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Manufacture, Pharmacological Analysis and Marketability Study of Cold
Rehydrating Beverage
K. Kishore1, M. Rahul
1, N. Priya
1*
1Department of Biotechnology, Sir M Visvesvaraya Institute of Technology, India
*Author for Correspondence: [email protected]
Abstract
With several aerated drinks and processed
beverages gaining popularity due to their
taste, the need to introduce healthier and
long-lasting beverages has simultaneously
risen. One such formulation constitutes
several natural ingredients that are
medicinally beneficial. The solid content
constitutes tamarind extract and palm
jaggery (Phoenix dactylifera) in a simple
ratio by weight, with a nominal percentage
of Aloe vera, all in water. The manufacture
protocol is similar to that of RTS (Ready-
to-serve) beverages. The drink possesses a
few important pharmacological properties
along with phytochemical properties. A
trifling amount of preservative has been
observed to substantially increase the
shelf-life of the beverage to around 3
months. Possibilities of packaging in
several types of material and maintenance
of freshness were studied. Taken together,
this nutritious drink with its potential
abundance of health benefits can stand in
as a commercial product that is rendered
valuable to its consumers.
Keywords: beverage, jaggery,
Phoenix dactylifera, Aloe vera,
pharmacological, shelf-life.
Introduction
The shift of the preferences of the current
population in several aspects of life is
worth noting. And such a shift in food
preferences does not just shift regularly,
but also shows continuously varying
distributions of such preferences over
different populations, in several regions of
the world. However, even in such a state of
vastness, there might exist some key points
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to note – some preferences that are
common to a majority, if not all, of a given
population. And one of them is –
preference to healthy alternatives. In the
food market, such novel techniques in the
form of value-addition, functional food
design, scouting for potential alternative
sources of food that are inexpensive and
readily available are in the process. These
approaches are also applied in the
beverage sector. As this sector continues to
prevail with several products offering
novel flavours and customer choices, the
potential of fully natural sources to provide
several health benefits to the same section
of consumers is becoming more visible.
There is now a need for healthy beverages,
that last long, and can be commercially
valuable, ensuring for the firm a long stay
in the market amidst all competition and
challenges [10]
.
The reasons for healthy beverages are: 1.)
Hydration – Water being the oldest source
of hydration, has posed a base for
development of several flavours of
hydrating beverages that customers can
choose from. The main advantage of
hydration is, obviously, prevention of loss
of body fluids. 2.) Nutrition – Along with
the hydration property, a beverage
accounted for by the right sort of
fortification, can be a good source of
energy, carbohydrates, certain vitamins,
and minerals. 3.) Energy balance – A
beverage must carry with it the right kind
of energy and also nutrient balance, so as
to be incorporated readily into the regular
diet of a consumer and to exert its benefits
consequently.
The beverage product that was intended to
be designed is backed up by its own set of
rational points: Zero Fat, High Energy,
Electrolyte Balance, Fully Natural
Sources, Zero Artificial Sugars.
Considering the fact that most cold
beverages do not possess high energy
values (the energy only being on account
of sugars present), do not comprise fully of
natural sources, contain artificial sugars,
etc., and most importantly, do not possess
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all of these properties as a single product.
The drink, as mentioned earlier, comprises
natural sources alone. The only artificial
element would be the very small amount
of Class II preservative added. The main
constituents include the sugar obtained
from the sap of the date palm (Phoenix
dactylifera), commonly known as palm
jaggery or “Karuppattivellam” in Tamil
Nadu, where the jaggery is popularly
consumed; and the extract of Aloe vera,
added in a nominal yet crucial percentage.
With other important ingredients coming
into play, the beverage shall possess
important pharmacological properties.
One question does arise – why would it be
difficult for such high-benefit beverages to
make it to the commercial sector? There
exist three plain reasons for this
1) The shelf-life of the product must be
analysed, and related issues need to be
addressed.
2) The most suitable storage and transport
conditions and setup need to be addressed.
3) Awareness, advertising, consumer
targeting must be done in a holistic
manner, therefore the origin of production,
the preferences of customers in a region,
the profit feasibility, need to be addressed.
This leads one to a general viewpoint of
the changing trends in the food-and-
beverage industry, the need for
manufacturers to develop novel techniques
and products, and the need for more and
more scientific involvement to validate the
beneficial properties of such food
products, to make them a profitable
venture intended for the goodwill of the
society and the general health of a
population.
Raw Materials
Palm jaggery: Cane jaggery, obtained
from sugarcane, is the most common sugar
source used in preparation of
perishable/non-perishable food products,
such as sweets, sweet milk, and some
value-added ready-to-eat foods. To
understand the efficacy of palm jaggery as
a more beneficial alternative to cane
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jaggery, comparative studies were carried
out by reference to suitable journals. The
most striking difference between these two
types is in the physical appearance. Palm
jaggery bears a darker brown colour than
the conventional cane jaggery. One
teaspoon of palm jaggery contains
approximately 4-5 mg calcium, 2-3 mg
phosphorus, 8 mg magnesium, 48 mg
potassium, 0.5 mg iron, as well as trace
amounts of zinc, copper, thiamine,
riboflavin, and niacin. The moisture
content of palm jaggery is 10.2% dry basis
[4]. It is worth noting that the
corresponding values for refined white
sugar are nearly negligible. Palm jaggery
also boasts a few other advantages – albeit
marginal – over cane jaggery. The former
has greater amount of reducing sugar –
87% dry basis as opposed to 80% in the
latter [13]
, and a lesser amount of non-
reducing sugar – no more than 68%, which
could make the beverage a suitable
consumable for diabetics. Jaggery is
already a known substitute to refined
sugar, benefitting people with such
ailments. Hence the incorporation of such
a component to our drink would enhance
the possibility of universal consumption.
Other factors worth noting is the presence
of a higher percent of protein in palm
jaggery – 2.80% as opposed to 0.77% in
the latter [12]
, and presence of a good
percentage of minerals, which could be
touted just enough for the daily intake of
the consumer.
Aloe vera: Aloe vera has a plethora of
properties – ranging widely from anti-
inflammatory, anti-oxidant, anti-cancer,
anti-diabetic to several other unexplored
properties [9]
. The gel of the plant is a well-
known component, in hand-sanitisers,
cosmetic products and topicals. A trifling
percentage, however, when added to our
beverage, is expected to account for the
pharmacological properties [1]
. Aloe vera is
also reported to remove toxicity and
balance hormonal problems. The benefits,
and the best methodology to secure highest
yield, were described in
detail [3]
. The
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methodology comprised of heating to
80°C, and then flash cooling. We
integrated this methodology into the
primary manufacturing process of the
drink.
Tamarind: Tamarind (known by the
scientific name Tamarindus indica) is
known to have several health benefits [11]
.
The polyphenols in tamarind have
antioxidant and anti-inflammatory
properties. These serve to protect against
cardiac diseases and are even touted to be
effective against cancer and diabetes. The
traditional beverage form of tamarind,
known as “Puli Saaru”, was used to treat
conditions such as diarrhoea, constipation,
fever, and peptic ulcers. Tamarind is a rich
source of several nutrients including
magnesium, potassium, Vitamin B1, B2
etc. The plant extract also possesses a good
number of calories.
Cardamom: Research work [2]
throws
light on the abundant properties of
cardamom, known by the scientific name
Elettaria cardamomum. The spice bears
diuretic properties. This, along with the
anti-oxidative property, is reported to
relieve blood pressure. Apart from anti-
cancer properties, cardamom is an
effective anti-inflammatory agent.
Methodology
Manufacture
Before beginning with the process of
manufacture, the raw material must be
lined up in detail. They include jaggery
(Phoenix dactylifera), tamarind
(Tamarindus indica), cardamom (Elettaria
cardamomum), Aloe vera, dried ginger,
sodium benzoate, and water. The standard
reference for ratio is 1 litre of water. The
weights of the ingredients need not vary,
however, for suiting selective regional
preferences, the ratios can be modified to
accommodate the ideal taste for the
population. This would include increasing
of sweetness, or increasing the cardamom
and dried ginger component for a spicy
tinge, and so on. Parameters such as Total
Soluble Solids (TSS), acidity and amount
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of preservative to be added are adhered to
[6]. The preparation of the beverage is
similar to that of the RTS (Ready-to-serve)
beverages [5]
.
Selection and Sorting
The process of selection involves scouting
for any below-par quality parts of the raw
material, discarding them, and keeping the
higher quality material ready for the next
step. The different materials must be kept
distinguishably apart from each other.
Figure 1: Manufacturing process
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Washing
This is done before the mixing of the
ingredients takes place. Usually the
washing is carried out in water baths, but
depending on the quantity of production,
this may be scaled up.
Extraction of Pulp
Especially for tamarind and aloe vera. For
tamarind, the pulp is extracted by prior
dipping in water for a period of 30
minutes, and using muslin cloth to filter.
For aloe vera, the extraction is carried out
by skinning off the rind and taking out the
gel.
Mixing
This is done in a kettle, usually the
blanching kettle unit. With a stirrer, the
contents may be mixed thoroughly in
water. The cardamom and dried ginger are
ground finely before mixing.
Addition of preservative
Sodium benzoate is added in the order of
120 ppm (parts per million) per litre of
beverage. This amount, usually a pinch in
simple words, is mixed with a small
amount of the beverage mixture, and then
added to the rest of the mixture.
Sterilisation
The process of pasteurisation is carried out
at 80°C for 20 minutes. During this
process, the TSS is measured using a
refractometer, to ensure the solubility
factor is not compensated for.
Cooling and Storage
The initial cooling is done to room
temperature, and later, may be stored in
conditions of refrigeration at 4°C. The
packaging may be done depending on the
demand for supply.
Sensory Evaluation
The sensory evaluation can be done in
various ways, ranging from simple ranking
tests to more complex action rating tests.
We employed a hedonic scale in a food
action rating test, to determine subject
likeability to the manufactured product [15]
.
The Food Action Rating Test, known as
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the FAR Scale Test, this test is done to
determine the attitude of the subject
towards the product presented to them.
The subject tastes the sample and gives
their opinion by ticking, or entering a
number according to a scale, an example
of which is given: 9 - Like extremely, 8 -
Like very much, 7 - Like moderately, 6 -
Like slightly, 5 - Neither like nor dislike, 4
- Dislike slightly, 3 - Dislike moderately, 2
- Dislike very much, 1 - Dislike extremely.
The hedonic rating scale test consisted of
40 subjects, out of which 15 were domain
experts in the food science sector, and the
other 25 were subjects from the general
public.
Qualitative Metabolite Analysis
The beverage was expected to bear the
presence of three major metabolites –
tannins, flavonoids, and alkaloids. To
detect the presence of these, subsequent
tests were carried out and the inferences
noted.
Ferric Chloride Test for Tannins
A few drops of ferric chloride solution
(10%) is added to the sample (here, the
drink). The appearance of an intense green,
blue, purple or black colour indicates the
presence of tannins.
Lead Acetate Test for Flavonoids
About 1ml of lead acetate is added to 5ml
of the sample. The formation of a distinct
yellow precipitate indicates the presence of
flavonoids.
Wagner’s Test for Alkaloids
Wagner’s reagent is prepared by adding
2.5g of iodine to 12.5g of potassium iodide
and mixing with 250ml of water. A few
drops of the reagent are added to the
sample. A reddish-brown precipitate
indicates the presence of alkaloids.
Apart from procedure references [14]
, it was
adhered to as a common step for all
qualitative tests – slow addition of the
reagents, slight mixing, and waiting for a
minimum of 30 seconds before observing
for the precipitate or colour formation.
Anti-Inflammatory Protocol
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The anti-inflammatory protocol was
carried out in an order of steps [16]
. Initial
work involves preparation of the Alsever’s
solution, which is nothing but the blood
preservative solution. It consists of 2%
dextrose, 0.8% sodium citrate, 0.5% citric
acid, and 0.2% NaCl. After addition of
equal volumes of fresh blood sample and
Alsever’s solution, centrifugation was
carried out for 20 minutes at 3000 rpm.
The pellet obtained was washed with
isosaline solution. Then,
1 ml of the sample (the drink), 1ml of
phosphate buffer to maintain the pH, 2 ml
of hyposaline solution and 0.5 ml of the
blot solution were taken in a test-tube. In
case of the control, 2 ml of distilled water
is added in place of the hyposaline. Second
centrifugation is carried out for 20 minutes
at a higher speed of 7000 rpm. The
absorbance or optical density (O.D) of the
sample was observed at 560nm. The
percentage of lysis prevention, or simply,
the anti-inflammatory activity, was
calculated using the formula given below:
Apart from the control (aspirin) the sample
was subjected to anti-inflammatory
property deduction in fresh blood sample
at two different temperatures – a colder
temperature of 10°C, and higher
temperature of 70°C (equivalent to the
temperature of a hot beverage like coffee
or tea). The results were tabulated.
Anti-Oxidant Activity
The antioxidant activity was evaluated
according to the method given in the
research work by [8]
. DPPH refers to 1,2-
diphenyl-picryl-hydrazyl, a dark-colored
crystalline powder composed of stable
free-radical molecules. DPPH is a well-
known scavenger or trap for other radicals.
% Protection = (OD of sample / OD of control) x 100
% Activity = [(Ac - A
T)/ A
c] X 100
Ac = Absorbance of control.
AT = Absorbance of sample.
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Rate of reduction of chemical reaction
upon addition of DPPH is indicative of
radical nature of the reaction. The reagent
is prepared by adding 97ml of the sample
(here, the drink) in 3 ml of the DPPH
reagent, which itself is prepared by adding
25mg of DPPH powder in 100ml of
methanol, producing a characteristic dark-
blue colour. Incubation is carried out in the
dark for 30 minutes. The absorbance is
observed at 579nm. The anti-oxidant
property is measured by calculating the
DPPH radical scavenging activity, which
is given by the formula below:
The beverage was manufactured in the
month of February 2020, and the DPPH
activity was tested every 14 days. Hence,
three trials were taken for the month and
the results tabulated.
Nutritional analysis
The sample was sent to the Indian Institute
of Food Processing Technology for
analysis. The components that were
analysed, keeping in mind the objective to
market the beverage as a natural energy
drink, were protein, fat, carbohydrate, ash,
and energy (kcal). The parameters were
evaluated, and values obtained in the order
of grams per 100 grams of sample. The
minerals tested for included sodium,
potassium, magnesium, and iron.
In labelling, the most important panel that
must be included, without doubt, is the
nutrition information panel. This
encompasses the nutrients present, along
with the values in accordance with the
Dietary Reference Intake (RDI) prescribed
by the World Health Organization (WHO).
tIn addition, other parameters such as
nutrient function claims, ingredient list,
logo, vegetarian/non-vegetarian marks are
to be added.
In the packaging optimisation, the various
possibilities that were considered were
simple plastic packets (inexpensive,
suitable for rural businesses only),
polyethylene bottles (suitable for urban
businesses, but expensive), retort pouches
(effective, but expensive as well),
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polyethylene spout pouches (suitable for
both urban and rural businesses,
inexpensive). To assess the best fit for
storage, four different conditions were
evaluated -refrigeration at 4°C, deep
freezing at -18°C, room temperature
conditions in a ripening chamber at 25°C,
and humid room temperature conditions at
about 30°C.
The marketability study was carried out on
the basis of a hypothetical business setup
of a home-scale industry, which is defined
as one consisting of a workforce of not
more than 5 members. With such a
scenario in mind, five points need to be
addressed before starting the business: 1)
Initial capital investment largely localized,
like the target region – The initial
investment must be carefully planned, and
depending on a survey of the target region,
be adjusted so as to not cause excess
investment. 2) Enough backup capital for
second round investment – In case a trial-
and-error scenario of region targeting
persists, a second round of sales must be
financed for the new region and cycle. 3)
To ensure slow but steady recovery of cost
– One must embark on the business only
on a, at least, hypothetical assurance that
the capital cost may be recovered. 4)
Careful analysis of consumer population
and market in the region – It must be made
sure that the data is collected in the right
manner, so as to not let any discrepancy
affect the estimated profit values. Hence,
the consumer population and their
likeability, and the potential for the
product to make its mark in the market,
and stay, must be scrutinized. 5) Adjust
selling price only according to demand
and supply – The initial selling price must
itself not be too high. The business must
start with the sole objective of only
recovering capital. The profit targeting
may be done once the product gains
popularity in the consumer market.
Viable Profit Evaluation
The viable profit evaluation is done by
evaluating the cost of raw material used
per unit (here, per litre of beverage) is
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deduced, and the total expenditure in this
section makes up the manufacturing
capital. Other miscellaneous costs include
those for consumption of energy (heat,
electricity), the packaging and labelling,
and local transport (if required). These
make up the total capital.
Results and Discussion
The Sensory Evaluation
The parameters that were to be ranked by
the subjects were appearance, aroma,
colour¸ taste and an overall score of what
the subjects felt about the drink.
Additional comments were welcome. The
rankings of the 40 subjects were compiled
and tabulated as average scores for every
parameter. The additional comments
included these, and corrections as
suggested were incorporated into the
further batches of production: The usage of
wild tamarind (grown in the hilly regions
of Tamil Nadu) in the sample proved not
to be very welcoming. General subjects
expressed the rather dominating presence
of the sour taste. When replaced with the
normal commercial tamarind, the taste
seemed to be enhanced. The aroma was
recognizable, although not from afar. It
was nevertheless rated well by the
subjects. The aroma of the jaggery and
tamarind together was touted to be
attractive. The dark-brownish colour of the
drink was also appreciated. The consumers
described the colour as a warm one, that
could, by itself, prompt a customer to
purchase it. Overall, the taste of the drink
was appreciated, and according to the
subjects, the overall appearance from a
distance was also worth noting. The final
score for the drink was 8 – liked very much
by the subjects.
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Table 1: Average Scores of Sensory Evaluation
Presence of Metabolites
The series of qualitative tests performed on
the beverage sample to detect the presence
of tannins, flavonoids, and alkaloids
brought positive inferences. A purple
colour in response to the ferric chloride
test indicated presence of tannins.
Formation of flocculating yellow
precipitate on addition of lead acetate
indicated presence of flavonoids. Reddish
brown precipitate on addition of Wagner’s
reagent indicated presence of alkaloids.
The Anti-Inflammatory Activity
For aspirin (the control), a lysis prevention
of 29% was observed. The values could
vary depending on factors such as the
individual’s disposition towards the drug.
When taken as a hot beverage, the drink by
itself exerts a considerable lysis prevention
of 20%. The value could be so because of
the presence of a high amount of sugars. It
can be deduced that the anti-inflammatory
property was on account of the tamarind
and cardamom present in the beverage.
However, like any other soft drink or cold
beverage/food not recommended to
patients with symptoms, a refrigerated
drink ONLY serves as a rehydrating one,
and cannot be used to treat conditions
involving fevers because of its negative
value of lysis prevention. Hence, it could
be recommended to serve the hot beverage
as an anti-inflammatory agent.
APPEARANCE COLOUR AROMA TASTE OVERALL
8 8 7 7 7.5 (~8)
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Table 2. Anti-Inflammatory Activity
The Anti-Oxidant Activity
On conduction of the DPPH protocol to
determine the anti-oxidant property of the
beverage, some positive inferences were
arrived at. It was observed that a
substantial DPPH activity (or rather, the
anti-oxidant activity) of 70-73% was
observed. Presence of the metabolites such
as tannins and flavonoids contribute
extensively to the anti-oxidant property of
the drink.
Table 3. Anti-oxidant activity
DAY FROM SHELF
LIFE
COMMENCEMENT
ABSORBANCE
OF CONTROL
(Ac)
ABSORBANCE
OF SAMPLE
(AT)
DPPH
ACTIVITY
(in %)
1 0.52 0.14 73.07
14 0.52 0.14 73.07
28 0.51 0.15 70.59
Nutritional analysis
The nutritional analysis was conducted at
the Department of Food Safety, Quality
and Testing at the Indian Institute of Food
Processing Technology, Thanjavur. The
minerals, namely – sodium, potassium,
magnesium, and iron, were analysed as
Test OD of
Control
@560nm
OD of Test
@560nm
% of Lysis
Prevention
Aspirin
0.21
0.15 29%
Heated Drink (@70°C) 0.17 20%
Refrigerated Drink (@10°C) 0.31 <0%
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well. The test results revealed no detection
of fat in the drink. Sufficient protein and
carbohydrate levels in one serving were
present. No excess value that can cause
side effects is liable to be claimed. A high
energy value of 50.96kcal was reported.
Sodium, potassium and magnesium were
detected in acceptable levels. This
indicates substantial electrolyte balance,
given the values of sodium and potassium
at 268.0mg/kg and 423.8mg/kg
respectively.
Table 4. Nutritional Analysis
Sr. No. Nutrient Results (g/100g)
1 Protein 1.62
2 Fat Not Detected
3 Carbohydrate 11.12
4 Moisture 86.51
5 Fibre 0.5
6 Ash 0.25
7 Energy (kcal) 50.96
Sr. No. Mineral Results (mg/kg)
1 Sodium (Na) 268.0
2 Potassium (K) 423.8
3 Magnesium (Mg) 69.03
4 Iron (Fe) Not Detected
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From the different possibilities of
packaging analysed, the two most feasible
types were looked into in more detail- 1.)
Retort pouch. The retort pouches are made
of aluminium, polyethylene, polyester,
polypropylene, and nylon stacked in five
layers [7]
. Although an effective packaging
material, it is highly expensive. Another
thing that had to be noted was that the
retort pouch is completely sealed from end
to end, which meant that a consumer
would find it inconvenient to consume the
beverage on purchase. Another major
disadvantage was that to perform retort
pouch packaging, sterilization is a
necessary step. And since pasteurization
was already carried out during the
manufacturing process, a second time
exposure to a higher temperature of 121°C
and pressure of 15 lbs would adversely
alter the property of the drink. 2.) Spout
pouch. Spout pouches are simpler in
structure, made entirely of polyethylene.
These have spout like protrusions making
it easy to fill the pouch with rack-and-rail
apparatuses in industries, or simply via
dispensers. These are fairly inexpensive
for the numerous choices of size and shape
they offer. Although they may be deemed
to be not so environment-friendly, the
advantage of reusability over a long period
of time may compensate for this concern.
Hence, spout pouches were deduced to be
the optimal packaging material for the
beverage.
Table 5. Storage condition evaluation
CONDITION TEMPERATURE SPOILAGE
Low temperature refrigerator 4°C No (over 4 months)
Lower temperature freezer - 18°C No (over 1 week)
Dry ripening chamber 25°C Yes, after 48 hours
Humid room temperature 30 °C Yes, in less than 48 hours conditions
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Labelling
The logo of the drink and the primary
nutritional information panel were also
designed consequently. These would be
incorporated into the material used for
packaging, either printed or wrapped
around as a sticker. The nutritional
information panel consists of the
information of the nutrients present in the
beverage, along with the reference intakes.
The label format is horizontally long. It
can, however, be modified depending on
the type of packaging material. The label
defining the ingredients, and the symbol
indicating that the product is completely
vegetarian, are also other additions to the
entire labelling contingent.
Viable Profit Evaluation
While performing a marketability study,
evaluation of possible profit is a must. The
profit was estimated per litre of the
beverage. The raw material made up for
most of the capital investment, defined as
the manufacturing capital. The other
factors adding to the capital included the
energy consumption, packaging, labelling,
and local transport (if required).
Figure 2: (from top) Nutritional label, ingredient list, vegetarian logo
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Table 6: Total capital evaluation
Conclusion
On account of the COVID-19 pandemic,
further marketability surveys had to be
stopped. However, few unofficial surveys
included discussions with retailers and
vendors, carrying out a very local sales
cycle, filling the beverage in packets and
selling them at a nominal price. Substantial
consumer preferability was observed, but
is yet to be recorded.
Some conclusions, however, were arrived
at, backed up by valid results:
establishment of a sophisticated
manufacturing process for the drink,
acquisition of the qualitative and
quantitative results indicating its
MATERIAL USED FOR
MANUFACTURE/MISCELLANEOUS
PARAMETERS
COST PER LITRE OF
BEVERAGE
(₹)
Date Palm Jaggery 62.5
Tamarind 1.8
Cardamom 2.4
Dried Ginger 0.55
Aloe vera Gel 0.015
Sodium Benzoate (Preservative) 0.01
Domestic Heating 1-3
Packaging and Labelling 8-10
Local Transport 1-2
TOTAL CAPITAL 80-85
Kishore et al., 2021. Journal of Agriculture Environment & Food Security. 2: 9-31
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Page | 31
nutritional value and health benefits,
experimentation of effective storage and
packaging fits, completion of base analysis
of the market feasibility with a home-scale
business model plan and profit target, and
registration of the beverage as a supplier
product under the state government.
“In sum, we expect to introduce to all a
nutritious drink whose abundance of
properties shall benefit the society in terms
of health and the retailers, manufacturers
in terms of income.”
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