CHAPTER ONE

INTRODUCTION

Over the years, Cardiometabolic syndromes such as hypertention, diabetes,

hypercholesterolamia, obesity, and cancer have become a prevalent public health predicament

in many countries and World Wide as a result of unhealthy life styles. By reason of this

prevalence, it has become a threat to human lives and eventually result to numerous deaths.

According to statistical analysis from the National Health and Nutrition Examination Survey, the

prevalence of these syndromes increases linearly with age. Cardiometabolic syndromes involve a

group of metabolic disorders that are risk factors of cardiovascular diseases. It occur due to

complicated factors such as High blood pressure, hyperlipidemia, diabetes, obesity, smoking,

excessive alcohol intake, imbalance diet and lack of physical activities (Paraveen et al., 2013).

Notwithstanding, Medicinal plants and vegetables continue to provide imperative therapeutic

agents in both modern and traditional system to prevent cardiometabolic syndromes as well as

other dieaeses (Raqueen et al.,1985). Amongst other vegetables, Onions has prominently been

used as food additives and as supplement for many centuries. Epidemiological studies had

proven the fact that onions extract consumption by humans and lab animals over a stipulated

period of time will positively affect cardiometabololic syndromes.

Onions is botanically called Allium cepa, it is also called Yabari, Alubosa, Albasa and Alubara

by the Igbo's, Yoruba's, Hausa's and Beni's respectively. Onions plant is a bulbous vegetable of

about 1.2m in height with ⁿ4 to 6 hallows, cylindrical leaves. It underground bulb consist of

fleshy leaf sheath forming a thin skinned capsule and differ in sizes. Onions are generally

consumed by people owing to it numerous phytochemical constituents which provide a

promising health benefits. Onions contains 89% water,1.5% protein, as well as Vitamin B, B2,

C, E and minerals like potassium, calcium, magnesium, phosphorus, and zinc (Block, 2010). It

also contains flavanoid such as quercetin and sulfur containing compound like alkyl propyl

disulphide which are of biological importance ( Griffita et all., 2002). Moreover, onions are rich

in sulfur compounds basically in the form of Cysteine derivatives, S-alkyl Cysteine sulfoxide

which are decomposed by the enzyme allinase into a volatile compounds such as thiosulfinate

and polysylfide during extraction. These compounds are responsible for the potential health

benefits onions extract provide which include; antidiabetic, hypocholesterolamic, fibrinolytic,

anticancer, immunological effect as well as other health benefits.

Also, the non volatile sulfur containing compounds in Alium cepa posses antibiotic effect and as

such improving the functionality of the white blood cells which boost the body immune system

(Augustii, 1996). Furthermore, studies has proven that onions juice exhibit antioxidant properties

which altogether reduces oxidative stress that may lead to cell and tissue damage . Being that

onions is generally consumed by people due to it nutritional constituents, there arise the need to

examine it effect on cardiometabolic parameters.

1.1 Aim of Study and objectives

This study was carried out to examine the effect of onions extract on cardiometabolic

parameters.

1. Oral administration of onions extract ( Alium Cepa) to evaluate it effect on glucose and

Haemoglobin A1C level on selected individuals.

 CHAPTER TWO

LITERATURE REVIEW

1.1.1 Cardiometabolic syndrome

Cardiometabolic syndrome is a combination of metabolic disorders basically characterized by

insulin resistance , impaired glucose tolerance , dyslipidemia, hypertention and central adiposity.

According to WHO, CMS is now known as a disease entity. People who are are at risk of

Cardiometabolic syndromes are prone to die from coronary heart diseases or stroke compared to

those who lack the syndrome. It is well known that hyperlipidemia is the major factor that

contribute to the rising Cardiometabolic risk ( Srivastana, 2012).

Nonetheless, several challenges have been encountered to bringing CMS risk factors under

control. A number of Cardiometabolic programmes and the use of therapeutic agents in plants

and vegetables are specifically employed in modern and traditional system of medicine to

maximize the goal in reducing Cardiometabolic risk factors.

Prevelance

In United States and World Wide, Cardiometabolic Syndrome has become a primary public

health predicament because of it prevalence. According to the data from the third National

Health and Nutritional Examination found that the age adjusted prevalence of CMS was defined

using the ATP111 criteria. The prevelance of CMS increases linearly with Age.
1.1.2 Pathophysiology and pathogenesis of Cardiometabolic syndromes

Several pathophysiological Cardiometabolic factors are associated with the risk of myocardial

infarction. Cardiometabolic syndrome is also known as insulin resistance syndrome because it

has been reported that insulin resistance is the major mechanism responsible for the metabolic

disorders of the syndromes (Reaven, 1988). The pathogenesis of hyperglycemia and

dyslipidemia associated with Cardiometabolic syndromes may result to the excessive release of

free fatty acid from adipose cells into plasma , increase in plasma free acid concentration,

thereby causing the inability of insulin to stimulate muscle glucose uptake ( Abate et all.,1996).

Moreover, elevated level of free fatty acid delivery to the liver can increase hepatic very low

density lipoprotein , triglyceride production, and plasma triacyglycerol concentration (Lewis et

al., 1993). These conditions eventually lead to increased transfer of TAG from VLDL to HDL,

which result to elevated HDL clearance and decreased plasma HDL concentration ( Hopkins and

Barter , 1986).

1.1.3 Risk factors of Cardiometabolic syndromes and treatment

1.1.3.1 Blood pressure

Hypertention commonly referred to as high blood pressure is a major contribution to

cardiovascular risk. The age and sex adjusted prevalence of hypertention was 28% in North

America Countries And 44% Europe ( Wolf et al.,2017). The European guideline defined

hypertention as a blood pressure ( BP 140/90mmHg). Hypertention affect more than 150 million

people and it's prevalence is predicted to rise by 15% to 20% by 2015. High blood pressure can

cause injury to the lining of blood vessels, called endothelium which can lay the groundwork for

atherosclerosis ( Bolad et all., 2018). Vegetable consumption is an independent factor in blood

pressure. Study showed that the consumption of raw vegetables reduces blood pressure to normal

( Chan et all., 2014).

1.1.3.2. Lipid profile

The lipid profile consists of total cholesterol, HDL, LDL, and triglycerides. It is a major risk

factor of heart disease . It has been reported that high cholesterol is responsible for 2.6 million

deaths ( WHO, 2020). Low density lipoprotein cholesterol ( LDL_ C) is typically the target of

cholesterol lowering therapy, as it is a major factor of the plagues that build up on blood vessels

wall leading to atherosclerosis, that prevent blood flow across the body ( Bantwal et al., 2012).

According to WHO, stated that a 40_ year_ old man can half his 5years heart disease risk with

10% reduction in LDL. However, plant rich diets was helpful in the treatment and prevention of

hyperlipidemia or elevated cholesterol.

1.1.3.4. Obesity/ Overweight/Adiposity

Universally, the prevalence of overweight and obesity has tripled since 1974 due to unhealthy

life style and was at 39% in 2016. The trend continue in children, with a 4% prevalence in 1975

and an overall 18% prevalence in 2016. As a result of the trend, becoming a risk factor of

Cardiometabolic disorder ( Aronne and Isoldi, 2007). Excessive fat storage in the adipose cells

can cause inflammation as a result of lack of vasculature in adipose tissue stores (Booth et all.,

2016).

Studies showed that plant based diet are effective tools in weight control. In the BROAD study,

researchers showed that a significant weight loss result with a low fat plant based diet.
1.1.3.5. Glucose Metabolism

Plant based diets are thought to improve glucose Metabolism by several mechanisms through the

reduction of insulin resistance in body tissues and the improvement of the functionality of rhw

pancreatic beta cells, which are basically responsible for the secretion of insulin that function as

glucose uptake into the cell ( Kahleova et all., 2018). Greater stores of fats within the muscle

cells are associated with greater insulin resistance as fat interferes with blood glucose uptake by

muscles and the storage of glucose as glycogen. Since glucose cannot enter the cell , it is left in

the circulation, promoting more insulin secretion by pancreatic beta cells. Persistent over

working of pancreatic beta cells can lead to hyperglycemia

1.1.3.6 Inflammation and Endothelial function

The endothelial refers to cells lining the inner portion of blood vessels, making direct contact

with circulating blood. These cells have vital roles in the prevention and genesis of heart disease.

The endothelium is important for heart health because it secrete substances that enable blood

vessels to constrict, dillate and form clots needed (Bolad et all.,2018). Several risk factors

influence endothelial health which include; hypertension, shear stress from increase s blood

pressure inside the blood vessel can injure endothelial cells, thus, inhibiting their ability to

maintain normal function of blood vessels ( Mudua et all., 2021). White blood cells stick to areas

of endothelial dysfunction and travel to deeper layers of blood vessels, initiating an

atherosclerotic plagues. Once in this layer , the WBC termed foam cells , at this point collect

LDL cholesterol, further increasing inflammation. Factors associated with endothelial injury

include elevated LDL cholesterol, High blood pressure and diabetes. However, study showed

that endothelial health can be improved with plant based diet.

1..1.4. Morphology of Onions

Onions also known as the bulb onions, is a vegetable that is the most widely cultivated species of

the genus Alium. The shallet is a botanical variety of the onions which was classified as a

separate species. A cepa is exclusively known from cultivation. Its ancestral wild original form

is known although escapes from cultivation have become established in some regions . Onions

is most frequently a biennial and a perenial plant, but is usually treated as an annual and

harvested in the first growing season. Onions are thought to have originated in Afghanistan and

are now produced in more than 175 Countries around the world. Onions belong to the Liliaceae

family. They are perennial crop that can be red , white or yellow and eaten raw, or powder in its

tender condition. Onions is highly regarded and consumed for its flavour and nutritional value.

.1.1.4.1. Specific classification of Alium Cepa

Kingdom : Plantae

Division: Magnoliophyta

Class: Liliopsida
Order: Asparagales

Family: Alliaceae

Order: A.cepa

Culinary uses of Onions

Onions are commonly chopped and used as ingredient In various hearty warm dishes,and may

also be used as a main ingredient gridien on it own. They are versatile and can be baked, grilled

or eaten raw 37.

Just like garlic, onions can show an additional colour after cutting, an effect caused by the

reactions of amino acid with sulfur compounds ( Lee et al , 2012)

NON CULINARY USES

Onions oil is used in Europe and UK as pesticide agent ( Sharp and Russell 2021). Onions also

contains one of the natural oil that can be use in hair oil. Onions are commonly use in science

education to teach the use of microscope for observing cell structures ( Huang et al.,2014).

Onions contain quercetin , a plant base pigment which contains antioxidant properties in vitro.

One Japanese study suggests that onions can help in alleviating oxidative stress caused by

diabetes ( Azuma et al., 2007)

Again, another study conducted in China occluded that onions juice reduces oxidative stress and

can be recommended to patients dealing osteoporosis ( Law et al., 2016)⁷

NUTRITIONAL COMPOSITION OF A. Cepa

1.1.5 Chemical Composition of Onions

Onions has a high dietary fiber and sugar content of about 90% water. Onions extract is low in

sodium and has an elevated concentration of folic acid , vitamin B6, magnesium, potassium ,

phosphorus as well as vitamins. Onions has a low lipid level , only glutamic and arginine are

exceptional in it amino acid content (Suleria et al., 2015). A good number of phytochemical

analysis of onions and several compounds are responsible for its unique aroma and medicinal

properties have been discovered. Phenolic compounds have gained enormous interest as they

contribute to the biological properties of medicinal plants. The purple onions has quercetin

content(Peter and Arts, 2000). Various experiments also found that onions contains

anthocyanins. In addition, methalonic extract of red onions yielded four anthocyanins. A study

conducted by Vazquez et al, showed that dipropyl trisulfide and dipropyl disulfide are the main

constitients (Varzquez et al, 2014). These compounds, S- alkyl-L -Cysteine sulfoxide is a class

of biologically active organo- Surfur compound. As the plant material is crushed, the aroma and

taste of fresh onions is caused by the release of methiinne, allicin, Iso-allin, propin, and lipid

soluble sulfur compounds. It has been assumed that the pungent lachrymal factor released from

cut onions is created spontaneously as a result of the enzyme allinase(Tomotake et al.,2002).

Thiopropal- S-oxide, the sulfide volatile, is a lachrymal element present in red onions, which

ultimately transforms it into methyl- pentanol (Thomas and Parkin, 1994). In red Onions, a thin

layer chromatography with dichloromethane extraction was observed to have many disulfide

radicals (methyl, alkyl, and propyl (Giffiths et al.,2002). Some organic acid was detected in red

onions by Dhumal et al. They were citric, tartaric, malic, oxalic, abscorbic and succinic acids.

Another compound isolated from onions was found to improve the possible fungicidal efficacy

of traditional bactericidal antibiotic polymyxin B(Borjihan et al.,2010)

1.1.6 Bioactive compounds in Onions

Onions is rich in a variety of phytochemicals with beneficial constituent, including organosulfur

compounds (Wang, 2004), phenolic compounds, polysaccharides and saponins (Munday, 2001).

The major bioactive compounds of onions are sulfur containing compounds, such as Onioninns

A and Cysteine sulfoxide, as well as the phenolic compounds, Such as rutin, quercetin and

quercetin glucoside. The 8bioactive content varies in species (A.J Chakraborty et al.,2021).

Purple onions has the highest content of anthocyanins and flavonols (Lanzoti 2006). Quercetin

was the major compound in the skin of purple onions , while quercetin - 4- glucoside was the

major compound in it bulb(Parkin and Xiao 2007)

1.1.7. Therapeutic potentials of onions extract

1.1.7.1Effect on Lipid Profile

Dyslipidemia which is characterized by a combination of four abnormalities in Serum lipid

profile include increase level of total cholesterol , triglycerides and low density lipoprotein and

low level of high density lipoprotein cholesterol increases the morbidity and mortality

rate(Nelson 2013).

A number of animal and human research projects on the lipid modifying effect of A. Cepa have

shown a decrease in serum and hepatic values of triglyceride and cholesterol (Srinvasan 2013).
Several in vitro studies has shown that S-propyl Cysteine act as a reducing g agent in the release

of apolipoprotein B100, triglyceride, cholesterol, and phospholipids from cells. Nevertheless ,

the inverse relationship between the rate of substitution ofì Cysteine was detected (Han SY et

al.,2002). Quercetin is found to provide prophylaxis of atherosclerosis. This flavanoid

stimulated cholesterol removal through the expression of peroxosome proliferation activated

receptor gamma and ABCA1 in humans THP-1 macrophages and thus prevent the formation of

foam cells, which is a factor in atherosclerotic plagues development (Sun.L et al.,2015).

Animal Studies; in cholesterol treated rabbits, administration of onions extract for three months

was effective in the reduction of plasma cholesterol and fibrinogen values and aorta lipid

content. Also, it induced the rise of blood clotting time and fibrinolytic activity. The blood

samples of high cholesterol fed fat rats showed a significant decrease in triglycerides, and

phospholipids as well as a rise in High density lipoprotein- cholesterol after six weeks treatment

with 5% and 10% dehydrated onions powder. Also, an increase in the concentration of

antioxidants compounds such as total thiols, gluthathione, alpha tecopherol and abscorbic acid

also increase during examination (Vidyavati et al., 2010)

Clinical Studies; oral daily administration of 100ml of Onions juice in volunteers with mild

hypercholesterolamia after 8 weeks restrained lipid peroxidation and extended the lag time

required for LDL oxidation by reducing oxidative stress. There was also a decrease in the plasma

values of total cholesterol, LDL- cholesterol, HDL- cholesterol ratio by 10.2%, 7% and 6.0%

respectively but did not influence triglyceride and HDL- cholesterol (Marrelli et al.,2018). Fifty

four patients were randomly divided into control and therapeutic groups. They received 10 to

15g of fresh red onions twice a day for two months. There was a considerable ⁷ within both

groups in comparison before the administration.

1.1.7.2 Effect on Obesity

Obesity is a common global health concern. In 2016, it was estimated that obese population

reach over 790 millions. The prevalence of overweight and obesity combined was nearly

31%(51).

Research projects have shown the anti- obesity characteristic of Onions and phytoconstituents

available in the plant such as quercetin and organosulfurs(Marrelli et al.,2018)

Animal studies; According to Yoshinari, he evaluated the properties of Onions extract against

body fat in male diabetic fatty rats. Four weeks of treatment with the extract led to a significant

decrease in the rate of body weight gain. Additionally, onions and it sulfur compounds could

prevent white adipocyte differentiation (Yoshinari et al.,2012). The addition of quercetin and red

onions extract to the high fat diet significantly reduced inguinal and epididymal adipose tissue

weight in mice.

Clinical studies; Onions peel extract capsule, consist of 50mg quercetin, were administered

Korean overweight and obese participants twice a day for 12 weeks in a doubles blind ,

randomized, placebo- control trial performed in 2013. A decline in Body Mass Index and body

fat mass was indicated (Lee JS et al.,2016). Studies suggested that several mechanisms such as

reduction of fatty acids and biosynthesis of triglycerides, antioxidants and antiinflammatory

activity, elevation of adiponectin and energy expenditure, decrease in plasma resistance,

prevention of white adipocyte differentiation are involved in anti- obesity properties of onions

and its active components

1 .1.1.7.3Effect on Hypertension

Hypertension is a leading metabolic risk factor for the development of life threatening

cardiovascular diseases, including strokes and heart failure (Blacher et al.,2016). Although, tjw

current anti- hypertensive medications such as angiotensin converting enzyme (ACE), inhibitors,

diuretic and calcium channel blockers have been proven to be beneficial in blood pressure

control but with side effects(Aslam et al.,2017). Blood pressure lowering phytochemicals are

gaining attention as preventive and curative agents , mainlynfor prehypertensive patients (Chen

et al.,2009). Several studies have confirmed the antihypertensive potential of onions and

quercetin.

In vitro studies; the inhibition of ACE by purple onions extract was assessed. According to the

result, the extract was able to reduce enzyme activity in a concentration dependent way (Oboh et

al., 2018). The binding energy of quercetin with the active site of ACE was calculated to be -

8.5kcal/mol in comparison to the standard ( - 7.0kcal/mol). So, this molecules can reduce blood

(Fatima 2015).

Animal studies; The ethanol extract of onions at dose 0.2, 0.6 and 6 mg/kg was used

intravenously in rats. According to the result, onions dose- dependently reduce heart rate and

blood pressure. The mechanism of lowering blood pressure can be due to a decrease in heart rate

(Brankovic et al.,2011).

A. Cepa skin hydro alcoholic extract decreased blood pressure in fructose- fed hypertensive rats

presumably by a reduction in oxidative stress and the inhibition of Calcium influx in the cells of

vascular smooth muscle (Naseri et al.,2008).

Clinical studies; In a controlled study in 22 obese and hypertensive patients, increased systolic

and diastolic blood pressure after consumption of a high energy meal was not reduced by the

intake of either placebo or 54mg quercetin (Brull V et al.,2017). The ingestion of 400mg

quercetin increased brachial artery diameter in 15 healthy volunteers based on dose and time.

This flavanoid also induced vasodilation in human arteries in vitro (Perez et al.,2014).

Antimicrobial Effect

Onions extract and their derived bioactive compounds, such as thiosulfinate compound, phenolic

compounds, polysaccharides and essential oils, have been reported to possess potent antibacterial

properties ( 30), antifungal activities (Lee et al.,) and antiviral effect (147). Onions based

composite materials combining with isolated flavonoids from onions were reported to exhibit

certain antibacterial activity against staphylococcus auerus and E-coli (149). Besides, the ethanol

extract of red onions was effective in preventing tinea pedis caused by the fungal infection of

Trichiphytan rubrum(66).

Therefore, onions have been demonstrated to inhibit the growth of microbes , showing great

potential to be used as natural preservative in food industry (155)

Other potential health benefit of onions include; antiinflammatory effect , cancer prevention

improve hematological parameters, neuroprotection, and digestive health.

Antioxidant Activity

Onions is a good source of natural antioxidant s(Aneh et al., 2012). Several studies have been

carried out to evaluate the antioxidant properties of onions, and found that onions exhibits strong

antioxidants properties by using a series of in vitro assays, including ( 3- methyl

benzolelthiawoline-6 sulfonic acid) , ferric reducing antioxidant power ( FRAP) , lipid

peroxidation, oxygen radical absorbance capacity (TAC) and trolax equivalent antioxidant
capacity ( TEAC) assays. Onions was effective for protection against oxidative stress by

enhancing the activity of antioxidant enzymes such as superoxide dismatase (SOD), CAT, and

gluthione peroxide (GPx), in hyper holesterolemic rats(Ayyanet et al.,2011). The oxifatt stress

in the liver and kidney was ameliorated by pre- treatment with red onions peel extract in carbon

tetrachloride challenged rats (Brue 1996). Overall , onions extract exhibit strong antioxidant

effect, and many factors could affect the antioxidant capability of onions. The antioxidant

activity of onions has been extensively investigated but still need more exploration

DIABETIC AND HYPOGLYCEMIC EFFECT

There has been a rising trend in the prevalence of age-standardized diabetes mellitus in adults

during 1980-2014. According to estimates, the number of diabetic adults elevated from 108 to

422 million worldwide over this period, and it is expected that it will exceed 700 million in 2025

(84). Diabetes mellitus and its micro-and macrovascular complications, including cardiovascular

and kidney diseases, retinopathy, loss of vision, neuropathy, foot ulcers and amputation are

important morbidity and mortality causes (85).

In vitro studies

Anti-protein glycation of bovine serum albumin with D-fructose and antioxidant properties of 25

herbs have been measured. Among them, the skin of A.cepa showed the most anti-glycating

potential and scavenging capacity of free radicals using 1,1-Diphenyl-2-picrylhydrazyl (DPPH)

test with IC50= 16.8±5.0 and 4.49±0.59 μg/ml, respectively (Kim 2003). The inhibitory activity

of quercetin and different parts and sizes of onion were seen on porcine pancreatic α-amylase.

Furthermore, it was found that outer layers of onions and smaller ones possess more enzymatic

activity inhibition (Jaiswal and Rizviz 2007). Rat intestinal α-glucosidase was inhibited with
IC50 values 1.27 and 0.15 mg/ml by ethanolic extract of onion skin and quercetin, respectively

(Kim et al., 2011). Sodium-glucose linked transporter (SGLT1) and glucose transporter 2

(GLUT2) play key roles. Onion extractgluited human SGLT1 expressed in Xenopus

laevisoocytes in a concentration-dependent manner with a maximum effect of 86% at 1 mg/ml.

The inhibition of expressed human GLUT2 in oocytes by 0.25 mg/ml concentration of onion

extract was 78%. Among the onion flavonols, the highest levels of SGLT1 and GLUT2

suppression were observed by quercetin-4′-Oglucoside and aglycone quercetin, respectively.

Onion extract was able to reduce glucose transport into mouse jejunal intestinal sections

competitivelyreversibly (Schulzel et al., 2012). Glucose transporter 4 (GLUT4), which is

regulated by insulin, transports glucose from the blood into fat and muscle cells. An in vitro

study revealed that the ethanolic extract of A. cepa bulbs stimulates glucose uptake through

GLUT4 in L6 myotubes in a dose and time reliant pattern. The insulin-like activities of the

extract were exerted by increasing phosphorylation of insulin receptor-β, insulin receptor

substrate-1, and protein kinase B (Akt) together with the elevation of GLUT4 content and

translocation of this protein to the cell surface (Gautam et al., 2015).

Animal studies

Several studies have evidenced the health beneficial hypoglycemic action of onion and its

functional constituents in animal models (Augusti 1977).Thirty mins after consuming sucrose

solution (2.0 g/kg) by five-week-old Sprague-Dawley male rats, the ethanolic extract of onion

skin and quercetin (both 0.5 g/kg) showed a significant reduction in blood glucose comparable

with acarbose (5.0mg/kg) as an efficient drug for postprandial hyperglycemia. One hr after

administration, the lowering blood glucose effect of the extract disappeared but acarbose

maintained glycemia near base value for about 2 hrs (88). In another animal study on alloxan
diabetic rats which treated by 100, 300 and 600 mg/kg daily doses of aqueous extract of A. cepa

for 21 days, a decrease in serum levels of glucose, LDL, TG, TC, AST, ALT, and ALP along

with an increase in HDL value was reported. The maximum dose of the extract and 2 mg/kg

glibenclamide exhibited approximately the same efficacy. The fraction of the plant containing a

kaempferol glycoside improved the diabetic conditioMDAIkexhukwu and Ifeanyi 2016). A

meta-analysis was performed in 2008 on antidiabetic activities of onion extract and SMCS

inexploration rats.

The findings showed that onion extract and SMCS significantly contributed to the control of

blood glucose and body weight (Kook et al., 2009). Both quercetin and red onion extract as a

food supplement in C57BL/6J mice on a high-fat diet for 9 weeks provoked a reduction in

methylation of PGC-1α promoter and the augmentation of NT-PGC-1α expression (Dervashi et

al., 2017). Quercetin (0.2% w/w almost equal to 1000 mg/day in humans according to dose

conversion factor provided by FDA (Food and Drug Administration)) was added to the high-fat

diet of C57BL/6J male mice for a period of 10 weeks. It reduced hyperglycemia,

hyperinsulinemia, creatinine, and inflammatory indicators such as C-reactive protein. Increase in

acyl-coenzyme A oxidase 1 (ACOX1) gene expression in the liver was also observed while no

significant alterations occurred in energy expenditure, body weight, and lipid profilenitriteee.

Because of the low bioavailability of curcumin, its beneficial antidiabetic properties in

combination with piperine and quercetin were evaluated on induced diabetes in rats. After 4

weeks of daily oral feeding by this mixture (CPQ) at the dose of 100 mg/kg, amelioration in

fasting plasma glucose, glucose tolerance, LDL, HD L, TG, Cholesterol, food intake and weight

loss was obtained in comparison with diabetic control and the animals receiving only curcumin.
Obtained results were similar to those of 10 mg/kg/day glibenclamide. It was concluded that

small amounts of quercetin and piperine in CPQ may exert their effects by lowering the

metabolism of cudifferen(Kaur et al., 2016). Thiosulfinate (20 and 40 mg/kg) in diabetic rats

possibly through non-competitive α-glucosidase inhibition and pancreatic beta cells stimulation

showed improvements in postprandial glycemic control, glucose tolerance, and insulin secretion

with an effect comparable to 10 mg/kg acaevidence (Al-Malki, 2016). S-methyl cysteine (100

mg/kg/day, orally for 60 days) along with a significant decline in blood glucose, insulin plasma

concentration, TNF-α, and HOMA-IR (homeostatic model assessment of insulin resistance)

showed anti-oxidant properties in rats receiving highfructose diet. For example, a significant

decrease in the serum amount of MDA and an increase in the levels of reduced glutathione

(GSH), glutathione peroxidase (Gx), and catalase (CAT) were detected (Thomas et al., 2015).

Some researchers have studied the effects of A. cepa and its bioactive constituents on diabetic

complications.

For example, Gomes et al. showed that quercetin could be useful for the management of diabetic

nephropathy. This polyphenol (10 mg/kg/day, orally for 28 days) showed improvement in renal

function by reducing proteinuria, the plasma concentration of creatinine, uric acid, and urea as

well as increasing creatinine clearance in streptozotocin induced diabetic nephropathy mice.

Among them, modifications in creatininerelated factors were significant. Treatment with

quercetin also significantly decreased kidney weight/body weight ratio, glomerulosclerosis, and

the formation of apoptotic renal cells. They demonstrated that the anti-oxidant behavior of

quercetin could lead to renoprotective results (Gomes ef al.,2013). The daily IP injection of

aqueous onion extract (500 mg/ml/kg) to diabetic rats for 28 days showed a significant lowering

effect on elevated serum amount of thromboxane B2 (TXB2). This extract also resulted in
reducing the synthesis of TXB2 and aggregation of platelets, which was induced by collagen and

arachidonic acid in vitro (Yung et al., 2002).

The ethanolic seed extract of A. cepa in streptozotocin-induced diabetic Wistar male rats (200,

400 mg/kg/day, orally for 4 weeks) improved reproductive system performance by affecting

factors such as seminiferous tubular diameter, luminal diameter, the volume density of lumen

together with raising the generation of primary spermatocytes and spermatids (Fallah et al.,

2017). In an experiment which was conducted on mice, onion exhibited neuroprotective benefits

in the prevention and therapy of diabetic neuropathy. In addition to the amelioration of

hyperalgesia and hyperglycemia, levels of thiobarbituric acid reactive substances (TBARS) and

serum nitrite decreased, and GSH level increased. According to the resofts, the Neroprotion

impact of onions might be correlated with it antioxidant and hypoglycemic effect (Bhanott and

shri 2010)

High blood glucose can be induced by taking some medications. JafarpourSadegh et al. in a

randomized, triple-blind, placebo-controlled

CLINICAL TRIALS

Study showed that dietary raw yellow onion at the doses of 100 to 160 g/day based on BMI

index for 8 weeks significantly improved doxorubicin-induced insulin resistance and

hyperglycemia in patients with breast cancer undergoing chemotherapy (JafarpourSadegh et al.,

2017,). In a clinical trial which was performed on 84 patients suffering from type 1 and 2

diabetes mellitus with an average age of 44±3.87 years, the intake of 100 g raw red onion

improved oral glucose tolerance and fasting blood sugar after (Association). An onion meal in

people with lactose intolerance diminished the maximum blood glucose to a greater extent than
those who could tolerate lactose (44.2% versus 19.3%, P<0.05). It can be due to the

decomposition of quercetin glucosides into quercetin in lactose-tolerant subjects and further

inhibition of GLUT2 by glucosylated compounds (Hoffman et al., 2016).

Overall, A. cepa and its active components may be regarded as prophylactic or therapeutic agents

against diabetes through different mechanisms including anti-oxidant, α-glucosidase and α-

amylase inhibitory effect, up-regulation of adiponectin receptors, reducing insulin resistance and

glucose absorption from intestine, elevation in the liver and muscle glycogen content, increasing

insulin secretion and phosphorylation of AMPK, insulin-mimetic actions and GLUT4

translocation in skeletal muscles. Moreover, they exhibit benefits in diabetic complications by

renal and neural protective effects, enhancing the function of the male reproductive system, and

prevention of atherosclerosis.

Unfortunately, there is a lack of clinical-based evidence to support improving complications of

diabetes

1.1.7.5 Anti oxidant Activity

Onions is a good source of natural antioxidants (Aneh et al., 2012). Several studies have been

carried out to evaluate the antioxidant properties of onions, and found that onions exhibits strong

antioxidants properties by using a series of in vitro assays, including ( 3- methyl

benzolelthiawoline-6 sulfonic acid) , ferric reducing antioxidant power ( FRAP) , lipid

peroxidation, oxygen radical absorbance capacity (TAC) and trolax equivalent antioxidant

capacity ( TEAC) assays. Onions was effective for protection against oxidative stress by

enhancing the activity of antioxidant enzymes such as superoxide dismatase (SOD), CAT, and

gluthione peroxide (GPx), in hyper holesterolemic rats(Ayyanet et al.,2011). The oxifatt stress
in the liver and kidney was ameliorated by pre- treatment with red onions peel extract in carbon

tetrachloride challenged rats (Brue 1996). Overall , onions extract exhibit strong antioxidant

effect, and many factors could affect the antioxidant capability of onions. The antioxidant

activity of onions has been extensively investigated but still need more exploration.

BLOOD GLUCOSE REGULATION

Glucose, also known as dextrose one of a group of carbohydrates known as simple sugars

(monosaccharides). Glucose (from Greek glykys; “sweet”) has the molecular formula C6H12O6.

It is found in fruits and honey and is the major free sugarcirculating in the blood of higher

animals. It is the source of energy in cell function, and the regulation of its metabolism is of great

importance . Molecules of starch, the major energy-reserve carbohydrate of plants, consist of

thousands of linear glucose units. Another major compoundcomposed of glucose is cellulose,

which is also linear. Dextrose is the molecule D-glucose.(Kara et al., 2009)Blood sugar, or

glucose, is the main sugar found in your blood. It comes from the food you eat, and is your

body's main source of energy. Your blood carries glucose to all of your body's cells to use for

energy. (Emily et al., 2017)The body tightly regulates blood glucose levels as a part of metabolic

homeostasis.Glucose is stored in skeletal muscle and liver cells in the form of glycogen;in

fasting individuals, blood glucose is maintained at a constant level at the expense of glycogen

stores in the liver and skeletal muscle. (Shiveta et al., 2008) Blood sugar levels for in humans

without who are not fasting should be below 6.9 mmol/L (125 mg/dL). However blood glucose

target range for diabetics, accordingto the American Diabetes Association, should be 5.0–7.2

mmol/l (90–130 mg/dL) before meals and less than 10 mmol/L (180 mg/dL)two hours after

meals (as measured by a blood glucose monitor

3.7. Blood Glucose Level

Glycemia also known as blood sugar level, blood sugar concentration, or blood glucose level is

the measure of glucose concentrated in the blood of humans or other animals. Approximately 4

grams of glucose, a simple sugar, is present in the blood of a 70 kg (154 lb) human at all times.

(Wesserman DH, 2009). The body tightly regulates blood glucose levels as a part of metabolic

homeostasis. (Weserman DH, 2009) Glucose is stored in skeletal muscle and liver cells in the

form of glycogen;in fasting individuals, blood glucose is maintained at a constant level at the

expense of glycogen stores in the liver and skeletal muscle( Walker et.al, 2006)

In humans, blood glucose level of 4 grams, or about a teaspoon, is critical for normal function in

a number of tissues, and the human brain consumes approximately 60% of blood glucose in

fasting, sedentary individuals.(Wesserman DH, 2009). A persistent elevation in blood glucose

leads to glucose toxicity, which contributes to cell dysfunction and the pathology grouped

together as complications of diabetes. Glucose can be transported from the intestines or liver to

other tissues in the body via the bloodstream. Cellular glucose uptake is primarily regulated by

insulin, a hormone produced in the pancreas.

Glucose levels are usually lowest in the morning, before the first meal of the day, and rise after

meals for an hour or two by a few millimoles. Blood sugar levels outside the normal range may

be an indicator of a medical condition. A persistently high level is referred to as hyperglycemia;

low levels are referred to as hypoglycemia. Diabetes mellitus is characterized by persistent

hyperglycemia from any of several causes, and it is the most prominent disease related to the

failure of blood sugar regulation. There are different methods of testing and measuring blood
sugar levels . The intake of alcohol causes an initial surge in blood sugar and later tends to cause

levels to fall. Also, certain drugs can increase or decrease glucose levels.(Walker et.al, 2006)

UNITS

There are two ways of measuring blood glucose levels: In The United Kingdom and

commonwealth countries (Australia, Canada, India, etc.) and ex-USSR countries molar

concentration, measured in mmol/L (millimoles per litre, or millimolar, abbreviated mM). In the

United States, Germany, Japan and many other countries mass concentration is measured in

mg/dL (milligrams per decilitre).[4]

Since the molecular weight of glucose C6H12O6 is 180, the difference between the two units is a

factor of 18, so 1 mmol/L of glucose is equivalent to 18 mg/dL.[5]

NORMAL RANGE

Normal value ranges may vary slightly between laboratories. Many factors affect a person's

blood sugar level. The body's homeostatic mechanism of blood sugar regulation (known as

glucose homeostasis), when operating normally, restores the blood sugar level to a narrow range

of about 4.4 to 6.1 mmol/L (79 to 110 mg/dL) (as measured by a fasting blood glucose test).[6]

Normal blood glucose level (tested while fasting) for non-diabetics is between 3.9 and 7.1

mmol/L (70 and 130 mg/dL). The global mean fasting plasma blood glucose level in humans is

about 5.5 mmol/L (100 mg/dL); (Danaei 2011).However, this level fluctuates throughout the

day. Blood sugar levels for those without diabetes and who are not fasting should be below 6.9
mmol/L (125 mg/dL).[8] The blood glucose target range for diabetics, according to the American

Diabetes Association, should be 5.0–7.2 mmol/L (90–130 mg/dL) before meals and less than 10

mmol/L (180 mg/dL) two hours after meals (as measured by a blood glucose monitor).(Schuster

2008)

Despite widely variable intervals between meals or the occasional consumption of meals with a

substantial carbohydrate load, human blood glucose levels tend to remain within the normal

range. However, shortly after eating, the blood glucose level may rise, in non-diabetics,

temporarily up to 7.8 mmol/L (140 mg/dL) or slightly more. For people with diabetes

maintaining "tight diabetes control", the American Diabetes Association recommends a post-

meal glucose level of less than 10 mmol/L (180 mg/dL) and a fasting plasma glucose of 3.9 to

7.2 mmol/L (70–130 mg/dL).[11]

The actual amount of glucose in the blood and body fluids is very small. In a healthy adult male

of 75 kg (165 lb) with a blood volume of 5 L, a blood glucose level of 5.5 mmol/L (100 mg/dL)

amounts to 5 g, equivalent to about a teaspoonful of sugar.[12] Part of the reason why this

amount is so small is that, to maintain an influx of glucose into cells, enzymes modify glucose by

adding phosphate or other group to it.

REGULATION OF BLOOD GLUCOSE

The body's homeostatic mechanism keeps blood glucose levels within a narrow range. It is

composed of several interacting systems, of which hormone regulation is the most important.
There are two types of mutually antagonistic metabolic hormones affecting blood glucose levels:

catabolic hormones (such as glucagon, cortisol and catecholamines) which increase blood

glucose;(Leighninger and Nelson 2017)

and one anabolic hormone (insulin), which decreases blood glucose.

These hormones are secreted from pancreatic islets (bundles of endocrine tissues), of which there

are four types: alpha (A) cells, beta (B) cells, Delta (D) cells and F cells. Glucagon is secreted

from alpha cells, while insulin is secreted by beta cells. Together they regulate the blood-glucose

levels through negative feedback, a process where the end product of one reaction stimulates the

beginning of another reaction. In blood-glucose levels, insulin lowers the concentration of

glucose in the blood. The lower blood-glucose level (a product of the insulin secretion) triggers

glucagon to be secreted, and repeats the cycle.( Tortora 2016)

In order for the blood glucose to be kept stable, modifications to insulin, glucagon, epinephrine

and cortisol are made. Each of these hormones has a different responsibility to keep blood

glucose regulated; when blood sugar is too high, insulin tells muscles to take up excess glucose

for storage. Glucagon responds to too low of a blood glucose level; it informs the tissue to

produce more glucose. Epinephrine prepares the muscles and respiratory system for activity in

the case of a "fight and flight" response. Lastly, cortisol supplies the body with fuel in times of

heavy stress.
GLUCOSE ABNORMALITIES

High blood sugar(Hyperglycemia)

If blood sugar levels remain too high the body suppresses appetite over the short term. Long-

term hyperglycemia causes many health problems including heart disease, cancer,[22] eye,

kidney, and nerve damage.[23]

Blood sugar levels above 16.7 mmol/L (300 mg/dL) can cause fatal reactions. Ketones will be

very high (a magnitude higher than when eating a very low carbohydrate diet) initiating

ketoacidosis. The Mayo Clinic recommends emergency room treatment above 16.7 mmol/L (300

mg/dL) blood glucose.[citation needed] The most common cause of hyperglycemia is diabetes.

When diabetes is the cause, physicians typically recommend an anti-diabetic medication as

treatment. From the perspective of the majority of patients, treatment with an old, well-

understood diabetes drug such as metformin will be the safest, most effective, least expensive,

and most comfortable route to managing the condition.[24] Diet changes and exercise

implementation may also be part of a treatment plan for diabetes.

Some medications may cause a rise in blood sugars of diabetics, such as steroid medications,

including cortisone, hydrocortisone, prednisolone, prednisone, and dexamethasone.[25]

Low blood sugar( Hypoglycemia)

If blood sugar levels drop too low, a potentially fatal condition called hypoglycemia develops.

Symptoms may include lethargy, impaired mental functioning; irritability; shaking, twitching,

weakness in arm and leg muscles; pale complexion; sweating; loss of consciousness.

Mechanisms that restore satisfactory blood glucose levels after extreme hypoglycemia (below

2.2 mmol/L or 40 mg/dL) must be quick and effective to prevent extremely serious consequences

of insufficient glucose: confusion or unsteadiness and, in the extreme (below 0.8 mmol/L or 15

mg/dL) loss of consciousness and seizures. Without discounting the potentially quite serious

conditions and risks due to or oftentimes accompanying hyperglycemia, especially in the long-

term (diabetes or pre-diabetes, obesity or overweight, hyperlipidemia, hypertension, etc.), it is

still generally more dangerous to have too little glucose – especially if levels are very low – in

the blood than too much, at least temporarily, because glucose is so important for metabolism

and nutrition and the proper functioning of the body's organs. This is especially the case for those

organs that are metabolically active or that require a constant, regulated supply of blood sugar

(the liver and brain are examples). In healthy individuals, blood glucose-regulating mechanisms

are generally quite effective, and symptomatic hypoglycemia is generally found only in diabetics

using insulin or other pharmacological treatment, and in starvation or severe malnutrition or

malabsorption (of various causes), and conditions such as anorexia.[dubious – discuss]

Hypoglycemic episodes can vary greatly between persons and from time to time, both in

severity of onset. For severe cases, prompt medical assistance is necessary so as to prevent

damage of tissues and brain as a result of low blood sugar.

CLINICAL CORRECTION

Fasting blood glucose level, which is measured after a fast of 8 hours, is the most commonly

used indication of overall glucose homeostasis, largely because disturbing events such as food

intake arefasting blood glucose level, which is measured after a fast of 8 hours, is the most

commonly used indication of overall glucose homeostasis, largely because disturbing events

such as food intake are avoided. Conditions affecting glucose levels are shown in the table

below. Abnormalities in these test results are due to problems in the multiple control mechanism

of glucose regulation.[citation needed]

The metabolic response to a carbohydrate challenge is conveniently assessed by a postprandial

glucose level drawn 2 hours after a meal or a glucose load. In addition, the glucose tolerance test,

consisting of several timed measurements after a standardized amount of oral glucose intake, is

used to aid in the diagnosis of diabetes.

Error rates for blood glucose measurements systems vary, depending on laboratories, and on the

methods used. Colorimetry techniques can be biased by color changes in test strips (from

airborne or finger-borne contamination, perhaps) or interference (e.g., tinting contaminants) with

light source or the light sensor. Electrical techniques are less susceptible to these errors, though

not to others. In home use, the most important issue is not accuracy, but trend. Thus if a meter /

test strip system is consistently wrong by 10%, there will be little consequence, as long as
changes (e.g., due to exercise or medication adjustments) are properly tracked. In the US, home

use blood test meters must be approved by the federal Food and Drug Administration before they

can be sold.

Finally, there are several influences on blood glucose level aside from food intake. Infection, for

instance, tends to change blood glucose levels, as does stress either physical or psychological.

Exercise, especially if prolonged or long after the most recent meal, will have an effect as well.

In the typical person, maintenance of blood glucose at near constant levels will nevertheless be

quite effective.

CAUSES OF ABNORMAL GLUCOSE LEVEL

The Reference range of Fasting Blood Sugar (FBS) is 70- 110mg/dl. A deviation from this value

may cause abnormalities.

Persistent Hyperglycemia: Diabetes millitus, Obesity, hyperthyroidism.

Transient hyperglycemia: Pheochromocytoma, severe liver disease, Acute stress, shock,

convulsion.

Persistent hypoglycemia: Insulinoma, Adrenal cortical insufficiency, Hypopituitarism,

Galactosemia.

Transient hypoglycemia: Drugs, antituberculosis agents, severe liver disease, severe Glycogen

storage diseases, Hereditary

GLYCATED HAEMOGLOBIN

Other than Fasting blood sugar , glycated Haemoglobin test can also be used to estimate the

average blood glucose level for the past two to three months. It is usually estimated in

percentage.

Glycated hemoglobin (glycohemoglobin, hemoglobin A1c, HbA1c, less commonly HbA1c,

HgbA1c, Hb1c, etc., also A1C informally with patients[1]) is a form of hemoglobin (Hb) that is

chemically linked to a sugar. Most monosaccharides, including glucose, galactose and fructose,

spontaneously (i.e. non-enzymatically) bond with hemoglobin, when present in the bloodstream

of humans. However, glucose is less likely to do so than galactose and fructose (13% that of

fructose and 21% that of galactose), which may explain why glucose is used as the primary

metabolic fuel in humans.(Bunn and Higgins 1981).

The formation of the sugar-hemoglobin linkage indicates the presence of excessive sugar in the

bloodstream, often indicative of diabetes. A1C is of particular interest because it is easy to

detect. The process by which sugars attach to hemoglobin is called glycation. HbA1c is a

measure of the beta-N-1-deoxy fructosyl component of hemoglobin.(Miedema 2005)

A1c is measured primarily to determine the three-month average blood sugar level and can be

used as a diagnostic test for diabetes mellitus and as an assessment test for glycemic control in

people with diabetes. The test is limited to a three-month average because the average lifespan of

a red blood cell is four months. Since individual red blood cells have varying lifespans, the test is

used as a limited measure of three months. Normal levels of glucose produce a normal amount of

glycated hemoglobin. As the average amount of plasma glucose increases, the fraction of
glycated hemoglobin increases in a predictable way. In diabetes, higher amounts of glycated

hemoglobin, indicating poorer control of blood glucose levels, have been associated with

cardiovascular disease, nephropathy, neuropathy, and retinopathy.

However, Glycated hemoglobin causes an increase of highly reactive free radicals inside blood

cells, altering the properties of their cell membranes. This leads to blood cell aggregation and

increased blood viscosity, which results in impaired blood flow.(Seleh and Jumana 2015).

Another way glycated hemoglobin causes damage is via inflammation, which results in

atherosclerotic plaque (atheroma) formation. Free-radical build-up promotes the excitation of

Fe2+-hemoglobin through Fe3+-Hb into abnormal ferryl hemoglobin (Fe4+-Hb). Fe4+ is

unstable and reacts with specific amino acids in hemoglobin to regain its Fe3+ oxidation state.

Hemoglobin molecules clump together via cross-linking reactions, and these hemoglobin clumps

(multimers) promote cell damage and the release of Fe4+-hemoglobin into the matrix of

innermost layers (subendothelium) of arteries and veins. This results in increased permeability of

interior surface (endothelium) of blood vessels and production of pro-inflammatory monocyte

adhesion proteins, which promote macrophage accumulation in blood vessel surfaces, ultimately

leading to harmful plaques in these vessels. (Selah and Jumana 2015)

Highly glycated Hb-AGEs go through vascular smooth muscle layer and inactivate

acetylcholine-induced endothelium-dependent relaxation, possibly through binding to nitric

oxide (NO), preventing its normal function. NO is a potent vasodilator and also inhibits

formation of plaque-promoting LDLs (i.e. “bad cholesterol”) oxidized form.

This overall degradation of blood cells also releases heme from them. Loose heme can cause

oxidation of endothelial and LDL proteins, which results in plaques.

This overall degradation of blood cells also releases heme from them. Loose heme can cause

oxidation of endothelial and LDL proteins, which results in plaques.

blood cells also releases heme from them. Loose heme can cause oxidation of endothelial and

LDL proteins, which results in plaques.

Normal, prediabetic, and diabetic range

The 2010 American Diabetes Association Standards of Medical Care in Diabetes added the

=HbA1c ≥ 48 mmol/mol (≥6.5 DCCT %) as another criterion for the diagnosis of diabetes.[38]

Diagnostic Standard for HbA1C in Diabetes[39]

HbA1C Diagnosis

<5.7% Normal

5.7–6.4% Prediabetes

>6.4% Diabet
1.1.8. ÀIM

The purpose of this study is to evaluate the effect of onions extract on fasting blood sugar and

Haemoglobin a1c level using selected individuals.

 CHAPTER THREE

MATERIALS AND METHODS

2.1. Materials

2.1.1. Tests materials

A bag of fresh purple onions was purchased at Lagos Street, Edo State, Nigeria. These onions

were large and healthy for consumption. They were stored under room temperature during the

period of use for the experimental purpose.

2.2. Apparatus and Equipment

Electronic Juicer

A bag of onions

Knives

Syringes and needles

Latex gloves

Cotton wool

Methylated spirit

Tourniquet

Spectrophotometer

Testubes
Specimen Bottles with heparin

Measuring Cups

Refrigerator

Micro pipettes

Incubator

Beakers.

Measuring bottles.

2.2.1. Chemical /Reagents

Reagents/ enzymes kit and other reagents used in this experiment were of standard quality and

were purchased from qualified and accredited suppliers in Nigeria. The chemicals which were

used were of analytical grades.

2.2.1.1. Reagents/ Chemicals used for the assessment of Glycated Haemoglobin.

Reagent 1: Ion Exchange Resin ( predisposed tubes)

Reagent 2: ( Lysing) lysing reagents

Control ( 10% GHb)

Distilled water( for dillution)

A Resin Seperator ( shouldn't be inhaled because it causes irritation).

2.2.1.2. Reagents / Chemicals used for the assessment of fasting blood glucose

Glucose oxidase reagent, hydrogen peroxidase, Phenol, 4 amino phenalzone.

2.3. Methods

2.3.1. Experimental design

The experimental findings were based on phase 1 clinical trials. It lasted for about a period of 8

weeks. Onions extract which was hygienically prepared was administered to the volunteers 3

times in a week . The administration of the extract was strictly based on dosage, some of the

Volunteers were given 30 ml while others were given 40 ml. A total number of 10 controls and

20 test were used for the experiment, amongst who were UNIBEN students. The limited

numbers of volunteers available was due to society ignorance and lack of awareness about the

effective use of medicinal plants and it importance. A written letter was given by my Project

Supervisor so as to enlighten the Volunteers and to validate the purpose and course of study.

However, a lot of challenges were actually encountered in the course of this experiment, a lot of

people were actually not fascinated about consuming the raw onions extract because of it arsh

taste and so had to be refrigerated to keep the taste refreshing. Also in other human studies , the

dosage of the onions extract used was quite more than the one used in this study and might have

minimal effect on the experiment.

Exclusion criteria

Volunteers who had history of Allergy to onions, smokers, ages below 16, alcohol addicts,

people on certain medications were excluded.

Inclusion criteria

Randomly selected individuals with unknown basal metabolic rate, ages between 17 to 30, non

smokers, non alcoholic individuals, administration of the onions extract was basically on dosage

and was categorised.

2.4. Blood Sample collection

5ml of Individual blood sample was collected from venous into labeled specimen bottles .

Anticoagulant containing lithium heparin/sodium flouride bottles were for the biochemical assay

of fasting blood sugar and Haemoglobin a1c. These specific specimen bottles were used to

prevent glycolysis in the blood samples before the test. Sample collection was done in the

morning after an overnight fast. Samples were collected fom the vein using sterilised syringes

and needles and then placed into the already labelled specimen bottles, and kept under room

temperature.

2.5 . Preparation of Onions Extract

Some large purple onions was peeled , diced and washed thoroughly. The diced onions were then

placed in an electric Juicer, the clear purple onion extract was obtained. The onions extract was

stored at room temperature prior to use.

Determination of Fasting Blood Glucose

Assays Principle of Fasting blood glucose.

Methodology: Glucose Oxidase method

Principle: Glucose is oxidized is oxidixed to glucoronic acid and Hydrogen peroxide in the

presence of the enzyme Glucose Oxidase. Hydrogen peroxide is further broken down to oxygen

and water in the presence of Hydrogen peroxidase. The oxygen released then react with Phenol

and 4 amino phenalzone to form a pinkish color which is read Spectrop.

Procedures

Add one ml of glucose oxidase reagent to a testube

Add 10 microlitre of the sample and treat standard as test.

Mix and incubate for 10 min at room temperature

Read at 490 nm wavelength using Spectrophotometer.

Calculation:

Concentration of test = Absorbance of Test/Absorbance of standard × concentration of stansard(

100 mg/dl)

Determination Of Glycated Hemoglobin ( HBA1C)

Assay Principle of HBA1C

Methodology: ION EXCHANGE RESIN METHODS

Principle: Glycosylated hemoglobin has been defined as the fast fract

ion of hemoglobin HbA1 ( HbA1a, A1b, A1c) which eluted first during colu chromatography.

The non glycosylated hemoglobin which consist of the bulk of hemoglobin , has been

designated as HbA1c. A hemosylated preparation of whole blood is mixed continuously for 5

mins with a weakly binding cation exchange resin. The Labile fraction is eliminated during the

hemosylated preparation and during the binding. During this mixing , H bAo binds to the io n

exchange resin leaving GHb free in the supernatant. The percent glycosylated hemoglobin is

determined by measuring the absorbance of the glycosylated hemoglobin ( GHb) and the total

hemoglobin fraction ( THb). The ratio of the absorbance of GHb and THb of the control and

test is used to calculate the percent of GHb of the sample.

PROCEDURE

(A) Hemolysate preparation

1. Dispense 0.5 ml lysing reagent into tubes labeled as control(C) and test (T)

2. Add 0.1 ml of reconstituted control and we'll mixed blood sample into the appropriate test

tubes . Mix until complete lysis is evident

3. Allow to stand for 5 mins

B. Glycosylated hemoglobin separation

1. Remove cap from the ion exchange resin tubes and label as control and test

2. Add 0.1 ml of hemo lysate from step A into the appropriately labeled ion- Exchange Resin

tubed.

3. Insert a resin Seperator into each tube so as the rubber sleeve is approximately 1 cm above

the liquid level of the resin suspension.

4. Mix the tubes on a rocker , rotator, or a vortex mixer continously for 5 mins
5. Allow the resin to settle , then push the resin seperator into the tubes until resin is firmly

packed.

6. Pour or aspirate each supernatant directly into a corvette and measure each absorbance

against Distilled water.

(C) Total hemoglobin fraction ( THb)

1. Dis pense 5.0 ml of Distilled water into tubes labeled as control and test

2. Add to it 0.2 ml of hemo lysate from step A into the appropriately labeled tube

3. Mix well

4. Read each absorbance against Distilled water.

Calculation

Ratio of control( Rc) = Abs of control GHb/Abs control THb

Ratio of Test (Rt) = Abs of tes GHb/Abs THb

GHb Ratio% = Ratio of test/ Ratio of control × 10 ( value of control)

STATISTICAL ANALYSIS

Data are represented as Mean and standard deviations, as well 0as Pearson' s correlation

analysis, were done using Microsoft Excel 2003. Significant differences of the resuls among the

Variances were statistically analysed by SPSS 14.0 software. Statistical significance was

accepted at a level of P < 0.0 5

 CHAPTER FOUR

RESULTS

BLOOD GLUCOSE LEVEL

GROUP 1 (30ML)

Parameter Control Test

FBS 86.8 ± 19.11a 82.8 ± 18.02b
HbA1C 3.61 ± .738a 3.01 ± .72b

The result in the table above for group 1 showed that GLU (82.8 ± 18.02 b) had a significant (P <

0.05) decrease as compared to the control group (86.8 ± 19.11 b). HbA1C (3.01 ± .72b) had a

significant (p < 0.05) decrease as compared to the control group (3.61 ± .738b).

GROUP 2 (40ML)

Parameter Control Test

FBS 86.8 ± 9.11a 82.8 ± 8.70b
HbA1C 3.61 ± .738a 3.23 ± .583b

The result in the table above for group 2 showed that GLU (82.8 ± 8.70 b) had a significant (P <

0.05) decrease as compared to the control group (86.8 ± 9.11 a). HbA1C (3.23 ± .583b) had a

significant (p < 0.05) decrease as compared to the control group (3.61 ± .738a).
 FBS

120

100

80

control
60
30ml
40 ml
40

20

0
1 2 3 4 5 6 7 8 9 10

Fig. 1 Bar chart showing the effect of Onion extract on fasting blood glucose of volunteers based on
dosage

HBAc

control
3
HBAc 30ml
HBAc 40ml
2

0
1 2 3 4 5 6 7 8 9 10

Fig. 2 Bar chart showing the effect of Onion extract on HbA1c on volunteers based on dosage
 CHAPTER FIVE

DISCUSSION OF RESULTS

Insulin resistance to blood glucose has been a major risk factor associated with cardeometalsolie

syndromes, which has become a prevalent cause of morbidity in both developed and developing

countries.

Insulin resistance to glucose has been reported to be a critical mediator in the association

between obesity and its commobidities such as type 2 diabetes, hypertension and CVDs. Insulin

resistance has been used in cardiovascular risk production model (Liy et al., 2013).

This study has shown that blood glucose level in terms of fasting blood sugar and HbAlC can be

regulated using the extract of onions (A cepa) when administered to volunteers.

The result observed in the present study with respect to blood glucose regulations in volunteers

with unknown basal metabolic rate, drastically decreased fasting blood sugar and improvement

in serum HbAlC when compared to the control group. Alium cepa was found to be effective in

improving the glycemic control at a specific close.

However, exceedingly, significant change was observed at a higher dose of 40ml/kg as compared

to when the onions extract was administered at a dose of 30ml/kg.

In a previous clinical trial-performed in 84 patients suffering from type 1 and 2 diabetes mellitus,

the intake of 100g of fresh onions improved oral glucose tolerance and fasting blood sugar after

4hours. (Taj Edim et al., 2010).

The mechanism of action in which A. cepa is regarded as a therapeutics agent against diabetes

includes; antioxidant, alpha glucose dose, and alpha analysis inhibitory effect, up regulation of a

diponection receptors, reducing insulin resistance and glucose absorption from the intestines,

elevation in the liver and muscle glycogen content, increasing insulin secretion and

phosphorylation of AMP kinase.

In accordance with this present study, some research have studied the effect of A. cepa and it

bioactive constituents in diabetic complications. Quercetin could be useful in the management of

diabetic nephropathy (Gomes et al., 2002).

HbAlC is considered to be a marker of long term blood glucose central (Sacks et al., 2002) and it

reflect both fasting plasma glucose and posrandial glucose. Because HbAlC has a strong

predictive value for diabetes complications (Knowler at al., 2002), reduced HbAlC value in

response to the administration of onions extract could contribute to a lower risk of diabetes

complications.

CONCLUSION

From the study it can be deduced that the label of fasting and blood glucose and glycated

hemoglobin showed a significant reduction. The reduction in glucose parameters was a result of

the onions extract that was administered to the volunteers.

Onions extract is therefore recommended as a therapeutic agent for the control of blood sugar

and therefore decreasing the risk of insulin resistance. However, this course of study still need

more exploration especially in clinical trials.

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