EFFECT OF PRETREATMENTS AND DRYING ON THE CHEMICAL

COMPOSITION OF OKRA (Abelmoschus esculentus)

by

Kopila Thapa Magar

Department of Food Technology

GoldenGate International College

Institute of Science and Technology

Tribhuvan University, Nepal

2021
 Effect of pretreatments and drying on the chemical composition of Okra
(Abelmoschus esculentus)

A dissertation submitted to the Department of Food Technology, GoldenGate International

College, Tribhuvan University, in partial fulfillment of the requirements for the degree of
B.Tech. in Food Technology

by

Kopila Thapa Magar

Department of Food Technology

GoldenGate International College, Kathmandu
Institute of Science and Technology
Tribhuvan University, Nepal
January, 2021
 Tribhuvan University
Institute of Science and Technology
Department of Food Technology
GoldenGate International College
Battisputali, Kathmandu

Approval Letter
This dissertation entitled Effect of pretreatments and drying on the chemical composition
of Okra (Abelmoschus esculentus) presented by Kopila Thapa Magar has been accepted
as the partial fulfillment of the requirements for the B.Tech. degree in Food Technology.

Dissertation Committee

1. Head of the Department __________________________

(Prof. Ganga P. Kharel, PhD)

2. External Examiner

(Mr. Pramod Koirala)

(Senior Food Research Officer,
MoALD)

3. Supervisor

(Dr. Alok Shrestha)

January, 2021

iii
 Acknowledgement
At the very beginning, with an immense pleasure and profound sense of gratitude, I take this
opportunity to thank my esteemed supervisor, Dr. Alok Shrestha, coordinator and faculty
member, Department of Food Technology, GoldenGate International College, for his inspiring
guidance, valuable suggestions and guidance throughout my research work. With the same
spirit, I express my fathomless gratitude to benevolent and ever generous Mr. Ramesh Silwal,
CEO and Prof. Dr. Ganga Prasad Kharel, HOD, Department of Food Technology, GoldenGate
International College for providing research and education oriented environment.

It is my privilege to express my sincere regard and heartfelt gratitude to Mr. Pravin Ojha,
Scientist, Food Research Division, NARC, Mr. Pradeep Kaji Poudel and Mr. Rajesh Shrestha,
Faculty members, Department of Food Technology, GoldenGate International College for
their prudent guidance, generous help, painstaking interest and valuable suggestions. I am
equally thankful to all my teachers and friends. I am highly obliged to Miss Srijana Duwal for
her immense support, guidance and valuable time that she has poured over me during the
research. I acknowledge her with respectful thanks. I would also like to thank Mr. Ravi
Shrestha for providing necessary facilities during my lab works.

I feel immense pleasure in expressing appreciation and indebtedness to my friends Saphal

Ghimire, Anita Shrestha, Kamla Rai and Puspa Khatiwoda for their forthright help,
constructive suggestions, inspiration and moral help which went a long way in successful
completion of this study.

My thanks endowed with the feeling of honor to my family members whose support always
motivated me.

Finally, thanking each and every person soliciting good wishes for my future.

Date of submission: January, 2021

…………………………..

(Kopila Thapa Magar)

iv
 Abstract
The present study entitled “Effect of pretreatments and drying on the chemical composition of
Okra (Abelmoschus esculentus)” was carried out with the objectives to estimate the chemical
composition of the fresh Okra and to study the effect of processing on them. At first,
proximate composition of the fresh samples were determined. Fresh Okra was then subjected
to different pretreatments (KMS blanching, NaCl blanching and water blanching) and then
dried at three different temperatures (40±2˚C, 50±2˚C and 60±2˚C). Untreated samples were
also dried at these temperatures and were taken as control samples. Experiments were carried
out to determine the phytochemicals (total polyphenol content, total flavonoid content,
ascorbic acid, total carotenoids and total chlorophyll), antioxidant activity and rehydration
ratio. Samples retaining highest amount of bioactive phytochemicals with better rehydration
ratio were considered to be of best quality and their proximate analysis were carried out.
Calcium content of the fresh and dried Okra was also determined and compared.

The mean values for total polyphenol content, total flavonoid content, ascorbic acid, total
chlorophyll, total carotenoids and antioxidant activity for fresh Okra were 1933.808 mg
GAE/100gm, 725.708 mg GAE/100gm, 156.422 mg/100gm, 24.27 mg/gm, 4.82 mg/L and
84.28%, respectively. After pretreatment and drying, highest retention for total polyphenol
content, total flavonoid content, ascorbic acid, total chlorophyll and antioxidant activity were
obtained for KMS blanched samples dried at 50±2˚C for 14h. Retention % observed were
72.36%, 60.27%, 39.66%, 56.21% and 72.73%, respectively. Rehydration ratio obtained was
5.52. However, highest carotenoid retention of 54.98% was reported by salt blanched samples
for same drying temperature. Samples took 10 h at 60±2˚C, 14 h at 50±2˚C and 18 h at
40±2˚C to attain final safe moisture level. Proximate analysis of dried sample showed slight
reduction in protein and fat whereas, slight increase in fiber, ash and carbohydrate after
treatment and drying. Reduction in mineral content was also observed. There were significant
differences in retention of phytochemicals among samples due to variation in pretreatments
and drying.

v
Content

s
Approval Letter........................................................................................................................iii
Acknowledgement.....................................................................................................................iv
Abstract......................................................................................................................................v
List of Tables.............................................................................................................................ix
List of Figures............................................................................................................................x
List of Plates..............................................................................................................................xi
List of Abbreviation................................................................................................................xii
1. Introduction........................................................................................................................1-5
1.1 General introduction......................................................................................................1
1.2 Statement of the problem...............................................................................................3
1.3 Objectives........................................................................................................................3
1.3.1 General objective.....................................................................................................3
1.3.2 Specific objectives...................................................................................................3
1.4 Significance of the study................................................................................................4
1.5 Limitations of the study.................................................................................................4
2. Literature review..............................................................................................................6-28
2.1 History and origin..........................................................................................................6
2.2 Classification..................................................................................................................6
2.2.1 Taxonomic classification.........................................................................................7
2.2.2 Various names of Okra............................................................................................7
2.2.3 Varieties grown in Nepal..........................................................................................8
2.3 Cultivation and production of Okra...............................................................................8
2.4 Chemical composition....................................................................................................9
2.4.1 Proximate composition............................................................................................9
2.4.1 Bioactive phytochemical composition..................................................................10
2.5 Nutritional potential of Okra........................................................................................15
2.5.1 Okra pod...............................................................................................................15
2.5.2 Okra seed...............................................................................................................15

vi
 2.5.3 Mucilage and its potential.....................................................................................16
2.5.4 Antioxidant activity of Okra..................................................................................17
2.6 Mineral composition of Okra.......................................................................................18
2.7 Drying..........................................................................................................................18
2.8 Blanching.....................................................................................................................19
2.9 Effects of processing variables on dried Okra.............................................................20
2.9.1 Effect of drying method........................................................................................20
2.9.2 Effect of drying time and temperature..................................................................21
2.9.3 Effect of pretreatments..........................................................................................21
2.9.4 Effect of slice thickness.........................................................................................22
2.10 Effects of pretreatment and drying on the chemical and nutritional composition.....23
2.10.1 Effect on proximate composition........................................................................23
2.10.2 Effect on bioactive components..........................................................................23
2.11 Rehydration................................................................................................................27
2.12 Storage of Dried okra.................................................................................................27
3. Materials and methods...................................................................................................29-35
3.1 Materials.......................................................................................................................29
3.2 Methods........................................................................................................................29
3.2.1 Sample preparation................................................................................................29
3.2.2 Pretreatments.........................................................................................................29
3.3 Analytical methods.................................................................................................31
3.3.1 Proximate analysis.................................................................................................31
3.3.2 Phytochemical analysis.........................................................................................31
3.3.3 Calcium determination..........................................................................................34
3.3.4 Rehydration Ratio..................................................................................................35
3.4 Data Analysis..............................................................................................................35
4. Results and discussion....................................................................................................36-48
4.1 Phytochemical composition of fresh Okra...................................................................36
4.2 Effects of temperature and treatment on phytochemicals............................................37
4.2.1 Effect on Total Polyphenol content.......................................................................37

vii
 4.2.2 Effect on Total Flavonoid content.........................................................................39
4.2.3 Effect on Total carotenoids...................................................................................40
4.2.4 Effect on Total chlorophyll...................................................................................41
4.2.4 Effect on Ascorbic acid.........................................................................................42
4.2.6 Effect on Antioxidant activity...............................................................................44
4.3 Rehydration Ratio........................................................................................................45
4.4 Proximate analysis of the fresh and dried samples......................................................46
4.5 Calcium content...........................................................................................................47
5. Conclusions and recommendations..............................................................................49-50
5.1 Conclusions..................................................................................................................49
5.2 Recommendations........................................................................................................50
6. Summary..............................................................................................................................51
References.................................................................................................................................52
Appendices................................................................................................................................67

viii
 List of Tables
Table No. Title Page No.
2.1 Various names of Okra 8

2.2 Proximate composition of Okra 10

4.1 Phytochemical composition of fresh Okra 36

4.2 Proximate composition of fresh and dried Okra 46

4.3 Mineral content of fresh and dried Okra 48

ix
 List of Figures
Figure No. Title Page No.
3.1 Flowchart for the preparation of dried Okra 30

4.1 Effect of different treatments and temperatures on Total 38

Polyphenol content of Okra
4.2 Effect of different pretreatments and drying on Total Flavonoid 39
content of Okra
4.3 Effect of different pretreatments and drying on Total Carotenoid 40
content of Okra
4.4 Effect of different pretreatments and drying on Total 41
Chlorophyll content of Okra
4.5 Effect of different pretreatments and drying on the Ascorbic 43
Acid content of Okra
4.6 Effect of different pretreatments and drying on Antioxidant 44
activity content of Okra
4.7 Effect of different pretreatments and drying on Rehydration 45
ratios of Okra

x
 List of Plates
Plate No. Title Page No.
1 Fresh samples brought for analysis 72

2 Samples treated and cut into sizes for drying 72

3 Extract prepared for phytochemical analysis 73

4 Dried samples preserved for further analysis 73

5 Dried v/s rehydrated samples 74

xi
 List of Abbreviation
Abbreviation Full form

Abs Absorption
AD Air dried
AD-MVD Air drying combined with microwave vacuum drying
ANOVA Analysis of variance
˚C Degree Celcius
CE Catecheul Equivalent
Chl Chlorophyll
DB Dry basis
DPPH 2, 2- diphenyl-1-1-picrylhydrazyl
FAO Food and Agriculture Organization
FD-MVD Freeze drying combined with microwave vacuum drying
FW Fruit weight
g gram
GAE Gallic acid equivalent
h hour
KMS Potassium metabisulphite
L Liter
m asl Meter above sea level
mL mililiter
MoALD Ministry of Agriculture and Livestock Development
MT/ha Metric ton per hectare
mm Millimeter
mins minutes
MVD Microwave vacuum drying
nm nanometer
No number
NaCl Sodium Chloride
PER Protein Efficiency Ratio
QE Quercetin Equivalent
RAE Retinol Activity Equivalent
TFC Total Flavonoid Content

xii
TPC Total Polyphenol Content
USDA United States Department of Agriculture
UV-VIS Ultraviolet-visible
v volume
w weight

xiii
xiv
 Part I

Introduction

1.1 General introduction

Okra (Abelmoschus esculentus) is a semi-woody, fibrous and herbaceous annual plant which
was originated probably from East Africa and today, is widely distributed in the tropics,
subtropics and the warmer temperate regions (Hussien et al., 2018). It suits the region with
moderate rainfall and grown during summer (Gulsen et al., 2007). Okra is one of the
economically important, most popular summer vegetable crop in Terai, inner Terai and lower
hills of Nepal (Acharya, 2004). The cultivation production and consumption of Okra have
been widely practiced due to its nutritional importance and medicinal value (Hussien et al.,
2018).
Okra is a rich source of vitamin C, vitamin A, thiamin, riboflavin and other B vitamins,
folic acid, calcium, zinc and dietary fiber (Audu et al., 2015). Okra has been described as a
‘storehouse’ of nutrients (Siemonsma and Kouame, 2004; Tidall, 1983). In spite of the health-
benefit potential, there is a dearth of information on antioxidant-phyto constituents present in
the vegetable (Ahiakpa et al., 2013). According to Adelakun et al. (2009) and Ahiakpa et al.
(2013), Okra possesses high amounts of total flavonoids as well as moderate amounts of total
phenolics, making it a good source of natural antioxidants. It has significant health benefits
which are well documented in the literatures (Tiwari and Ahamad, 2018), (Dubey and Mishra,
2017), (Gemede et al., 2014), (Sindhu and Puri, 2016), and (Gemede, 2018). Pods and seeds
are rich in catechin oligomers, flavonol derivatives, quercetin derivatives and
hydroxycinnamic derivatives (Arapitsas, 2008). Okra also contains significant amount of
minerals. Potassium, Sodium, Magnesium and Calcium are the principal elements in pods,
which contain about 17% seeds. Presence of Iron, Zinc, Manganese and Nickel also has been
reported (Moyin-Jesu, 2007). Sindhu and Puri (2016) has also mentioned the phytochemical,
nutritional and pharmacological evidences of Okra in their research.
Okra is seasonal in nature and about 90% of the total Okra is produced within four months
(May to August) only and the farmers have to sell their vegetables with low price at the
harvesting time due to lack of processing and preservation knowledge (Hosian et al., 2010). It
is harvested at green and tender stage: hence it cannot be stored for longer period (Wankhade
et al., 2012). It is highly perishable because of its high moisture and respiratory activities
(Falade and Omojola, 2008). Quality deterioration begins immediately after harvesting
resulting in chemical, physical and biological changes (Adedeji et al., 2008). Different
technologies can be applied to preserve these losses so as to ensure supply of this vegetable
throughout the year (Hosian et al., 2010). Okra can be preserved by freezing whole pod,
drying, canning and pickling.

Drying is one of the oldest and most effective methods of preservation since it has a great
effect on the quality of the dried product. The main objective of drying is the reduction of the
moisture content to a level, which allows safe storage over an extended period (Wankhade et
al., 2013). There are several drying techniques available viz. sun drying, solar drying, hot air
drying, microwave drying, oven drying, etc. (Hussien et al., 2018;Shams El-Din and Shouk,
1999).

Traditionally, Okra have been processed by slicing and sun drying on the ground, racks,
trays, concrete floor, etc. till they become brittle. This method requires little investment,
however, this technique is not devoid of problems including lack of pretreatments, slow drying
rate, direct exposure to sunlight, dust, dirt, insects and other pests, thus, affecting the
nutritional and sensory qualities of the final product (Adom et al., 1997). The drawbacks of
sun drying have also been documented (Doymaz, 2005).

Currently, hot air drying is the most widely used method in post-harvest technology of
agricultural products. A more uniform, hygienic and attractively colored dried product can be
obtained (Wankhade et al., 2012). However, drying can accelerate some reactions that can
adversely affect the product quality (Hussien et al., 2016). It may lead to the depletion of some
nutrients as well as undesirable color and textural changes. Pretreatment of some foods prior
to drying has been reported to help reduce some of these undesirable changes (Daly-Koziel
and Fudeko, 1997). Blanching is the most important treatment conducted prior to food
processing methods to reduce enzymatic activity. It helps in modifying the texture while
maintaining the nutritional value of fruits and vegetables (Corcuera et al., 2004).

2
1.2 Statement of the problem
About 25- 30 % of the fruits and vegetables are being wasted during handling from point of
production to consumer’s plate (Wankhade et al., 2012). In spite of enormous benefits,
farmers and marketers encounter several challenges regarding the keeping quality of these
vegetables. Okra has a poor shelf-life due to its quick degeneration and decomposition after it
is harvested. There is a need to develop improved methods for maintaining the product quality.
Thus, adequate processing, preservation and utilization of Okra is necessary to arrest the
wastage being experienced during the peak season. Drying offers an alternative way of using
Okra, thus, preventing the post-harvest loss and making them available throughout the year at
comparatively lesser cost (Hussein et al., 2018). Drying of okra can be an alternative method
for long-term storage and to avoid any kind of wastage (Pendre et al., 2012). During the lean
season, Okra is produced in low quantities, thus they are scarce and expensive (Bamire and
Oke, 2003).

There are no proper evidences regarding the significant benefits and not much research
have been conducted regarding its richness in bioactive phytochemicals and antioxidant
property of Okra in Nepal. So, this study can be useful for providing evidences of bioactive
components present in Okra and also to select proper pretreatment conditions for the
production of dried Okra.

1.3 Objectives

1.3.1 General objective

The general objective of the work is to study the effect of pretreatments and drying conditions
on the nutritional quality of okra.

1.3.2 Specific objectives

The specific objectives are follows:

 To determine the proximate composition and calcium content of fresh Okra.

 To carry out different pretreatments and drying of Okra at different temperatures.
 To determine the rehydration ratios of the samples.
 To carry out the phytochemical analysis of the raw and dried samples.

3
  To determine proximate composition and calcium content of best sample obtained on
the basis of retention of their bioactive phytochemicals after pretreatments and drying.
 To statistically compare the influence of different pretreatments and drying
temperatures on the quality of dried okra.

1.4 Significance of the study

Okra is used as fresh vegetable, but better as dried products as they are of great importance
and delicacy for people, especially to provide proper nutrients for those times when the
products are not fresh in season. Its seasonality means that its availability is limited to those
lean periods of the year when they cannot be available as fresh, but it’s drying and re-
composition from the dry state does not mean its nutritional quality has changed drastically
(Tsado, 2015). There is considerable demand for the processed vegetables, especially during
the off-seasons, and the dried products have tremendous potential to meet this demand.
Although okra is a vegetable crop of considerable economic value, processing of okra has not
received adequate attention (Shivhare et al., 2000).

The effect of some drying methods and pretreatments has been studied worldwide by many
researchers, but there appears to be little or no information on the drying and preservation of
okra in Nepal. This study was aimed at preservation of okra for the prolongation of shelf-life
and making the nutritional benefits of okra available during the off seasons in a more
concentrated form.

Nepal, being an agro-based country, still depends on the imported fruits and vegetables to
fulfill the demand during the off seasons. The result of this study might help in the
establishment of effective and optimized processing method for the production of dried okra
which would preserve highest quality. The study focuses on the introduction of an effective
method of preservation of Okra which could also encourage the local farmers to increase the
production. The findings of the research might also form a basis for further studies related to
Okra. Though not knowingly practiced in Nepal, the medicinal as well as health benefits of
Okra and nutritional values of dried Okra seems promising.

4
1.5 Limitations of the study
The limitations of the study are:

 Only one variety of Okra could be studied.

 The anti-nutritional factors such as Phytate, Tannin and Oxalate content were not
studied.
 Though high in iron, iron content could not be determined.
 Storage stability of samples were not studied.

5
 Part II

Literature review

2.1 History and origin

The geographical origin of okra is disputed (Abidi et al., 2018). Okra is believed to be
originated in tropical Africa and was grown in Mediterranean region and its wild forms are
found in India (Hussain et al., 2006). It is also believed to have been originated in the south
Asian subcontinent, chiefly India, Pakistan and Burma. However, researches also suggest its
origin to be Ethiopia and Nepal (Acharya, 2004). Lamont (1999) suggested that Okra was
originated somewhere around Ethiopia, and was cultivated by the ancient Egyptians by the 12th
century B.C. Its cultivation spread throughout Middle East and North Africa.

According to Saifullah and Rabbani (2009), Okra originated in Ethopia and was then
propagated in North Africa, the Mediterranean, Arabia and India by 12 th century B.C (Nzikou
et al., 2007). The name Okra probably derives from one of the Niger-Congo group of
languages (Benjawan et al., 2007). The term okra was in the use of English by the late 18 th
century (Arapitsas, 2008).
A theory that it is native to India was dismissed by DeCandolle (1886) because it does not
have a Sanskrit name that an important native Indian plant would be expected to have. The
possibility of an American origin was also dismissed because there is a record in Arabi of the
plant being cultivated in the Egypt in 1216, long before the voyages of Columbus. De
Candolle concluded that the okra originated in Africa. It is now widely cultivated in the
tropics, subtropics and warmer temperate zones. It is particularly popular in Brazil, India,
Spain, Thailand, the Philippines, southern USA, Turkey, and West Africa (Tong, 2016).

2.2 Classification
Okra plant or lady’s finger was previously included in the genus Hibiscus, section
Abelmoschus in the family Malvaceae. The section Abelmoschus was subsequently proposed
to be raised to the rank of distinct genus (Linnaeus, 1753).
2.2.1 Taxonomic classification
According to USDA (2020), the Taxonomy hierarchy of Okra is given below:

Kingdom Plantae

Subkingdom Tracheobionta

Superdivision Spermatophyta

Division Magnoliopsida

Class Magnoliopsida

Subclass Dillenidae

Order Malvales

Family Malvaceae

Genus Abelmoschus

Species esculentus

2.2.2 Various names of Okra

Okra is known by many local names is different parts of the world. The various names of Okra
are:

7
Table 2.1 Various names of Okra

Countries Names

Nepal Bhendi/Ramtoriya

India Bhendi
United states Okra
Caribbeans Okra
China Qui-kui
Taiwan Qui-kui
Europe Quiabo
Portuguese Guigambo
Spanish Gombo
French Gombo
Japan Okura
Thailand Krajiab kheaw

Source: (Abidi et al., 2018; Fekadu, 2014)

2.2.3 Varieties grown in Nepal

The main varieties of Okra grown in Nepal are Pusasawami, Parbhanikranti, Pusamakhmali,
Pusa A-4, Varsha Uphar, Kalimpong, Hissarunnat, Arkaanamika, Arka Abhaya, Red bhindi,
Hybrids:- DVR-1, DVR-2, Varsha and Vijaya, Adhanik and Panchali (Parajuli, 2015).

2.3 Cultivation and production of Okra

Okra is grown throughout the tropical and warm temperature regions of the world for its
fibrous pods full of round, white seeds which, when picked young are eaten as vegetable
(Doymaz, 2005). Okra is grown in many parts of the world. This crop can be grown on a large
commercial farm or as a garden crop. Okras have been a commercial plant in many countries
such as India, Japan, Turkey, Iran, Western Africa, Yugoslavia, Bangladesh, Afghanistan,
Pakistan, Myanmar, Malaysia, Thailand, Brazil, Ethiopia, Cyprus and in the Southern United
States (Benjawan et al., 2007). According to Varmudy (2011), India is largest producer

8
(67.1%), followed by Nigeria (15.4%) and Sudan (9.3 %). In 2009/10, the total world area
under cultivation was 0.43 million hectares and the production stood at 4.54 million tons
(Anonymous, 2011). Kumar et al, (2015) also reported India as the leading okra producing
country with 72.9 % share in world okra production and produces okra in an area of 532.7
thousand hectares with production of 6346.4 thousand tonnes and productivity of 11.9 tonnes/
ha. Highest productivity is reported from Ghana (20.0 tonnes/ha) followed by Egypt (14.0
tonnes/ha).

According to MoALD (2020), okra was cultivated in 9,531 ha of land in Nepal with a total
production of 110,344 MT and an average productivity of 11.58MT/ha covering 3.21% of
total vegetable cultivating area in Nepal.

In Nepal, okra is being cultivated upto 1500 m asl. It cannot thrive frost and snowfall. It
prefers hot and humid climate for proper growth, however, it can develop well in hot and dry
climate provided there is ample supply of irrigation. The best temperature range for seed
germination is 25˚to 35˚C and below 17˚C the germination is severely hampered. Okra can be
cultivated in wide range of soil condition. However, it performs well on sandy loam to loamy
soil that is well drained (Acharya, 2012).

2.4 Chemical composition

2.4.1 Proximate composition

The proximate composition of okra is given in table 2.2

9
Table 2.2 Proximate composition of Okra

Parameter g/100g
Moisture 87.11
Carbohydrate 55.7
Crude protein 18.62
Crude fat 2.64
Crude fiber 15.83
Total ash 7.21

Source: (Shams El-Din and Shouk, 1999)

2.4.1 Bioactive phytochemical composition

Phytochemical is a collective term for plant chemicals with varied structure and function.
‘Phytochemical’ is a broad term meaning plant (phyto) chemical referring to a wide variety of
compounds that occur naturally in plants. Guaadaoui et al. (2014) defined bioactive
compounds as those that have the ability to interact with one or more component(s) of a living
tissue presenting a wide range of probable effects. Generally, phytochemicals have been
classified into six major categories based on their chemical structures and characteristics.
These categories include carbohydrate, lipids, phenolics, terpenoids and alkaloids and other
nitrogen-containing compounds (Harborne and Baxter, 1993; Campos-Vega and Oomah,
2013). The most common sources of phytochemicals are fruits, vegetables, whole grains, nuts
and seeds and other plant foods. Huang et al. (2016) has defined bioactive phytochemicals as
non-nutrient compounds derived from plants that have biological activity in humans. These
include phenolic compounds, terpenoid compounds, and alkaloids.

2.4.1.1 Polyphenols
Polyphenols are naturally occurring bioactive compounds found largely in the fruits,
vegetables, cereals and beverages. Polyphenols are secondary metabolites of plants which are
generally involved in defense against ultraviolet radiation or aggression by pathogens.
Polyphenols may contribute to the bitterness, astringency, color, flavor, odor and oxidative
stability in fruits and vegetables. Studies suggest that long term consumption of diets rich in
plant polyphenols offered some protection against development of cancers, cardiovascular

10
diseases, diabetes, osteoporosis and neurodegenerative diseases. Polyphenols and other food
phenolics are the subject of increasing scientific interest because of their possible beneficial
effects on human health (Graf et al., 2005).
Polyphenols may be classified into different groups based on the number of phenol rings
they contain and the structural elements that bind these rings to one another. The main classes
include phenolic acids, flavonoids, stilbenes and lignans (Spencer et al., 2008). Distribution of
phenolics in plants at the tissue, cellular and sub cellular levels is not uniform. Insoluble
phenolics are found in cell walls, while soluble phenolics are present within the plant cell
vacuoles (Wink, 1997). Certain polyphenols like quercetin are found in all plant products;
fruit, vegetables, cereals, fruit juices, tea, wine, infusions etc., whereas flavanones and
isoflavones are specific to particular foods. In most cases, foods contain complex mixtures of
polyphenols. The outer layers of plants contain higher levels of phenolics than those located in
their inner parts (Simon et al., 1992).
Ahiakpa et al. (2013) had assessed the total flavonoid, phenolic and antioxidant activity in
25 accessions of Okra. He found that mean TPC in Okra ranges from 63.22mg/g/GAE to
6.82/g/GAE in the aqueous extract and 25.83±5.30mg/g/GAE to 8.0±0.37mg/g/GAE in the
ethanol extracts depending upon the variety. Sekar (2016) has reported 159.7 mg GAE/100g
TPC in methanolic extract. Liao et al. (2012) has confirmed fruitful presence of phenolics in
different organs (flower, fruit, leaf and seed) of the A. esculentus plant. Ribarova and
Atanassova (2005) has listed Okra among the vegetables rich in Polyphenols. His results
showed that total phenolics in Okra to be 153.7mg GAE /100g fw. Agbangnan et al. (2018)
and Xia et al. (2015) has also reported the presence of polyphenols in Okra indicating its
antioxidant and anti-fatigue potential.

2.4.1.2 Flavonoid
Favonoids comprise the most studied group of polyphenols. This group has a common basic
structure consisting of two aromatic rings bound together by three carbon atoms that form an
oxygenated heterocycle. More than 4,000 varieties of flavonoids have been identified, many of
which are responsible for the attractive colours of the flowers, fruits and leaves (de Groot and
Rauen, 1998). Based on the variation in the type of heterocycle involved, flavonoids may be

11
divided into six subclasses: flavonols, flavonesflavanones, flavanols, anthocyanins and
isoflavones (Pandey and Rizvi, 2009).

In the research by Ahiakpa et al. (2013), the mean TFC values for Okra ranged from
871.57mg QE/g to 5159.21mg QE/g in the ethanolic extract depending upon the variety.
However, in the aqueous extract, values obtained were lower. Khomsug et al. (2010) obtained
TFC of 1075±0.02mg/g and 1424.8±0.02mg/g for pulped okra and okra seeds, respectively,
while Adelakun et al. (2009) obtained 32.54±32.42mg/g for blanched okra seeds,
48.3±0.00mg/g for raw okra seeds, and 51.28mg/g for soaked okra seeds. Similarly, Ribarova
and Atanassova, (2005) has reported 41.1mg CE /100g fresh mass TFC in Okra in his research
whereas 26.3 mg QE/100g has been recorded by (Sekar, 2016). Thus, Okra contains
significant amount of flavonoids although different values has been reported by different
studies due to varietal and genotypic differences.

2.4.1.3 Total carotenoids

Carotenoids are the lipid-soluble pigments found in all photosynthetic organisms which
contain more than 700 compounds responsible for the red, orange, and yellow colors. Most
carotenoids are hydrocarbons containing 40 carbon atoms and two terminal rings (Bell, 2000).
Two classes of carotenoids are found in nature: (a) the carotenes such as 𝛽-carotene, which
consist of linear hydrocarbons that can be cyclized at one end or both ends of themolecule, and
(b) the oxygenated derivatives of carotenes such as lutein, violaxanthin, neoxanthin, and
zeaxanthin, known as xanthophylls (Botella-Pav´ıa and Rodr´ıguez-Concepci´on, 2006). The
system of conjugated double bonds gives these pigments high chemical reactivity that can be
easily isomerized and oxidized (Oliver and Palou, 2000). According to Olson (1999),
carotenoids quench singlet oxygen, remove peroxy radicals, modulate carcinogen metabolism,
inhibit cell proliferation, stimulate communication between cells (gap junctions), and increase
the immune response. Tests in vitro and in vivo suggest that carotenoids are excellent
antioxidants, scavenging and inactivating free radicals. Krinsky (2001) has also described the
antioxidant properties of carotenoid. Carotenoids are natural pigments that occur widely in
nature. Plants and some microorganisms produce these pigments; however, animals must
obtain them through diet (de Quir´os and Costa, 2006).

12
 Akpapunam (1984), had assessed the total carotenoid content in Nigerain fresh Okra, and had
recorded 56.4 mg/100g of total carotenoids. In a recent study by Kumari (2016), who accessed
phytochemical dynamics in 20 genotypes of Okra, she found mean carotenoid content in Okra
was 1.24 mg/ 100g FW. Highest carotenoid content was obtained in KashiKranti (1.71 mg/100
g FW) variety while lowest was shown by VRO-109 (0.25 mg/g FW). Shams El-Din and
Shouk (1999), reported 32.9 mg/100g carotenoid in Okra in dry weight basis. Gemede et al.
(2014), reported 185 μg /100g FW 𝛽-carotene for Okra pod, whereas Rai and
Balasubramanian (2009), have reported significantly higher values (10 mg/ 100 gFW).
Petropoulos et al. (2017), has studied the nutritional composition of 8 varieties of Okra in
relation to its harvest stage. According to findings, 𝛽-carotene in Okra ranged from0.079-
0.317 mg/g FW whereas, Lycopene content ranged from 0.0311-0.170mg/g FW. The
differences in reported values may be due to differences in genotype, maturity or growing
conditions.

2.4.1.4 Chlorophyll
Chlorophyll are the most widely distributed plant pigment responsible for the characteristic
green color of the fruits and vegetables (Almela et al., 2000). The major chlorophylls in food
are chlorophyll a, which has a methyl group at C-3 carbon, and chlorophyll b, where a formyl
group is bounded to the same carbon atom. Chlorophyll a and b are generally found in higher
plants and occurs approximately in the ratio of 3:1,in the chloroplast, in fruits and vegetables.
Chlorophyll c and d are found, often with Chlorophyll a, in different algae; chlorophyll e is a
rare type found in some golden algae; bacterio-chlorophyll are found in certain bacteria.
Chlorophyll a appears blue-green while Chlorophyll b gives yellow-green appearance. They
also differ in their thermal stability. Chlorophyll a is reported to be thermally less stable than
chlorophyll b (Erge et al., 2008).

Determination of chlorophyll is important since it is related to the esthetic quality of fruits

and vegetables. It is also an important parameter in studying the degree of maturation and
ripening (Rai and KC, 2012).

According to the findings by Kumari (2016), chlorophyll content in Okra varies depending
on the genotypes and ranges from0.30 mg/100g FW to 5.75 mg/100g FW. Shivhare et al.

13
(2010), found the chlorophyll content of fresh Okra to be 2.46 mg/L of extract. He has also
studied the degradation of chlorophyll with the drying time and pretreatment. Shams El-Din
and Shouk (1999), reported 59.7 mg/100g DB of Chlorophyll in Okra.

2.4.1.5 Ascorbic Acid

Vitamin C (also referred as L- ascorbic acid) is an odourless, white solid having the chemical
formula C6H8O6 and a molecular weight 176.12 g/mol. Vitamin C is the L-enantiomic form of
ascorbic acid which also encompasses the oxidation product of dehydroascorbic. Ascorbic
acid is a colourless and odourless crystalline substance, slightly sour in taste and optically
active. This water soluble vitamin contributes to iron absorption, cold tolerance, antioxidizing
activity, metabolism of tryptophan, phenylalanine and tyrosin and body growth. Ascorbic acid
acts as an antioxidant, a nutrient that chemically binds and neutralizes the tissue-damaging
effects of substances in the environment known as free radicals. As a result, ascorbic acid is
vital for the growth and maintenance of healthy bones, teeth, gums, ligaments and blood
vessels (El-Ishaq and Obirinakem, 2015). Humans cannot synthesize vitamin-C, and must be
solely obtained from the diet, mainly through fruits and vegetables to maintain a normal
metabolic functioning of the body (Daud et al., 2016).

Ascorbic acid in Okra ranges from 19.63 (Kashi lalima) to 10.32 (BO-13) mg/100g FW
(Kumari, 2016). However, Shams El-Din and Shouk (1999), has reported higher ascorbic acid
(207 mg/100g) in fresh Okra pods brought from local market of Egypt. According to the
USDA (2020), National Nutrient Database, one cup of raw Okra, weighing around 100 g,
contains 23mg of vitamin C. The concentration of ascorbic acid has been reported to decrease
with maturity and it is also strongly influenced by the season, shelf life, time of storage and
cooking practices (Lee and Kader, 2000). All fruits and vegetables undergo progressive and in
some cases rapid changes if stored untreated under ambient temperature. These changes may
have profound effect on the overall nutritive values. Most prominent amongst the nutrients
affected is L-ascorbic acid, so it is often used as a ‘marker’ for post-harvest deterioration
(Steskova et al., 2006) .

14
2.5 Nutritional potential of Okra
Okra is more a diet food than staple. Okra is considered as a multipurpose crop due to its
various uses of the fresh leaves, buds, flowers, pods, stems and seeds (Mihretu et al., 2014).
Okra plays an important role in the human diet by supplying carbohydrate, minerals and
vitamins (Sindhu and Puri, 2016). The enormous nutritional and other biological activities in
the pods and seeds have been reported by many researchers. A review on nutritional quality of
Okra by Gemede et al. (2014), shows the potential nutritional importance of Okra and its role
in improving nutrition and health. They concluded Okra as an affordable source of protein,
carbohydrates, minerals and vitamins, dietary fiber and health promoting fatty acids. Okra is
also high in antioxidants activity with different parts of the plant (Shui and Peng, 2004; Liao
et al., 2012). Besides being a rich source of various nutrients and antioxidants, okra pods are
also a source of dietary medicines. Anti-diabetic activity of okra has been reported by many
researchers (Dubey and Mishra, 2017).

2.5.1 Okra pod

Consumption of young immature Okra pod is important as fresh fruit (Ndunguru and Rajabu,
2004). Fresh pods are low in calories (20Kcal per 100 g), practically no fat and high in fiber.
Besides this, it also contains several valuable nutrients, which includes about 30% of the
recommended levels of vitamin C, 10 to 20% of folate and about 5% of vitamin A Pods are
also rich in several minerals including K, Na, Mg and Ca. The presence of Fe, Zn, Mn, Ni and
Zn also has been reported (Dubey and Mishra, 2017). Immature pods are usually consumed as
vegetable in different forms. They can be used in salad, soup and stews, fresh or dried, fried or
boiled (Ndunguru and Rajabu, 2004). Okra provides an important source of vitamins, calcium,
potassium and other mineral matters which are often lacking in the diet in developing
countries (IBPGR, 1991). Seven days old fresh okra pods have been reported to have the
highest concentration of nutrients (Agbo et al., 2008). The 100 g edible portion of Okra also
contains Ca (84 mg), P (90 mg), Fe (1.20 mg), β-carotene (185 μg), riboflavin (0.08 mg),
thiamin (0.04 mg), niacin (0.60 mg) and ascorbic acid (47 mg) (Singha et al., 2018).

2.5.2 Okra seed

Okra seed is a rich source of essential nutrients and could serve as a source of protein and
crude fiber (Adelakun et al., 2009). Benchasr (2012), reported Okra seeds as a potential source
15
of oil, its concentration varying from 20% to 40%. The seed oil mainly consists of linoleic
acid (up to 47.4%) (Andras et al., 2005). Okra has been called “a perfect villager’s vegetable”
because of its robust nature, dietary fiber, and distinct seed protein balance of both lysine and
tryptophan amino acids (Holser and Bost, 2004; Sanjeet et al., 2010) and according to
Ndangui et al. (2010), this amino acid pattern renders it an adequate supplement to legume or
cereal based diet. Okra seed flour could also be used to fortify cereal flour (Adelakun et al.,
2009). For example, supplementing maize ogi with okra meal increases protein, ash, oil and
fiber content. It is also used to supplement corn flour for a very long time in countries like
Egypt to make better quality dough and to increase protein, ash, oil and fiber contents
(Akingbala et al., 2003). The amino acid composition of okra seed protein is comparable to
that of soybean, PER being higher than that of soybean (Adetuyi et al., 2011).
Okra seeds can be dried and the dried seeds are a nutritious material that can be used to
prepare vegetable curds, or roasted and ground to be used as coffee additive or substitute
(Moekchantuk and Kumar, 2004). Antioxidant properties of Okra seeds have also been
reported. It is mainly composed of oligomericcatechins (2.5 mg/g of seeds) and flavonol
derivatives (3.4 mg/g of seeds) (Arapitsas, 2008). Gemede et al. (2015) has mentioned that
promoting the consumption of okra could provide cheap sources of nutrients that can improve
the nutritional status and reduce the prevalence of malnutrition especially among resource-
constrained households and can also be used as a means of dietary diversification. Thus, okra
seed plays a vital role in the human diet. However, Okra seeds are considered as underutilized
crop. One of the major drawbacks limiting the nutritional qualities of seeds is the presence of
anti-nutritional factors, which have adverse effects on bioavailability of some minerals.
However, there are no published studies available on nutritional compositions and anti-
nutritional factors of Okra seeds.

2.5.3 Mucilage and its potential

Okra offers mucilaginous consistency after cooking. Okra mucilage refers to the thick and
slimy substance found in fresh as well as dried pods. These mucilaginous substances are
usually concentrated in the pod walls and are chemically acidic polysaccharides associated
with proteins and minerals (Woolfe et al., 1977). The variations in polysaccharides found in
the mucilage are higher. However, neutral sugars rhamnose, galactose and galacturonic acid

16
have been reported often (Hirose et al., 2004; Sengkhamparn et al., 2009). Its physical and
chemical properties include high water solubility, plasticity, elasticity and viscosity. Often the
extract obtained from the fruit is added to different recipes like soups, stews and sauces to
increase the consistency (Gemede et al., 2014).

Okra mucilage has potential for use as food, non-food products, and medicine. Food
applications include use as a whipping agent for reconstituted egg whites, as an additive in the
formulation of flour-based adhesives, and as an additive in India for clarifying sugarcane
juice. Polysaccharides can be combined with acrylamide to develop new biodegradable
polymeric materials (Mishra et al., 2008). Potential of mucilage for medicinal applications
includes uses as an extender of serum albumin, as tablet binder (Ofoefule et al., 2001) and as
suspending agent in formulations (Kumar et al., 2011). Okra mucilage is used in Asian
medicine as a protective food additive against irritating and inflammatory gastric diseases.
The mucilage of okra is believed to binds cholesterol and bile acid carrying toxins dumped
into it by the liver (Lengsfeld et al., 2004).

2.5.4 Antioxidant activity of Okra

Okra fruit presents significant antioxidant properties, mostly due to their high content of
vitamin C, carotenoids and phenolic compounds, especially flavonoids (Gemede et al., 2014).
Arapitsas, (2008); Khomsug et al. (2010); Liao et al. (2012) has reported the richness of okra,
especially seeds, in Phenolic compounds, flavonol derivatives and oligomericcatechins.
Ahiakpa et al. (2013) has reported that antioxidant activity of Okra ranges from 55.97-16.74%
in ethanolic extract depending on the genotypes. Khomsug et al. (2010) have found
procycanidin B2 as predominant phenolic compound followed by procycanidin B1 and rutin in
seeds. In pulped seed catechin, procycanidin B2, epicatechin and rutin are reported to be
present. Liao et al., (2012) has done a comparative analysis of total phenolics and total
flavonoids and antioxidant ability of different organs (flower, fruit, leaf, and seed).These data
suggests Okra as a good contributor to the antioxidant status and promising chemopreventive
agent as described in several traditional medicines for human race. Seed and pod extracts
have been used to treat Mice for many pharmacological evidences which is well described in
(Sindhu and Puri, 2016). Antioxidant assays, including 1-diphenyl-2-picrylhydrazyl
scavenging, ferric reducing antioxidant power and reducing power test, and weight-loaded
17
swimming test showed okra seeds possessed significant antioxidant and anti-fatigue effects
(Xia et al., 2015).

2.6 Mineral composition of Okra

Okra pods are rich in minerals such as Ca, K and Mg, while they also contains significant
amount of trace elements such Fe, Zn and Mn. Minerals content differs significantly among
the genotypes and fruit sizes (Olivera et al., 2012; Petropoulos et al., 2017). According to
Petropoulos et al. (2017), depending upon the genotypes and fruit size, Ca content in Okra
ranges from 142 g/100g -393 g/100g FW, Mg content from 36 g/100g - 109 g/100g FW and K
content ranges from 204 g/100g - 836 g/100g FW. According to him Okra also contains Na,
Fe, Zn and Mn but in lower amount. In the research by Gemede et al. (2015), eight accessions
of dried Okra Pods were investigated. His concluded that Okra contains considerable amount
of minerals: calcium (111.11–311.95 mg/100g), Iron (18.30–36.68 mg/100g), potassium
(122.59–318.20 mg/100g), zinc (3.83–6.31 mg/100g), phosphorus (25.62–59.72 mg/100g),
and sodium (3.33–8.31 mg/100g) on dry weight bases.

2.7 Drying
Drying is the most widely used, oldest and a primary method for preservation of food
practiced by humans. The removal of moisture during drying prevents the growth and
reproduction of microorganisms which cause decay, and minimizes many of the moisture-
mediated deteriorative reactions. It brings about substantial reduction in weight and volume
thus, minimizing packing, storage and transportation cost and enables storability of the
product under ambient temperatures. Drying offers efficient processing and long-term storage
of okra by reducing moisture content to a suitable level by various drying methods
(Wankhade et al., 2012). Audu et al. (2015), had recommended that, for nutritious value such
as protein, fat and fibre, drying is the most beneficial processing method. But for vitamin, ash
and carbohydrate, it is advisable to maintain okra fresh. Various drying methods reported for
Okra are Sun, Solar, Cabinet, Microwave and Foam-mat drying. Sun drying is the cheapest
method of drying but its drawbacks has been well documented in literature (Doymaz, 2005).
Solar drying could be a better option in areas with good sunshine at the harvest season but, it
requires high manpower and the temperature control is bit difficult (Wankhade et al., 2012).
Currently, hot-air drying is the most widely used method. It has an advantage of low
18
complexity and simplicity of equipment with better temperature control. When well applied,
it can give good quality products at relatively lesser cost. However, it may result in loss of
some heat sensitive nutrients (Francislaine et al., 2019).

Various drying methods and their effect on quality of dried Okra have been studied by
many researchers. Drying methods employed to dry different agricultural products has its own
advantages and limitations. Care must be taken in choosing the drying system. Studies
comparing traditional drying and other drying methods for the reduction of the drying time
and to improve the product quality in terms of quality, color, texture and taste has been already
performed (Wankhade et al., 2012).

Drying is an advantageous process; however, it results in the changes on the physical,

biological and chemical properties of food (Hussien et al., 2016). It slightly affects the
chemical composition but it severely results in the loss of bioactive compounds (Huang et al.,
2016; Shams El-Din and Shouk, 1999). Heat treatment generally leads to loss of some
vitamins and other nutritional components that possess antioxidant properties (Adelakun et al.,
2009). These researches have provided evidence regarding the higher nutrient retention in
pretreated Okras in comparison to those untreated samples. Treatments prior to drying
minimize the undesirable changes. A number of pretreatments can be applied depending on
the food to be dried, its end use, and availability. Out of which blanching is one of the most
widely used pretreatment techniques in the food industry (Baloch et al., 1977).

2.8 Blanching
Blanching is a unit operation prior to freezing, canning and/or drying in which fruits or
vegetables are heated in order to inactivate enzymes responsible for browning; modify texture;
preserve color, flavor, and nutritional value; and remove trapped air. Hot water and steam are
the most commonly used heating media for blanching in industry (Corcuera et al., 2004). It
can be performed by exposing vegetables to hot water (the most common method), hot and
boiling solutions containing acids and/or salts, by steam or by microwaving product dipped in
water or for several seconds or minutes (Severini et al., 2005) and then spraying with cold
water immediately, or by conveying them to a flume of cold water. However, blanching
produces a decrease in the nutritional value of foods. Nutrient leaches out from the product,

19
especially during water blanching. Additionally, vitamins are also destroyed by heat. But,
storage of untreated fruits and vegetables is more vulnerable (Viña et al., 2007). Stone et al.
(1986), Shams El-Din and Shouk, (1999), Shivhare et al. (2010), Severini et al. (2005) have
reported the advantages of blanching Okras prior to dehydration.

2.9 Effects of processing variables on dried Okra

Adom et al. (1997), worked on the combined effect of drying time and slice thickness on the
quality of Okra. Sobukola (2009), studied the effect of air temperature and slice thickness on
the drying characteristics and kinetics of okra. According to other similar researches, drying
method, drying time, drying temperature, the slice thickness, pre-drying treatments and the
combined effect of these parameters are very important factors to be considered in producing
dried okra. All of these parameters significantly affects the drying rate and quality of Okra.

2.9.1 Effect of drying method

Shams El-Din and Shouk, (1999), made a comparative study between microwave and
conventional dehydration of Okra. He concluded that the Okra samples dehydrated in
microwave oven resulted in higher retention of Ascorbic acid, chlorophyll and carotenoid as
well as the chemical composition and sensory scores obtained for microwave were
significantly higher. Tsado (2015), studied the effect of three methods of drying (oven, open
sun and an improvised solar drying) on the proximate composition of Okra. According to him,
although oven dried okra appeared darker in color, direct sun dried sliced okra fruits produced
best quality and higher acceptance. Falade and Omojola (2008), studied the effect of
freezing/thawing, sun drying, solar drying and foam-mat drying on the physical, chemical,
rheological, and sensory attributes of Okra. He found the quality attributes of foam-mat dried
Okra comparable to that of the fresh Okras whereas the solar and sun dried Okra were poorer
in color, aroma, taste and overall acceptability. Hussien et al. (2018), worked on the effect of
three different methods using solar, sun and oven drying on the chemical properties of Okra.
His result showed that okra samples dried by hot air oven took minimum time for drying with
maximum removal of moisture. Similar results were also reported by (Wankhade et al., 2012).
However, Hussien et al. (2018), suggests solar drying method and Wankhade et al. (2012)

20
suggests Hot-air drying method for the best quality dried Okras. They concluded that the
drying method used have a significant effect on the chemical composition of Okra.

2.9.2 Effect of drying time and temperature

Quality parameters of dried Okras are significantly affected by the time-temperature regime.
Irrespective of other conditions, higher drying air temperature will take lesser time to obtain
the final constant weight whereas lower drying air temperature will require higher drying time.
An increase in drying temperature caused an increase in the drying rate thus reducing drying
time (Doymaz, 2005; Sobukola, 2009). Higher drying temperature results in more loss of
bioactive components (Huang et al., 2016). Temperature of drying also affects the textural
quality. According to the research by Wankhade et al. (2012), higher temperature (90˚) had
poor textural quality and dried Okra were shriveled and burnt, whereas slices dried under hot
air oven at 40˚C remained their texture well. Adom et al. (1997) worked on the combined
effect of drying time and slice thickness on the solar drying of Okra. He found that the effect
of drying time was highly significant on all the parameters he studied. Vitamin C was most
affected. He observed chromatic shift of the sample color from green to red (a * values), which
was due to the temperature-dependent degradation of chlorophyll to pheophytins, which was
in turn affected by the drying time. L*values showed a general decrease during the 1 st 48 h of
drying followed by an increase after 48 h. The decreasing trend was due to enzymatic
browning occurring at the cut surfaces of the samples, as well as possibly the degradation of
chlorophyll to olive-brown pheophytins, resulting in darkening and hence the decrease in L*
values. The increasing trend after 48 h may be due to the fact that, with increasing dryness,
browning becomes less severe after it has reached a maximum at48 h resulting in higher L*
values. Different researchers have suggested different time-temperature regime for the drying
of Okra. Rindiani et al. (2018) has also reported the effect of drying temperature on water,
protein, fat, carbohydrate of Okra flour.

2.9.3 Effect of pretreatments

Different pretreatments like Ultrasound (Tüfekçi and Özkal, 2017), water blanching,
microwave and Pulse Electric Field (Adedeji et al., 2008), blanching in SO2 (Stone et al.,
1986), blanching in salt water (Shivhare et al., 2000) etc. prior to dehydration has been
reported to decrease the drying time of Okra. These researchers have also reported the
21
enhancement of rehydration characteristics by these treatments. Kocabay and Ismail (2017)
reported that, for open-sun drying, drying of okra samples occurred as blanched< salted <
natural. Stone et al. (1986), has found better color retention in blanched Okra samples with or
without addition of SO2 than in unblanched samples because of the inhibition of enzymatic
browning by heat treatment and/or by addition of SO 2. He also found that the unblanched
samples retained less ascorbic acid than blanched samples after storage. The unblanched
samples continued losing its ascorbic acid, whereas the blanched product had minimal
additional losses. Also, blanching resulted in protection of the characteristic flavor of okra
identified as “cooked green vegetable”; prevention of excessive development of hay-like
aroma and flavor; and more sweeter okra samples. Similar, results on color was obtained by
Severini et al. (2005) in potato cubes. Shams El-Din and Shouk (1999) has reported higher
ascorbic acid retention, higher retention of pigments percentages (chlorophyll and carotenoid),
higher rehydration ratios, general appearance and color scores in samples blanched in 0.1 %
SO2.
Shivhare et al. (2010), found best quality in the samples blanched at 95˚C in 0.5% NaCl
solution for 5 min and dried at 55˚C. In his research, parameters like bulk density,
reconstitutability, chlorophyll content, appearance, texture and flavor were computed. The
effect of blanching on chemical changes of bioactive phytochemicals depends on several
factors, including the method of blanching, thermal stability of different phytochemicals,
enzyme activity, and location of phytochemicals in the plant structure. Loss of bioactive
phytochemicals into water medium is a critical point for control during water blanching.
Intense heat from the boiled water can lead to disruption of cellular structure increasing
release of soluble bioactive phytochemicals from plant cellular compartments into the water
medium (Rungapamestry et al., 2007).

2.9.4 Effect of slice thickness

Sobukola (2009), reported the drying time required to lower the moisture content of okra
slices from 88.24% wb to <10% wb increased from about 60mins for 2 mm slice thickness and
drying air temperature of 70˚C to about 138mins when the slice thickness increased to 6mm.
Drying time decreases with the decrease in slice thickness can be attributed to a larger surface

22
area exhibited by the samples which facilitates effective moisture diffusion from the samples
thus increasing drying rate. Adom et al. (1997), reported that the slice thickness had a
significant effect on moisture, crude fiber and ash contents but not on vitamin C content,
viscosity, color and microbial load. He worked on three slice thickness (5mm, 10mm and
15mm) and dried in solar dryer. He found slice thickness of 10mm to be better in quality for
the solar drying of okra.

2.10 Effects of pretreatment and drying on the chemical and nutritional composition

2.10.1 Effect on proximate composition

Adom et al. (1997); Audu et al. (2015); Shams El-Din and Shouk, (1999), has reported an
increase in the chemical composition of Okra which may be attributed to the concentration
effect due to the loss of water. However, it has been seen that the dehydration method and the
time-temperature regime affects the final moisture content. According to the results obtained
by Shams El-Din and Shouk (1999), drying caused a slight decrease in protein and fat as well
as slight increase in ash, fiber and carbohydrate on dry weight basis. Untreated samples had
more protein, fat and ash but less fiber and carbohydrate than treated samples. Loss of protein
might be due to the loss of some water-soluble proteins and minerals during blanching, and
loss of fat may be due to oxidative reaction during or leaching.

2.10.2 Effect on bioactive components

Temperature has significant effect on the bioactive compounds and thus, the antioxidant
activities (Huang et al., 2016)

2.10.2.1 Effect on total polyphenols and flavonoid content

Phenolics are molecules sensitive to temperature, and may increase or decrease with different
drying temperatures (Jiang et al., 2017). The reduction of phenolic compounds during drying
may be the result of the activation of oxidative enzymes (polyphenol oxidase and peroxidase)
(An et al., 2016) and change in their structures via binding to proteins (Toor and Savage,
2006). Reid et al. (2017), has reported an increment in TPC in mushroom due to the better
extractability of bound polyphenols as a result of cell wall destruction after drying. Choi et al.
(2006) and Jeong et al. (2004), has also reported that heat treatment might distrupt the cell
wall and liberate phenolic compounds from the insoluble portion of the plant. The disruption

23
of cells may also result in release of oxidative and hydrolytic enzyme, which are capable of
oxidizing endogeneous polyphenols. However, drying at above 60˚C is regarded unfavorable
due tothe possibility of inducing oxidative condensation or decomposition of thermolabile
compounds (Asami et al., 2003). A study on effect of drying on TPC in jujube fruits showed
that phenolics were rather stable at 55˚C and only small reduction was observed, but drying
and storage at ambient temperature elevated the TP content (Pu et al., 2018). Heat treatment
like cooking has been reported to increase the phenolic content in vegetables due to improved
extractability of phenolics from the food. In contrast Ornelas-Paz et al. (2013), reported that
decrease in antioxidant activity of cruciferous vegetables during aqua thermal processing were
observed due to the loss of water soluble antioxidant; polyphenols.

Jiang et al. (2017) performed different drying methods on the Okra snacks and evaluated
the antioxidant activities. He found that the different drying treatments (air drying, freeze
drying, microwave vacuum drying, combined freeze and microwave vacuum drying and
combined air and microwave vacuum drying) had significantly different effects on the TFC
and TPC of okra samples. Freeze dried samples had the highest TFC and TPC values (17.46 ±
0.23 mg rutin/g d.w. and 11.59 ± 0.02 mg GAE/g in DB respectively), followed by the FD-
MVD, MVD, AD-MVD, and AD samples. These results could be ascribed to the effects of
temperature and oxidation (Lou et al., 2015). KMS blanched AD samples retained about
60.49% TPC and 58.79% of its original TFC. This accounts for the lowest retention which is
due to the high temperature and long drying time required, which can accelerate oxidation.
Hamrouni-Sellami et al. (2013) reported similar observation in sage. The low-oxygen and
temperature environment during drying effectively minimizes the loss of phenolic acids and
flavonoids. Less drying time and low exposure to oxygen reduces the thermal degradation of
phenolic compounds.

2.10.2.2 Effect on total carotenoids

The degeneration of carotenoids, with concomitant loss of color and vitamin A value, is a
common problem in the processing and storage of vegetables and fruits. Shams El-Din and
Shouk (1999), reported similar observation for carotenoid as he had reported for total
chlorophyll. Dehydrated okra samples by microwave oven retained the highest percentages of
carotenoids, i.e. (48.94%-59.27%) followed in a decreasing order by conventional oven
24
(22.8%-29.48%) and sun drying methods (18.54%). The involved short time for dehydration
by microwave reduced the destruction of carotenoids. Immersion of okra samples in
0.1%sodium metabisulphite solution at 92-95 °C before dehydration by either conventional or
microwave oven retained greater amounts of carotenoids than other all dried okra samples.
Nezam El-Din and El-Ashwah (1991), had also reported similar observations. According to
Kapoor and Kaur (2001), sulphite had a beneficial action as an antioxidant in stabilizing the
carotenoid in dehydrated vegetables. Blanching treatment also results in loss of carotenoid.
Akpapunam (1984) had reported 29.5% loss of carotenoid in Okra during blanching. But,
Jorge et al. (2014), had reported that blanching prior to drying promotes the stability of some
carotenoid compounds like lycopene and β-carotene. The fact behind this phenomenon could
be explained by the reason that heating promotes the change from the cis to trans
conformation form, which intensifies the detection of these compounds.

2.10.2.3 Effect on total chlorophyll

Chlorophylls are known to be easily degraded by conditions such as dilute acids, heat, light
and oxygen. Color is the major sensory characteristics in determining product acceptability, so
its loss must be minimized during thermal processing in food industry. The main reason
behind the color loss is attributed to the conversion of chlorophyll to pheophytins by the
influence of pH. In green peas, chlorophyll a degraded 12 to 18 times faster than chlorophyll b
depending on temperature (Erge et al., 2008). Dehydration of Okra causes an increase in
acidity, which let to more degradation of chlorophyll (Sweeney and Martin, 1961). The same
authors reported that, the destruction of chlorophyll a was found to be the principal factor
responsible for color loss. Degradation of chlorophyll is affected by the dehydration period.
Mabesa and Baldwin (1979), observed that the retention of chlorophyll components was
significantly greater after cooking in a microwave oven.

As reported by Shams El-Din and Shouk (1999), dehydrated okra samples by conventional
oven had lower retention percentages of total chlorophyll (37.02%%-44.39%) than those in
dehydrated okra samples by microwave oven (40.03%-49.25%) depending upon the
pretreatments, which was probably due to the faster dehydration period in microwave. Sun
drying showed highest loss of chlorophyll (65.66%). His results showed that, immersion of
okra samples in 0.1 % sodium metabisulphite solution and dehydration by either conventional
25
or microwave oven retained higher percentages of chlorophyll than other all dried okra
samples, which was due to the preservative action of sulfur dioxide and its effect on
chlorophyllase inhibition. Nezam El-Din and El-Ashwah (1991), had reported very low
chlorophyll retention in dried okra samples when compared to frozen ones, this could be
related to the effect of drying on changes and degradation of chlorophyll by the reactivation of
enzyme chlorophyllase. The heat of drying (60-65 °C) had main role in the activation of
chlorophyllase and pectin methylesterase. Shivhare et al. (2010), found that chlorophyll
retention significantly depends upon the pretreatments and dying temperature. He reported that
dehydration at 55˚C after blanching in 0.5% NaCl retained highest chlorophyll (61.92%).
Higher temperature i.e. 70˚C resulted in substantial loss of chlorophyll.

2.10.2.4 Effect on Ascorbic acid

Since, ascorbic acid is vulnerable to heat and oxidation, it is easily destroyed during
dehydration. Rapid drying retains greater amount of Ascorbic acid than slow drying.
Immersion of okra samples in 0.1 % sodium metabisulphite solution at 92-95 °C and
dehydration by microwave had the highest ascorbic acid contents when compared with other
treatments and dehydration methods. In general, the reduced dehydration time required for
microwave dehydration produced far less destruction of ascorbic acid (Shams El-Din and
Shouk, 1999). He reported 55.56% retention in microwave and 40.58% in conventional drying
in samples blanched in 0.1% SO2. However, sun dried samples retained only 26.09% of
original ascorbic acid. Similar findings were reported by (Mabesa and Baldwin, 1979). They
found that, microwave cooked peas had greater retention of total ascorbic acid than peas
cooked conventionally. Krutman (1981) reported that, okra blanched in sodium sulfite and
cabinet-dried had the highest ascorbic acid content when compared with other pretreatments.
Effects of blanching and use of sulfur dioxide as pretreatment before dehydration were more
evident after storage (Stone et al., 1986).

2.10.2.5 Effect on antioxidant activity

The ability to scavenge free radicals is highly correlated to the TPC and TFC (Šumić et al.,
2017;An et al. 2016). So, antioxidant activity reduces with the degradation of TPC and TFC,
and increases with the corresponding increase in both. The findings of Khomsug et al. (2010),
also support the above statement. They concluded that phenolic compounds are the main
26
reason for antioxidant power of plant. Reduction in antioxidant potency is due to the loss of
antioxidant compounds (polyphenols, ascorbic acid and carotenoid) (Petropoulos et al., 2017).
According to Chong et al. (2013), phytochemical compounds can oxidize during hot-air
drying because the food material has greater contact with oxygen. Longer drying time also
promotes reduction in antioxidant capacity. This is also supported by the findings of (Jiang et
al., 2017).

2.11 Rehydration
Rehydration is the measure of the injuries to the material caused by drying and treatments
preceding dehydration. It is generally accepted that the degree of rehydration is dependent on
the degree of cellular and structural disruption (Kocabay and Ismail, 2017). Thus, higher the
rehydration ratio, the less damage to the tissues and the greater product hydration (Goula and
Adamopoulos, 2009). Rehydration of fruits and vegetables follow the first order kinetics. The
water temperature influences the rehydration rates and the equilibrium moisture content in a
positive way. Most of the dehydrated products are usually rehydrated during their use
(Krokida and Marinos-Kouris, 2003). Rehydration capability of the dried product is the most
important quality required in the dried product and it is level of convertibility to former state
with the given water. If the product recovers the fresh state’s level of water then it is admitted
as qualified product. Rehydration capability is impressed with various factors such as drying
conditions, type of product, warmth, ratio of rehydration water quantitative to dried product
(Kocabay and Ismail, 2017).

Rehydration ratio is affected by the drying time and drying method. It is found better with
reduced drying time. However, no significant difference in the rehydration ratios of samples
were reported for either treated or untreated (Shams El-Din and Shouk, 1999).

2.12 Storage of Dried okra

In a research done by Stone et al. (1986), dried okra slices were packaged in laminated
polyester, polyethylene Seal-a-Meal bags (63.5 micron) and then stored at ambient
temperature (22-25°C) for 6 weeks. He studied the effect of pretreatments, drying and storage
in moisture content, ascorbic acid, thiamin, color and sensory characteristics of dried Okra. He
found that, blanching with or without SO2 produced samples with higher water activity which

27
remained unchanged after 6 weeks of storage. Whereas, unblanched samples had low water
activity after dehydration but increased after storage. For ascorbic acid, there was no
significant difference in retention for treated and untreated samples. But the effect of
blanching and use of sulfur dioxide as pretreatment were more evident after storage. Untreated
samples underwent greater loss during storage while the SO 2 blanched samples exhibited
higher ascorbic acid regardless of the drying temperature and duration. Six weeks of storage
resulted in 9.1% loss of ascorbic acid, 0.2% increase in moisture and 9.8% loss of thiamin.
Eze and Akubor (2012), studied the effect of blanching, drying and storage on the proximate
composition and micro-nutrients of Okra. He observed that for oven dried samples stored in
dark cool dry place, crude protein contents decreased. This may be attributed to Maillard
browning that probably occurred during protein hydrolysis. Crude fat content also decreased
and ash content increased during storage. Changes in mineral content were not expected since
minerals are generally stable during storage but slight changes were observed which was due
to the changes in ash content. Vitamin C was most affected during storage. Contact with light
and air during storage has vulnerable effect on phytochemical component.

28
 Part III

Materials and methods

3.1 Material
Green fresh okra pods, cultivar “Kalimpong” (18-22 mm in diameter, 130-140mm in length)
were collected from “Krishi Bikash Bahuudeshye Shakari Sanstha”, Tikathali, Kathmandu and
brought into the laboratory of GoldenGate International College. Sound pods were separated,
washed with clean water, strained and kept in refrigerator until used.

3.2 Method

3.2.1 Sample preparation

Okra samples brought were sorted according to their size followed by washing in potable
water and strained to remove adhered water. The process variables considered were: treatment
prior to drying and drying air temperature.

3.2.2 Pretreatments
For each time-temperature regime the samples were treated prior to dehydration. The
pretreatments were:

a. Green fresh pods were dipped in 0.5% NaCl solution in boiling water for 5 min and
strained.
b. Green fresh pods were dipped in 0.1% KMS (Na 2S2O5) solution in boiling water for 5
min and strained.
c. Green fresh pods were blanched in boiling water for 3 min and strained.
d. Green fresh pods without any treatments.

After treatments, tails and butts of treated pods were removed by a stainless steel knife. It was
then sliced to approximately 10mm length and dehydrated using cabinet dryer in below
mentioned drying conditions.

1. At 40±2˚C for 18 h.
2. At 50±2˚C for 14 h.
3. At 60±2˚C for 10 h.
The general outline of the process is shown below:

Fresh Okra

Washing and sorting

Unblanched

Blanching

Water blanching 0.1% KMS blanching 0.5%NaCl blanching

(3min) (5min) (5min)

Dipping in cold water

Straining

Cutting (10mm)

Cabinet drying

40±2˚C, 18 h 50±2˚C, 14 h 60±2˚C, 10 h

Chemical analysis

Fig 3.1 Flowchart for the preparation of dried Okra

Source:- Modified from Adom et al. (1997); Shams El-Din and Shouk (1999); Shivhare et al.
(2010)
30
3.3 Analytical methods

3.3.1 Proximate analysis

3.3.1.1 Determination of moisture

Moisture content was determined by using hot air oven as described by (Rai and KC, 2012).

3.3.1.2 Determination of Carbohydrate

Total carbohydrate was determined by difference method.

Total carbohydrate% = 100 - (moisture + protein + fat + crude fiber + ash)

3.3.1.3 Determination of Crude Protein

Crude protein was determined by macro Kjeldahl method as described by Rai and KC (2012).

3.3.1.4 Determination of Crude Fat

Crude fat was determined by using Soxhelt apparatus as described by Rai and KC (2012).

3.3.1.5 Determination of Crude Fiber

Crude fiber was determined by the procedure as described by Rai and KC (2012).

3.3.1.6 Determination of Total ash

Total ash was determined by dry ashing as described by Rai and KC (2012).

3.3.2 Phytochemical analysis

3.3.2.1 Extract preparation

The extract of Okra was prepared according to the method described by Dimitrijevic et al.
(2014) with some modification. 5 g of raw/dried sample was ground with 80% methanol (30
mL) and was kept under continuous shaking for 20 minutes and then filtered through
Whatmann no. 1 filter paper. The residue was again submitted to two more extraction cycle
for 20 minutes each totalizing 60 mins of extraction time. The filtrate was combined in
volumetric flask, and the volume was made up to 100mL. The extracts were stored in
refrigeration for analysis of polyphenol, flavonoids, carotenoids and antioxidant activity.

31
3.3.2.2 Polyphenol content
The total phenol content of sample extracts was measured by using Folin-Ciocalteau method,
as described by Madhavi et al. (2010). 1mL of extract or standard solution of gallic acid
(100µg/mL-1000µg/mL) was decanted in 25 mL volumetric flask, which contained 9mL of
distilled water. 1mL of Folin-Ciocalteau reagent was added to the mixture and shaken. After 5
mins, 10mL of 7% Na2CO3 solution was added and the solution was diluted to volume with
distilled water and mixed. After incubation for 90 min at room temperature, the absorbance
against prepared reagent blank (distilled water) was measured using an automated UV- VIS
spectrophotometer at wavelength of 765 nm. Standard solution of gallic acid was used to
obtain standard curve. The results were expressed as solution of gallic acid equivalents (GAE)
per 100g of sample.

3.3.2.3 Flavonoid content

The total flavonoid content (TFC) of beans was determined as Samatha et al. (2012) and
Walvekar and Kaimal (2014) using the aluminium chloride assay through colorimetry.

Aluminum chloride forms acid stable complexes with the C-4 keto group and either the C-3
or C-5 hydroxyl group of flavones and flavonols. In addition, aluminum chloride forms acid
labile complexes with the orthodihydroxyl groups in the A- or B- ring of flavonoids. Thus,
determining the total flavonoids by using aluminum chloride method is based upon the
formation of stable complex between aluminum chloride and keto and hydroxyl groups of
flavones and flavonoids (Hassan et al., 2013).

An aliquot (0.5 mL) of extracts were taken in different test tubes then 2mL of distilled
water was added followed by the addition of 0.15 mL of sodium nitrite (5% NaNO 2, w/v) and
allowed to stand for 6 min. Later 0.15 mL of aluminum trichloride (10% AlCl 3) was added
and incubated for 6 min, followed by the addition of 2 mL of sodium hydroxide (NaOH, 4%
w/v) and volume was made upto the 5mL with distilled water. After 15 min of incubation the
mixture turned to pink color whose absorbance was measured at 510 nm using a colorimeter.
Distilled water was used as blank. The calibration standard curve was prepared by preparing
gallic acid solutions and results were expressed as mg of Gallic acid equivalents per 100 g o f
sample.

32
3.3.2.4 Total carotenoid
Total carotenoid content of the Okra samples was determined as described by Rainha et al.
(2011). Methanolic solutions of sample extracts were analyzed in UV/VIS spectrophotometer
at 470, 653 and 666 nm. The concentrations of carotenoids and chlorophylls a and b were
determined according to the equations reported by Lichtenthaler and Wellburn (1983) as
follows:

Total carotenoids (mg/L) = 1000|¿ 470|−2.860 C a−129.2C b ¿

Chlorophyll a (mg/L) = 15.65|¿666|−7.340|¿653|¿ ¿

Chlorophyll b (mg/L)= 27.05|¿ 653|−11.21|¿666|¿ ¿

3.3.2.6 Total Chlorophyll

Total chlorophyll was determined according to the procedure described by Rai and KC (2012).
Chlorophyll was extracted in 80% acetone and the absorbance at 663 and 645nm were noted
in a spectrophotometer. Then, using the absorbance coefficients, the amount of chlorophyll
was calculated using the empirical formula:

V
Chl a, mg/g tissue =12.7 ( A663 )−2.69 ( A 645 ) ×
1000 ×W

V
Chl b, mg/g tissue = 22.9 ( A645 )−4.68 ( A 663 ) ×
1000 ×W

Total chlorophyll, mg/g tissue = Chl a + Chl b

Where, A= absorbance at specific wavelengths; V= final volume of chlorophyll extract; W=

fresh weight of tissue extracted.

3.3.2.5 Ascorbic acid

The ascorbic acid of the Okra samples were determined by the 2,6-dichlorophenol indophenol
titration method as described by Rai and KC (2012).

33
3.3.2.7 Antioxidant activity by DPPH Radical Scavenging Assay
The antioxidant activity of beans were determined by the DPPH radical scavenging method as
described by Walvekar and Kaimal (2014) and Roy (2011).

DPPH assay is a stable free radical method. It is an easy, rapid and sensitive way to survey
the antioxidant activity of a specific compound or plant extract. The principle of DPPH is
based on the reduction of DPPH in the presence of a hydrogen donating antioxidant due to the
formation of diphenyl picryl hydrazine. Extracts reduce the color of DPPH from purple to
yellow due to hydrogen donating ability. The degree of discoloration indicates the scavenging
potential of the antioxidant compounds. DPPH solution (0.004% w/v) was prepared in 95%
methanol. The samples were mixed with 95% methanol in 1: 9 ratio so as to make final
volume 10mL, thus the extract was prepared. Equal volume of extract and freshly prepared
DPPH (0.004% w/v) were mixed and the tubes were incubated at room temperature in dark for
10 minutes, the absorbance was taken at 517 nm using a UV-Vis spectrophotometer. 95%
methanol was used as blank.

The scavenging activity of the extract against the stable DPPH was calculated using the
following equation,

( A−B)
Scavenging activity (%) =
A × 100

Where A is the absorbance of DPPH and B is the absorbance of DPPH and extract
combination.

3.3.3 Calcium determination

Calcium content was determined by volumetric method as described by Rai and KC (2012).
Calcium content was determined using the formula:-

Titer × 0.2×V T (ml) ×100 ×100

Ca++ (mg/100gm, DB) =
V E (ml)× Wt . of sample ( g) × Dry matter (%)

Where, VT = total volume of ash solution

VE = volume taken for estimation

34
3.3.4 Rehydration ratio
Rehydration ratio was obtained by dividing mass of the rehydrated sample by mass of the
dried sample. Rehydration of the dried sample was carried out by adding 80 mL distilled water
to 5 g dried okra contained in a 500 mL beaker. The beaker was covered with a watch glass
and the contents were brought to boiling point within 3 min and the boiling was continued for
25 min. Excess water was removed by placing the sample on a screen and mass of the
rehydrated sample was determined (Shivhare et al., 2000).

weight of rehydrated sample

Rehydration ratio =
weight of dried sample

Rehydration ratio can also be measured in terms of fresh weight. It is often expressed as
percentage and can be determined as (Berk, 2009):-

weight of rehydrated sample

Rehydration Ratio = ×100
weight of fresh sample

3.4 Data Analysis

GeneStat (12th Edition developed by VSN International Limited) and Microsoft Office Excel
2010 were used for the statistical analysis and data interpretation. The values for
phytochemical analysis were calculated using Microsoft Office Excel 2010 and were analyzed
by two-way Analysis of variance (ANOVA) using GeneStat at 5% level of significance.

35
 Part IV

Results and discussion

Okra samples were collected from Tikathali, Kathmandu, and brought in the laboratory of
GoldenGate International College. Proximate, bioactive components (Ascorbic acid, total
polyphenols, flavonoids, total chlorophyll and carotenoid) and mineral (calcium) content of
the fresh samples were studied. Samples were subjected to different blanching treatments
(water, KMS and NaCl) and then dried at different temperatures (40±2˚C for 18h, 50±2˚C for
14h and 60±2˚C for 10h) in a cabinet dryer. Untreated samples were also dried under the
above mentioned conditions. The bioactive phytochemicals of the obtained dried samples were
determined and compared. All the values were interpreted in dry weight basis. From the
results, the processing method which retained higher amount of nutrient was considered to be
best. The best samples were again subjected to proximate analysis along with calcium content
determination. The results were statistically analyzed to study the effect of different
processing techniques on the nutrient retention of Okra.

4.1 Phytochemical composition of fresh Okra

The phytochemical composition of fresh Okra is presented in the table 4.1

Parameter Value
Total polyphenols (mg 1933.81±84.67
GAE/100g)
Flavonoid (mg GAE/100g) 725.71±20.43
Total carotenoid (mg/L) 4.82±0.46
Total chlorophyll (mg/g) 24.27±0.74
Ascorbic acid (mg/100g) 156.42±1.91
Antioxidant activity (%) 84.28±2.23

The values in the table are the means of triplicate ± standard deviation. All parameters are
expressed on dry weight basis.

The mean polyphenol content in the Okra samples in the present study was found to be
1933.81mg GAE/100g which was found to be in range justified by Ahiakpa et al. (2013). The
mean flavonoid content was found to be 725.71mg GAE/100g . The values were higher than
those reported by Sekar (2016) and Ribarova and Atanassova (2005). Similarly, the mean
carotenoid content was found to be 4.82 mg/L which agrees with Shams El-Din and Shouk
(1999) but lower values were reported by Kumari (2016). The chlorophyll content of the
samples did not agree with any of the previously reported researches. The reason may be
attributable to the considerable variation of chlorophyll depending upon the genotype and
harvest stage. Mean ascorbic acid in the samples was 156.42 mg/100 g. Results obtained were
higher than those reported by Kumari (2016) but lower than Shams El-Din and Shouk (1999).

The current findings are inconsistence which may be attributed to the source and
differences in the genetic makeup of the accession, which is one of the major factor
influencing the synthesis of phenolic compounds in plant (Hanson et al., 2004), maturity stage
of the raw material, agricultural treatments, unknown external factors such as time,
temperature, presence of oxygen, and/or light, or it might just be a consequence of extent of
extraction of the phytochemicals to be analyzed and the method of determination (Kamiloglu
et al., 2016).

4.2 Effects of temperature and treatment on phytochemicals

4.2.1 Effect on Total Polyphenol content

Fig. 4.1 shows the effect of different pretreatments and drying temperature on the total
polyphenol content of the dried Okra samples. It showed that the mean polyphenol content in
the dried samples ranged from 1399.24 mg GAE/100g to 856.65 mg GAE/100g. Highest
amount of polyphenol was recorded by the samples blanched in 0.1% KMS solution and
cabinet dried at 50±2˚C (72.36% retention) for 14 h whereas lowest retention was found in the
water blanched samples dried at 40±2˚C (44.29%) for 18h.

Statistical analysis showed that there was significant effect (p<0.05) on the polyphenolic
content of Okra due to the difference in temperatures. There was considerable decline in the
TPC with the drying conditions since these compounds are relatively unstable. Higher loss
was observed at 40±2˚C for 18 h for all treated samples. The extended drying period at this
temperature attributed to longer exposure of samples to oxygen during air drying, which might
have caused higher loss of polyphenols as explained by Jiang et al. (2017). Similar

37
 observation was reported by
1600 S.B K.B W.B N.T
1399.24 1385.66 Mbondo et al. (2018) in African
1400 1314.36
1230.25
1216.03 1217.74 eggplant, where higher loss of
1153.17
TPC (mg GAE/100gm)

1200 1078.12 1100.07

1019.71 992.27 phenols was observed at 50˚C as
1000 856.65
compared to 70˚C. This may also be
800
600 associated with the inactivation of

400 polyphenol oxidase enzymes at

200 higher temperature and delayed

0 inactivation at lower temperature.
40˚C/18h 50˚C/14h 60˚C/10h However, in the present study
samples dried at 50±2˚C for 14 h were found better than samples dried at 60˚±2C for 10 h.
This might be due to the degradation of some phenolics at higher temperature. Explanation for
this phenomenon could be better ascribed by the relation between length of drying and drying
temperature.

SB= Salt blanched

KB= KMS blanched

WB = Water blanched

NT= No treatment

Fig.4.1 Effect of different treatments and temperatures on Total Polyphenol content of Okra

The present findings showed that, pretreatments also caused a noticeable reduction (p<0.05)
on the TPC. Samples blanched in KMS and NaCl were better in TPC for all dehydration
temperatures. Water blanched samples had highest loss ranging from 55.7% to 44.25%, which

38
 might be due to the leaching of
SB KB WB NT
500
437.39 426.17 phenolic compounds during
450 405.94
385.93
TFC (mg GAE/100gm)

400 353.14 362.8 blanching (Danesi and Bordoni,

350
274.74 293.77 2008). However, KMS and NaCl
300 251.04 259.84
250 232.37 blanched samples retained higher
200 165.56
TPC due to the antioxidant activity
150
100 of KMS and prevention of loss of
50
phenols by salt (Kaur et al., 2018) .
0
40˚C/18h 50˚C/14h 60˚C/10h In contrast to the finding of Jiang et
al. (2017), considerable loss was
observed in the control samples which might be due to the enzymatic browning in non-treated
samples. The activation of oxidative enzyme, polyphenol oxidase may lead to higher
degradation of phenolics (Kamiloglu et al., 2016; Lim and Murtijaya, 2007)

4.2.2 Effect on Total Flavonoid content

Fig 4.2 illustrates the total flavonoid content retained in the dried Okra samples after
pretreatment and drying.

SB= Salt blanched

KB= KMS blanched

WB = Water blanched

NT= No treatment

Fig 4.2 Effect of different pretreatments and drying on the Total flavonoid content of Okra

The results expressed above showed that maximum flavonoid retention of 60.27% was seen
in samples dried at 50±2˚C after KMS blanching followed by samples dried at 60˚C (58.72%)

39
for same pretreatment. Total flavonoid retained in the study ranged from 437.49 mg
GAE/100g to 165.56 mg GAE/100g.

Different drying treatments had significantly different effects on the TFC of okra samples
(p<0.05). Two-way ANOVA showed that samples dried at 50±2˚C and 60±2˚C had no
differences, but were significantly different from those dried at 40±2˚C. Similarly, WB and
NT samples had no significant differences but, TPC for SB and KB were significantly higher.

Samples dried at 40±2˚C for 18 h has shown greater loss of TFC which can be explained
with the extended drying period at this temperature which leads to longer exposure of samples
to oxygen during drying that accelerates oxidation, which might have caused higher loss of
polyphenols as explained by Jiang et al. (2017). This result is also supported by Mbondo et al.
(2018) and Zaro et al. (2015). There was concomitant loss of TFC with the loss of TPC
because polyphenols are rendered thermolabile by prolonged heat treatment, leading to the
destruction of flavonoid. On average, KMS treated samples had better retention for all drying
temperature and water blanched had the lowest. The antioxidant effect of KMS prevented the
oxidation of flavonoid during drying. Leaching of flavonoids from the cellular structures
during blanching and oxidation during drying might be the reason for significant loss for WB
samples. However, the retention of flavonoid on drying after different treatment is not
completely studied till now. Thus, more deep research would be needed.

4.2.3 Effect on Total carotenoids

The carotenoid content retained in the dried samples ranged from 2.65 to 0.88 mg/L. Samples
blanched in NaCl and then dried at 50±2˚C retained the highest carotenoid content (54.98%)
whereas water blanched samples retained only 18.23% of the original carotenoid for same
temperature.

40
 3 S.B K.B W.B N.T
2.65 SB= Salt blanched
2.5 2.43
Total Carotenoids (mg/l)

2.04 KB= KMS blanched

2 1.79
1.67 1.58 WB = Water blanched
1.54
1.5 1.31
1.17 1.21 1.17 NT= No treatment
1 0.88

0.5

0
40˚C/18h 50˚C/14h 60˚C/10h

Fig 4.3 Effect of different pretreatments and drying on the Total carotenoid content of Okra

Statistical analysis showed that there was no significant effect (p>0.05) on carotenoid
content due to the drying temperature. Also, there was no significant difference between the
NT, KB and WB samples, whereas the values obtained for SB were significantly higher.

Samples dried at 50±2˚C for 14 h showed the highest values for total carotenoid for all
treatments except for WB. Values for samples dried at 60±2˚C for 10 h were almost
comparable to those of 50±2˚C but slightly lower values were observed for 40±2˚C for 18 h.
This could be explained to the relation between the drying time and temperature. Short time
required for dehydration reduced the destruction of carotenoid as reported by Shams El-Din
and Shouk (1999). Long exposure of samples to heat and air stimulates the oxidation reaction,
which activates the enzymes, which in turn stimulates the oxidative reactions that result in
carotenoid loss (Kamiloglu et al., 2016). When drying is conducted, the carotenoid inside the
materials is concentrated and becomes more vulnerable to destruction mainly due to the
oxidation of conjugated double bond in the carotenoid molecule.

NaCl blanched samples had highest carotenoid content for all drying temperature followed
by the KMS treated samples. Salt and sulphite might have antioxidative effect in preventing
the oxidation of carotenoids. This may be the reason for preventing degradation of carotenoid
and retaining more carotenoids as compared to the control samples. Water blanched samples
had highest loss which might be due to leaching, aided by oxidation during drying.

41
 4.2.4 Effect on Total chlorophyll
16 S.B K.B W.B N.T
13.65 The total chlorophyll retained in the
14 12.93
12.01 12.03 treated and dried Okra samples is
Chlorophyll (mg/100gm)

12 11.56
10.9
10 9.36 shown in the Fig 4.5.
8.59
7.64
8 7.4
6.66 7.1

6
4 SB= Salt blanched
2
KB= KMS blanched
0
40˚C/18h 50˚C/14h 60˚C/10h WB = Water blanched

NT= No treatment

Fig 4.5 Effect of different pretreatments and drying on the Total Chlorophyll content of Okra

From the analysis, it was revealed that the chlorophyll content were found to be decreased
for all samples and higher loss was observed for longer drying time. Thermal processing
induced structural and chemical variation to the tissue which results in loss of color
components (Canjura et al., 1991). Samples treated in KMS and then dried at 50±2˚C for 14 h
retained about 56.21% of original chlorophyll followed by NaCl blanched samples dried at
60±2˚C for 10 h (53.24%). Lowest retention was observed in the water blanched samples dried
at 40±2˚C for 18 h (27.42%).

Statistical analysis shows that, no significant differences were observed in the chlorophyll
content for the samples dried at 50±2˚C and 60±2˚C, but were significantly different than
those dried at 40±2˚C. For the pretreatments, no significant differences were seen for KB and
SB whereas NT and WB showed significant differences on the chlorophyll retention of dried
Okra.

42
 The results shown above cleared that the samples dried at 40±˚C for 18 h had undergone
highest degradation of total chlorophyll. Shams El-Din and Shouk (1999), had concluded that
the method of dehydration, especially the time and temperature involved had significant effect
on the chlorophyll degradation of Okra. Studies have shown that longer treatment time results
in rapid loss of chlorophyll. Nezam El-Din and El-Ashwah (1991) also came to same
conclusion.

In general, highest chlorophyll content was observed in the KMS blanched samples
followed by NaCl blanched samples which might be due to the preservative action of salt and
sulphites and its effect on chlorophyllase inhibition. Control and water blanched samples were
severely affected by the drying temperature. The results were in close agreement in with the
findings of Shivhare et al. (2010) and Shams El-Din and Shouk (1999). The result was also
supported by Kaur et al. (2018) who has reported that KMS treated samples retained higher
chlorophyll than NaCl treated broccoli samples.

4.2.4 Effect on Ascorbic acid

Since, ascorbic acid is vulnerable to heat and oxidation, the blanching treatment and drying
temperature both had significant effect on the total ascorbic acid content. The significant
reduction of ascorbic acid on drying has been reported by Stone et al. (1986) in Okra, Joshi et
al. (2011) in apple, García-Martínez et al. (2013) in apricot, Yang et al. (2010) in sweet potato
and Kaur et al. (2018) in brocoli. The dominate factor affecting the retention of ascorbic acid
are time, method and storage/processing temperature (Padayatty et al., 2003). The mean
ascorbic acid in dried Okra ranged from 25.741mg/100g (at 40±2˚C, NT) to 62.04 mg/100g (at
50±2˚C, KB) which indicates only 16.46% to 39.66% retention on dry weight basis, which
concurs with the results reported by Shams El-Din and Shouk (1999).

43
 70 S.B K.B W.B N.T
62.04 59.98
58.76 56.67
Ascorbic acid (mg/100gm) 60 56.05 SB= Salt blanched
50.12
50 KB= KMS blanched
40 37.92
33.25 30.82 33.18 WB = Water blanched
29.29
30 25.74
NT= No treatment
20
10
0
40˚C/18h 50˚C/14h 60˚C/10h

Fig.4.4 Effect of different pretreatments and drying on the Ascorbic acid content of Okra

Statistical analysis shows that there was significant differences (p<0.05) on the ascorbic
acid retention among the pretreatments. Whereas, no significant differences were observed in
the values for the drying temperatures 50±2˚C for 14 h and 60±2˚C for 10 h but 40±2˚C for 18
h had vulnerable effect on the Ascorbic acid degradation. This can be ascribed to the relation
between length of drying and drying temperature. A longer drying time facilitates faster
oxidation of ascorbic acid (Nindo et al., 2003). The reduced dehydration time at 60±2˚C
produced least destruction. The ascorbic acid content retained after the pretreatment during
drying has been presented in the Fig.4.2. From the data it can be depicted that, ascorbic acid
retention was comparatively higher in KMS treated samples for all drying temperature which
is due to antioxidant effect of sulphites that minimizes the loss of ascorbic acid. The result in
this study agreed with the findings of Shams El-Din and Shouk (1999) and Kaur et al. (2018).
Of all the parameters studied, highest loss was observed for ascorbic acid. Unblanched
samples resulted up to 83.54% loss in ascorbic acid. Blanching slows down or stops the
enzymatic activity and helps to protect the vitamin content (Korus, 2011), so blanched
samples comparatively showed higher values for ascorbic acid. Depletion of ascorbic acid
might also be due to the utilization of the compound for protecting the oxidation of
polyphenols during drying (Kamiloglu et al., 2016).

44
 4.2.6 Effect on Antioxidant
S.B K.B W.B N.T
80 72.74 activity
69.81 70.99
67.94
70 From Fig 4.6, it can be depicted that
59.33 56.89 highest antioxidant activity is shown
55.71
60 53.21
Antioxidant activity%

50.97 50.53
50 47.47
42.61 by the samples dried at 50±2˚C for
40 14 h after KMS blanching. The
30
sample showed 72.74% antioxidant
20
activity. Lowest antioxidant capacity
10
of 42.61% was recorded by the
0
40˚C/18h 50˚C/14h 60˚C/10h water blanched samples dried at
40±2˚C for 18 h.

SB= Salt blanched

KB= KMS blanched

WB = Water blanched

NT= No treatment

Fig 4.6 Effect of different pretreatments and drying on the Antioxidant activity of Okra

Statistical analysis shows that there was no significant difference (p<0.05) in antioxidant
activity for drying temperatures 50±2˚C and 60±2˚C. Similarly, no significant differences
were observed for KB and SB samples. However, values for NT and WB samples were
significantly lower.

45
 Antioxidant activity depends on
SB KB WB NT
the retention of antioxidant
6 5.51
components on different processing
5 4.67 4.65 4.45
4.35
4.23 4.15 4.37
methods. In turn, retention depends
Rehydration ratio

4 3.75
3.75 3.66
3.54
on the temperature and length of
3
drying process. Usually, temperature
2 has negative effect on the
1 antioxidant capacity, which can be
0 attributed to irreversible oxidative
40˚C/18h 50˚C/14h 60˚C/10h
process occurring during drying.
However, no differences were observed for temperatures 60±2˚C and 50±2˚, which was
probably due to the formation of new compounds with antioxidant activities at high
temperature (López-Vidaña et al., 2017). Samples were exposed to oxygen for a longer time at
lower temperature which leads to greater oxidation of phytochemicals resulting in the
reduction of antioxidant activity. Our result indicated that the observed decrement in
antioxidant activity resulted from the degradation of biologically active compounds during
drying (Mbondo et al., 2018). This can also be correlated to the pretreatment.

4.3 Rehydration Ratio

The rehydration capabilities of the dried samples were computed as rehydration ratios. The
rehydration ratios of the dried Okra samples as affected by different drying process are shown
in the Fig 4.6.

SB= Salt blanched

KB= KMS blanched

WB = Water blanched

NT= No treatment

46
 Fig.4.7 Rehydration ratios of the dried samples

Statistical analysis showed that there were no significant differences among the samples
dried at 40±2˚C for 18 h and 60±2˚C for 10 h whereas the values showed significant
differences with the samples dried at 50±2˚C for 14 h. For different treatments, no significant
differences were seen for salt blanched and KMS blanched samples whereas these samples
were significantly different and were superior to water blanched and control samples. Samples
blanched in KMS solution and then dried at 50±2˚C for 14 h showed highest rehydration ratio,
so it can be concluded that theses samples had undergone least structural disruption, due to
which it retained highest nutritional quality.

4.4 Proximate analysis of the fresh and dried samples

The proximate analysis of the fresh and best obtained dried sample is tabulated in the Table
4.2

Table 4.2 Proximate composition of fresh and dried Okra

Parameter Fresh sample (%) Dried samples (%)

Moisture 91.02±0.27 8.727±0.02
Carbohydrat 56.08±0.96 56.56±1.52
e
Protein 10.91±0.03 8.35±0.44
Fat 3.87±0.012 3.15±0.22
Fiber 22.047±0.88 23.77±1.06
Ash 7.073±0.08 7.77±0.05

The values in the table are the means of triplicate ± standard deviation. All parameters are
expressed on dry weight basis except for moisture content.

The analysis of fresh samples in the present study showed that the mean moisture content
of Okra was found to be 91.02%. The result was in accordance with the finding of Kumari,
(2016), where she reported values from 79.46-93.12%. Also, this was in accordance with the

47
finding of Petropoulos et al. (2017) (80.1-91.3%). The mean utilizable carbohydrate in Okra in
this study was found to be 56.08% which is in line with the results reported by Shams El-Din
and Shouk (1999). However, Audu et al. (2015) has reported very lower (10.24%)
carbohydrate in Okra. The average crude protein was found to be 10.91% which was lower
than the values reported by Shams El-Din and Shouk (1999). The values were nearer to the
findings of Audu et al. (2015) (10.24 g/100g) while higher than the value reported by
Petropoulos et al. (2017) (1.37-3.44%) and lower than Adetuyi et al. (2011) (13.61–16.27%).
Similarly, the mean fat content was found to be 3.87±0.0124%. This value was slightly higher
than the findings of Shams El-Din and Shouk (1999). However, Nwachukwu et al. (2014),
reported very low fat content (0.18 g/100 g) and Adetuyi et al. (2011), had reported 9.22–
10.57 g/100 g fat in Okra Mean Fiber content was found to be 22.047%. The value was found
to be higher than the values obtained by Shams El-Din and Shouk (1999), Kumari (2016) (1.3-
4.40%) and Adetuyi et al. (2011) (10.15-11.63%). Present finding showed that the average ash
content was 7.073%. The results obtained are in accordance with the findings of Shams El-Din
and Shouk (1999) and Adetuyi et al. (2011) (7.19–9.63 g/100g).

This variation in proximate components might be due to the variation in moisture content,
difference in maturity and genetic potentials among the studied genotypes which was
explained by Petropoulos et al. (2017).

From the above results obtained in the phytochemical analysis of treated Okra, it can be
concluded that the samples dried at 50±2˚C after blanching in 0.1% KMS were better in
nutritional quality than any other samples. So, proximate analysis of those samples was carried
out. The data in the Table 4.2 showed the effect of pretreatment and dehydration on the
proximate composition of Okra.

The results showed that the final moisture was reduced up to 8.727% on drying.
Dehydration caused slight decrease in protein and fat as well as slight increase in fiber, ash
and carbohydrate when compared to the fresh samples. Loss of protein might be due to heat
denaturation or loss of some water-soluble proteins during blanching and drying. Loss of fat
might be due to oxidative reactions and also leaching during the process. The increase in
carbohydrate can be expected due to changes that occurred on other proximate compositions

48
since carbohydrate was obtained by difference. The heat treatment degraded protein and fat
content to some extent which resulted in concomitant increase in carbohydrate content of the
sample. The results were in agreement with the findings of Shams El-Din and Shouk (1999).

4.5 Calcium content

Calcium content of the fresh and the best quality sample were analyzed which is tabulated in
Table 4.3

Table 4.3 Calcium content of fresh and dried Okra

Sample Calcium content (mg/100g)

Fresh 435.846±19.917
Dried 249.072±5.56
The values in the table are the means of triplicate ± standard deviation. All parameters are
expressed on dry weight basis.

The mean calcium content in the fresh Okra sample was found to be 435.846 mg/100g.
The result obtained in the present study was found to be lower than the values obtained by
Petropoulos et al. (2017). After drying there was significant reduction in the Ca content, this
might be due to the mineral loss or outflow during blanching. Blanching and drying had
caused 42.709% reduction on Ca content of Okra.

49
 Part V

Conclusions and recommendations

5.1 Conclusions
In this study, Okra samples were subjected to different pretreatments (SB, KB and WB), and
then dried at different temperatures (40±2˚C, 50±2˚C, 60±2˚C). Untreated samples were taken
as control. The effect of different pretreatment and drying temperature on the chemical
composition of Okra were analyzed in the lab. Within the scope of present work following
conclusions can be drawn.

i. From the phytochemical analysis (total polyphenol, total flavonoid, total chlorophyll,
total ascorbic acid and total carotenoids), KMS blanched samples were found to be
superior in quality.
ii. There were significant differences (p<0.05) in the phytochemical composition of Okra
due to the difference in drying temperature and pretreatments except for carotenoid.
iii. The role played by KMS and NaCl in the prevention of oxidation of chemical
components were almost comparable except for carotenoid.
iv. Okra samples dried at 50±2˚C were better in quality. The longer drying time required
at 40±2˚C had vulnerable effect on the chemical components of Okra. Shortest time
was taken for 60±˚C, but this temperature lead to higher destruction of some sensitive
components. Thus, time and temperature regime played vital role in quality of dried
Okra.
v. Drying also had effect on the proximate composition of Okra. On drying, there was
slight decrease in protein and fat whereas increase in carbohydrate, fiber and ash
content was observed due to concentration effect.
5.2 Recommendations
In Nepal, there is almost no research done in Okra regarding its chemical composition. So, an
intense effort must be invested so as to obtain further information about Okra and effect of
different treatments and dehydration on it.

Based on the present study, following recommendation could be drawn for further studied.

i. Further studies on the nutrients like thiamin, β-carotene and antinutrients like oxalate,
phytate and tannin can be carried out.
ii. Variation in the drying methods and comparison of different varieties can be done.
iii. Sensory analysis of dried Okra can be carried out.

50
 Part VI

Summary
Okra is a vegetable with enormous nutritional benefits. It is an affordable source of natural
antioxidants containing good amount of polyphenols, carotenoids, vitamins, minerals and most
importantly, dietary fibers. It is high in moisture content, thus undergoes deterioration quickly.
During the lean seasons, it is scarce and expensive. Thus, drying of Okra can be of great
importance, since it renders the availability of this vegetable throughout the year. In the
present study, Okra samples were dried at three different temperatures (40±2˚, 50±2C˚and
60±2˚C) in cabinet drier after subjecting to different blanching pretreatments (KMS blanching,
NaCl blanching and water blanching). The retention of nutrients on above processing were
compared taking untreated samples as control.

The effect of pretreatments and drying on the chemical composition of Okra was estimated.
All the phytochemicals were reduced on processing, but the temperature of 50±2˚C and KMS
blanching was found to be optimum for drying Okra since, it retained comparatively higher
amount of bioactive phytochemicals. The highest average values for TPC, TFC, Ascorbic acid,
total chlorophyll, antioxidant activity, rehydration ratio and moisture content was found to be
1399.24 mg GAE/100g, 437.39 mg GAE/100g, 62.04 mg/100g, 13.65 mg/g, 72.74%, 5.51and
8.72% respectively, obtained for KB samples dried at 50±2˚C for 14h. However, highest total
carotenoid of was recorded by NB samples for same drying temperature. Time and
temperature regime had significant effect on the retention of phytochemicals. Samples dried at
40±2˚C for 18h were poor in nutrients due to the longer drying time required to attain the final
safe moisture level. The proximate analysis of the fresh and best quality samples shows slight
changes due to blanching and drying treatment.

The above results showed that a product with safe moisture level and higher self-life with
better nutrient, serving with reduction of volume and ease of handling can be obtained so as to
make the nutritional benefits of okra available during the off seasons in a more concentrated
form. This helps in minimizing the loss encountered throughout the world.
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Lebensmittel-Wissenschaft und-Technologie. 64, 802-808. 10.1016/j.lwt.2015.06.061.

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 Appendices
Appendix A

Two way ANOVA table for phytochemical content of fresh Okra.

Table A.1 Two way ANOVA results for Total Polyphenol retention at different
temperatures and treatments

Variate: Total Polyphenol

Source of variation d.f. s.s. m.s. v.r. F pr.

Temperature 2 32432.2 16216.1 34.44 <.001
Treatment 3 220651.9 73550.6 156.20 <.001
Residual 6 2825.2 470.9
Total 11 255909.3

Table A.2 Two way ANOVA results for Total flavonoid retention at different
temperatures and treatments

Variate: Flavonoid

Source of variation d.f. s.s. m.s. v.r. F pr.

Temperature 2 7249.9 3625.0 10.14 0.012
Treatment 3 75840.5 25280.2 70.74 <.001
Residual 6 2144.1 357.4
Total 11 85234.5

Table A.3 Two way ANOVA results for Total carotenoid retention at different
temperatures and treatments

Variate: Carotenoid

Source of variation d.f. s.s. m.s. v.r. F pr.

Temperature 2 0.00156 0.00078 0.01 0.989

67
Treatment 3 2.73197 0.91066 12.46 0.005
Residual 6 0.43834 0.07306
Total 11 3.17188

Table A.4 Two way ANOVA results for Total ascorbic acid retention at different
temperatures and treatments

Variate: Ascorbic acid

Source of variation d.f. s.s. m.s. v.r. F pr.

Temperature 2 74.819 37.410 17.66 0.003
Treatment 3 2044.302 681.434 321.74 <.001
Residual 6 12.708 2.118
Total 11 2131.829
Table A.5 Two way ANOVA results for Total chlorophyll retention at different
temperatures and treatments

Variate: Chlorophyll

Source of variation d.f. s.s. m.s. v.r. F pr.

Temperature 2 4.4401 2.2200 5.38 0.046
Treatment 3 60.6840 20.2280 48.98 <.001
Residual 6 2.4777 0.4129
Total 11 67.6017

Table A.6 Two way ANOVA results for Total antioxidant activity retention at different
temperatures and treatments

Variate: Antioxidant activity

Source of variation d.f. s.s. m.s. v.r. F pr.

Temperature 2 431.864 215.932 41.05 <.001
Treatment 3 644.443 214.814 40.84 <.001
Residual 6 31.561 5.260
Total 11 1107.868

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 2.5
Absorbance at 765 nm

2 f(x) = 0 x + 0.16 Absorbance Table A.7 Two way ANOVA

R² = 0.98 Linear
1.5 (Absorbance) results for Rehydration ratios at

1 Linear different temperatures and

(Absorbance)
0.5 treatments
Linear
0 (Absorbance)
Variate: Rehydration ratio
0 100 200 300 400 500 600 700 800
Standard Gallic acid (µg/ml)

Source of variation d.f. s.s. m.s. v.r. F pr.

Temperature 2 1.43032 0.71516 14.91 0.005
Treatment 3 1.63220 0.54407 11.34 0.007
Residual 6 0.28784 0.04797
Total 11 3.35036

Appendix B

The standard curves of gallic acid for the determination of polyphenol and flavonoid content
of the dried Okra samples.

69
 0.5
0.45
0.4 f(x) = 0.01 x − 0
Absorbance at 510nm

0.35 R² = 0.99
0.3 Absorbance
0.25 Linear
(Absorbance) Fig B.1 Standard Gallic acid
0.2
Linear
0.15 (Absorbance) Equivalent Calibration Curve at
0.1 765 nm
0.05
0
0 10
Standard20
Gallic30 40
acid (µg/ml) 50 60

Fig. B.2 Standard Gallic acid Equivalent Calibration Curve at 510 nm

70
 Appendix C

Values obtained for different parameters. All values are expressed in dry basis.

1. Total polyphenol content (mgGAE/100gm)

Temperatur
e 40˚C/18h 50˚C/14h 60˚C/10h
S.B 1216.029±8.34 1314.361±16.5 1217.744±16.8
K.B 1230.245±30.1 1399.24±22.3 1385.661±8.4
W.B 856.65±68.1 1078.123±8.41 992.266±8.59
N.T 1019.715±22.9 1153.167±31.2 1100.071±23.1

2. Total flavonoid

Temperatur
e 40˚C/18h 50˚C/14h 60˚C/10h
SB 353.138±8.37 405.941±13.17 362.797±6.18
KB 385.929±13.11 437.391±4.1 426.168±30.96
WB 165.558±2.39 251.041±15.85 259.839±15.74
NT 232.374±9.44 274.745±22.54 293.767±6.19

3. Total carotenoids (mg/L))

Temperatur
e 40˚C/18h 50˚C/14h 60˚C/10h
S.B 2.044±0.012 2.654±0.034 2.433±0.036
K.B 1.675±0.023 1.786±0.034 1.578±0.035
W.B 1.535±0.026 0.88±0.028 1.168±0.029
N.T 1.166±0.022 1.209±0.025 1.313±0.019

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 4. Total Chlorophyll (mg/100gm)

Temperat 60˚C/10
ure 40˚C/18h 50˚C/14h h
10.898±10 12.014±10 12.93±9.
S.B .54 .56 97
11.557±0. 13.650±0. 12.03±0.
K.B 274 32 34
7.397±083 8.585±0.5 9.358±0.
W.B 4 6 37
7.643±0.4 7.103±0.
N.T 6.66±0.29 6 49

5. Ascorbic acid (mg/100gm)

Temperatu
re 40˚C/18h 50˚C/14h 60˚C/10h
50.115±0.7 58.758±1.0
S.B 48 21 56.674±0.576
56.047±0.8 62.039±1.8
K.B 6 7 59.976±0.65
33.254±0.8 37.916±1.0
W.B 5 9 33.183±0.87
25.741±0.9 30.823±1.0
N.T 2 32 29.286±0.82

6. Antioxidant activity (%)

Tempera
ture 40˚C/18h 50˚C/14h 60˚C/10h
53.213±0. 69.807±0. 67.935±0.
S.B 311 63 922
55.708±1. 72.739±0. 70.992±0.
K.B 92 54 34
42.608±0. 50.967±0. 50.530±0.
W.B 45 133 462
47.473±0. 59.326±1. 56.893±0.
N.T 86 32 521

7. Rehydration ratio

Temperature 40˚C/18 50˚C/14h 60˚C/10h

72
 h
SB 4.35 4.67 4.45
KB 4.232 5.51 4.37
WB 3.75 4.65 3.66
NT 3.75 4.15 3.54
Appendix D

Plate 1. Fresh samples brought for

analysis

73
 Plate 2. Samples treated and cut into
sizes for drying

Plate 3. Extract prepared for phytochemical analysis

74
 Plate 4. Dried samples
preserved for further
analysis

Plate 5. Dried v/s rehydrated

samples

75