ABSTRACT
Some plants are known to be sources of food with significant medicinal properties that can combat the deleterious effects of some chemicals incorporated into foods and Mucuna sloanei is one of such plants and that led to the evaluation of the nutritional properties of Mucuna sloanei seed endosperms and some selected biochemical effects of the ethanol extract of the seed endosperms on Monosodium glutamate (MSG)-intoxicated rats. Thirty (30) albino female rats used were divided into 6 groups, n= 5. The groups were; Group 1; given feed and water, Group 2; given MSG only at the dose of 8000 mg/kg, Group 3; given 400 mg/kg of the extract only and Groups 4, 5 and 6 were given different concentrations (200, 400 and 800 mg/kg) of the extract with MSG at the dose of 8000 mg/kg respectively. They were treated for 28 days and their blood, liver and kidneys were collected for analysis and Histology. The evaluations were done using standard analytical procedures. The results were as follows: moisture (5.20%), ash (3.50%), crude protein (22.75%), crude fat (7.20%), crude fiber (2.20%), and carbohydrate (59.15%). The mineral analysis revealed sodium (146.35 mg/100 g), potassium (252.80 mg/100 g), calcium (44.80 mg/100 g), magnesium (125.80 mg/100 g), iron (9.60 mg/100 g) and zinc (3.00 mg/100g). The vitamin analysis showed that it contained thiamine (vitamin B1) (2.14 mg/100 g), vitamin A (4.75mg/100 g), vitamin C (14.40 mg/100 g), vitamin E (0.15 mg/100 g) while quantitative phytochemical analysis showed that the endosperm contained alkaloid (0.16%), saponins (0.06%), flavonoids (0.20%), tannin (0.03%), oxalate (0.40%) and phenols (0.02%). The kidney functioning capacity showed that MSG significantly increased the urea (29.42±1.44) (mg/dl) and creatinine (1.30±0.18) (ml/min) values in Group 2, while the extract in Group 3 significantly decreased the urea and creatinine values (P<0.05) when compared to Group 2. Group 6 gave the highest sodium (130.03±3.59) (mEq/L) and chloride (116.90±2.44) (mEq/L) values significantly different (P<0.05) from Groups 1, 2 and 3. Group 3 gave the highest values (5.85±0.48) (mEq/L) of potassium when compared to the rest of the groups. Biochemical evaluation of the live’ r functional indices showed that Group 3 significantly reduced the AST, ALP, ALT and bilirubin values (P<0.05) when compared to Group 2 that increased them. The heamatological parameters showed that Group 3 at the dose of 400 mg/kg had the highest significant RBC level of 8.29±0.04 (x1012/L) together with Group 6 that had a value of 8.23±0.33(x1012/L). Group 2 had the least values of RBC 5.89±0.07(x1012/L), PCV 45.33±1.45%, Hb 12.44±0.33 g/dl and WBC 12.44±0.33 (x109/L). The antioxidant result showed that Group 3 had a significantly increased glutathione value (P<0.05) of 63.08±3.98 (U/L) when compared to the other groups while Group 2 had the least glutathione value of 37.37±2.27 (U/L). The photomicrograph of the liver showed that Group 1 and Group 3 had a well preserved liver architecture while Group 2 showed a moderate to severe portal inflammation. The kidney plates showed no significant difference in the Histology of the different groups since all their glomeruli were evenly distributed, with normal mesangial cellularity and normal endothelia. Thus, Mucuna sloanei seed endosperms could be said to combat the oxidative stress caused by MSG through its high antioxidant properties
TABLE OF
CONTENTS
Title page i
Declaration ii
Certification iii
Dedication iv
Acknowledgments v
Table of contents vi
List
of tables x
List of figures xi
List of Plates xii
Abstract xiii
CHAPTER 1: INTRODUCTION
1.1 Medicinal Plants 1
1.2
Statement of the Problem 2
1.3
Justification of the Study 2
1.4
Aim and Objectives of the Study 3
CHAPTER 2: LITERATURE
REVIEW
2.1 The Mucuna Plant 4
2.1.1 Description of mucuna sloanei 4
2.1.2 Uses of mucuna sloanei 7
2.1.3 Empirical studies on mucuna sloanei 7
2.1.4 Medicinal properties
of mucuna sloanei 8
2.2 Monosodium Glutamate 8
2.2.1 History of monosodium
glutamate 9
2.2.2 Production of
monosodium glutamate 9
2.2.3 Physical and chemical
properties of monosodium glutamate 10
2.2.4 Uses of monosodium
glutamate 10
2.2.5 Safety and adverse
effects of monosodium glutamate 10
2.3 Nutraceutical Properties 11
2.3.1 Alkaloids 11
2.3.2 Flavonoids 12
2.3.3 Saponins 12
2.3.4 Tannins 13
2.3.5 Phenolic compounds 14
2.3.6 Cyanogenic glycosides 14
2.4 The Liver 15
2.4.1 Diseases of the liver 16
2.4.2 The liver function
tests (LFT) 16
2.4.2.1 The Transaminases 17
2.4.2.2 Aspartate
aminotransferases (AST/SGOT) 17
2.4.2.3 Alkaline phosphatase
(ALP) 17
2.4.2.4 Bilirubin 18
2.4.2.5 Total protein 18
2.5 The Kidney 19
2.5.1 Urea 19
2.5.2 Creatinine 19
2.5.3 Protein 20
2.5.4 Calcium 20
2.5.5 Phosphorus 20
2.6 Vitamins 20
2.6.1 Vitamin A 21
2.6.2 Vitamin B1 (Thiamine) 21
2.6.3 Vitamin C 22
2.6.4 Vitamin E 23
2.7 Minerals 23
2.7.1 Sodium and Potassium 23
2.7.2 Calcium and
Phosphorus 24
2.7.3 Iron 24
2.7.4 Zinc 24
2.8 Serum Electrolytes 25
2.8.1 Chloride 25
2.8.2 Bicarbonate (HCO3) 25
2.8.3 Potassium (K+) 25
2.9 Haematological Profile 26
2.9.1 Red blood cells (RBC) 26
2.9.2 White blood cells
(WBC) 26
2.9.3 Packed cell volume
(PCV) 27
2.9.4 Hemoglobin (Hb) 27
2.9.5 Platelet count 28
2.9.6 Mean corpuscular volume
(MCV) 29
2.9.7 Mean corpuscular hemoglobin (MCH) 29
2.9.8 Mean corpuscular
hemoglobin concentration (MCHC) 29
2.10 Antioxidant Parameters 30
2.10.1 Superoxide dismutase 30
2.10.2 Catalase (CAT) 31
2.10.3 Glutathione (GSH) 31
CHAPTER 3: MATERIALS AND METHODS
3.1
Collection of Plant Materials 33
3.2
Extraction 33
3.3
Animals 33
3.4
Reagents and Apparatii Used 34
3.5
Experimental Design 35
3.6
Treatment 35
3.7
Proximate Content Determination 36
3.7.1
Moisture content 36
3.7.2
Ash content 36
3.7.3
Crude fibre 37
3.7.4
Fat content 38
3.7.5
Protein content 38
3.7.6
Carbohydrate 40
3.8
Determination of Minerals 40
3.8.1
Sodium and potassium contents 40
3.8.2
Calcium and magnesium 41
3.8.3
Extraction of iron and zinc 42
3.9
Determination of Vitamins 43
3.9.1
Vitamin A 43
3.9.2
Vitamin B1 44
3.9.3
Vitamin C (Ascorbic Acid) 44
3.9.4
Determination of Vitamin E 45
3.10
Determination of Nutraceutical Properties 46
3.10.1
Saponin 46
3.10.2
Alkaloids 47
3.10.3
Flavonoids 47
3.10.4
Tannins 48
3.10.5
Phenols 49
3.11
Assay of Biochemistry Parameters 50
3.11.1
Quantitative estimation of serum alanine transaminase (ALT)
and aspartate transaminase (AST) 50
3.11.1.1
Procedure for determination of ALT and AST 51
3.11.2
Quantitative estimation of serum alkaline phosphatase (ALP) 52
3.11.2.1
Procedure for determination of serum alkaline phosphatase (ALP) 52
3.11.3
Quantitative estimation of serum total bilirubin 53
3.11.3.1
Procedure for determination of serum total bilirubin 53
3.11.4
Procedure for determination of serum total protein 54
3.12
Determination of Some Antioxidant Parameters 55
3.12.1
Catalase 55
3.12.1.1
Procedure for determination of catalase 56
3.12.2
Superoxide Dismutase (SOD) 56
3.12.2.1
Procedure for determination of superoxide dismutase (SOD) 56
3.12.3
Determination of reduced glutathione 57
3.12.3.1
Procedure for determination of reduced glutathione 57
3.13
Determination of Haematological Indices 58
3.13.1
Determination of blood cells count (WBC and RBC)
3.13.1.1
Procedure for the determination of blood cells count (WBC and RBC) 58
3.13.2
Packed cell volume (Packed Cell Volume) 59
3.13.2.1
Procedure for the determination of packed cell volume (PCV) 59
3.13.3
Procedure for the determination of hemoglobin 59
3.13.4
Procedure for the determination of mean cell volume (MCV) 60
3.13.5
Procedure for the determination of mean cell haemoglobin (MCH) 60
3.13.6
Procedure for the determination of mean cell hemoglobin concentration (MCHC) 60
3.14
Determination of Serum Electrolytes 59
3.14.1
Potassium ion 59
3.14.1.1
Procedure for the determination of potassium ion 60
3.14.2
Chloride ions 60
3.14.2.1
Procedure for the determination of chloride ions 61
3.14.3 Statistical analysis 61
CHAPTER 4
4.1 Results 62
4.2 Discussion 75
CHAPTER 5
5.1 Conclusion 83
5.2 Recommendation 83
REFERENCES 84
APPENDIX 98
LIST OF TABLES
4.1: Proximate composition of toasted Mucuna sloanei seeds
4.2: Mineral contents of toasted Mucuna sloanei seeds
4.3: Some vitamin contents of toasted Mucuna sloanei seeds
4.4: Some phytochemical components of
toasted Mucuna sloanei seeds
4.5: Kidney parameters and
electrolytes of all the groups
4.6: Liver parameters of all the
groups
4.7: Haematological parameters of all
the groups
4.8: Antioxidant parameters of all the
groups
LIST OF FIGURES
2.1:
Mucuna sloanei seeds
2.2: Mucuna sloanei seed endosperm after removing their shells
2.3: Mucuna
sloanei pods hanging in their natural habitat
2.4: Picture of Mucuna
sloanei pods containing seeds
2.5:
Monosodium glutamate crystals
LIST OF PLATES
4.1: Histology of the control group
liver
4.2: Histology of the MSG only group
liver
4.3: Histology of the extract only
group liver
4.4: Histology of the MSG and 200mg/kg
extract group liver
4.5: Histology of the MSG and 400mg/kg
extract group liver
4.6: Histology of the MSG and 800mg/kg
extract group liver
4.7: Histology of the control group
kidney
4.8: Histology of the MSG only group
kidney
4.9: Histology of the extract only
group kidney
4.10: Histology of the MSG and
200mg/kg extract group kidney
4.11: Histology of the MSG and
400mg/kg extract group kidney
4.12: Histology of the MSG and
800mg/kg extract group kidney
CHAPTER 1
INTRODUCTION
1.1 MEDICINAL PLANTS
Plants
with medicinal properties have been in use since the existence of man as both
food and medicine because they produce many chemical compounds that are used
for treating man and animals. Chemical compounds in these plants have effects
that resemble those seen with conventional drugs. They are able to exert such
effects on man and animals because they contain bioactive constituents such as
steroids, phenols, terpenoids, carotenoids, flavonoids, alkaloids, tannins and
glycosides in their parts such as the leaves, barks, roots, fruits and seeds which
can be gotten through processing. (Shad et
al., 2014; Dhan et al., 2011) Medicinal
plants are directly used as medicines by most world cultures for example, the
Chinese and Indian medicinal practitioners. (Bhushan et
al., 2006) These
plants include ginger, green tea, walnuts, garlic, neem, bitter leaf, bitter
kola, turmeric etc. (Si-Yuan et al.,2013)
which are mainly used as raw materials for drugs since they serve as active
ingredients used in the synthesis of useful drugs such as the antibiotics,
antifungals and antimalarials that are used against pathogenic infections.
The
efficacies of these plants came to human notice by observing how sick animals utilize
them to treat themselves. (Ekor, 2013). Today, scientific methods are employed to
isolate active compounds from medicinal plants. (Madubuike et al., 2016; Igwe et al.,
2016)
Food additives
Food
additives such as monosodium glutamate (MSG) commonly called white maggi are
added to foods and food spices to enhance their taste. These chemicals when in
large quantities elicit free radicals in the body which can be produced in
excess beyond the body’s capability thus, leading to cell, tissue or organ
damage due to oxidation of biomolecules seen in these cells which lead to
degenerative symptoms that are manifested. (Aiyegoro
and Okoh 2010; Sen et al., 2010).
Mucuna sloanei
Mucuna sloanei a medicinal plant which
must be processed before it can be eaten as vegetable after soaking its leaves
in water to leach out its L-dihydroxyphenylalanine (L-dopa) content or as a
soup thickener after toasting or boiling the seeds to denature the anti-nutritive
substances embedded in them. It is used traditionally in various parts of the world for the
treatment of various ailments such as diarrhea, hemorrhoids,
(Oladipo et al., 2013; Burkill, 1995)
hernia and treatment of filaria worms infestation (Quattrocchi, 2000;
Waryekeche et al., 2003)
1.2 STATEMENT OF PROBLEM
Over
the years, the Foods and Drugs Administration Agencies have received many complains
of reactions to foods with monosodium glutamate (MSG). These reactions, known as
MSG symptom complex, include headache, sweating, facial pressure or tightness, skin
insensitivity, tingling of the body, shaky hands known as Chinese restaurant
syndrome, rapid fluttering heartbeats (heart palpitations), chest pain, nausea
and weakness. So far, no known remedy is in place to prevent these adverse
effects of MSG without losing its exquisite flavouring property.
1.3
JUSTIFICATION OF THE STUDY
Mucuna
sloanei seed endosperms are used in the preparation of food
as vegetables when immature and as soup thickeners when mature and dried, they
also possess medicinal properties while monosodium glutamate (MSG) is added
to majority of the seasonings sold in the market are often abused by commercial
cooks and in private homes in attempt to improve food taste. This led to side
effects such as Chinese restaurant syndrome and
atremulous effects
acclaimed to be experienced by the regular consumers in foods. Thus the need to evaluate other food additives for their
potentials in ameliorating the effects of MSG.
This study therefore wants to evaluate the nutritional properties
of Mucuna sloanei seed endosperms and
selected biochemical effects of its ethanol extract in monosodium glutamate-intoxicated
rats.
1.4.1 AIM
OF THE STUDY
To
evaluate the nutritional properties of Mucuna
sloanei seed endosperm and some biochemical effects of its ethanol extract
in monosodium glutamate-intoxicated rats.
1.4.2 OBJECTIVES OF THE STUDY
(i)
Determination of the Proximate content including vitamin
contents, mineral contents and phytochemical contents of the Mucuna sloanei seed endosperm.
(ii)
Effects of the ethanol extract of Mucuna sloanei seed endosperm on the
(a) Biochemical biomarkers of normal and
MSG-intoxicated rats.
(b) Kidney function markers of normal
and MSG-intoxicated rats.
(c) Hematological parameters of normal
and MSG-intoxicated rats.
(d) Antioxidant parameters (catalase,
Superoxide dismutase, Glutathione) of normal and MSG-intoxicated rats.
(e) Histology of the kidney and liver of
normal and MSG-intoxicated rats.
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