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ABSTRACT

In this present work, the phytochemical constitution of Borreria stachydea , Cassia absus and Aspillia Kotschyi plants belonging to rubiacea , leguminosae and Compositae families respectively were investigated. The three plants methanolic and petroleum ether extracts were analyzed for alkaloids, tannins, saponins, steroids, terpenoids, flavonoids, glycosides and phenolic compounds. The study results showed that the petroleum ether extract of the three plants contained glycosides, steroids carbohydrates and alkaloid while the methanolic extracts of the three plants revealed the presence of carbohydrates, glycosides, cardiac glycosides, saponins, and flavonoids, triterpene, and alkaloids. Tannins were also observed to be present in the methanolic extract of Borreria stachydea only. The proximate analysis of the two plant materials Borreria stachydea and Cassia absus (whole) showed moisture content of 8.67 % and 6.67 % and ash content of 8.5 % and 8.0 % in Borreria stachydea and Cassia absus respectively. The crude protein was 8.08 % and 9.81 %, crude fibre 7.11 % and 3.17 %, crude fat 2.15 % and 2.11 %, and Nitrogen Free Extract (NFE) 65.59 % and 70.36 % in Borreria stachydea and Cassia absus methanolic extracts respectively. The petroleum spirit extracts of Borreria stachydea and Cassia absus showed crude protein of 5.63 % and 4.88 %, crude fibre 0.11 % and 2.65 %, crude fat 3.13 % and 1.94 % and NFE 89.27 % and 88.39 % in the two plants respectively On the other hand Aspilla kotschyi Proximate composition analysis showed moisture content of 5.7%, total ash of 4.03% ,crude protein 10.94%, fibre 9.06%,Fat value 0.83% and Nitrogen free extract of 70.19%. This study also assesses the presence of Calcium, Magnesium, Zinc, Lead, Chromium, Cadmium, in the three plants using AAS in the plants extracts average elemental concentrations for Borreria stachydea were 0.4272, 0.42625, 0.0193, 0.0003, 0.1425 and 0.0008 mg/Kg for Ca, Mg, Zn, Pb, Cr, and Cd respectively, while those for Cassia absus were 0.5093, 0.6423, 0.0320, 0.0010, 0.1616 and 0.0028 mg/Kg for Ca, Mg, Zn, Pb, Cr, and Cd respectively. Aspilla kotschyi extract showed the following elemental concentrations Calcium 0.3099, Magnesium 0.1380, Zinc 0.2449,Lead 0.0020,Chromium 0.0018, and Cadmium 0.0026. Other metals ; Se, Ag, Fe, Cu, Ni, As, Co, Mn and Al were determined from both the methanolic and petroleum spirit extracts of the three plants using micro wave plasma atomic emission spectroscopy technique. From the result, Silver, Copper, Nickel and Cobalt are of very negligible
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concentrations in all the three plants, however Seleniun is found to be 0.780 (mg/kg) in Casssia absus methanolic extract , 0.556 (mg/kg) in Borreria stachydea methanolic extract, and 0.530 (mg/kg) in Aspillia kotschyi methanolic extract. Iron on the other hand was found to be 3.712 (mg/kg), in Aspillia Kotschyi metahanolic extract, while in Borreria stachydea petroleum spirit extract was 3.674 (mg/kg) . Arsenic was found to be 0.506 and 1.301 (mg/kg) in Aspillia Kotschyi extracts, 2.373(mg/kg) and 0.789(mg/kg) in Borreria Stachydea extracts, and in Cassia absus extract was found to be 3.315 (mg/kg). The concentration of Aluminium was found to be of 3.050(mg/kg) in Aspillia extract and 2.99(mg/kg) in Borreria stachydea extract. Sodium and potassium were determined in the three plants extracts using flame photometry. Sodium was found to be 50, 78 and 99 mg/l in Borreria stachydea, Cassia absus and Aspillia kotschyi respectively. The potassium level in the three plants were 870,860 and 895 mg/l respectively. Functional group analysis of the three plants extracts were also carried out using Fourier Transform Infrared Spectroscopy (FTIR) after the extract has been purified by column chromatography. The functional groups determined in the three extracts showed that the plant samples contained the following functional groups; O-H Alcohols, phenols , C-H for Alkanes , C=O Unsaturated aldehydes and ketones and C=O Aliphatic amines. The results of this study suggest some merit in the popular use of these plants in herbal medicine as these functional groups are known to have therapeutic effectiveness against wide array of microorganis

TABLE OF CONTENTS

Title page……………………………………………………………………………1
Declaration…………………………………………………………………………ii
Certification……………………………………………………………………………..iii
Dedication………………………………………………………………………….iv
Acknowledgement………………………………………………………………….v
Table of Contents …………………………………………………………………vi
Abstract……………………………………………………………………………vi
Table of Contents………………………………………………………………….vii
List of Tables………………………………………………………………. …….xiii
List of Figures…………………………………………………………………… .xv
List of Plates………………………………………………………………………xvi
List of Appendices………………………………………………………………..xvii
List of Abbreviations………………………………………………………………xvii
Abstract……………………………………………………………………………xix
CHAPTER ONE
1.0 Introduction…………………………………………………………..1
1.1 Mineral Elements ……………………………………………….…..……7
1.2.0 Proximate Analysis………………………………………………………..7
1.3 Borreria stachydea [(Dc)Hutch &Dalziel]…………………………………9
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1.4 Cassia absus Linn…………………………………………………………9
1.5 Aspilla kotschi …………………………………………………. ………….9
1.7 Statement of Problem…………………………………… …… …………10
1.8 Aims of the Research………………………………………………………10
1.9 Objectives of the Research…………………………………………………11
1.10 Justification of the Research……………………………………………….12
CHAPTER TWO
2.0 Literature Review…………………………………………………………….13
2.1 Description of Borreria stachydea …………………………………………………13
2.1.1 Brief Taxonomy of Borreria stachydea………………………………………………….13
2.1.2 Local names of Borreria stachydea………………………………………..13
2.2 Description of Cassia absus Linn…………………………………………….17
2.2.1 Brief Taxonomy of Cassia absus Linn……………………………………..18
2.2.2 Local names Cassia absus Linn…………………………………………….18
2.3 Description of Aspillia kotschyi(sch.Bipex,Hochst)Oliv ………………………19
2.3.1 Brief Taxonomy of Aspillia kotschyi………………………………………..21
2.3.2 Local names of Aspillia kotschyi……………………………………………..21
2.4.0 Proximate Analysis………………………………………………………….21
2.4.1 Nitrogen and nitrogenous constituents……………………………………..23
2.4.2 Total Nitrogen………………………………………………………………23
2.4. 3 Total Fat…………………………………………………………………….24
2.4.4 Fat (-acylglycerols)…………………………………………………………..24
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2.4.5 Protein……………………………………………………………………….27
2.4.6 Carbohydrates……………………………………………………………….29
2.4.7 Total ash…………………………………………..…………………………30
2.4.8 Dietary fibre………………………………………………………………….31
2.5.0 Mineral Elements……………………………………………………………33
2.5.1 Classification of Minerals………………………………..…………………..34
2.5.2 The importance of the mineral elements……………….…………….……..35
2.5.3 Biochemical functions of mineral elements in humans and animals……….36
2.5.4 Calcium …………………………………………………………………….37
2.5.5 Zinc…………………………………………………………………….…..37
2.5.6 Lead………………………………………………………………………….38
2.5.7.Manganese …………………………………………………………………39
2.5.8 Copper …………………………………………………………………….40
2.5.9 Chromium………………………………………………………………….41
2.5.10 Selemium………………………………………………………………….42
2.5.11 Cadmium…………………………………………………………………….43
2.5.11.1 Cadmium Toxicity………………………………………………………45
2.5.12 Cobalt………………………………………………………………………45
2.5.13 Iron…………………………………………………………………………47
2.6.0 FTIR Spectroscopy for Material Identification…………………………….48
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2.6.1 Valuable Analysis with FTIR Spectroscopy…………………………………..48
2.6.2 Analyzing Samples in a Variety of Forms…………………………………….49
2.6.3 Advantages of FTIR………………………………………………………………49
CHAPTER THREE
3.0.0 Materials and Method…………………………………………………………51
3.1.0: Sample Collection and Identification…………………………………………51
3.2.0 : Sample Treatment……………………………………………………………53
3.3.0 : Extraction of Plant Materials………………………………………………..53
3.3.1 Materials for phytochemical analysis………………………………………..53
3.3.2 Preliminary phytochemical screening……………………………………….55
3.3. 2.1 General test for alkaloids………………………………………………….55
3.3.2.2 Extraction of Alkaloids from the Plant Material…………………………55
3.3.2.3 Test for Alkaloids in the Extracts………………………………………….56
3.3.2.4 Test for alkaloids in the Extract of Borreria Stachydea, Cassia absus Linn, and Aspillia kotschyi plants……………………………………………………………………..56
3.3.3 Test for tannins……………………………………………………………..56
3.3.4 Test for glycosides…………………………………………………………..57
3.3.5 Test for saponins…………………………………………………………….57
3.3.6 Test for cardiac glycosides…………………………………………………..58
3.3.7 Test for steroids/triterpenes…………………………………………………58
3.3.7.1 Salkoski Test …………………………………………………………..58
3.3.7.2 Keller killiani Test…………………………………………………………59
3.3.8 Test for carbohydrates……………………………………………………….59
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3.3.8.1 Fehlings Test ……………………………………………….……………..59
3.3.8.2 Molisch Test …………………………………………………..……………59
3.3.9 Test for flavonoids……………………………………………………………..59
3.3.9.1 Shinoda Test…………………………………………………………………59
3.3.9.2 Sodium hydroxide Test………………………………………………………60
3.3.10 Test for anthra quinines……………….……………………………………..60
3.3.11 Test for tannins………………………………………………………………60
3.3.12 Chemical Test for fixed oils and fats………………………………………..60
3.4.0 Determination of Anti nutritional Components……………………………….61
3.4.1 Determination of total phenols by spectrophotometric methods…………… 61
3.4.2 Determination of Oxalate…………………………………………………… 61
3.4.3 Determination of Crude hydrogen cyanide……………………………………62
3.4.4 Determination of Phytates…………………………………………………….62
3.5.0 Determination of nutritional content………………………………………….62
3.5.1 Determination of moisture content……………………………………..……..63
3.5.2 Determination of Ash content………………….……………………..….……63
3.5.3 Determination of Crude Protein………………..……………………….……..63
3.5.4 Determination of Crude Fibre………………..…………………………..……64
3.5.5 Determination of Crude Fat……………………………………………….…..64
3.5.6 Determination of Free nitrogen extract (NFE) (Dutcher et al, 1951) …………65
3.6.0 Determination of Mineral Content………………………………………………65
3.6.1 Determination of other Minerals Using EAS…………………………………. 66
3.6.1.1 Experimental Parameters for MP-EAS……………………………………….68
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3.6.1.2 Blank and Standards Preparation……………………………………………..,………..68
3.6.1.3 Ultrasonic Nebulizer (USN) Setup, Calibration, and Instrument Detection Limits…….69
3.7.0 Determination of Sodium and Potassium using Flame Photometry………………………70
3.8.0 Thin layer chromatography………………………………………………………………..70
3.8.1 Chromatographic Separation………………………………………………………………71
3.9.0 Functional Group Analysis using Fourier Transform Infrared Spectroscopy
( FTIR)…………………………………………………………………………….…………72
CHAPTER FOUR
4.0 RESULTS…………………………………………………………………………………..74
4.1 Yields of crude extracts……………………………………………………………………..74
4.2 Preliminary phytochemical screening of methanolic and petroleum spirit extracts of Borreria stachydea, Cassia absus , and Aspillia kotschyi Plants………………………………..74
4.3 Quantitive phytochemical screening of methanolic extracts of Borreria stachydea, Cassia absus , and Aspillia kotschyi Plants………………………………………………………………78
4.4 Proximate composition of Methanolic extracts of Borreria stachydea, Cassia absus , and Aspillia kotschyi Plants…………………………………………………………………………..79
4.5 Elemental Analysis of Borreria stachydea, Cassia absus and Aspillia kotschyi methanolic and petroleum spirit extracts using Atomic Absorption spectrometry………………………….81
4.6 Determination of other minerals in Borreria stachydea, Cassia absus and Aspillia kotschyi methanolic extract using Micro Wave Plasma Atomic Emission Spectroscopy technique ( MP- EAS)……………………………………………………………………………………………..864.7 Determination of Sodium and Potassium using Flame Photometry………………………..91
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4.8 Determination of Anti nutritional factors in Borreria stachydea, Cassia absus , and Aspillia kotschyi Plant………………………………………………………………………………….…92
4.9 Determination of nutritional factors in Borreria stachydea, Cassia absus , and Aspillia kotschyi Plants……………………………………………………………………………………93
4.10 Column Chromatography ………………………………………………………………….95
4.11 Functional Group Analysis using Fourier Transform Infrared Spectroscopy (FTIR)…………………………………………………………………………………………….96
CHAPTER FIVE
5.0 Yields of crude extracts…………………………………………………………………….106
5.1 Preliminary phytochemical screening of methanolic and petroleum spirit extracts of Borreria stachydea, Cassia absus , and Aspillia kotschyi Plants………………………………106
5.1.1 Phytochemical screening of methanolic and petroleum spirit extracts of Borreria stachydea, Cassia absus ………………………………………………………………………..107
5.1.2 Phytochemical screening of methanolic and petroleum spirit extracts of Aspillia kotschyi plant …………………………………………………………………………………………….108
5.1.3 Quantitaive phytochemical screening of methanolic extracts of Borreria stachydea, Cassia absus , and Aspillia kotschyi Plants…………………………………………………….108
5.2 Proximate composition of methanolic extracts of Borreria stachydea, Cassia absus , and Aspillia kotschyi Plants………………………………………………………………………….110
5.3 Elemental Analysis of Borreria stachydea, Cassia absus and Aspillia kotschyi methanolic and petroleum spirit extracts using Atomic Absorption spectrometry…………………………110
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5.4 Determination of other minerals in Borreria stachydea, Cassia absus and Aspillia kotschyi methanolic extract using Micro Wave Plasma Atomic Emission Spectroscopy technique ( MP- EAS)…………………………………………………………………………………………….111
5.5 Sodium and Potassium contents in the analyzed plants using Flame Photometry………….113
5.6 Anti nutritional factors of the analyzed plants Borreria stachydea, Cassia absus , and Aspillia kotschyi Plant……………………………………………………………………………………114
5.7 Vitamin content in the analyzed plants (Borreria stachydea, Cassia absus , and Aspillia kotschyi )………………………………………………………………………………………..117
5.8 Functional Group Analysis using Fourier Transform Infrared Spectroscopy( FTIR)………117
5.9 Conclusion …………………………………………………………………………………119
5.10 Recomendation……………………………………………………………………………120
References: ………………………………………………………. ……………………………122 Appendix……………………………………………………………………………………..…140

CHAPTER ONE

Project Topics

1.0 Introduction
Plants in general, contribute to the energy/calorie, mineral, vitamin and fibre contents of diets. Among plants, vegetables are excellent sources of minerals and contribute to the recommended dietary allowance (RDA) of the essential nutrients. Minerals are very important ingredients for normal metabolic activities of body tissues. They are constituents of bones, teeth, blood, muscles, hair and nerve cells. Vitamins cannot be properly assimilated without the correct balance of minerals (Sonni, 2002). Plants have occupied very important position in human life for a very long time. They provide food, medicine, fibre and fodder for man and his animals. A variety of plants have promising nutritive value which could nourish the ever increasing human population. They however remain underutilized due to lack of awareness and technologies for their utilization.
Currently, there is a gradual revival of interest in the use of medicinal plants in developing countries because herbal medicines have been reported safe and without any adverse side effect, especially when compared with synthetic drugs. Thus, the search for new drugs with better and cheaper substitutes from plant origin is a natural choice. The medicinal value of these plants lies in some chemical substances that induce a definite physiological action in the human body (Edeoga et al., 2005).
In Nigeria, numerous edible wild plants are exploited as sources of food, which provide adequate levels of nutrients for inhabitants. Some edible plants are also used in traditional medicine, some of these herbs are indigenous to Africa, America and India. The use of herbs is common element in ayurvedic, homoeopathic, naturopathic traditional oriented medicine
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(Ogundipe, 2008). The pharmacological actions of crude drugs are determined by the nature of their constituents (Mukherjee, 2002).
Plants based drugs have been used against various diseases since time immemorial. The primitive man use herbs as therapeutic agents and medicaments, which they were able to produce easily. Nature has provided abundant plant wealth for all living creatures, which possess medicinal virtues. The most important values of some plants have been published in many books and journals but a large number of them remain unexplored. So there is need to explore their uses and conduct pharmacognostic and pharmacologicacal studies to ascertain their therapeutic properties (Mushtaq,et al., 2009).
Traditional medicine as a major African socio cultural heritage, obviously in existence several hundred years, was once believed to be primitive and wrongly challenged with animosity, especially by the conventional or orthodox medical practioners. However today traditional medicine have been brought into focus for meeting the goals of a wider coverage of primary health care delivery, not only in Africa but also to various extents, in all countries of the world. Traditional medicine is the first choice health –care treatment for at least 80% of Africans who suffer from high fever and other common ailments(Eliyoba et al., 2005).
Each medicinal plant species has its own nutrient composition besides having pharmacologically important phytochemicals. These nutrients are essential for the physiological function of human body. Such nutrients and biochemical‟s like carbohydrates, fats, and proteins play important roles in satisfying human needs for energy and life processes (Novac and Hasiber, 2000).
Plants generally contain compounds (such as saponins, tannins, oxalates, phylates, trypsin inhibitors and cynogenic glycosides) known as secondary metabolites which are biologically active (Soetan and Oyewale, 2009). Secondary metabolites may be applied in nutrition and as
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pharmacologically active agents (Soetan and Oyewale, 2009).Plants are also known to have high amounts of essential nutrients, vitamins, minerals, fatty acids and fibre(Gafar and Itodo, 2011).
Phytochemical screening of plants materials makes detailed pharmacological and other academic studies possible as it offers information of a particular chemical compound responsible for some said activities in a given plant (Sofowora, 1982).
The phytochemical investigation of a plant may thus involve the following: extraction of the plant material, separation and isolation of the constituents of interest, characterization of the biosynthetic pathways to particular compounds and quantitative evaluation (Trease and Evans, 2000).
Not all the chemical compounds found in plants are of equal interest to the pharmacognosist. The so called “active principles” are frequently alkaloids or glycosides and these therefore deserve special attention. Other groups such as carbohydrates, fats and proteins are of dietic importance, and many others such as starches and gums are used in pharmacy but lack any marked pharmacological action. Other substances such as calcium oxalate, silica, lignin and coloring matters may be of assistance in the identification of drugs and the detection of adulteration (Trease and Evans, 2000).
Results from phytochemical analysis have shown that the general classes of compounds found in plants which are of medicinal interest include alkaloid, cardiac glycosides, saponins, tannins and anthraquinones (Sofowora, 1982).
Plant materials contain several chemicals which act against diseases and infections of human and animals when properly used. Plants contain different types of compounds such as resins, rubbers,
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gums, waxes, dyes, flavours, fragrances, proteins, amino acid, bioactive peptides, sugars, flavonoids and biopesticides (Muhammed et al., 2006).
Today according to the World Health Organization (WHO) as many as 80% of the world people depend on traditional medicine for their primary health care needs (Leila et al., 2011). High plants are sources of drugs, which have made important contribution to the welfare and quality of life in the urban as well as rural communities, especially in tropics and subtropics (Sofowora,1993).
In the ancient days of human existence, by instinct, intuition, or trial and error, man was able to identify various plants used to combat various ailments. In fact, it is the knowledge derived from the active components of these plant extracts that guided man to synthesize and use modern drugs in health care delivery. Presently, there is a renewed interest in the study of medicinal plants such that much percentage of pharmaceutical preparations are based on natural products from plants. Over the years, people have passed down knowledge of the types and applications of medicinal plants from generation to generation, often orally. The compilation of useful drugs derived from medicinal plant is impressive; these include; heart drugs, analgesics, anesthetics, antibiotics, anti-cancer and anti-parasitic compounds, anti-inflammatory drugs, oral contraceptic hormones, as well as laxative diuretics (Morris, 2004).
Available medical literature revealed that plants have been used for curative purposes (Ming,1999). There are many plants whose green leaves, roots and stems are used in herbal preparations for the treatment of various ailments(Ekop, 2007).Most of the times the potency of those plants is accounted for in terms of their organic constituents but, it is an established fact that there is a high relationship between the chelation of metals and some chemotherapeutic agents. The role of inorganic elements in animal and plant metabolism has long been established
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but the effect and an influence of these elements on administration of medicinal plants has received relatively little attention (Gwarzo et al., 2006).
It is a common knowledge that administration of plants claimed to have medicinal properties has largely been indiscrimate without due regard to possible side effects. It is thus important to determine the chemical content of these plants for pharmacological assays and other probable roles in the curative process. In addition to that, plants are generally the major sources of food and minerals to both man and animals. The different species of plants play important roles in the diet of inhabitants of arid and semi –arid savanna countries of Africa. They contain minerals in quantities compared favorably with World Health Organization (W.H.O.) standard (Baminas et al., 1998).
From the environmental, pollution and toxicological point of view, many minerals including those that are essential to life can be toxic at high doses or in certain compound formulation. So the amount of minerals in living organism normally correlate significantly with amounts in the environment. Also, heavy metals do not degrade but accumulate in food and are a serious threat to human health if consumed. In fact, there are certain plants known to have special ability which enable them to take elements with the tendencies of accumulating same to dangerous levels, thus making the plants toxic (WHO, 1980, IRI 1989, Szefer and Szefer 1994 and Last, 1995). Likewise it is a known fact that different parts of plants contain trace elements that are harmful to the body of man at a certain concentrations (Sani, 2014).
Sewage from homes, industrial effluents into aquatic environment, smoke from industries and vehicles,parents materials from which the soil is formed; types of agrochemicals used in farming and waste disposal systems all contribute to the elemental composition of plants.
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It is estimated that today, plants materials are present in or have provided the models for 50% of industrially manufactured drugs (Iwu,1994). Therefore there is greater tendency for the accumulation of a particular element as suggested by Kovac (1979) and Whitton (1989) and the extent of accumulation is a reflection of the concentration of such materials in the environment as well as species of the plant in use. It is an established fact that metals in particular are present in both plants and animals as components of simple salt and complexes performing various functions (Yalwa, 2002).They are required by plants in amounts that result in their classification as essential, beneficial and non-essential for proper growth and development (Daniel, 1990).
It is a common knowledge that administration of plants claimed to have medicinal properties has largely been indiscriminate without due regard to possible side effects, or long term accumulation of toxic compounds in the plants tissue which may accumulate and became harmful to human tissues. It is therefore important to determine the chemical content of these plants for pharmacological assays and other probable roles in the curative process.
In recent times focus on plant research has increased all over the world and a large body of evidence has accumulated to show immense potential of medicinal plants used in various pharmaceutical, cosmetic, agrochemical applications. Plants have been the subject of human curiosity and use for thousand of years ( Ram et al., 2004). These plants have played important roles in many centuries by providing food, shelter, clothing, agrochemicals, flavours and fragrances and more importantly medicines ( Gurib- Fakim, 2006).
1.1 Mineral elements
Minerals are inorganic substances, present in all body tissues and fluids and their presence is necessary for the maintenance of certain physicochemical processes which are essential to life. Minerals are chemical constituents used by the body in many ways. Although they yield no
7
energy, they have important roles to play in many activities in the body (Malhotra, 1998; Eruvbetine, 2003).
In addition, plants are the major sources of food and minerals to both man and animals. The different species of plant play important roles in the diet of the man and his animals. They contain minerals in quantities compared favorably with foods of World Health Organization (W.H.O.) standard (Barminas et al., 1998). Of particular importance among the parts of the plant are the leaves and the seeds containing trace elements at varying levels depending on many factors such as species, variety, stage of maturity and environment (Daun and Mc Gregor, 1991).
Moreover metals and in particular trace metals are major class contaminants in our present world arising principally from natural and anthropogenic sources. The anthropogenic sources are primarily from industrialization and mining activities which are paramount importance to ecosystem sustainability (Okorie and Egila, 2012).
1.2.0 Proximate Analysis
A method was developed for quantitative analysis of the different macronutrient in plant samples and feed known as proximate analysis and the method was developed in the year 1860 by Henneberg and Stohmann in Germany (Greenfield and Southgate, 2003).
Proximate Analysis is a partitioning of compounds in a feed into six categories based on the chemical properties of the compound. The six categories are;
i) Moisture
ii) Ash
iii) Crude protein (or Kjeldahl protein )
iv) Crude lipid
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v) Crude fibre
vi) Nitrogen free extract (digestible carbohydrates)
In the proximate analysis of feed stuff‟s kjedahl nitrogen, ether extract, crude fibre and ash are determined chemically.
The determination of Nitrogen allows the calculation of the protein content of the sample. It is important to remember that proximate analysis is not a nutrient analysis rather it is a partitioning of both nutrients and non nutrients into categories based on common chemical properties.
Proximate and nutrient analyses of edible plant and vegetables play a crucial role in assessing their nutritional significance (Pandey et al., 2006). As various medicinal plant species are also used as food along with their medicinal benefits, evaluating their nutritional significance can help to understand the worth of these plant species (Pandey et al., 2006) For herbal drug‟s standardization, WHO also emphasize on the need and importance of determining proximate and micronutrients composition of the herbal plants. Such herbal formulations must pass through standardization processes (Niranjan and Kanaki, 2008).
1.3 Borreria stachydea [(Dc)Hutch &Dalziel]
This plant belongs to the phylum magnoliophyta and its class is manoliopsida and a member of the Rubiaceae family. It is found in Nigeria, Ghana, Sudan, Malaysia, India and several other nations of the world. A poultice of the whole plant is used to heal leg ulcer, wounds, urinary tract infections (Neoh, 2010). In Northern Nigeria the Hausa popularly know and call Borreria stachydea as “alkamar tururuwa” while Fulanis called it “fairare”. The plant is used in Northern
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Nigeria medicinally for urinary troubles, gonorrhoea ,etc and for women in childbirth, or to regulate the menses (www.wikipedia) June 2014.
1.4 Cassia absus Linn
This plant belongs to the family leguminosae and sub family caesalpinonodeae. It is found in tropical Asia, Australia, and Africa .It is a useful medicinal plant. It is regarded as a blood tonic, bitter astringent for the bowels and applied locally to heal ulcers (Hussain, et al, 2008). It is useful in the disease of eyes such as purulent conjunctivitis and opthalmia ( Ghani, et al, 1997). Cassia abus Linn is popularly known as “fidili” in Hausa and in English is called “jasmeejaz”
1.5 Aspilla kotschyi
Aspilla kotschyi belongs to the family compositae which is also known as Asteracea. The common names of this plant in Nigeria are “Ja majina” (draw up mucus in hausa) or “Jinin barewa”. Historically Aspillia africana was used in Mbaise and most Igbo speaking parts of Nigeria to prevent conception suggesting potential contraceptive and anti-fertility properties (Oluyemi, et al., 2007).
1.7 Statement of Problem
The continuous increase in the use of plants in traditional medicine has become a cultural heritage for several hundred years in many developing nations including Nigeria. A knowledge of the available metabolites, nutrients in these plants sources and the establishment of the presence of the level of toxic anti nutritional substances in the plants will help to determine the suitability of using these plants as remedies for various diseases.
This study underlook the task of determining the nutrient composition, the secondary metabolites and anti nutritional substances in three locally used medicinal plants in Nigeria namely;
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Borreria stachydea, Cassia absus and Aspillia kotschyi plants. The study highlighted the presence and suitability of using these plants locally as remedies for various diseases and sources of food.
1.8 Aims of the Research
The study aimed at achieving the following:
i. Phytochemical screening of the various extracts of the three different plants, Borreria stachydea {(D C Hutch & Daziel}, Cassia absus Linn and Aspillia kotschyi plants to determine the bioactive agents responsible for their physiological activity or otherwise.
ii. Evaluation of the nutritive components (Proteins, Lipids, carbohydrates, fibre and vitamins) and anti nutritive components (Cyanides, tannins, oxalates and phytates) in Borreria stachydea {(D C Hutch & Daziel},Cassia absus Linn and Aspilia kotschyi plants.
ii. Determine qualitatively and quantitatively the major, minor, trace and ultra trace elements in the above listed plants and
iii. Make comparative analysis, and come out with appropriate recommendations on their toxicity or otherwise.
The above aims are individually of special problem-solving relevance. They are also complimentary in helping to provide the much needed information on the scientific data of the plants extracts of Borreria stachydea {(DC Hutch & Daziel},Cassia absus Linn and Aspilia kotschyi which may find use as patentable and industrially exploitable compounds for drugs development, food and cosmetics industries among others..
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1.9 Objectives of the Research
The objectives of the research are as follows:
i. Collection of the three plant samples
ii. Proper botanical identification of the plant at the herbarium
iii. Air drying, segregating and pulverizing the plants.
iv. Extracting using appropriate solvent of the pulverized plant parts
v. Phytochemical screening of the extracts of the three different plants
vi. Conducting proximate analysis of the three plants
vii. Determination of anti nutritive factors in the three plants
viii. Determination of major, minor, trace & ultra trace elements from the three different plants
ix. Purifying the three plants extract
x. Determination of Functional groups present in the three plants by Fourier Transform Infrared (FTIR) spectroscopy.
xi. Conducting statistical analysis on the results obtained so as to draw conclusions and recommendations on the use of the three plants.
1.10 Justification of the Research
Right from time immemorial, wild plants have occupied very important position in human life. They provide food, medicine, fibre and fodder for domestic animals. So many plants of promising nutritive values, which could nourish the ever increasing human population, remain underutilized due to lack of awareness and practicalization of technologies for their utilization. Many of them are even more resilient, adaptive and tolerant to adverse climatic conditions than the conventional foods plants. The need to explore these underutilized (lesser) wild foods in
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order to enhance availability of foods and create a balance between population growth and agricultural productivity, particularly in developing countries like Nigeria is imperative. Exploration of the nutro-chemical potentiality of the wild lesser known plants through analysis of proximate composition will alleviate the problem of nutrient deficiency and reduce degenerative diseases in Nigerian populace. In addition, the in-depth understanding of the nutro-chemical characteristics of these plants under investigation will ensure their judicious use as food in addition to their role in traditional medicine.

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