OPTIMIZATION OF RICE STARCH HYDROLYSIS

FOR THE PRODUCTION OF RICE MILK USING PIGMENTED

RICE (Oryza sativa) IN RETORT PACKAGING

A Thesis Proposal
Presented to Thesis Committee and Faculty of
University of Science and Technology of Southern Philippines
Department of Food Science and Technology
College of Science and Mathematics
Cagayan de Oro City

In Partial Fulfilment of the Requirements for the

Degree of Bachelor of Science in
Food Technology

Presented by

Ampo, Rowie Bagayna

Ancog, Aldrian Dela Victoria

Remotin, Jennilyn

NOVEMBER 2019

Chapter 1
 INTRODUCTION

Background of the study

Rice (Oryza sativa)have been considered as a staple

food especially in the east and southeast Asian countries
which catered almost half of the world population and 95
percent of the yielded rice is being consumed by humans
(Britannica, 2019). It is also the second most important
cereal grain consumed worldwide after corn and provides a
major source of calories for the consumers in the
Philippines. Rice plays a vital role in the lives of every
Filipinos. In 2017, Philippine production of rice was 19.3
million metric tons, to feed the growing population, the
Philippines need to import 3 million metric tons this year
and is considered as the World’s biggest rice importer,
(Simeon, 2019). Despite of this criticization Dr. William
Padolina, former deputy director general for operations at
the International Rice Research Institute (IRRI), said "The
Philippines has enthusiastically taken up rice science
technologies that have helped farmers dramatically increase
their yields."

Rice milk is a plant-based milk used to be a substitute

for Dairy milk for a lactose free milk and also a vegan’s
milk for health-conscious consumers, (Sethi, Tyagi, &
Anurag, 2016). This research focuses on the development of
the rice milk formulation as well as the process techniques
to optimize the sensory characteristics as well as the
nutritional value of the rice milk. In order to meet the
consumers changing needs, food product development have been
an essential leap in meeting the demands of the consumers by
creating healthier alternatives to current food
trends(Vanga, 2018). Traditionally, whole kernels of white
and brown rice were used in the production of rice milk.
However, due to the growing demand of consumers for
healthier options, pigmented rice can be utilized in rice
milk production which are believed to have the potential for
health benefits due to their phenolic compounds which has
antioxidant characteristic.
 Starch is a key component in rice milk, the rheological
property or consistency of rice milk is greatly dependent on
the properties of rice starch(Vanga, 2018). Starch is
composed of amylose and amylopectin but their value and
ratio are entirely dependent on the variety of rice which
greatly affect the functional properties of starch. This
will also show different physicochemical properties in each
variant and this research aimed to single out the most
preferred by the consumer.

Conceptual Framework

This part shows the schematic presentation of the

conceptual framework of the study.

Phase 1
Dependent

Variables
 Independent
Variables

Pigmented Rice
Test Method
- Roasting Time
- 5 minutes Total Soluble Solids
- 25 minutes Optimized process
Response: Highest
o
- Soaking Ratio Brix
(rice:water)
- 1:1
- 1:5

- Soaking Time
- 3 hours
- 15 hours

Statistical
Analysis: Face
centered central
composite design

Figure 1.1 Schematic Presentation between independent and

dependent variables in phase 1.

Phase 2

Independent Dependent
Variables
Variables
Sensory Analysis
Factors
Response:Acceptabi Most Accepted
-Optimized lity test Formulation
process of Rice -Sensory
Attributes
Sugar -Color
-3%-5% -Appearance
-Odor
Statistical
-Texture
Design: Simplex
-Taste
Lattice Design
-Sweetness
-Overall
Acceptability
Figure 1.2 Schematic presentation between the independent
and dependent variable in phase 2.

Phase 3
Independent Dependent
Variables
Variables
Commercial Sterility
Rice milk with
Test
the most
accepted Retort
Nutritional Facts
formulation analysis

Physico-chemical test

-pH determination
-Water activity
-Colour
determination
-Rheological Test
-Total soluble
solids

Acceptability test
-Sensory Evaluation
Figure 1.3 Schematic presentation between the independent
and dependent variable in phase 3.

Statement of the Problem

This study aims to improve the processing parameters

and formulation of pigmented rice milk

Specifically, this study is required to answer the following

questions:

1. What is the optimized processing parameter to hydrolyze

starch in the production of rice milk?

2. What is the optimized formulation of rice milk with

additional sweetener?

3. What are the physico-chemical and nutritional

properties of pigmented rice milk?

Significance of the study

Rice milk has paved the way to solve many sorts of

common problems. As a non-dairy source of milk, it can be
recommended to lactose intolerant consumers. It is also free
of nuts, and beans so its application could be significant
to the masses where in such allergies is common. In terms of
its Nutritional aspect, Rice milk is high in calories which
came from its rich carbohydrates and also its sugar content.
Pigmented rice is particularly rich in anthocyanins, a group
of flavonoid plant pigments that have powerful antioxidant
and anti-inflammatory properties (He & Giusti, 2010).
Flavonoids can help decrease inflammation in your body, keep
free radical levels in check, and may reduce your risk of
chronic conditions, such as heart disease and type 2
diabetes. Entrepreneurs will also benefit from this study by
setting an idea to add value to rice by product development.
Farmers could also benefit from this by growing more
pigmented rice and also providing them the knowledge to make
this kind of technology could also benefit them by adding
extra income and increasing value added price for pigmented
rice.

Scope and Limitation

This study will only use one type of pigmented rice as

a substitute for the existing white and brown rice used in
the market and to optimize its starch hydrolysis. The
researchers will be purchasing the pigmented rice at the
local supplier located in Balingasag where the pigmented
rice is available. This application will only be limited to
Rice Milk alone.

Definition of terms

1. Rice
Rice (Oryza sativa) is the most important cereal crop
in the developing countries and is known to be a staple
food of over half the world's population. It is
generally considered a semi-aquatic annual grass
plant(Juliano, 1993). It is of special importance for
the nutrition of large reaches of the population in
Asia parts of Latin
America and the Caribbean and, increasingly so, in
Africa. As a result, it plays a pivotal role for the
food security of over half the world population. It is
also a central component of the culture of a number of
Communities (Calpe, 2006).

2. Plant-based Milk
Plant-based Milk is a liquid extracted from plants that
have similar characteristics of an animal’s milk. Nuts,
seeds, soy or grains like rice are the most popular in
plant-based milk production. it is now the trend in the
market especially to consumers who are changing
lifestyle’s into a vegan which focuses on what’s better
for the environment and healthier options or consumers
that are lactose-intolerant(Robbins, 2019).

3. Rice milk
 Rice Milk is a plant-based milk or non-dairy milk. It
is usually made from milled rice and water. Despite the
fact that it can be a substitute for dairy milk, it
lacks the property of a dairy milk especially in the
nutrient level where it lacks lactose which are
beneficial to consumers that are lactose intolerance.
The energy that can be gain from rice milk is much
similar to dairy milk due to its high carbohydrate
source and there are other benefits that can be gain
from drinking rice milk(Krans, 2019).

4. Starch
Starch is a chief storage constituent of rice grains
and made up with macromolecules of amylose and
amylopectin; the ratio of amylose and amylopectin in
the starch granule governs the physico-chemical
properties including thermal, texture, rheological and
cooking properties of rice(Koteswara Reddy, Kimi, &
Haripriya, 2016). It is a type of carbohydrate, also
referred to as a complex carbohydrate since it is made
up of long chains of sugar molecules called
polysaccharide.

5. Emulsion
Based on Anne Marie Helmenstine, (2019) Definition of
emulsion “An emulsion is a colloid of two or more
immiscible liquids where one liquid contains a
dispersion of the other liquids. In other words, an
emulsion is a special type of mixture made by combining
two liquids that normally don't mix. The word emulsion
comes from the Latin word meaning "to milk" (milk is
one example of an emulsion of fat and water). The
process of turning a liquid mixture into an emulsion is
called emulsification.” One of the challenges of Rice
milk is that particles settle in the bottom giving the
appearance of separating liquids this is due to high
starch content that gives poor emulsion stability(Sethi
et al., 2016).
 Chapter 2

Review of related Literature

Rice

Rice is an annual grass plant in the Poaceae family. It

is one of the most common food crops throughout the world
and the most common food crop in Asia, Africa including in
developing countries. About 65% of the population on the
country of China, they consider rice as one of the most
important staple part of their diet (Yang et al, 2018). The
rice word comes from in two Latin names: Oryza sativa for
Asian rice and Oryza glaberrima for African rice. Most rice
plant comes in two varieties the Oryza sativa indica and
Oryza sativa japonica. The variety of japonica is grown in
the countries with low temperature and high relative
humidity like Japan, China and Korea. While indica variety
grown both in tropical and subtropical countries throughout
South-East Asia, and Southern China (Agropedia?). White rice
is the most common rice consumed by humans and the rest are
pigmented rice (3 Deng, Xu et al, 2011). Pigmented rice
comprises of many color mainly black, red, purple rice and
black purple grains. It has been reported that this type of
rice contains anthocyanin compounds which belong flavonoids
family. Anthocyanin is naturally occurring pigments and are
water soluble. They are responsible for the colors purple,
red and blue. They can be found in numbers of food like in
the purplish blue-colored leafy vegetables, grains and
berries. Anthocyanin found in pigmented rice are the
cyanidin-3-glucoside and peonidin-3-glucoside, pelargonidin-
3, 5-diglucoside, cyanidin-3-glucosid, cyanidin-3, 5-
diglucoside, cyanidin-3-glucoside and pelargonidin-3-
glucoside (Yodmanee, 2011). Pigmented rice was extensively
consumed in countries of Japan and China due to its high in
polyphenols and anthocyanin contents. Consuming pigmented
rice found to have a good effect in human diet. It contains
bioactive compounds including anthocyanin and its high with
antioxidant which may prevent from chronic diseases caused
by oxidative damage to cells (Nam Chio et al, 2004). It is
also reported by (Murdifin et al, 2015) that pigmented rice
has gammaoryzanol, phytic acid, vitamins, minerals amino
acids. It also has anti-lipid peroxide, anti-inflamatory
and anti-hepatic steatosis. In addition, pigmented rice also
contains carotenoid which contain lutein, zeaxanthin,
lycopene and beta-carotene which can be found in the black-
purple rice (6 Caro, Watanabe et al, 2013). However the
composition of the nutrients available in the rice grain
depends on how it was cultivated (Seechamnan , Karilla,
2018).

Pigmented Rice

The pigmented rice is mostly dark, red, and dim purple

rice, and contains an assortment of flavones, tannin,
phenolics, sterols, tocopherols, γ-oryzanols, amino acids,
and basic oils. Anthocyanins are thought as major useful
parts of pigmented rice. A few anthocyanins have been
secluded and recognized from the pigmented rice, including
cyanidin 3-glucoside, cyanidin 3-galactoside, cyanidin 3-
rutinoside, cyanidin 3,5-diglucoside, malvidin 3-
galactoside, peonidin 3-glucoside, and pelargonidin 3,5-
diglucoside. The pigmented rice has been expended in China,
Japan, and Korea for quite a while. It has been utilized for
reinforcing kidney work, treating paleness, advancing blood
dissemination, expelling blood balance, treating diabetes,
and improving sight in conventional Chinese drug. The
concentrates from pigmented rice are utilized as
characteristic nourishment colorants in bread, frozen
yogurt, and alcohol just as practical nourishment (Deng
et.al, 2012). According to Chonbuk National University,
Jeonju (2013) pigmented rice brans were extracted by using
an aqueous mixture of acetone, ethanol, and methanol to
determine the most effective extraction solvent. The result
indicated pigmented rice bran might be used as natural
antioxidant.

Health Benefits of Pigmented rice

Since quite a while ago grained white rice is

generally a decent wellspring of vitality, sugars, calcium,
iron, thiamine, pantothenic corrosive, folate and vitamin E,
contrasted with maize, wheat and potatoes. It contains no
vitamin C, vitamin A, beta-carotene, or lutein and
zeaxanthin, and is prominently low in fiber. Dark colored
rice holds the wheat layer (containing numerous nutrients
and minerals just as fiber), as this has not been finished
to create white rice. Red rice is known to be wealthy in
iron and zinc, while dark and purple rice are particularly
high in protein, fat, unrefined fiber. Red, dark and purple
rice get their shading from anthocyanin colors, which are
known to have free radical searching and cancer prevention
agents limits, just as medical advantages.

Antioxidants

Pigmented rice’s color is created by a flavonoid called

anthocyanin pigment. This same pigment gives blueberries,
eggplants, and other healthy fruits and vegetables their
deep color. Anthocyanins are phytochemicals found in plants,
(He & Giusti, 2010).
 They may also have anti-inflammatory and anti-
carcinogenic properties. A powerful antioxidant, anthocyanin
has been linked to reducing cases of diabetes, obesity, and
heart disease. One study Conducted by (Summart & Chewonarin,
2014)linked pigmented rice to cancer prevention in rats.

Table 1 shows the Nutritional value of Rice milk per 100g

(“FoodData Central,” 2009).

Name Amount Unit

Water 89.28 g Lycopene 0 µg
Energy 47 kcal Lutein + zeaxanthin 0 µg
Protein 0.28 g Vitamin E (alpha-tocopherol) 0.4 mg
Total lipid (fat) 0.97 g 7
Carbohydrate, by difference 9.17 g Vitamin E, added 0.1 mg
Fiber, total dietary 0.3 g 3
Sugars, total including NLEA 5.28 g Vitamin D (D2 + D3) 1 µg
Calcium, Ca 118 mg Vitamin K (phylloquinone) 0.2 µg
Iron, Fe 0.2 mg Fatty acids, total saturated 0 g
Magnesium, Mg 11 mg
Phosphorus, P 56 mg
Potassium, K 27 mg
Sodium, Na 39 mg
Zinc, Zn 0.13 mg
Copper, Cu 0.037 mg
Selenium, Se 2.2 µg
Vitamin C, total ascorbic acid 0 mg
Thiamin 0.027 mg
Riboflavin 0.142 mg
Niacin 0.39 mg
Vitamin B-6 0.039 mg
Folate, total 2 µg
Folic acid 0 µg
Folate, food 2 µg
Folate, DFE 2 µg
Choline, total 2.1 mg
Vitamin B-12 0.63 µg
Vitamin B-12, added 0.63 µg
Vitamin A, RAE 63 µg
Retinol 63 µg
Carotene, beta 0 µg
Carotene, alpha 0 µg
Cryptoxanthin, beta 0 µg
Plant-based Milk

Plant based milk is also called as non-dairy milk. It

is an alternative milk for those people who are lactose-
intolerant individuals or have difficulty in digesting
lactose that is present in the cow’s milk. Non-dairy milk
also provides similar nutrients that can be found in dairy
milks. (Rai. et al, 2018) reported that the undigested
lactose will result to gastrointestinal symptoms including
diarrhea, bloating, nausea and distention. Almond milk,
coconut milk, oats milk, rice milk and soy milk are examples
of plant-based milk. Soy milk and rice milk are the most
common plant-based milk but due to allergen presents in soy,
rice milk serves as the most common alternative for dairy
milk. According to Latifha and Warganagegara (2018), rice
milk that was being prepared from brown rice using brown
algae as emulsifier, in every 100 mL of brown rice milk it
contains 18.0 mg and 13.1 mg fiber.

Compared the cow’s milk, almond milk, soy milk, rice

milk and coconut milk as milk alternatives in terms of their
nutritional characteristics(Krans, 2019). The Table 2 shows
the comparison in each milk.

Table2. Milk and milk alternatives: Nutrition comparison

per 8 fluid ounces (Krans, 2019)

Carbohydrates Sugar Fat

Calories Protein
(total) s (total)
 Cow's milk (whole) 150 12 g 12 g 8g 8g

Cow's milk (1%) 110 12 g 12 g 2g 8g

Cow's milk (skim) 80 12 g 12 g 0g 8g

Almond milk
40 1g 0g 3g 2g
(unsweetened)

Soy milk (unsweetened) 80 4g 1g 4g 7g

Rice milk (unsweetened) 120 22 g 10 g 2g 0g

Coconut milk beverage

50 2g 0g 5g 0g
(unsweetened)

Protein and Fiber content of Pigmented Rice

Sticky pigmented rice is a whole grain, meaning the

outer bran layer is intact. This makes it high in fiber, as
well as slightly nutty in flavor. Fiber is important for
regular bowel movements and overall bowel health. Fiber may
also help with losing weight and lowering cholesterol and
blood pressure(Whelan, 2019).

Pigmented rice is a good source of protein, making it

an excellent addition to a vegetarian diet. Protein helps
reduce muscle loss by helping the body build and repair
muscle tissue. It also helps with cell growth and keeps
bones strong.

Pigmented rice is a significant source of iron. Iron is

a mineral that is necessary for helping to make red blood
cells and transport oxygen. It also supports the
transmission of nerve impulses, which control body
movements. Without enough iron, anemia can result,(Whelan,
2019).
Hydrolysis of Rice Starch

The hydrolysis of starch to low molecular weight

products catalyzed by an α-amylase is one of the most
important roles of commercial enzyme process. Hydrolyzed
products are widely used for food, paper, textile industry.
The degree of hydrolysis is expressed as dextrose equivalent
(DE) which the percentage of reducing sugar calculated as
dextrose on dry weight basis (Bucke, 1999). According to
Ann-Charlotte Ellison (2004) an important element in process
of hydrolysis is to maintain the appropriate balance between
the amount of enzyme, temperature and incubation time.
 Chapter 3

Methodology

Research Design

The study conducted is an experimental type of

research. This study is sought to determine the level of
acceptability and stability of the pigmented rice milk in
retort packaging.

Research Setting

The development of formulation was conducted at

Northern Mindanao Food Innovation Center (NMFIC). The
physico-chemical analysis was conducted at Food Science and
Technology Laboratory. Both of NMFIC and Food Science and
Technology Laboratory are to be found inside the University
of Science and Technology of Southern Philippines (USTP)
Cagayan de Oro Campus. The pigmented rice was purchased from
a supplier residing in Balingasag, Misamis Oriental.

Sampling Scheme

The Pigmented Rice Samples that were used in this study

were obtain and delivered to the Northern Mindanao Food
Innovation Center in a good condition packaging material.
Prior to the preparation of the ingredients, the working
area and the materials used were cleaned and sanitized. The
pigmented rice was subjected to washing, drying, roasting,
soaking, blending and filtering. The blended pigmented rice
was subjected to TSS determination. The optimize rice was
then followed by addition of sugar. The different sample
formulation has been subjected to Sensory Evaluation in
order to identify the most preferred formulation; undergone
commercial sterility testing. The preparation and sensory
evaluation sessions were conducted at Science Complex
Building at USTP.
Sample Preparation

One kilogram of pigmented rice was obtained from a

local supplier from Balingasag and was transported to
Northern Mindanao Food Innovation Center (NMFIC). The
pigmented rice was washed properly by soaking the pigmented
rice with water in a ratio of 1:3 at room temperature (25 oC)
with occasional stirring for 5 minutes to remove unwanted
dirt, dust, and contaminants and was repeated three times.
After, the pigmented rice was washed then filtered using a
strainer. Then the strained pigmented rice was subjected to
drying using the Dryer (Memmert UF750) with a temperature of
60oC until the moisture content wet basis reach 13%.

1 kg of pigmented Rice

Wash and soak 1:3 for 5 minutes

Repeat wash and soak three times

Filter using strainer

Dry in the convection dryer (60⁰C) until moisture

content reach 13%
Phase 1: Process Optimization

Process

The dried pigmented rice was subjected to roasting in

low to medium heat (100-120oC) for about 5-25 minutes with
constant stirring. The fragrance immitted from the roasted
rice would give the end process. After roasting, the roasted
pigmented rice was then soaked in water for 3-15 hours and
with a ratio of 1:1,1:2 and 1:3. After which, the soaked
pigmented rice was blended for 10 minutes then filtered
using a nut filter bag. The samples are then subjected to
TSS determination.

Roast in low to medium heat (100-120⁰C) for 5-25 minutes

with constant stirring

Soak in water for 3-15 hours with the ratio of 1:1,

1:2, and 1:3

Blend for 10 minutes

Filter using a nut filter bag

TSS determination
Measurement of Total Soluble Solids (TSS)

Total soluble solids (TSS) indicate the thickness of

sample. % TSS of sample was determined using a digital brix
refractometer (Model: MA871; Make: USA), having a range of
0- 85% according to the methods proposed by Ranganna (1991).
Before measurement of TSS of sample, the refractometer was
calibrated using double distilled water. A drop of the rice
milk was placed on the sample slot of refractometer and the
TSS of the sample was recorded and expressed in ˚Brix.

Statistical Analysis

Optimization condition for rice milk process was

determined with the help of commercial software (Design
Expert Version 7.0.0) to obtain optimum sample with maximum
TSS value using face-centered central composite design.

Phase 2: Formulation

Using the simplex lattice design with 2 factors, rice

milk and sugar, formulation will be determined. These run
samples will be evaluated by 9 point Hedonic Scale reading.

Phase 3: Profiling

Formulated rice milk will undergo thermal processing

through water retort to create a shelf stable rice milk.
Final product will be subjected to commercial sterility
test, nutritional facts analysis and sensory evaluation for
acceptability.
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