THE EFFECT OF ALLIUM CEPA EXTRACT ON

CARDIOMETABOLIC SYNDROME

BY

Anietie Daniel Jeremiah

(BMS1701994)

A PROJECT SUBMITTED TO THE DEPARTMENT OF MEDICAL

BIOCHEMISTRY, SCHOOL OF BASIC MEDICAL SCIENCES, IN PARTIAL
FULLFILLMENT OF THE REQUIREMENT FOR THE AWARD OF
BACHELOR OF SCIENCE, B.Sc. (HONS) MEDICAL BIOCHEMISTRY, OF
THE UNIVERSITY OF BENIN, BENIN CITY.

JANUARY, 2023.

i
 CERTIFICATION

We the undersigned hereby certify that Mr. Anietie Daniel Jeremiah carried out this
work, in the department of Medical Biochemistry, University of Benin, Benin City
and we approve same as adequate in scope and quality for the award of Bachelors of
Science Degree (B.Sc) in Medical Biochemistry.

Signed:

........................................ …………………………

DR. OMOROWA Date

(Project Supervisor)

………………………..... …………………………

DR. A. OMOAKHUA Date

(Ag. Head of Department)

…………………………. ……………………….

External Supervisor Date

ii
 DEDICATION

This work is dedicated to my mum, Mrs. Ruth Anietie.

iii
 ACKNOWLEDGEMENT

My gratitude goes to my affable Supervisor Dr. Omorowa for his guidance and
directions in the course of this research work. My gratitude also goes to the Head of
Department, Dr. Akuekegbe Omoakhua and other lecturers of the Department.

iv
 TABLE OF CONTENTS

TITLE PAGE - - - - - - - - -i

CERTIFICATION - - - - - - - -ii

DEDICATION - - - - - - - -iii

ACKNOWLEGDEMENT - - - - - - -iv

ABSTRACT - - - - - - - - -v

TABLE OF CONTENT - - - - - - -.v

CHAPTER ONE:INTRODUCTION

1.1 BACKGORUND OF STUDY - - - - - -1

1.2 STATEMENT OF PROB - - - - -2
1.3 JUSTIFICATION OF PROBLEM - - -
1.4 AIMS & OBJECTIVES - - - - - - -4

CHAPTER TWO: LITERATURE REVIEW

2.1 MALARIA PARASITE - - - - - - -5

2.2 LIFE CYCLE OF MALARIA - - - - - -6

2.3 DIAGNOSIS AND TREATMENT - - - - -7

2.3.1 MICROSCOPY - - - - - - -8

2.3.2 ANTIGEN DETENTION - - - - - -9

2.3.3 TREATMENT - - - - - - - -10

2.4 FULL BLOOD COUNT - - - - - - -13

2.4.1 RED BLOOD CELLS - - - - - - -10

2.4.2 WHITE BLOOD CELLS - - - - - -11

2.4.3 PLATELETS- - - - - - - - -12

v
2.4.4 PACKED CELL VOLUME - - - - - -8

2.5 INCREASED RESISTANCE TO MALARIA DRUGS - - -9

2.5.1 CLASSIFICATION OF ANTI-MALARIA DRUG RESISTANCE- -9

2.5.2 HISTORY OF MALARIA DRUG RESISTANCE - -9

2.6 ALPHA BETA ARTEETHER - - - - - -11

2.6.1 DOSAGE - - - - - - - -12

2.6.2 MECHANISM OF ACTION - - - - - -13

2.6.3 PHARMACOKINETIC PROPERTIES - - -9

CHAPTER THREE: MATERIALS AND METHODS

3.1 SUBJECT OF STUDY - - - - - - -10

3.2 INSTRUMENTS AND MATERIALS USED- - - - -12

3.3 CHEMICALS AND REAGENTS USED - - - -10

3.3 EXCLUSION CRITERIA - - - - - -11

3.5 INCLUSION CRITERIA - - - - - - -12

3.6 NORMAL PATIENTS - - - - - - -12

3.7 METHOD OF SAMPLE COLLECTION - - - -10

3.8 LABOURATORY INVESTIGATION - - - - -11

3.8.1 PROCEDURE FOR MALARIA PARASITE TEST - - -12

3.8.2 PROCEDURE FOR FULL BLOOD COUNT - - - -10

3.9 TREATMENT - - - - - - - -11

3.10 POST LABOURATORY INVESTIGATION- - - - -12

4.0 STATISTICAL ANALYSIS- - - - - - -11

vi
 ABSTRACT

A study was conducted to investigate the effects of onion extract on various factors
related to cardiometabolic syndrome in healthy individuals aged 16-30. Participants
were given a specific dosage of onion extract for three months and their blood
samples were analyzed in comparison to a control group. The results revealed that the
test group exhibited varying levels of improvement in factors that mitigate
cardiometabolic syndrome, indicating that the antioxidants and other dietary
components in onions can aid in managing cardiometabolic syndrome risk factors.

vii
 CHAPTER ONE

INTRODUCTION AND LITERATURE REVIEW

1.1 Background Information

Onion, (Allium cepa L.), is one of the most consumed and grown vegetable crops in
the world. Onion bulb, with its characteristic flavor, is the third most essential
horticultural spice with a substantial commercial value. Apart from its culinary
virtues, A. cepa is also used traditionally for its medicinal virtues in a plethora of
indigenous cultures. Several publications have been produced in an endeavor to
validate such traditional claims. Nonetheless, there is still a dearth of up-to-date,
detailed compilation, and critical analysis of the traditional and ethnopharmacological
propensities of A. cepa. (Joaheer D Teshika et al., 2013).

The cardiometabolic syndrome represents a cluster of metabolic abnormalities that

are risk factors for cardiovascular disease. The mechanism(s) responsible for
developing the cardiometabolic syndrome is not known, but it is likely that multi‐
organ insulin resistance, which is a common feature of the cardiometabolic syndrome,
is involved. Insulin resistance is an important risk factor for type 2 diabetes and can
cause vasoconstriction and renal sodium reabsorption, leading to increased blood
pressure. Alterations in adipose tissue fatty acid and adipokine metabolism are
involved in the pathogenesis of insulin resistance. Excessive rates of fatty acid release
into the bloodstream can impair the ability of insulin to stimulate muscle glucose
uptake and suppress hepatic glucose production. Noninfectious systemic
inflammation associated with adipocyte and adipose tissue macrophage cytokine
production can also cause insulin resistance. In addition, increased free fatty acid
delivery to the liver can stimulate hepatic very low‐density lipoprotein triglyceride
production, leading to dyslipidemia.(Isomaa B et al., 2001).

This study was intended to show how the phytochemical composition of the
Allium cepa which includes S-methyl cysteine sulfoxide (SMCS), S-propyl cysteine
sulfoxide, cycloalliin, thiosulfinates, and sulfides which possesses an an array of
biochemical substances with numerous pharmacological properties including anti-
oxidant, anti-inflammatory, anti-diabetic effects and could therefore crucial in the

1
treatment or research for treatment pertaining to several cardio-metabolic
dysfunctions.

1.2 Aims and Objectives of the Study

This research thesis aims at studying the antioxidant effect of Onion (Allium cepa)
extract on hematological variables in relation to the Cardiometabolic effect (Full
Blood Count for this Thesis) of said extract on Human subjects.

2
 CHAPTER TWO

LITERATURE REVIEW

2.1 Onion Bulb (Allium cepa)

2.2.1 Taxonomy

The onion plant (Allium cepa), also referred to as the bulb onion or common onion, is
the most widely cultivated species of the genus Allium. Its scientific name Allium
cepa was given by Carl Linnaeus in his 1753 work Species Plantarum.A. cepa is
known exclusively from cultivation, but there are related wild species that grow
mainly in Central Asia and Iran. The most closely related species include A. vavilovii
from Turkmenistan and A. asarense from Iran. The vast majority of cultivars of A.
cepa belong to the common onion group (A. cepa var. cepa) and are usually referred
to simply as onions. The Aggregatum Group of cultivars (A. cepa var. aggregatum)
includes both shallots and potato onions. The genus Allium also contain a number of
other species variously referred to as onions and cultivated for food, such as the
Japanese bunching onion (A. fistulosum), Egyptian onion (A. × proliferum), and
Canada onion (A. canadense). (Fritsch et al., 2002).

2.2.2 Specific Classification of Allium cepa

(1) Kingdom: Plantae

(2) Division: Magnoliophyta

(3) Class: Liliopsida

(4) Order: Asparagales

(5) Family: Alliaceae

(6) Genus: Allium

(7) Species: A. cepa

3
2.2.3 Description

The onion is a plant that has a tall, leafless stalk that can grow up to 2.5-6 feet in
height. At the top of the stalk, there is a cluster of small, greenish-white flowers.
Some of these clusters also produce small bulbs, called bulbuls, which can be used to
propagate new plants. The onion's edible bulb is located underground and develops
from the concentric leaf bases. Onions can be grown from small bulbs or transplants,
but most commercially grown onions are started from small, black seeds that are
planted directly into the ground. Onions can grow in a variety of conditions and
climates, and the size, shape, color, and pungency of the bulbs can vary depending on
the location. Although warmer climates tend to provide onions with a milder, sweeter
flavor than do other climates (Abdissa et al., 2011).The leaves of the onion are long,
flattened, and have a fan-like shape. They are green to blue-green in color and have a
white sheath at the base that emerges from the bulb's basal plate. The onion's bulb
develops as the plant matures, and the crop is usually harvested in the fall when the
leaves begin to die back and the outer scales of the bulb become dry and brittle. The
onion has a distinctive pungency due to the sulfur-rich volatile oil it contains. The
onion's pH is around 5.5.

2.2 Etymology and Evolutionary History

"Onion" is somewhat a generic term that refers to several pungent members of the
genus Allium (Lilaceae family) including common (bulbous) onion, garlic, leek and
others. The word was derived from the Middle English union which, in turn, came
from the Latin unio. The latter means "one" or "unity" and refers to the onion's single
bulb consisting of concentric rings. The pungency of onions is due to volatile sulfur
compounds (thiosulfinates) which, in turn, are produced from sulfur-containing flavor
precursors released when onion cells are ruptured or cut.Onion is thought to have
originated more than 5000 years ago in Central Asia and is one of the most ancient of
food sources. Its consumption by humans can be traced back to the Bronze Age. A
staple in the diet of many early civilizations, it was especially important in ancient
Egypt. In addition to being consumed as a food, Egyptians worshiped onion thinking
its concentric rings symbolized eternal life. Indeed, it was often buried along with
their dead. Ancient Greek athletes consumed large quantities of them thinking it

4
would "balance" their blood and improve their athletic prowess. Later, after
conquering Greece, Romans ate onions regularly and also rubbed it on their gladiators
to tone their muscles.Throughout antiquity the medicinal properties of onion were
widely avowed. As a result, it was used by ancients to treat a wide array of conditions
ranging from irregularity to hair loss. Early Americans used wild onions to treat colds,
coughs, asthma and breathing problems. Today, onion is still considered a health
food. Its consumption has been associated with a reduced risk of cancer, heart disease
and diabetes because of its high level of phenolic and flavonoid compounds with high
antioxidant activity. In general, onions with greater pungency have higher antioxidant
activity han milder types.(David Trinklein,.2011).

2.3 Nutritional components of Allium cepa

2.3.1 Nutrients

Most onion cultivars are about 89% water, 4% sugar, 1% protein, 2% fibre and 0.1%
fat. Onions contain low amounts of essential nutrients (see table), are low in fats, and
have an energy value of 166kJ (40 kcal) per 100 g (3.5 oz) serving. They contribute
their flavor to savory dishes without raising caloric content appreciably.

Nutritional value per 100 g (3.5 oz) of Raw Onions:

Carbohydrates 166KJ (40 kcal)

Dietary fiber 9.34g
Fat 4.24g
Protein 1.7g
Thiamine(B1) 1.1g
Riboflavin (B2) 0.046mg (4%)
Niacin (B3) 0.027mg (2%)
Pantothenic acid (b5) 0.116mg (1%/)
Vitamin B6 0.123mg (2%)
Folate (B9) 0.12(9%)
Vitamin C 19 μg (5%)
Calcium 23mg(2%)
Iron 0.21mg(2%)
5
Magnesium 10 mg (3%)
Manganese 0.129 mg (6%)
Phosphorus 29 mg (4%)
Potassium 146 mg (3%)
Zinc 0.17 mg (2%)
Water 89.11 g
Fluoride 1.1 µg

Note!

+ Units: μg = micrograms • mg = milligrams * IU = International units

+ Percentages are roughly approximated using US recommendations for
adults.
Source: USDA Nutrient Database - Link to USDA Database entry.

2.4 Bioactive compounds in onions

Onion is rich in a variety of phytochemicals with beneficial functional, including

organosulfur compounds, phenolic compounds, polysaccharides, and saponins
(Lanzotti, 2012). The major bioactive compounds of onion are sulfur-containing
compounds, such as onionin A and cysteine sulfoxides, as well as the phenolic
compounds, such as rutin, quercetin, and quercetin glucosides (Figure 1). It is
different for the contents of bioactive compounds among different onion varieties to
differ onion had the highest contents of anthocyanins and flavonols, followed by the
yellow onion, but the white onion contained the lowest amount (Zhang, 2016).

Processing can change the bioaccessibility and content of bioactive compounds in

onion. The bioaccessibility of total flavonols in onion was not affected by high-
pressure processing, but the onion matrix could improve the bioaccessibility of its
flavonol. It has been found that onion skin quercetin aglycone was more bioavailable
than pure quercetin dihydrate in humans. The content and bioaccessibility of phenolic
compounds, especially quercetin-derivatives, were found increased by cooking, such
as baking, grilling, and frying (Cattivelli, et al., 2021). Besides, the contents of the
6
cysteine sulfoxides, including cycloalliin, isoalliin, methiin, and propiin, were
changed differently in onion by heat processing, depending on the cooking methods,.
for instance, their contents were decreased during boiling, but increased during frying,
microwaving, and steaming. Furthermore, the flavonoid content was decreased during
the processing of black onion, while the contents of isoalliin and fructose were
significantly increased (Moreno-Ortega, et al., 2021).

2.5 Therapeutic Potential of Onion Extract

Effects on lipid profile

Dyslipidemia which is characterized by a combination of four abnormalities in serum

lipid profile including elevated levels of total cholesterol (TC), triglyceride (TG), and
low-density lipoprotein cholesterol (LDL-C) and low level of high-density lipoprotein
cholesterol (HDL-C) raises the incidence of cardiovascular morbidity and
mortality.Numerous animal and human research projects on the lipid-modifying effect
of A. cepa have exhibited an important decrease in serum and hepatic values of TG
and cholesterol (Srinivasan K,.2013).

Effects on obesity

Obesity is a common global health concern. It was estimated that in 2016, the obese
population reached over 790 million, and an extra 1.5 billion were overweight. In fact,
the prevalence of overweight and obesity combined was nearly 31
percent(Lancet.,2017).Research projects have shown the anti-obesity characteristics
of onion and phytoconstituents available in this plant such as quercetin and
organosulfurs ( Marrelli M.,2018.).

Effects on hypertension

High blood pressure is a significant contributor to the development of serious heart

and blood vessel diseases, including coronary artery disease, stroke, atrial fibrillation,
vascular dementia, and heart failure. Lowering systolic blood pressure by as little as
10 mm Hg can significantly reduce the risk of these major cardiovascular events.
While medications such as ACE inhibitors, diuretics, and calcium channel blockers
are effective at controlling blood pressure, they may also cause unwanted side

7
effects.Phytochemicals that reduce blood pressure are receiving increasing attention
as both preventative and treatment options, particularly for individuals with
prehypertension.Several studies have confirmed the antihypertensive potential of
onion and quercetin.

Anti-diabetic Activity

Research has shown that onions have the ability to improve diabetes and treat related
complications. Studies have found that onion extracts, specifically from the skin or
waste, can effectively inhibit enzymes that contribute to diabetes, such as α-
glucosidase and α-amylase. Additionally, onion-derived silver nanoparticles have
been found to have strong inhibitory effects on these enzymes. Additionally, studies
have shown that onion fiber concentrates can decrease the digestion of starch and
production of glucose by suppressing α-amylase activity.

Cardio-metabolic Syndrome

Cardiometabolic syndrome is a group of metabolic issues that increase the risk of

cardiovascular disease. The exact cause of this syndrome is unknown, but it is
believed to be related to multi-organ insulin resistance, which is a common feature of
this syndrome. Insulin resistance can lead to type 2 diabetes, high blood pressure, and
other issues. It is thought that problems with fatty acid and other chemical levels in fat
tissue play a role in the development of insulin resistance. Additionally, inflammation
in the body can also contribute to insulin resistance. These issues can lead to an
imbalance in lipids (fats) in the blood, which is called dyslipidemia.The
cardiometabolic syndrome represents a constellation of metabolic abnormalities that
are risk factors for cardiovascular disease. The diagnosis of cardiometabolic
syndrome lacks a universally accepted definition, with multiple groups proposing
their own clinical criteria for diagnosis .The most widely used clinical criteria for
diagnosing the cardiometabolic syndrome are those proposed by the World Health
Organization and the National Cholesterol Education Program Adult Treatment Panel
III (NCEP ATP III) .The common characteristics of the cardiometabolic syndrome
among all groups include abdominal obesity (high body mass index and/or large waist
circumference), insulin‐resistant glucose metabolism (hyperinsulinemia, impaired
fasting glucose, impaired glucose tolerance, type 2 diabetes), dyslipidemia (high
8
serum triglyceride and low serum high‐density lipoprotein cholesterol concentrations),
and increased blood pressure.

2.5.1 Pathophysiology / pathogenesis of Cardio-metabolic syndrome

Increased Blood Pressure.

It is well known that there is a connection between insulin resistance and high blood
pressure. Fat can cause the blood vessels to narrow, and when the body becomes
resistant to insulin, it can lead to an increase in blood pressure. Insulin typically helps
to widen blood vessels, but in people who are resistant to it, this effect is diminished.
However, the ability of insulin to increase the amount of sodium retained in the
kidneys remains intact in these individuals. This can contribute to high blood
pressure, particularly in those with cardiometabolic syndrome.

Ectopic Fat

Ectopic accumulation of fat in liver and muscle cells is associated with insulin
resistance in those tissues, increased intrahepatic fat content is associated with hepatic
insulin resistance in the liver and impaired insulin‐mediated suppression of hepatic
glucose production, and increased intramyocellular fat content is associated with
skeletal muscle insulin resistance and impaired insulin‐mediated glucose disposal.
(Krssak M et al.,1992).

Fatty Acid Metabolism

The cardiometabolic syndromen is also referred to as the insulin resistance syndrome,

as it is thought to be caused by insulin resistance as the primary mechanism behind
the metabolic abnormalities seen in the syndrome. The metabolism of free fatty acids
is believed to play a significant role in the development of hyperglycemia and
dyslipidemia associated with the cardiometabolic syndrome. High levels of free fatty
acids in the blood, which can be caused by excessive release from fat tissue, can
decrease the effectiveness of insulin in controlling blood sugar and increase the
production of lipids in the liver.

Abdominal Adipose Tissue

9
Visceral fat, specifically abdominal fat, is linked to insulin resistance, however, it is
uncertain whether it is the cause or just a correlation. Although it only makes up a
small percentage of total body fat, it has been suggested that the fatty acids released
during the breakdown of visceral fat contribute to insulin resistance as they directly
reach the liver via the portal vein.

Full Blood Count (FBC)

The full blood count (FBC) is a type of blood test, which when carried out provides
important information about the kinds and numbers of cells in the blood: red blood
cells, white blood cells and platelets. Abnormalities in any of these types of cells can
indicate the presence of important medical disorders.

2.5.2 Red Blood Cells

Red blood cells(RBC) also known as erythrocytes,are the most common type of blood
cell and the vertebrate's principal means of delivering oxygen to the body tissues.An
Increase RBC count was observed in animals treated with onion juice extract as
revealed by Griffiths, et al. 2002, stated that; Onion is also a popular folk remedy, rich
in flavonoids such as quercetin and sulfur compounds, such as allyl propyl that have
perceived benefits to human health (Griffiths, et al., 2002). Conversely, Banerjee and
Maulik, 2002 observed that Allium species have been implicated in the induction of
haemolytic anaemia. They contain toxic components that may damage red blood cells
and provoke haemolytic anaemia accompanied by Heinz bodies in erythrocytes of
animals such as cattle, water buffalos, sheep, horses, dogs and cats (Banerjee and
Maulik, 2002).

2.5.3 White Blood Cells

White blood cells also referred to as leucocytes, are a vital component of the immune
system. They play a key role in defending the body against infections and illnesses.
There are three main types of WBCs: granulocytes, lymphocytes, and monocytes.
Studies have shown that compounds found in Allium plants, such as garlic and
onions, have antimicrobial properties and may also aid in increasing the number of
circulating white blood cells, which are necessary for protecting the body from
harmful pathogen, this was confirmed by the significant increase in the Lymphocytes
10
count in the animal treated with onion juice extract.Monocytes count was seen to be
significantly lower in the treated animals when compared with the control animals this
might have resulted from Anti-microbial effects and Antioxidant effects of onion
(Banerjee and Maulik. 2002). Microbial products can directly activate monocytes and
this leads to production of pro-inflammatory and with some delay of anti-
inflammatory cytokines. Typical cytokines produced by monocytes are TNF tumor
necrosis factor, interleukin-1 and interleukin-12 (Swirski et al; 2009). Eosinophils- a
type of granulocyte, demonstrated a significant decrease in onion treated animals
when compared with the control animals. The decrease in the Eosinophils count in
the treated animals demonstrate the antioxidant property of onion juice extract as the
free radical agent generate production of more Eosinophil this is in agreement with
(Rietz et al., 1995). Basophils- another type of granulocyte also showed significant
decrease in the treated animals compared with the control animals. Recent studies in
mice suggest that basophils may also regulate the behavior of T cells and mediate the
magnitude of the secondary immune response (Nakanishi et al., 2010).

2.5.4 Platelets

Platelets also known as thrombocytes is a types of Blood cell, they are small, colorless
cell fragments in our blood that form clots and stop or prevent bleeding. Platelets are
made in our bone marrow. When normal blood vessels are damaged, platelets are
activated by stimuli which are present in the walls of blood vessels, and induce
aggregation (Nieswandt and Watson 2003). Platelets are activated by agonists such as
collagen, thrombin and ADP, and it induces the signals by activating multiple G
protein–mediated pathways to activate platelet-shape change, degranulation and
aggregation (Nieswandt and Watson 2003). But in many cardiovascular diseases such
as acute coronary syndromes, atherosclerosis, stroke and peripheral vascular diseases,
excessive platelet activation is regarded as the cause of thrombosis. Abnormal platelet
aggregation leads to excessive TXA2 formation interacts with other platelets,
inducing thrombotic disorders (FitzGerald et al. 1987). Therefore, the development of
an anti-platelet agent could be a fundamental therapeutic approach to cardiovascular
diseases. Onion (Allium cepa) has been reported to have beneficial effects, including
preventing stroke, coronary thrombosis, atherosclerosis, hyperlipidemia and

11
hypertension. Especially, it has been reported to inhibit platelet aggregation induced
by various agonists in vitro and in vivo (Moon et al., 2000).

12
 CHAPTER THREE

MATERIALS AND METHODS

3.1 Methodology

Reagents used

Lyse

FDT Lyse

FDO Lyse

Diluent

Probe cleanser

Apparatus

Anticoagulant bottles (EDTA)

Syringes and needles

Tourniquet

Latex examination gloves

3 part Automated Hematological Analyzer

3.2 Allium cepa Procurement and preparation

Onion(Alluim cepa L.) was obtained from a local market in benin city, edo state,
nigeria. It was washed, pilled and transferred into a juicer extractor machine, in
which, it juice was extracted. There was no water added in the process.
Method of Collection of Blood Samples (Specimen)

This study was carried out within a period of eight weeks. All subject were given a
letter of recognition so as to cooperate with the process. Blood samples were collected
for both subject and control for a series of laboratory investigations using standard
protocol for the various tests done. A total of two hundred and ten subjects were

13
involved in the project research. Seventy of them were used as standard control and
One hundred and fourty subjects were used for test. Both control and test group, had
their blood samples collected.

Volume of Blood Collected and Site of Collection

10ml of blood was collected through venepuncture from both the control and test
groups.

Method of Onion Juices Administration

Onion juice (Allium cepa.) was administered orally to the Test group in which they
were divided into two group in respect of dosage. 30ml and 40ml respectively. While
the control group were not administered onion juice. Onion juice was administered
three times per

week for Eight weeks and blood sample was retaken to evaluate any observed
changes.

Determination of level of Full Blood Count

Procedure

Blood samples were obtained from the jugular vein by venepuncture before feeding
(fasted) on two sequential days after 8 weeks of treatment. Blood was also collected
from each person 3h post-feeding (postprandial), since the absorption of dietary
flavanols from onions in human blood reaches peak values between 0·7 and 2·9h after
feeding (Hollman et al. 1996, 1999; Aziz et al. 1998). On the first sequential sampling
day, blood was drawn into EDTA, clot activator and sodium citrate vacutainer tubes,
which were dedicated to blood coagulation tests (platelet count, prothrombin time,
activated partial prothrombin time and fibrinogen counts) and blood biochemistry
assays (cholesterol, TG and glucose).Blood samples were placed in ice and
transported for automated analysis (University of Benin teaching hospital
Haematology laboratory.)

14
Principle

On board the analyzer, the sample is agitated to evenly distribute the cells, then
diluted and partitioned into at least two channels, one of which is used to count red
blood cells and platelets, the other to count white blood cells and determine the
hemoglobin concentration. Some instruments measure hemoglobin in a separate
channel, and additional channels may be used for differential white blood cell counts,
reticulocyte counts and specialized measurements of platelets.The cells are suspended
in a fluid stream and their properties are measured as they flow past sensors in a
technique known as flow cytometry (Kottke-Marchant et al.,2012).Motocrit are used
to calculate the average amount of hemoglobin within each red blood cell, the mean
corpuscular hemoglobin (MCH); and its concentration, the mean corpuscular
hemoglobin concentration (MCHC). Another calculation, the red blood cell
distribution width (RDW), is derived from the standard deviation of the mean cell
volume and reflects variation in cellular size.Hemoglobin concentration is measured
by adding a reagent to the sample to break down red cells. This is done in a separate
channel from the one used for counting red blood cells. This allows for easier
counting of white blood cells on analyzers that perform both hemoglobin
measurement and white blood cell counts. Hemoglobin is measured using a process
called spectrophotometry, which is based on the relationship between the amount of
light absorbed and the amount of hemoglobin present.

15
 CHAPTER FOUR

RED BLOOD CELL

GROUP 1 (30ML)

Parameter Control Test

Red Blood Cells 5.12 ± .67b 4.01 ± 1.25b

count(RBC)

Haemoglobin(HGB) 13.5 ± 1.53b 10.9 ± 3.40b

Hematocrit 41.3 ± 4.89b 34.38 ± 9.93a

test(HCT)

Mean Corpuscular 76.47 ± 8.12b 86.17 ± 7.37a

Volume (MCV)

Meaning 26.52 ± 2.50b 27.11 ± 3.08b

Corpuscular
Haemoglobin
(MCH)

MCHC(Mean 33.33 ± 1.595a 31.43 ± 1.86b

Corpuscular
Haemoglobin
Concentration)

RBC Distribution 14.59 ± 1.705a 16.03 ± 3.683a

width(RDW)

The result in the table above for group 1 showed that RBC (4.01 ± 1.25 b) had a
significant (P < 0.05) decrease as compared to the control group (5.12 ± .67 b). HGB
(10.9 ± 3.40b) also showed a significant (p < 0.05) decrease as compared to the
control group (13.5 ± 1.53b). The result also showed that HCT (34.38 ± 9.93 a) had a

16
significant (p < 0.05) decrease compared to control group (41.3 ± 4.89 a). MCV (86.17
± 7.37a) had a significant (p < 0.05) increase compared to the control group (76.47 ±
8.12b). More so, MCH (27.11 ± 3.08a) had a significant (p < 0.05) increase as
compared to control group (26.52 ± 2.50b). MCHC (31.43 ± 1.86b) had a significant (p
< 0.05) decrease as compared to control group (33.33 ± 1.595 b). RDW (16.03 ± 3.68a)
had a significant (p < 0.05) increase as compared to control group (14.59 ± 1.705b).

GROUP 2 (40ML)

Parameter Control Test

RBC 5.12 ± .67b 4.97 ± .751b

HGB 13.5 ± 1.53b 12.43 ± 1.09b

HCT 41.3 ± 4.89b 39.57 ± 4.58a

MCV 76.47 ± 8.12b 83.59 ± 9.77a

MCH 26.52 ± 2.50b 31.30 ± 6.84b

MCHC 33.33 ± 1.595a 35.17 ± 4.01b

RDW 14.59 ± 1.705a 12.84 ± 1.521a

The result in the table above for group 2 showed that RBC (4.97 ± .751 b) had a
significant (P < 0.05) decrease as compared to the control group (5.12 ± .67 b). HGB
(12.43 ± 1.09b) also showed a significant (p < 0.05) decrease as compared to the
control group (13.5 ± 1.53b). The result also showed that HCT (39.57 ± 4.58 a) had a
significant (p < 0.05) decrease compared to control group (41.3 ± 4.89 a). MCV (83.59
± 7.37a) had a significant (p < 0.05) increase compared to the control group (76.47 ±
8.12b). More so, MCH (31.30 ± 6.84a) had a significant (p < 0.05) increase as
compared to control group (26.52 ± 2.50b). MCHC (31.30 ± 6.84b) had a significant (p
< 0.05) decrease as compared to control group (33.33 ± 1.595 b). RDW (12.84 ± 3.68a)
had a significant (p < 0.05) decrease as compared to control group (14.59 ± 1.705b).

RBC

17
HGB

18
HCT

19
MCV

MCH

20
MCHC

RDWC

21
RDWS

22
 CHAPTER FIVE

DISCUSSION AND CONCLUSION

In general, consumption of allium cepa extract was found to moderately modulate

Haematological variables in test subjects, but the responses varied doses of allium
cepa extract. Changes in blood cell counts were evident. The Allium cepa extract
consumption reduced both erythrocyte numbers and Haemoglobin concentrations as
indicated by the test results above which showed a decrease in RBC and HCT- the
measurement of the percentage of RBC in the blood. Reductions in haematocritic
measurements have also been observed in placebo-controlled studies with human
subjects using encapsulated onion (Mayer et al. 2001).There was an increase in Red
Blood Cells Distribution width (RBW) which measures the amount of red blood cells
variation in volume and size, this might indicates that there's anaemia. Excessive
onion Consumption has been shown in other studies to induce acute haemolytic
anaemia in sheep (Kirk & Bulgin, 1979), cattle (van der Kolk, 2000), dogs (Spice,
1976), largely due to the presence of onion disulfides: these generate H2O2 in the
presence of Hb and glutathione S-transferase within intact erythrocytes (Munday et al.
2003). It has been suggested that the resulting haemolysis is of the oxidative type and
occurred due to removal of damaged erythrocytes by cells of the reticulo-endothelial
system (Munday et al. 2003). . It has been suggested that the resulting haemolysis is
of the oxidative type and occurred due to removal of damaged erythrocytes by cells of
the reticulo-endothelial system (Munday et al. 2003). Although the erythrocyte
concentrations were decreased linearly by onion consumption in this present study,
there was a compensatory increase in cell volume and the amount of Hb per cell as
evident in the increase in MCH which measures the amount of Haemoglobin per red
blood cells , this implies an increased O2-carrying capacity per erythrocyte. The
excessive consumption of onions should be avoided, especially by individuals with
anaemia or those whose erythrocytes are unusually vulnerable to oxidative damage
(Munday et al., 2003).

23
Conclusion

The present study has shown that Allium cepa has functional properties with the
ability to modify hematological components and stimulate the immune system.
However, the differences in the responses observed in the present study demonstrate
the complex interaction between the onion extract and the dose which may be due to
the differences in the amount of active compounds. The side effects of Onion
Consumption was also observed, as it can induce acute haemolytic anemia as such,
excessive Consumption of onions should be avoided by those with anemia. There are
several more researches to be conducted to discover new potentialities of onions as
a remedy for treatment of Cardio-metabolic diseases.

24
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