean of pH=3.27 and TA=1.13%) than those of Washed and Non-
washed (mean of pH=3.36 and TA=0.99%), while no significant differences were
observed in TSS among the treatments (Fig. 2). In respect to the storage time, pH
stayed stable during the first 6 days of room temperature storage, but increased from 6
to 12 days, while TA gradually decreased throughout the storage period (Fig. 2).
Similar to ‘Duke’, washing itself didn’t show significant impact on pH, TA and TSS of
‘Elliott’.
141
1.6
4.0 Elliott Non-w ashed
Elliott Non-w ashed Washed
Washed Non-w ashed CC
3.8 Non-w ashed CC 1.4 Washed CC
Washed CC
3.6
1.2
TA
pH
3.4
1
3.2
3.0 0.8
2.8 0.6
0 3 6 9 12 0 3 6 9 12
16 3.0 Non-w ashed

Elliott Elliott
Washed
15 Non-w ashed CC
2.5 Washed CC
14
Firmness (N)
TSS (%)
13 2.0
12
1.5
11
10 1.0
0 3 6 9 12 0 6 9 12
Fig. 2. Physicochemical properties of ‘Elliott’ fruit during room temperature storage. Since no significant differences
(P>0.05) in TSS and weight loss among different treatments were identified, mean values of all treatments were
reported.
142
18
Elliott
16
14
Weight loss (%)
12
10
8
6
4
2
0
0 3 9 12
60 Non-w ashed
Elliott
Washed
50 Non-w ashed CC
Washed CC
Decay rate (%)
40
30
20
10
0
0 3 6 9 12
Fig. 2. Physicochemical properties of ‘Elliott’ fruit during room temperature

storage. Since no significant differences (P>0.05) in TSS and weight loss among
different treatments were identified, mean values of all treatments were reported.
(Continued)
143
During post-harvest storage, the acid metabolism as a result of fruit ripening
continued by converting starch and acid to sugar, resulted in decrease in TA and
increase in pH and TSS (Thompson, 1996; Verma and Joshi, 2000). The results on CC
coated fruit confirmed previous studies that CC coating helped control the changes of
posr-harvest physicochemical properties, such as pH and TA. This may be explained as
the protein-based CC delayed post-harvest respiration of fruit by providing a strong gas
barrier on the surface of fruit (Khwaldia et al., 2004; Letien et al., 2001; Lin and Zhao,
2007).
FIRMNESS. FN of all the ‘Duke’ samples except Non-washed control was reduced
(p<0.05) after 1-week cold storage, with reduction of 5% for Washed, 1% for Non-
washed SF, 7% for Washed SF, 9% for Washed CH, and 6% for Washed CC,
respectively (Table 4). There were no significant differences (P>0.05) in FN of ‘Duke’
samples subjected to different treatments during both cold and room temperature
storage (Table 2 and 3). Mean of FN in ‘Duke’ increased initially (P<0.05) during the
first 3 days of room storage (from 1.67N to 1.82N), then remained stable afterwards
(1.82-1.84N) (Fig. 1).
FN of ‘Elliott’ didn’t show significant (P>0.05) change after 1-week cold storage
(Tables 1 and 3). Mean of FN of Non-washed control (1.94N) ranked higher than all
other ‘Elliott’ samples (1.80-1.88 N) after 1-week cold storage. During room storage,
CC coated ‘Elliott’ had significantly (P<0.05) higher FN (mean of 1.99 N) than those
non-coated ones (mean of 1.75 N), and FN of coated samples slightly increased during
the first 9 days of room storage, then dropped, while those uncoated ‘Elliott’
144
continuously decreased throughout the storage period (Fig. 2). Washing did not affect
FN of ‘Elliott’ during room storage (P>0.05).
Fruit softening, one of the important quality deteriorations during post-harvest
storage, is generally caused by the hydrolysis of starch to sugar and the degradation of
pectin in the fruit cell wall associated with fruit ripening (Thompson, 1996). In
contrast, water loss of fruit may lead to the hardening of fruit. Both fruit softening and
hardening affect the measured fruit firmness. In this study, we demonstrated that CC
coating helped maintain the firmness of ‘Elliott’ during room storage. Calcium
caseinate (CC) has been shown to provide calcium ions which may be chelated by
adjacent acidic pectin polymers in the cell wall through non-covalent linkage forming
an “egg box” model at the biochemical level during storage (Seymour et al., 1993), thus
enhanced the firmness of ‘Elliott’ fruit.
WEIGHT LOSS. Overall, weight loss of both blueberry cultivars increased
(P<0.05) throughout the room storage period (Fig. 1 and 2). In ‘Duke’, no significant
differences were observed at the first 3 days of room storage, but at 6 days, Washed
(2.28%) and Non-washed SF (4.85%) had less weight loss than other ‘Duke’ samples
(6.38-6.66%) (Fig. 1). Weight loss of Non-washed SF and Washed SF ‘Duke’ (9.83%,
9.31%) was significantly lower than other ‘Duke’ samples at 12 day (Fig. 1). Similar to
‘Duke’, WL of ‘Elliott’ increased during room storage, up to a mean of 13% at the end
of 12 days (Fig. 2). Washed non-coated ‘Elliott’ (mean of 7.89 %) had the highest WL
among all samples (mean of 6.82-7.19%) over the 12 days of room storage (P<0.06).
145
Migration of water from the fruit to the environment is the major cause of weight
loss of fruit during storage. Our results were consistent with previous studies that a
hydrophobic coating material, such as SF, had high barrier to water loss (Morillon et al.,
2002), while CH and CC were hydrophilic coating materials with relatively high
moisture permeability. Therefore, it may be necessary to incorporate lipids into
hydrophilic coating formulae for better control of water loss of coated blueberries if this
is a major goal when applying coatings.
DECAY RATE. Fruit decay rate in both blueberry cultivars increased along with
the time under room temperature storage (Fig. 1 and 2). Although there was no overall
treatment effects on decay rate of ‘Duke’ (P>0.05) (Table 2), Washed CH ‘Duke’
showed a lower value (9.98% at 12 days) than other samples (20.57%-30.55% at 12
days) throughout the room storage (P<0.069), and Washed CC had lower decay rate
than Non-washed CC at 9 and 12 days (Fig. 1). In respect to ‘Elliott’, CC coating again
showed significant (P<0.05) effect on decay rate (P<0.05), but a opposite way, where
CC coated ‘Elliott’ showed higher value (mean of 34.82 at 12 days) than non-coated
ones (14.73% at 12 days) throughout the 12 days of room storage, and Non-washed CC
(24.67-37.66%) had higher number than Washed CC (17.92-29.97%) during 9-12 days
(P<0.05) (Fig. 2).
The anti-fungal function of CH coating to prevent fruit decay has been well
reported in several studies (Chien et al., 2007; Han et al., 2004; Park et al., 2004; Zhang
and Quantick, 1998;). However, this anti-fungal property may be limited by several
factors. Dipping and washing in chlorinated water may remove the natural waxy layer
146
on the surface of blueberries, thus weakening adhesion and durability of coatings.
Moreover, residual water on the surface of blueberries, after washing, possibly diluted
or dissolved applied coating materials, made it difficult to form a uniform and durable
layer of edible films on the fruit surface (Lin and Zhao, 2007). However, CC, as a
protein based material, may provide additional nutrients for fungi to grow when the fruit
is contaminated, potentially leading to increased decay rate. Washing using chlorine
solution has shown the benefit of decreasing fruit decay.
ANTIOXIDANT CONTENT. AC of Non-washed ‘Duke’ (3.38 mg AAE/g FW)
was significantly (P<0.05) higher than those of other ‘Duke’ samples (1.83-2.81 mg
AAE/g FW) right after coating treatment (Table 4). After cold storage, AC degraded
22% in Non-washed ‘Duke’ (P<0.05) while kept stable in Washed, Non-washed SF,
and Washed CH, but increased 32% and 25% in Washed SF and Washed CC,
respectively (p<0.05) (Table 4).
There was almost a linear increase in AC of ‘Duke’ over room storage, from initial
2.59 mg AAE/g FW to 3.29 mg AAE/g FW after 12 days (Fig. 3). Weight loss of
‘Duke’ (up to 14%) during room temperature storage might have caused the increase in
AC calculated on the basis of fresh fruit weight. Washed CH and Washed CC ‘Duke’
(3.46 and 3.67 mg AAE/g FW, respectively) had significantly (P<0.05) higher AC than
Non-washed (3.03 mg AAE/g FW) and Washed SF (3.25 mg AAE/g FW) at the end of
storage.
147
5.0
Antioxidant content (mg 4.5 Duke
AAE/g FW) 4.0
3.5
3.0
2.5
2.0
1.5
1.0
0 6 12
4.0
Duke
3.5
Total phenolics (mg
3.0
GAE/g FW)
2.5
2.0
1.5
1.0
0 6 12
Fig. 3. Antioxidant content and total phenolic content of ‘Duke’ fruit during room
temperature storage. Since no significant differences (P>0.05) among different
treatments were identified, mean values of all treatments were reported.
148
AC in Non-washed and Washed CC ‘Elliott’ declined 7% and 4%, respectively after
cold storage (p<0.05) (Table 4), but showed no change in Washed and Non-washed CC.
No differences were found among treated ‘Elliott’ with mean of 7.10 and 6.94 mg
AAE/g FW pre- and post- cold storage, respectively. Connor et al. (2002) reported that
fresh ‘Elliott’ harvested before turning fully blue showed an increase in antioxidant
activity after harvest and during cold storage. Therefore, changes in antioxidant
activities are likely affected by storage conditions and stage of fruit ripeness at harvest.
AC of all ‘Elliott’ samples declined at the first 3 days of room temperature storage,
from mean of 6.94 to 5.48 mg AAE/g FW, but kept stable from 6 to 12 days (Fig. 4).
149
8
Elliott
Total phenolics (mg 7
GAE/g FW) 6
5
4
3
2
1
0
0 6 12
9.0
8.5 Elliott
Antioxidant content (mg
Non-washed
8.0 Washed
7.5 Non-washed CC
AAE/g FW)
7.0 Washed CC
6.5
6.0
5.5
5.0
4.5
4.0
0 6 12
Fig. 4. Antioxidant content and total phenolic content of ‘Elliott’ fruit during room
temperature storage. Since no significant differences (P>0.05) in total phenolic
content among different treatments were identified, mean values of total phenolic
content were reported.
150
TOTAL PHENOLIC CONTENT. TPC in all ‘Duke’ samples decreased (P<0.05)
during cold storage, resulting in 27-49% loss depending on specific treatment (Table 4).
According to Verma and Joshi (2000) and Seymour (1993), the oxidation of phenolics
in ‘Duke’ catalyzed by polyphenoloxidase and peroxidase during post–harvest storage
is a major reason for the quick drop of TPC. However, TPC of ‘Duke’ increased at the
begin and then kept stable during room temperature storage (Fig. 3). At the 12 days of
room storage, Washed CH (3.20 mg GAE/g FW) and CC (3.48 mg GAE/g FW), and
Non-washed SF (3.41 mg GAE/g FW) had significantly higher TPC than Non-washed
control (2.94 mg GAE/g FW). This result indicated that the intermolecular binding
effect of CH, CC and SF coatings on ‘Duke’ may help stabilize the phenolics, thus
effectively slowing the degradation of phenolic compounds associated with fruit
ripening at the end of room storage.
Unlike ‘Duke’, TPC of ‘Elliott’ didn’t change (P>0.05) in Non-washed (mean=5.26
mg GAE/g FW) and Washed control (mean= 5.91 mg GAE/g FW) during cold storage,
but increased 25% in Non-washed CC and 14% in Washed CC (p<0.05) (Table 4),
which agreed with the results on fresh ‘Elliott’ reported by Connor (2002). During
room temperature storage, mean of TPC in ‘Elliott’ decreased from 0 day (6.19 mg
GAE/g FW) to 6 days (5.39 mg GAE/g FW), but increased from 6 days to 12 days (6.90
mg GAE/g FW) (Fig. 4). Specifically, TPC of Non-washed increased over storage time
and that of Washed remained stable and then increased significantly after 6 day.
Whereas, coated ‘Elliott’ decreased initially and then increased at the end of storage.
151
Washed, uncoated ‘Elliott’ had higher TPC than other ‘Elliott’ samples (P<0.05) at the
end of room temperature storage.
COMPARISON OF TWO BLUEBRRY CULTIVARS. Significant differences
between the two blueberry cultivars in post-harvest quality and response to coating
treatment were observed in this study. ‘Elliott’ is a late–season cultivar harvested
starting in August, compared to the early-season ‘Duke’ where harvest starts at the end
of June in the Willamette Valley, Oregon. ‘Elliott’ fruit had lower pH and higher TA
than those of ‘Duke’, while ‘Duke’ had a larger fruit size. In comparison with other
reported studies, the fruit pH of ‘Duke’ in our study, for Washed or Non-washed fruit
was slightly higher than that reported by Zheng et al. (2003), but the pH of ‘Elliott’ was
similar to that reported by Connor et al. (2002). TA and TSS of both cultivars were
slightly lower and higher than those reported by Connor (et al., 2002) and Zheng (et al.,
2003), respectively, which may have been a result of differences in macroclimate or
harvest time.
In respect to antioxidant capacity of the 2 cultivars, AC of ‘Elliott’ was significantly
higher than that of ‘Duke’. This result agreed with the previous report that the later
harvested ‘Elliott’ had higher antioxidant activity than earlier harvested ones such as
‘Bluecrop’ and ‘Jersey’ (Connor, 2002).
CONCLUSIONS
In conclusion, different coatings had various effects on the post-harvest quality of
fresh blueberries. Chitosan coating showed the potential to control the decay incidence
152
of ‘Duke’ during storage under room conditions. SemperfreshTM coating helped control
weight loss of ‘Duke’, while calcium caseinate coatings tended to improve the firmness
of ‘Elliott’ during room storage, but not weight loss and decay rate. In general, washing
the fruit using chlorine solution showed the benefit of reducing decay incidence in both
cultivars. All three coatings, SemperfreshTM, chitosan and calcium caseinate
demonstrated the potential to help retain antioxidant and total phenolic content of
‘Duke’ during room temperature storage, but calcium caseinate coating did not show
this benefit on ‘Elliott’.
Results from this study indicated the possibility of using edible coatings to develop
ready-to-eat fresh blueberries with extended shelf-life. The key for success is using an
appropriate coating material, formulation, and the method of applying coating. New
coating materials with enhance microbial barrier and antimicrobial functionality may be
evaluated, or modification on the existing coating formulation may be studied to obtain
high quality and nutritional value of fruit. The current dipping method of applying
coating might have caused the loss of some coating benefits; thereby new coating
application methods should be investigated. The use of electrostatic spraying for
coating application is under the way to be evaluated by the authors for achieving more
uniform coating and avoiding the removal of the natural waxy layer on the surface of
blueberries for maximizing the benefits of coatings.

153
LITERATURE CITED
Abidov, M., D.R.M. Jimenez, A. Ramazanov, O. Kalyuzhin, and I. Chkhikvishvili.

2006. Efficiency of pharmacologically-active antioxidant phytomedicine Radical
Fruits in treatment hypercholesteremia at men. Georgian medical news. 140: 78-83.
Avena-Bustillos, R.J., J.M. Krochta, M.E. Saltveit. 1997. Water vapor resistance of red
delicious apples and celery sticks coated with edible caseinate-acetylated
monoglyceride films. J. Food Sci. 62:351-354.
Baldwin, E.A., J.K. Burns, W. Kazokas, J.K. Brecht, R.D. Hagenmaier, R.J. Bender,
and E. Pesis. 1999. Effect of two edible coatings with different permeability
characteristics on mango (Mangifera indica L.) ripening during storage. Postharv.
Biol. Tech. 17:215–226.
Baldwin, E.A., M.O. Nisperos-Carriedo, and R.A. Baker. 1995. Edible coating for
lightly processed fruits and vegetables. Hortscience. 30:35-38.
Ballinger, W.E., E.P. Maness, and W.F. McClure. 1978. Relationship of stage of
ripeness and holding temperature to decay development of highbush blueberries. J.
Amer. Soc. Hort. Sci. 103:130-134.
Beaudry, R.M., C.E. Moggia, J.B. Retamales, and J.F. Hancock. 1998. Quality of
‘Ivanhoe’ and ‘Bluecrop’ blueberry fruit transported by air and sea from Chile to
North America. HortScience 33:313-317.
Brand-Williams, W., M.E. Cuvelier, and C. Berset.1995. Use of a free radical method to
evaluate antioxidant activity. Lebensmittel-Wissenschaft Technol. 28:25-30.
Chiabrando, V., C. Peano, G. Beccaro, G. Bounous, and L. Rolle. 2006. Postharvest

quality of highbush blueberry (Vaccinium corymbosum L.) cultivars in relation to
storage methods. Acta Hort. 715:545-551.
Chien, P.J., F.H. Yang, and F.Sheu. 2007. Effects of edible chitosan coating on quality
and shelf life of sliced mango fruit. J. Food Eng. 78:225-229.
Connor, M.A., J.J. Luby, F.J. Hancock, S. Berkheimer, and J.E. Hanson. 2002. Changes
in Fruit Antioxidant Activity among Blueberry Cultivars during Cold-Temperature
Storage. J. Agric. Food Chem. 50:893-898.
Fisk, L.C., M.A. Silver, B.C. Strik, and Y. Zhao. 2008. Postharvest quality of hardy
kiwifruit (Actinidia arguta ‘Ananasnaya’) associated with packaging and storage
conditions. Postharv. Biol. Tech. 47:338-345.
154
Fornes, F., V. Almela, M. Abad, and M. Agustí. 2005. Low concentration of chitosan
coating reduce water spot incidence and delay peel pigmentation of clementine
mandarin fruits. J. Sci. Food. Agric. 85:1105-1112.
Galletta, G.J., W.E. Ballinger, R.J. Moore, and L.J. Kushman. 1971. Relationship
between fruit acidity and soluble solid levels of highbush blueberry clones and their
keeping quality. J. Amer. Soc. Hort. Scie. 96:756-762.
Han, C., Y. Zhao, S.W. Leonard, and M.G. Traber. 2004. Edible coatings to improve
storability and enhance nutritional value of fresh and frozen strawberries (Fragaria ×
ananassa) and raspberries (Rubus ideaus). Postharv. Biol. Tech. 33:67-68.
Hancock, J., P. Callow, S. Serce, E. Hanson, and R. Beaudry. 2008. Effect of cultivar,
controlled atmosphere storage, and fruit ripeness on the long-term storage of
highbush blueberries. HortTech. 18:199-205.
Kahlon, T.S. and G.E. Smith. 2007. In vitro binding of bile acids by blueberries
(Vaccinium spp.), plums (Prunus spp.), prunes (Prunus spp.), strawberries (Fragaria
× ananassa), cherries (Malpighia punicifolia), cranberries (Vaccinium macrocarpon)
and apples (Malus sylvestris). Food Chem. 100:1182–1187.
Kalt, W., D.A. J. Ryan, J.C. Duy, L.R. Prior, M.K. Ehlenfeldt, and S.P.V. Kloet. 2001.
Interspecific variation in anthocyanins, phenolics, and antioxidant capacity among
genotypes of highbush and lowbush blueberries (Vaccinium Section cyanococcus
spp.). J. Agric. Food Chem. 49:4761-4767.
Kaynas, K. and I.S. Ozelkok. 1999. Effect of Semperfresh on postharvest behavior of

cucumber (Cucumis sativus L.) and summer squash (Cucubita pepo L.) fruits. Acta
Hort. 492:213-220.
Khwaldia, K., C. Perez, S. Banon, S. Desobry, and J. Hardy. 2004. Milk proteins for
edible films and coatings. Crit. Rev. Food Sci. Nutr. 44:239–251.
Letien, C., C. Vachion, M.A. Mateescu, M. Lacroix. 2001. Milk protein coatings
prevent oxidative browning of apples and potatoes. J. Food Sci. 66:512-516.
Lin, D.and Y. Zhao. 2007. Innovation in development and application of edible coatings
for fresh and minimally processed fruits and vegetables. Comp. Rev. Food Sci. Food
Safety. 6:1-15.
155
Martineau, L.C., A. Couture, D. Spoor, A. Benhaddou-Andaloussi, C. Harris, B.

Meddah, C. Leduc, A. Burt, T. Vuong, L.P. Mai, M. Prentki, S.A. Bennett, J.T.
Arnason, and P.S. Haddad. 2006. Anti-diabetic properties of the Canadian lowbush
blueberry Vaccinium angustifolium Ait. Inter. J. Phytotherapy Phytopharmacology
(Phytomedicine). 13:612-623.
Mei, Y. and Y. Zhao. 2003. Barrier and mechanical properties of milk protein-based
edible films containing nutraceuticals. J. Agric. Food Chem. 51:1914-1918.
Morillon, V., F. Debeaufort, G. Blond, M. Capelle, and A. Voilley. 2002. Factors

affecting the moisture permeability of lipid-based edible films: a review. Cri. Rev.
Food Sci.Nutri. 42: 67-89.
Nimitkeatkai, H., V. Srilaong, and S. Kanlayanarat. 2006. Effect of edible coating on

Pineapple fruit quality during cold storage. Acta Hort. 712:643-647.
Park H.J. 1999. Development of advanced edible coatings for fruits. Trends Food Sci.
Tech. 10:254-260.
Park, S.I., S.D. Stan, M.A. Daeschel, and Y. Zhao. 2005. Antifungal coatings on fresh
strawberries (Fragaria x ananassa) to control mold growth during cold storage. J.
Food Sci. 70:202-207.
Pen, L.T. and Y.M. Jiang. 2003. Effects of chitosan coating on shelf life and quality of
fresh-cut Chinese water chestnut. Lebensmittel-Wissenschaft Und-Technologie-
Food Sci Technol. 36:359-364.
Ribeiro, C., A.A. Vicente, A.J. Teixeira, and C. Miranda. 2007. Optimization of edible
coating composition to retard strawberry fruit senescence. Postharv. Biol. Tech.
44:63–70.
Rodriguez-Saona, L.E. and R.E. Wrolstad. 2001. Extraction, isolation and purification
of anthocyanins. Current Protocols in Food Analytical Chemistry. Hoboken, NJ:
John Wiley & Sons, Inc. F1.1.1-F1.1.11.
Seeram, N.P., L.S. Adams, Y. Zhang, R. Lee, D. Sand, H.S. Scheuller, and D. Heber.
2006. Blackberry, black raspberry, blueberry, cranberry, red raspberry, and
strawberry extracts inhibit growth and stimulate apoptosis of human cancer cells in
vitro. J. Agric. Food Chem. 54:9329-9339.
Seymour, G.B., J.E. Taylor and Tucker G.A. 1993. Biochemistry of Fruit Ripening.
Chapman & Hall, New York.
156
Singleton, V.L. and J.A. Rossi. 1965. Colorimetry of total phenolics with
phosphomolybdic-phosphotungstic acid reagents. Ameri. J Enol. Viticulture.
16:144-158.
Srivastava, A., C.C. Akoh, W. Yi, J. Fischer, and G. Krewer. 2007. Effect of storage
conditions on the biological activity of phenolic compounds of blueberry extract
packed in glass bottles. J. Agric. Food Chem. 55:2705-2713.
Thompson, A.K. 1996. Postharvest Technology of Fruit and Vegetables. Harlow,

Blackwell Science Ltd., Oxford.
Torri, E., M. Lemos, V. Caliari, C.A. Kassuya, J.K. Bastos, and S.F. Andrade. 2007.
Anti-inflammatory and antinociceptive properties of blueberry extract (Vaccinium
corymbosum). J. Pharm. Pharmacol. 59:591-596.
Trigo, M.J., M.B. Sousa, M.L. Botelho, and G. Veloso. 2006. Quality of gamma
irradiated blueberries. Acta Hort. 715:573-577.
Vachon, C, G. D’Aprano, M. Lacroix, and M. Letendre. 2003. Effect of edible coating

process and irradiation treatment of strawberry Fragaria spp. on storage-keeping
quality. J. Food Sci. 68:608-612.
Vargas, M., A. Albors, A. Chiralt, and C. Gonzalez-Martınez. 2006. Quality of cold-

stored strawberries as affected by chitosan–oleic acid edible coatings. Postharv.
Biol. Tech. 41:164–171.
Verma, L.R. and V.K. Joshi. 2000. Postharvest technology of fruits and vegetables:
handling, processing, fermentation, and waste management. Indus Pub. Co., New
Delhi, 1222 pp.
Willis, L., P. Bickford, V. Zaman, A. Moore, and A.C. Granholm. 2005. Blueberry
extract enhances survival of intraocular hippocampal transplants. Cell Transplant
14:213-23.
Wilson, M.A., B. Shukitt-Hale, W. Kalt, D.K. Ingram, J.A. Joseph, and C.A. Wolkow.
2006. Blueberry polyphenols increase lifespan and thermotolerance in
Caenorhabditis elegans. Aging Cell. 5:59-68.
Yaman, Ö. and L. Bayoindirli. 2002. Effect of an edible coating and cold storage on
shelf-life and quality of cherries. Lebensmittel-Wissenschaft Und-Technologie-
Food Sci Technol. 35:146-150.
157
Yi, W., C.C. Akoh, J. Fischer, and G. Krewer. 2006. Absorption of anthocyanins from
blueberry extracts by caco-2 human intestinal cell monolayers. J. Agric. Food
Chem. 54:5651-5658.
Zhang, D. and P. C. Quantick. 1998. Antifungal effects of chitosan coating on fresh

strawberries and raspberries during storage. J. Hort. Sci. Biotech. 73:763-767.
Zheng, W. and Y.S. Wang. 2003. Oxygen radical absorbing capacity of phenolics in
blueberries, cranberries, chokeberries, and lingonberries. J. Agric. Food Chem.
51:502-509.
Zheng, Y., Y.C. Wang, Y.S. Wang, and W. Zheng. 2003. Effect of high-oxygen
atmospheres on blueberry phenolics, anthocyanins, and antioxidant capacity. J.
Agric. Food Chem. 51:7162-7169.

Ruyi Wu Thesis

Загружено:

Сведения о документе

Авторское право

Доступные форматы

Поделиться этим документом

Поделиться или встроить документ

Параметры публикации

Этот документ был вам полезен?

Это неприемлемый материал?

Авторское право:

Доступные форматы

Ruyi Wu Thesis

Загружено:

Авторское право:

Доступные форматы

AN ABSTRACT OF THE THESIS OF

presented on December 12, 2008.

Title: Effects of Refrigeration Storage and Processing Technologies on the Bioactive

Compounds and Antioxidant Capacities of Blackberries (‘Marion’ and ‘Evergreen’).

The objective of this project was to investigate the effects of refrigeration

storage and processing technologies on the quality and nutraceutical benefit of

to different processing treatments including freeze drying, hot-air drying, canning-in-

products were stored at room temperature at 25 ± 2 oC and 30 ± 2% relative humidity

for up to 6 months. Physicochemical properties including pH, titratable acidity (TA),

respectively in ‘Evergreen’ (P<0.05) while remained stable in ‘Marion’ during

to 70% (P<0.05) in canned products except TPC in ‘Marion’ canned-in-water. ORAC

blackberries to jams resulted in extensive losses of bioactive compounds and

canned blackberries, ORAC in hot-air dried and canned-in-water blackberries of both

varieties were also observed. Two varieties of blackberries varied in physicochemical

properties, content of bioactive compounds and antioxidant capacities, with higher

Oregon State University

Presented December 12, 2008

Commencement June 2009

Major Professor, representing Food Science and Technology

Head of the Department of Food Science and Technology

Dean of the Graduate School

State University libraries. My Signature below authorizes release of my thesis to any

reader upon request.

Ruyi Wu, Author

This comprehensive work can not be completed successfully by me alone. Here

is my sincere acknowledgement to all the individuals generously contributing to my

research and giving support in my everyday life in Corvallis.

I would like to express my foremost appreciation to my advisor, Professor

grateful to her patient guidance, insightful advices, and constant encouragement

throughout my study at OSU.

I greatly appreciate my graduate committee members, Drs. James Kennedy,

would not have the blackberry products for my thesis research.

I am also very grateful to my colleagues in Dr. Zhao’s research group, Dr.

helps during my experiments.

Corvallis for their continuous care and friendship.

Finally, I would like to deliver my deepest appreciation to my parents and

2. CHAPTER 2 Literature Review............................................................................. 6

2.2 Overview of blackberries……………………………………………............... 7

2.2.1 History and botanical classification of blackberries………………….…... 7

2.3 Post-harvest handling technologies………………………………………....... 25

2.3.4 Edible coatings…………………………………………………............... 27

2.4 Processing of blackberries…………………………………………................ 29

2.6 Literature cited................................................................................................. 39

3. CHAPTER 3 Effects of Refrigeration Storage and Processing Technologies

MATERIALS AND METHODS………………………………………………... 57

RESULTS AND DISCUSSION……………………………………………….... 65

LITERATURE CITED………………………………………………………….. 100

4. CHAPTER 4 General Conclusion……………………………………................. 105

2.1 Diagrammatic scheme of juice processing…………………………………… 36

3.1 Flow diagram of experimental design………………………………............... 58

3.2 Changes of selected physicochemical properties of two blackberry

3.3 Changes of bioactive compounds in two blackberry varieties

3.4 Changes of antioxidant capacities in two blackberry varieties

3.5 Physicochemical properties of two blackberry varieties (‘Marion’

3.6 Bioactive compounds of two blackberry varieties (‘Marion’ and

3.7 Antioxidant capacities of two blackberry varieties (‘Marion’ and

2.1 Types and cultivars of blackberries…………………………………………… 9

2.2 Carbohydrate composition of fresh blackberry fruit (100g)………...…...….... 11

2.3 Vitamin composition of fresh blackberry fruit (100g)……………......………. 16

2.4 Mineral composition of fresh blackberry fruit (100g)………………..……….. 16

2.5 Average fruit weight of fresh blackberries……………………...…………….. 18

2.6 Range of colorimetric parameters of thornless blackberries………………….. 18

3.1 ANOVA results of two blackberry varieties (‘Marion’ and ‘Evergreen’)

3.2 ANOVA results of two blackberry varieties (‘Marion’ and ‘Evergreen’)