Republic of the Philippines

BATANGAS STATE UNIVERSITY

Pablo Borbon Main II
Alangilan, Batangas City

COLLEGE OF ENGINEERING, ARCHITECTURE & FINE ARTS

Chemical & Food Engineering Department

CONCEPT PAPER NO. 2

DEVELOPMENT OF PEARLS FROM MALUNGGAY (Moringa oleifera) ROOT

STARCH AND COMPARATIVE ANALYSIS TO TAPIOCA PEARLS

Prepared by:

ROOVIC F. FERNANDEZ
MARK KEVIN D. MARQUEZ

Submitted to:

ENGR. MARY ROSE F. PERSINCULA

Associate Professor II

NOVEMBER 2021
 I. RATIONALE

When a person looks around, he or she may notice a youth carrying his or her phone or
hard plastic long cups stuck with oversized straws and a brown appealing liquid inside with
floating dark circles or pearl milk tea in short. This drink craze all around the world happened
unexpectedly. It originated in Taiwan during the 1960s. The milk tea started at Chen Shui Tang
tea shop at Taichung city selling high mountain oolong tea. The owner Liu Han-chien wanted his
tea shop to stand out. He added milk tea in a cocktail shaker with ice. It revolutionizes the
industry because people thought it was crazy yet young people loved it. The “bubble tea” became
known because of the thick layer of foam after it is shaken. Then in 1987, Liu held a competition
among his staff to come up with creative concoction. During this time, the store manager likes to
savor on fen yuan or tapioca balls. The owner’s daughter added them to the milk tea. Thus, this
is the birth of pearl milk tea.

In year 2008, milk tea was officially introduced in the Philippine market by Dr. Juliet D.
Herrera-Chen and Peter L. Chen. In 2009, milk tea starts to invade the universities and this was
the start of booming industry in the country.

As we all know before, tea drinks are not popular in the Philippines because Filipinos are
more coffee drinkers but milk teas and other tea drinks captured the attention of young
generation today. That is why milk tea became popular and many people tend to have these kinds
of business. Milk tea was easily accepted by our society because of its satisfying taste and aroma.
Common milk tea consists of milk, sugar, tea and its highlight tapioca pearl that gives the chewy
liked mouth feel on consumers. Milk tea came out with add ons like popping boba, nata and
pudding. But it will be better if these add-ons can be altered with a new product that is healthier
and more nutritious.

Tapioca pearls are translucent spheres produced from tapioca, a starch extracted from the
cassava root. They are most referred to as boba or pearls in bubble tea. These are typically five-
to ten-millimeter starch balls, consisting of sweet potato powder, potato powder, or jelly. The
various forms of tapioca balls include the original black, flavored, popping, mini, and clear. By
adding different ingredients, like water, sugar, and seasoning, tapioca balls can be made to vary
in color and in texture. The most common, and original, tapioca balls are usually soaked in a
simple sugar syrup to give it a little bit of sweetness while keeping it fresh. They typically lack
flavor and gel easily, and are therefore often used as a thickening agent in foods like pudding.
The tapioca pearls are simply there to provide texture, the flavor really comes from the drink
itself.
 Cassava is a significant crop in the Philippines because of its many uses and increasing
demand from feed milling industries. However, the supply for this crop in the country, compared
to other ASEAN countries, remains low and relatively less competitive.

Malunggay or moringa oleifera is one of the most useful trees in the Philippines. Almost
all of the parts of this tree are used for food and traditional medicine. The moringa industry has
grown significantly for the past 10 years as researches are focused in this crop. It takes up to 8
months to grow after planting. It tolerates a wide range of soil conditions but prefers neutral and
slightly acidic, well drained sandy or loamy soil. It is easy to plant and grow.

Despite of having its general acceptability taste, cassava bears a natural toxin that could
harm consumers if taken above moderately. With an increasing demand of cassava and its
products, the researcher came up to the development of moringa pearls as a substitute for tapioca
pearls. These products would give the consumers a good health and can also contribute to
decrease the demand for cassava for tapioca pearl.

II. OBJECTIVES

This study aims to compare the tapioca pearls with the developed moringa pearls.
Specifically, it sought to answer the following questions:

1. Is there any significant difference in the properties of tapioca pearls and moringa pearls
which were prepared in similar ratios and conditions, in terms of:

A. Physicochemical Properties

a) Amylose content

b) Swelling power

c) Gelatinization temperature

d) Moisture content

e) Water activity

B. Microbiological Properties

a) Aerobic Plate Count (APC)

b) Coliform

c) Yeast and Mold

2. Is there any significant difference in the nutritional content of tapioca pearls and moringa
pearls which were prepare in similar ratios and conditions?
 3. Is there any significant difference in the sensory attributes of tapioca pearls and moringa
pearls which were prepared in similar ratios and conditions, in terms of:

a) Appearance

b) Aroma

c) Mouth feel

d) Taste

e) Over-all acceptability

III. METHODOLOGY
A. Materials
a. Raw materials
i. Moringa roots
ii. Cassava roots
iii. Brown sugar
iv. Distilled water
b. Extraction of Starch
i. Blender
ii. Knife
iii. Chopping board
iv. Basin
v. Cotton cloth
vi. Distilled water
vii. Oven
c. Production of Pearls
i. Pot
ii. Brown sugar
iii. Distilled water
iv. Stove
v. Ladle
vi. Bowl
d. Amylose Content
i. Moringa starch
ii. Analytical balance
iii. Spectrophotometer
iv. Iodine solution
v. Beaker
 vi. Distilled water
vii. Acetic acid
e. Swelling Power
i. Moringa starch
ii. Distilled water
iii. AgNO3
iv. Test tubes
v. Water bath
vi. Centrifuge
vii. Analytical balance
f. Gelatinization Temperature
i. Moringa Starch
ii. SDT 2960 DSC/TGA
iii. Aluminum pans
iv. Cahn C-30 microbalance
v. Lindberg/Blue gravity oven
g. Moisture Content
i. Analytical balance
ii. Oven
iii. Moringa pearls
h. Water Activity
i. Capacitance Sensors
ii. Moringa pearls
i. Aerobic Plate Count
i. Pipet
ii. Food homogenate (moringa starch)
iii. Beakers
iv. Test tubes
v. Petri dish
vi. Plate count agar
vii. Distilled water
viii. Moringa starch
j. Yeast and Mold
i. Peptone water broth
ii. Beaker
iii. Stirring rod
iv. Sucrose/glycerol
v. Dichloran Rose Bengal Chloramphenicol agar (DRBC)
 vi. Incubator
k. Sensory evaluation
i. Cooked moringa pearls
ii. Hedonic Scale Rating Sheets

B. Procedures

Gathering of Raw Materials

The moringa roots which are the main raw material will be harvested from a selected
moringa farm that allowed the researchers to gather roots. The cassava roots will be gathered in
the same manner. Brown sugar, water, and other raw materials will be gathered from the local
market.

Extraction of Starch

The starch from the moringa and cassava were extracted by the method utilized by Benesi
et al. (2004) called the wet method. Fresh tubers were washed, peeled, chopped into
approximately 1 cm cubes and then pulverized in a high-speed blender for 5 min. The pulp was
suspended in ten times its volume of water, stirred for 5 minutes and filtered using double fold
cotton cloth. The filtrate was allowed to stand for 2 hours for the starch to settle and the top
liquid was decanted and discarded. Water was added to the sediment and the mixture was stirred
again for 5 minutes. Filtration was repeated as before and the starch from the filtrate was allowed
to settle. Kaur, Ahluwalia and Singh (2016) suggested that after decanting the top liquid, the
sediment (starch) was oven dried for 1 hour at 55°C.

Production of Pearls

The method preparation and cooking of starch into pearls was set out by De Bradandere
(2019). Based from the instruction, moringa pearl and tapioca pearl will be made using the same
method and will be done separately.

First, brown sugar will be dissolved of boiling water. It will be continuously stirred until
it forms a syrup then it will be set aside to cool. The starch will be mixed with boiling water.
After mixing well, the dough will be kneaded with hands. Then, the dough will be rolled into
cylinders of small diameter and cut into smaller pieces. Each piece will be rolled into balls.

An enough amount of syrup prepared will be reheated to boil. The remaining syrup will
be used for storing the cooked pearls. The balls will be cooked to the boiling syrup. At a certain
time, the balls will float. It will take at around 20 minutes to cook the balls. The researcher will
carefully check the balls for doneness. Once cooked, the pearls will be placed to the cold
remaining syrup to cool.

PHYSICOCHEMICAL PROPERTIES

1. Amylose Content
In order to determine the amylose content of the moringa pearl, amylose-iodine
colorimetry will be employed. This will be performed using a spectrophotometer. To prepare the
amylose-iodine solution, 100mg of the moringa root starch will be added to 100mL of distilled
water. To 5mL of this solution, 1mL of 1M acetic acid and 2mL of 0.2% iodine solution will be
added. Linear amylose will produce a deep blue complex when it reacts with iodine. The
chromophore can be easily monitored by light absorption spectroscopy.

2. Swelling Power
The method used by Matsuki (1998) in his study, “Effect of Wheat Starch Structure on Swelling
Power,” to determine the swelling power of the moringa pearls. Swelling power will be
determined using 0.32 g of wholemeal or 0.16 g of isolated starch by the modified method of
McCormick et al (1991), in which 0.1% AgNO3 will be used instead of distilled water to inhibit
α-amylase activity (Yasui et al 1996). Wholemeal or isolated starch will be weighed into glass
tubes with coated screw caps to which 5 mL of 0.1% AgNO3 will be added. The tubes will be
placed in a shaking water bath at 70°C for 10 min and will be transferred into a boiling water
bath. After boiling for 10 min, the tubes will be cooled in cold water for 5 min and centrifuged at
1,700 × g for 4 min. The supernatant will be removed carefully and swelling power will be
determined as sediment weight (g/g).

3. Gelatinization Temperature

The methods employed by Bertrand et al. on their study, “Rapid Estimation of Parameters
for Gelatinization of Waxy Corn Starch will be used in this study to determine the gelatinization
temperature of the moringa root starch. According to the study, the experiments will be carried
out on a SDT 2960 DSC/TGA (Thermogravimetric analysis) (TA Instruments, New Castle, DE,
USA), which will be calibrated using sapphire calibrant at 40–150˚C. the TA Universal Analysis
software will be used to analyze the resulting Differential Scanning Calorimetry (DSC) traces
and curves. Hermetically sealed aluminum pans (4 × 6 mm) with flat bottoms will be used to
contain samples. A Cahn C-30 microbalance with a precision of 0.001 mg will be used to
measure starch and water, and a Lindberg/Blue gravity oven will be used to treat or cook the
samples in the gelatinization time study. All experimental runs were performed in triplicate, with
results reported as an average with an error bar equivalent to 1SD (one standard deviation).
Starch gelatinization is characterized by several endotherms which can be identified by DSC.
These endotherms appear as peaks on DSC curves and are dependent on water content and the
type of starch.

4. Moisture Content

Moisture content will be determined using the loss on drying method as provided by
Mermelstein (2009) in his article, “Measuring Moisture Content and Water Activity.” In this
method, a wet sample is weighed on a balance, placed in an oven, and heated until the end of the
drying period, i.e., until the sample reaches equilibrium. The weight loss is the moisture content
of the sample. In moisture balances, this process is sped up by superheating the sample by a
variety of means, including infrared, halogen, and microwaves.

5. Water Activity

Water activity will be measured using capacitance sensors. These sensors consist of two
charged plates separated by a hygroscopic polymer membrane. As the membrane adsorbs water,
its ability to hold a charge increase. The resulting capacitance change is approximately
proportional to the water activity as determined by a sensor calibrated against known saturated
salt standards. The sensor gives a signal relative to the ERH and thus the water activity (as
ERH/100). Commercially available instruments measure over the entire water activity range with
an accuracy of ±0.015 aw. this method was adapted from Mermelstein (2009) in his article,
“Measuring Moisture Content and Water Activity.”

MICROBIOLOGICAL PROPERTIES
1. Aerobic Plate Count (APC)
4. Aerobic Plate Count is used as an indicator of bacterial populations on a sample. It is also
called aerobic colony count, standard plate count, Mesophilic count or Total Plate Count. The
conventional plate count method will be used to determine the aerobic plate count of the moringa
pearls. Using separate sterile pipets, decimal dilutions of 10-2, 10-3, 10-4, and other as appropriate,
of the food homogenate (moringa pearl preparation) by transferring 10mL of the previous
dilution to 90mL of diluent. All the dilutions will be shook for 25 times in 30cm arc within 7s. 1
mL of each dilution will be pipetted into separate, duplicate, and properly labeled petri dishes.
The dilution bottle will be re-shook 25 times in 30cm arc within 7s if it stands more than 3
minutes before it is pipetted into the petri dish. 12-15mL of plate count agar (cooled to 45 ± 1°C)
will be added to each plate within 15 minutes of the original dilution. Since the sample diluent
contains hygroscopic materials in the form of moringa starch, the agar should be added
immediately to the petri dishes. Then, agar and dilution water control plates will be poured for
each series of samples. The sample dilutions and agar medium will immediately be mixed
thoroughly and uniformly by alternate rotation and back-and-forth motion of plates on flat level
surface. The agar will be left to solidify. Therafter, the solidified petri dishes will be inverted and
incubated promptly 48 ± 2 h at 35°C. Plates should not be stacked when pouring agar or when
agar is solidifying.
2. Yeast and Mold
To determine the yeast and mold count of the moringa pearls, conventional colony count
methods will be applied, based on the procedures employed by Maturin and Peeler (2001). An
initial suspension will be prepared by mixing the sample with a suitable diluent such as peptone
water broth. It may be beneficial to add a small quantity of a suitable surfactant to the diluent
(e.g. polysorbitan 80) to help break up clumps of mould spores and conidia. Since the product to
be tested is the dried version of the moringa pearls and will be tested for xerophilic moulds and
yeasts, 20-30% sucrose or glycerol will be added to the diluent to prevent osmotic shock. Surface
plating technique will be employed wherein a known quantity of the sample or the initial
suspension will be spread over the surface of a suitable selective agar medium. ISO 21527-1
recommends Dichloran Rose Bengal Chloramphenicol agar (DRBC), while the preferred
medium specified in 21527-2 is Dichloran 18% Glycerol agar (DG18). Other media used include
Oxytetracycline Glucose Yeast Extract agar (OGYE) and Yeast Extract Dextrose
Chloramphenicol agar (YGC). Selective agars for yeasts and moulds usually contain antibiotics
to help suppress bacterial growth. The plates will be incubated at 25˚C for 5 to 7 days and then
examined for the presence of yeast and mould colonies.

SENSORY EVALUATION
Sensory attributes are often used to evaluate the quality of food. These characteristics can
be the basis on the selection foods.

l. Appearance

The appearance of moringa pearls will be analyzed through visualization inspection and
characterized qualitatively. It can be evaluated by its color or the color temperature and the
nature of the structure f the product affect the appearance.
m. Aroma

Aromas greatly influence what we select to eat. By aroma you can easily think that that
food is pleasing to taste. Through smelling, the aroma of the moringa pearls will be evaluated.

n. Mouthfeel
 Mouthfeel can be generalized as taste of food, but it is different. Mouthfeel refers to the
physical sensations in the mouth caused by some food and drinks. Like for example; big, sweet,
tannic, chewy etc.

o. Taste

Taste on the other hand is a chemical sense due to taste stimuli falling on taste receptors
located on the tongue called taste buds.

p. Over- all acceptability

Over-all acceptability is the result of all sensory attributes mention above. It is the overall
grade or marking to a certain food.

Hedonic Scale

The Hedonic scale will be used to measure the sensory attributes of the moringa pearls,
specifically the appearance, aroma, mouthfeel, taste and overall acceptability.

Table 1. Hedonic Scale

9-Point Hedonic Scale

9 Extremely Like
8 Like Very Much
7 Like Moderately
6 Like Slightly
5 Neither Like or Dislike
4 Dislike slightly
3 Dislike Moderately
2 Dislike Very Much
1 Extremely Dislike
This table and its interpretation will be used to analyze the results obtained from the sensory
evaluation of the moringa pearls from 50 participants. This will measure the appearance, aroma,
taste, mouthfeel and overall acceptability of the pearls according to the participants’ personal
opinions base on their evaluation.

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