ABSTRACT
The proximate content; moisture, crude protein,
crude ash, crude fat, crude fibre and carbohydrate, of Simarouba glauca were determined, along with other properties to
include the glucose and amino acid contents, and the qualitative macromolecular
content of the plant. The proximate properties of the plant were determined
based on the A.O.A.C (1990) methods for moisture, crude protein and crude ash;
A.O.A.C (1984) for crude fibre and carbohydrates; and the acid base extraction
technique described by Phillips et.al. (2001).
The concentration of different amino acids were determined using the ninhydrin,
while the glucose content was determined Nelson-Simogy’s method. The moisture
contents of the
leaves and the roots were quite high, over 60%
in both. The root of the plant was also found to be rich in fibre and low in
fat, meanwhile the leaves recorded a higher fat concentration than fibre. With
the moisture and fibre content of the roots taking a large percentage, it was
found to be pretty low in crude ash, carbohydrates, crude protein and crude
fat. The leafs however was much higher in ash, carbohydrates and protein, and a
little higher in fat. Among the essestial amino acids, methionine was the
highest while phenyl alanine was negative. The highest amino acid was cysteine,
which is a conditionally essential amino acid. From these results, it is
obvious that Simarouba glauca roots
and leaf extract might not enough nutritional supplements while being used
medicinally. They can serve as a good source of animal feed, however, proteins
and fat would need to be supplemented.
TABLE OF CONTENTS
TITLE PAGE - - - - - - - - - ii
CERTIFICATION - - - - - - - - - iii
DEDICATION - - - - - - - - - iv
ACKNOWLEDGEMENT - - - - - - - v
TABLE OF CONTENTS -
- - - - - - -
vi
ABSTRACT - - - - - - - - - viii
CHAPTER
ONE
1.1 INTRODUCTION - - - - - - - 1
1.2 MOISTURE CONTENT - - - - - - 2
1.3 CARBOHYDRATES - - - - - - - 3
1.4 PROTEINS - - - - - - - - - 7
1.5 CRUDE ASH - - - - - - - - - 8
1.6 FAT AND FIBRE - - - - - - - - 9
1.7 AIM OF THE RESEARCH WORK - - - - - 9
CHAPTER
TWO
2.1 LITERATURE REVIEW - - - - - - - 10
2.2 PHYSICAL QUALITIES OF SIMAROUBA GLAUCA: - - 11
2.3 MEDICINAL IMPORTANCE OF SIMAROUBA GLAUCA: - - 12
2.4 BIOFUEL PRODUCTION: - - - - - - - 15
2.5 CONSTITUENTS OF SIMAROUBA SEEDS: - - - - 16
CHAPTER
THREE
MATERIALS AND METHODOLOGY - - - - - 20
3.1 MATERIALS - - - - -
- - - - 20
3.1.1 PLANT - -
- -
- - - - - - 20
3.2 CHEMICALS/REAGENTS - - - - - - - 20
3.3 APPARATUS - - - - - - - - - 21
3.4 EQUIPMENTS - - - - - - - - - 22
3.5 METHODS - - - - - - - - - 22
3.5.1 DETERMINATION OF MOISTURE CONTENT - - -
23
3.5.2 QUANTITATIVE DETERMINATION OF PROTEIN
CONTENT - 24
3.5.3 QUANTITATTIVE DETERMINATION OF ASH
CONTENT - 27
3.5.4 DETERMINATION OF CRUDE FIBRE CONTENT
- - - 28
3.5.5 DETERMINATION OF CRUDE FAT CONTENT - - - 28
3.5.6
QUANTITATTIVE DETERMINATION OF CARBOHYDRATE CONTENT- - - - - - - - - -
29
3.5.7
QUANTITATTIVE DETERMINATION SOME MACROMOLECULES - - - - - - - - - - - - 30
3.5.8 QUALITATIVE DETERMINATION OF PROTEIN
CONTENT - 32
3.5.9 QUANTITATIVE DETERMINATION OF GLUCOSE
CONTENT - 33
3.5.10 QUALITATIVE DETERMINATION OF SOME AMINO
ACIDS (NINHYDRIN METHOD) - - - - - - - - 24
CHAPTER
FOUR
4.1 PROXIMATE
ANALYSIS ON SIMAROUBA LEAF
AND ROOT
EXTRACT - - - - - - - 35
4.2
QUALITATIVE TEST FOR MACROMOLECULES IN LEAVES AND ROOTS - - - - - - - - - -
35
4.3
RESULT FOR QUANTITATIVE ANALYSIS OF GLUCOSE IN SIMAROUBA LEAVES AND ROOT
- - - - - 37
CHAPTER
FIVE
5.1 DISCUSSION - - - - - - - - 38
5.1.1
MOISTURE CONTENT - - - - - -
38
5.1.2
CRUDE PROTEIN AND AMINO ACIDS - - - - 38
5.1.3
CRUDE ASH - - - - - - - - 39
5.1.4
CARBOHYDRATE, GLUCOSE AND CRUDE FIBRE - - 40
5.2 CONCLUSSION - - - - - - - - 41
REFERENCES - - - - - - - - 42
APPENDIX
1 - - - - - - - - 45
CHAPTER ONE
1.1 INTRODUCTION
Plants undergo
photosynthesis and they constitute a primary resource of carbon, vitamins,
minerals, protein, essential fatty acids, and utilizable energy for food
production (Young and Pelett, 1994). Plants have played a significant role in
maintaining the health and improving the quality of human life for thousands of
years. (Mishra, 2010). They provide a
major source of food and nourishment for man and animal.
Nutrition is a science of food and its
relationship to health. Nutrition refers to nourishment that sustains life. The
study of nutrient requirements and the diet providing these requirements is
also known as ‘nutrition’ (Chutani, 2008). Pike and Brown, 1984 defined it as
“the science that interprets the relationship of food to the functioning of
living organism. It includes the uptake of food, liberation of energy,
elimination of wastes and all the processes of synthesis essential for
maintenance, growth and reproduction (Chutani, 2008). Apart from maintaining normal body
functioning, nutrition is important in fighting infections and in the
recuperation of an ill person. Nutrition interacts with infections in a
synergistic manner, such that recurrent infections lead to a loss of body
nitrogen and worsen nutritional status; the resulting malnutrition, in its
turn, produces a greater susceptibility to infection (Kurpad, 2005). Aristotle
(384-322 B.C.) was the first to suggest that the composition of foods in the
normal diet might contribute to health.
In an 1897 literature
on metabolic investigations, Atwater divided food composition into five
classes; protein, fat carbohydrate, energy and water. However, today, proximate
composition is the term usually used to describe six components of food namely;
moisture, crude protein, crude ash, crude fibre, crude fat and carbohydrate
(nitrogen free extract) which are all expressed in percentage (%) or gram per
100 grams (g/100g). The study of proximate analysis on foods was devised over a
hundred years ago by two German scientists, Henneberg and Stohmann, and even
though new techniques have been introduced, their system of proximate still
forms the basis for the statutory declaration of the composition of foods.
(Dublecz, 2011).
1.2 MOISTURE CONTENT
Water is essential for
every living organism. In the human body, water content ranges from 50-70% in
different tissues. It is present in different fluid compartments of the human
body- Intracellular (fluid inside the cells) and extra cellular. Plasma,
interstitial fluid, cerebrospinal fluid, ocular fluid, lymph, peritoneal,
pericardial, pleural and synovial fluids are part of the extra cellular fluid
(Chutani, 2008). However, the moisture
content of a feed is seldom of interest nutritionally as water is usually taken
on its own.
The active ingredients
from the view of feed nutrition are present in the part of dry matter (solid
matter); therefore the level of moisture content is an important factor in both
economy and storage. At high temperature
and humidity the risk of putrefaction is
predicted due to the proliferation of molds, etc., or self-digestion by enzymes
in the feed when moisture in the feed is not less than about 15%. As the assay for moisture in the feed
measures loss on drying by heating at normal pressure as moisture, the result
includes most of volatile substances other than H2O. Therefore, it may be more appropriate to be
referred to as volatile matter rather than moisture for accuracy. Organic acids such as acetic acid and butyric
acid in silage as well as ammonia and flavor components in feed materials are
also vaporized and thus measured as moisture.
Because the content of these in most feed is extremely low, there has
hardly been a need to consider their influence on the measured value. (Chutani,
2008).
1.3 CARBOHYDRATES
Photosynthesis is a
process used by plants and other organisms to convert light energy from the
sun, into chemical energy that can be later released to fuel the organisms'
activities. The light energy harnessed from the sun drives the reduction of
carbon from CO2 to produce O2 and fixed carbon in form of
carbohydrate.
Early in the twentieth
century, it was mistakenly thought that light absorbed by photosynthetic
pigments directly reduced CO2 which then combined with water to form
carbohydrate. In fact, photosynthesis in plants is a two stage process in which
light energy is harnessed to oxidise H2O:
2H2O → O2 + 4 [H+]
The electrons thereby
obtained subsequently reduce CO2:
4H+
+ CO2 → (CH2O)n + H2O
The two stages of
photosynthesis are traditionally referred to as the light reactions and the
dark reactions:
1. In the light reactions, specialised pigment
molecules capture light energy and are thereby oxidized. A series of electron-
transfer reactions which culminate with the reduction of NADP+ to
NADPH, generate ATP from ADP + Pi. The oxidised pigment molecules are reduced
to H2O, thereby generating O2.
2. The dark reactions use NADPH and ATP to reduce
CO2 and incorporate it into the three-carbon precursors of
carbohydrate.
The light reactions
takes place in the thylakoid membrane of chloroplasts in leaves and green parts
of plants. The inside of the thylakoid
is referred to as the lumen. The light reactions are catalysed by enzymes
located in the thylakoid membrane, whereas the dark reactions take place in the
stroma. The principal photoreceptor of light is chlorophyll. These chlorophyll
molecules do not participate directly in photochemical reactions but function
to act as light harvesting antennas. The absorbed photons are transferred from
molecule to molecule until it reaches the photosynthetic reaction centre.
In the respiratory
chain, electrons flow from NADH + H+ to O2, with the
production of water and energy. However in photosynthesis, electrons are taken
up from water and transferred to NADP+, with an expenditure of
energy. Photosynthetic electron transport is therefore energetically “uphill
work.” To make this possible, the transport is stimulated at two points by the
absorption of light energy. This occurs through two photo systems protein
complexes that contain large numbers of chlorophyll molecules and other
pigments Another component of the transport chain is the cytochrome bf
complex, an aggregate of integral membrane proteins that includes two
cytochromes (b563 and f). Plastoquinone, which is comparable to ubiquinone, and
two soluble proteins, the copper containing plastocyanin and ferredoxin,
function as mobile electron carriers. At the end of the chain, there is an
enzyme that transfers the electrons to NADP+. Because photosystem II
and the cytochrome b/f complex release protons from reduced plastoquinone into
the lumen, photosynthetic electron transport establishes an electrochemical
gradient across the thylakoid membrane, which is used for ATP synthesis by an
ATP synthase.
ATP and NADPH + H+,
which are both needed for the dark reactions, are formed in the stroma. (Voet et al., 2013).
1.3aCalvin cycle
The actual CO2 fixation i.e., the
incorporation of CO2 into an organic compound is catalysed by
ribulosebisphosphate carboxylase/oxygenase (“rubisco”). Rubisco, the most
abundant enzyme on Earth, converts ribulose 1,5-bis-phosphate, CO2
and water into two molecules of 3-phosphoglycerate. These are then converted,
via 1,3-bisphosphoglycerate and 3-phosphoglycerate, into glyceraldehyde
3-phosphate. In this way, 1,2-glyceraldehyde 3-phosphates are synthesized from
six CO2. Two molecules of this intermediate are used by
gluconeogenesis reactions to synthesize glucose 6-phosphate. From the remaining
10 molecules, six molecules of ribulose-1,5-bisphosphate are regenerated, and
the cycle then starts over again. In the Calvin cycle, ATP is required for
phosphorylation of 3-phosphoglycerate and ribulose-5-phosphate. NADPH + H+,
the second product of the light reaction, is consumed in the reduction of
1,3-bisphosphoglycerate to glyceraldehyde-3- phosphate.
Carbohydrates are the most abundant
biomolecules produced on earth; photosynthetic plants and algae convert over
100 billion metric tons of CO2 and water into sugars, starches, and
cellulose like substance. Carbohydrates supply energy for the human body to
function. They are the most abundant bulk nutrients and form the major source
of biological energy through their oxidation in the tissues. They also furnish
organic precursors for the biosynthesis of many cell components. Carbohydrates
are not essential in the human diet, but because carbohydrate rich foods are
abundant and cheap, compared with fats and protein, they naturally form a major
part of the diet in most of the world. (Voet et al., 2013).
1.4 PROTEINS
The requirements for total protein, at various
stages during the life cycle of humans, were reviewed and evaluated by a joint
panel of the Food and Agriculture Organization, the World Health Organization,
and the United Nations University (FAO/ WHO/UNO).
The requirement for dietary protein consists of
two components:
1) the
requirement for the nutritionally indispensable amino acids (histidine,
isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan,
and valine) under all conditions and for conditionally indispensable amino
acids (cysteine, tyrosine, taurine, glycine, arginine, glutamine, proline)
under specific physiological and pathological conditions and
2) the requirement for nonspecific nitrogen for
the synthesis of the nutritionally dispensable amino acids (aspartic acid, asparagine,
glutamic acid, alanine, serine) and other physiologically important
nitrogen-containing compounds such as nucleic acids, creatine, and porphyrins.
With respect to the first component, it is
usually accepted that the nutritive values of various food protein sources are
to a large extent determined by the concentration and availability of the
individual indispensable amino acids. Hence, the efficiency with which a given
source of food protein is utilized in support of an adequate state of
nutritional health depends both on the physiological requirements for the
indispensable amino acids and total nitrogen and on the concentration of
specific amino acids in the source of interest (Young and Pelett, 1994).
Proteins, are synthesized from a complex
series of steps which involves the transcription of DNA already present in each
cell of an organism, and its consequent transcription into a polypeptide chain.
This chain is modified by other inherent mechanisms in the cell to yield
protein.
1.5 CRUDE ASH
Ash is the inorganic
residue remaining after the water and organic matter have been removed by
heating in the presence of oxidizing agents, which provides a measure of the
total amount of minerals within a food. The ash content is a measure of the total amount of minerals present
within a food. Minerals are required for many purposes like forming the frame
and rigid structure of the body, as part of the body/cell fluids and for number
of cellular and sub cellular physiological functions (Chutani, 2008). The mineral
content includes specific inorganic components present within a food,
such as Ca, Na, K and Cl. Determination of the ash and mineral content of foods
is important for a number of reasons. The most important reason in regards a
plant like S. glauca is the
nutritional importance. Some minerals are essential to a healthy diet (e.g.
calcium, phosphorous, potassium and
sodium) whereas others can be toxic (e.g. lead, mercury, cadmium and
aluminum).
1.6 FAT AND FIBRE
The importance of fat
and fibre in nutrition cannot be underestimated. Crude fat contains fat, complex lipid, sterols, fatty
acids and fat soluble dyes; while crude fibre contains cellulose,
hemicellulose, and lignin.
1.7 AIM OF THE RESEARCH WORK
This study is designed to screen the proximate
constituents of the leaf extracts of Simarouba
glauca which includes moisture, protein, carbohydrates, ash fibre and lipid
content of Simarouba glauca. In
addition to this, to determine the quantitative carbohydrate and amino acid
constituents of Simarouba glauca.
Thus, the nutritional value of Simarouba roots and leaves with some emphasis on
their possible use both as a medicinal and nutritional food for the sick or
convalescent.
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