Comparative Study of White and Yellow Sorghum Seed Ground in Aliero Metropolis

CRTIFICATION

This project report titled “Comparative Study of White and Yellow Sorghum Seed Ground in Aliero Metropolis” by IBRAHIM HARUNA MUHAMMAD (1410203008) has met one of the requirements for the award of Bachelor of Science (B.Sc. Hons) Degree in Biochemistry of the Department of Biochemistry, Kebbi State University of Science and Technology, Aliero and is hereby approved for its contribution to knowledge.

……………… …………………

Dr. Jude Nwaogu Date

(Project Supervisor)

………………… ……….………

Dr. Ibrahim B. Abubakar Date

(Head of Department)

………………

Prof. M. S. Sule ………………… (External Examiner) Date

DEDICATION

I dedicate this work to my beloved parent Late. Haruna Muhammad and Amina Musa.

ACKNOWLEDGEMENT

All praise and glorification be to Almighty Allah the most beneficient the most merciful, He who spared m life to this moment and grant me knowledge, wisdom, guidance, protection, health, ability, and success to see the beginning as well as the end of this project research work. May peace and blessings of Allah be upon his prophet Muhammad (S.A.W).

I would like to extend my sincere gratitude and appreciation to my project supervisor, Dr. Jude Nwaogu for her patience, guidance, suggestion, encouragement to the success of this work.

I would also thank the Head of Department, Dr. I. Babangida and the entire academic staff and laboratory staff of the Department of Biochemistry, Kebbi State University of Science and Technology Aveiro for their contributions towards my academic upliftment.

I am deeply extending my special thanks and appreciation to my beloved parents, Haruna Muhammad and Amina Musa for their love, encouragement, and financial assistance toward the success of my program.

My special thanks to my aunty, Jamila Haruna, and my brothers, Kabiru Haruna, Bashar Haruna, Bello Haruna, Buhari Usman, Abubakar Usman, and my wife Suwaiba Prof. F.M. Tambuwal and Mal. Husain Adamu Zuru for their assistance and encouragement towards the success of my study.

Finally my special thanks to my friends; Abdullahi Hassan, Suleiman Yahuza, Qamar Ahmed, Kabiru U. Danko, Akah O. Sunday for their words of encouragement during my studies. My other friends and colleges, especially those in my Department, I say thank you all. Other people that rendered help in one way or the other but could not be mentioned. I remain grateful

TABLE OF CONTENT

TITLE PAGE ………………………………………………………………………………………i

CERTIFICATION ii

DEDICATION iii

ACKNOWLEDGEMENT iv

TABLE OF CONTENT vi

LIST OF TABLES x

LIST OF FIGURES xi

ABSTRACT xii

CHAPTER ONE 1

1.0 INTRODUCTION 1

1.2. AIM AND OBJECTIVES 4

1.2.1 To access the elemental composition of sorghum seed 4

1.2.2 To determine the vitamin content of sorghum seed 4

1.3 LITERATURE REVIEW 5

1.3.1 Taxonomy 5

1.3.2 Centre of Diversity and Domestication 6

1.3.3 Commercial uses 6

1.3.4 Food 7

1.3.5 Biofuel 8

1.3.6 Commercial propagation 8

1.3.7 Morphology of Sorghum 9

1.3.8 Anti-nutritional Factors 11

CHAPTER TWO 14

2.0 METHODOLOGY 14

2.1 Materials 14

2.1.1 Chemicals and Equipment used 14

2.1.2 Collection of Sample 14

2.2 Methods 14

2.2.1 Proximate analysis 14

2.2.2 Determination of Moisture Content (AOAC, 1998) 14

2.2.3 Determination of Ash Content (AOAC, 1998) 15

2.2.4 Determination of Crude Protein (Kjeldahl, 1883) 15

2.2.5 Determination of crude lipid (AOAC, 1998) 17

2.2.6 Determination of crude fibre 17

2.2.7 Estimation of Available Carbohydrate and Energy Value 18

2.3 Mineral analysis 18

2.3.1 Determination of potassium (K) 19

2.3.2 Determination of sodium (Na) 20

2.3.3 Determination of calcium (Ca) 20

2.3.4 Determination of magnesium (Mg) 20

2.3.5 Determination of phosphorus (P) 21

2.3.6 Determination of vitamin 22

CHAPTER THREE 23

3.0 RESULT 23

3.1 Proximate analysis 23

CHAPTER FOUR 25

4.0 Discussion, Conclusion and Recommendation 25

4.1 Discussion 25

4.2 Conclusion 27

4.3 Recommendation 27

REFERENCES 28

LIST OF TABLES

Table 3.1 Proximate parameters (S.D.M) of white and yellow sorghums seed…………….21

Table 3.2 Mineral contents in (Mg/Kg) of white sorghum and yellow sorghum…………...22

Table 3.3 Vitamin content of white and yellow sorghum…………………………………..22

LIST OF FIGURES

Figure 1: Sorghum plants showing upright stalk growth and alternate leaf pattern………… 4

ABSTRACT

White and Yellow sorghum plays an important role in the diet of Africans, it serves as a major source of carbohydrate in both human and livestock diet for the release of energy needed for normal body metabolism. In this research white maize and white sorghum (Guinea corn), both of the grass family were analyzed for their proximate contents.

The white and yellow sorghum (Guinea corn) were found to be nutritious both contain carbohydrates, protein, fats, and oil, crude fiber content, and minerals (Ash). In the research carried out it was found that white sorghum contained moisture content of 14.14%, Protein 6.89%, Fats and oils 10.42%, Crude fiber 2.90%, carbohydrates 67.16%, Ash content was found to be 1.68%.

While yellow sorghum had a moisture content of 9.75%, Protein at 6.56%, Fats and oils at 9.40%, Crude fiber at 2.76%, carbohydrates at 74.52%, and Ash content 1.75%. From the analyses carried out both white and yellow sorghum are good nutritional foods with yellow sorghum having a higher percentage of carbohydrates and white sorghum a higher percentage of protein.

CHAPTER ONE

1.0 INTRODUCTION

There is a wide distribution of biologically-active constituents throughout the plant kingdom, particularly in plants used as animal feeding stuff and in human nutrition (Igile, 1996). The knowledge that these compounds elicit both toxic and advantageous biological responses has given rise to several investigations in recent times as to their possible physiological implications in various biological systems. It is well known that plants generally contain anti-nutrients acquired from fertilizer and pesticides and several naturally-occurring chemicals (Igile, 1996).

Some of these chemicals are known as ‘‘secondary metabolites’’ and they have been shown to be highly biologically active (Zenk, 1991). They include saponins, tannins, flavonoids, alkaloids, trypsin (protease) inhibitors, oxalates, phytates, haemagglutinin (lectins), cyanogenic glycosides, cardiac glycosides, coumarins, and gossypol among others. The list is inexhaustible. Some of these plant chemicals have been shown to be deleterious to health or evidently advantageous to human and animal health if consumed at appropriate amounts (Kersten et al., 1991; Sugano et al; 1993). Most of these secondary metabolites elicit very harmful biological responses, while some are widely applied in nutrition and as pharmacologically-active agents (Oakenfull and Sidhu, 1989; Soetan, 2008).

It is worthy of note that plant poisons can either be accumulated in the animal or in certain organs or they are metabolized and excreted in milk (Liener, 1969). Reports showed that ruminants may convert cyanide to less toxic thiocyanate, which is goitrogenic (Jones et al., 1997). By this food chain, toxins or their metabolites thereof may become harmful to man (Habermal, 1987). Nutrition education should emphasize adequate and thorough preparations of human foods and animal feed, especially in humans where there are increases in reported cases of renal diseases (Salako, 2005).

The availability of these nutrients in wild seeds after ingestion also depends on the anti-nutrients present in the seeds. The anti-nutrients tend to bind to mineral elements thereby forming indigestible complex (Ladeji et al., 2004).

Oxalate for instance binds to calcium to form complexes (calcium oxalate crystals). These oxalate crystals formed to prevent the absorption and utilization of calcium. The calcium crystals may also precipitate around the renal tubules thereby causing renal stones (Ladeji et al., 2004). Cyanogenic glucoside is an organic compound containing sugar and is capable of yielding cyanide on hydrolysis (Clark, 1989).

The presence of anti-nutritional factors such as tannin, phytate, and oxalic acid, polyphenols leads to the formation of insoluble complexes with vitamins, protein, minerals rendering them unavailable to the body (Arab et al., 2010). Hence, removal of anti-nutritional factors prior to consumption is a better way of handling the problem (Khattab O. & Arntfield, H. 2009).

The nutritional acids, such as citric acid, lactic acid, and malic acid, have an indirect effect on the bioavailability of minerals and nutrients. Certain acids act as chelating competitive with phytate. It’s known that acid phytic is more sensitive to the physic activity than salts of phytate. The presence of the nutritional acids reduces the formation of salts of phytate and can facilitate the degradation of phytate during biological processes (Maenz et al., 1999).

Hydrocyanic or prussic acid (HCN) is one of the most toxic and rapidly act as common poisons; its sodium and potassium salts are only slightly toxic (Clark, 1989). Consumption of food substances containing HCN may cause death within a few minutes to 3h, depending on the concentration consumed in the food (Leiner, 2000).

Death has also occurred in young children after eating apricot and other seeds containing cyanogenetic glucoside (Sayre and Kamkal, 2004). The toxic action of HCN is due to the cyanide ion whose toxic properties are shared by all the soluble inorganic cyanide salts present in the samples (Smith et al., 2003).

Nitrates are present in all plants and are an essential source of nitrogen for normal growth, a review of the occurrence of nitrate in unprocessed foods showed that high concentrations are frequently found in vegetables (Walker, 1975). Cooking treatment significantly improves the destruction, inactivation, or reductions of anti-nutritional factors (Anuonye et al., 2012). While anti-nutritional factors decrease the bioavailability of food nutrients, organic acids increase them (Adewusi et al., 1999).

1.1 JUSTIFICATION

In the last few decades or more, large numbers of scientific data have emerged, linking diet and food selection patterns to the maintenance of health and the prevention of some chronic diseases (Oyewole and Atinmo, 2008). Nutrients, as though required in minute quantities in the diet, their absence in the diet leads to malnutrition and other detrimental health consequences such as marasmus, kwashiorkor, marasmic-kwashiorkor, cachexia, cancer, hypertension, cardiovascular diseases, obesity, and diabetes which have increased in the last decades as a result of changes of diet and lifestyle habits, among other possible causes. The presence of anti-nutritional factors such as phytate and oxalic acid leads to the formation of insoluble complexes with vitamins, protein, minerals rendering them unavailable to the body (Arab et al., 2010). When these happen, minerals and vitamin deficiencies such as dental caries, osteoporosis, rickets, scurvy, growth retardation, electrolyte imbalance among many others may result.

There is a need to establish data on the nutrient and non-nutrient composition of Sorghum bicolor which is of great importance in solving nutritional problems in society. This research is therefore aimed at determining both nutritional and antinutritional factors in Sorghum bicolor and establish data for Human Nutritionists, Biochemical Nutritionists, and Food Chemists.

1.2. AIM AND OBJECTIVES

Aim: To study the nutritional properties of methanolic seeds extract of Sorghum bicolor

The specific objectives are to:

⦁Conduct proximate analysis of methanolic seeds extracts of Sorghum bicolor.

⦁To access the elemental composition of sorghum seed

⦁To determine the vitamin content of sorghum seed

1.3 LITERATURE REVIEW

1.3.1 Taxonomy

The genus Sorghum belongs to the grass family Poaceae (Gramineae), subfamily Panicoideae, tribe Andropogoneae, subtribe Sorghinae (Clayton and Renvoize, 1986). The Andropogoneae also contains important crops such as sugarcane (Saccharum spp.) and maize (Zea mays). The genus Sorghum is a very diverse group which has made the classification of domesticated and wild sorghums difficult (Wiersema and Dahlberg 2007). It consists of 25 recognized species that are classified morphologically into five subgenera: Chaetosorghum, Heterosorghum, Parasorghum, Stiposorghum, and Eusorghum (Price et al. 2005).

Kindom; Plants

Clade; Angiosperms

Clade; Monocots

Clade; Commelinda

Order; Poaleac

Family; Poaceae

Subfamily; Panicoideae

Subtribe; Andnopogoradae

1.3.2 Centre of Diversity and Domestication

The center of origin and domestication for cultivated sorghum is considered to be the north-eastern part of Africa, most likely in the modern Ethiopia and Sudan regions where cultivation started approximately 4000 - 3000 BC (Dillon et al. 2007). Cultivated sorghums of today arose from the wild Sorghum bicolor subsp. arundinaceum (Doggett 1988). Early domestication occurred via a process of disruptive selection. Initially, selection efforts are likely to have concentrated on replacing the small-seeded, shattering, open panicles of wild types with the large-seeded, non-shattering, and compact panicles of domesticated lines (Doggett 1965).

These changes contributed to improved yields over the original landrace varieties (Dillon et al. 2007) where several traits advantageous to cultivation were favored (Doggett 1988). In addition to disruptive selection, geographic isolation and recombination in different environments led to the creation of a large number of types, varieties, and races of sorghum.

As a result, three broad groups of S. bicolor were generated; cultivated and improved types; wild types; and intermediate types (Kimber 2000). Cultivated sorghums developed with diverse morphological traits including height and inflorescence characters, and for numerous uses including food, fodder, fiber, and as a building material (Dillon et al. 2007).

1.3.3 Commercial Uses

Sorghum is the fifth largest and most important cereal crop in the world after wheat, maize, rice and barley (Doggett 1988; Ejeta and Grenier 2005). Annual global production of sorghum is estimated at approximately 60 million tonnes. Uses of sorghum are diverse and a number of in-depth reviews are available (Doggett 1988).

Sorghum is an important crop that serves as a human staple and is a major livestock feed in intensive production systems. Sorghum may be seen as one of the crops best suited to future climate change due to its ability to adapt to conditions such as drought, salinity, and high temperatures. Different races or cultivars of Sorghum bicolor may be described as grain sorghum, fodder sorghum, or sweet sorghum depending on their morphology or end-use. In some cases, sorghum is used as a dual-purpose crop; after the grain is harvested, cattle are grazed on the stubble. It's potential as a biofuel crop has been identified and is gaining in importance(Purseglove 1972).

1.3.4 Food

Sorghum is an important dietary staple for more than 500 million people in 30 countries of Africa and Asia. In Africa, sorghum underpins food security due to its drought tolerance and its ability to withstand periods of high temperatures and waterlogging. It is well suited to the semi-arid and sub-tropical climatic conditions of much of Africa where intense rainfall often occurs in short periods (Doggett 1988).

Cultivation in Africa is predominantly part of subsistence agriculture systems as opposed to the industrialized production methods used in most other regions of the world. Africa produces about one-third of the world’s sorghum but has the lowest yields per hectare (Taylor 2003). Worldwide over 50% of the sorghum produced is used for animal feed, however, in some regions, particularly sub-Saharan Africa, the vast majority of sorghum production is for human food use (FAO 1995).

Sorghum grains are prepared for a variety of food products including use as a boiled food similar to rice; roasting or popping like maize; threshing and grinding into flour to make bread, porridges, pancake, muffins, dumplings, breakfast cereals or couscous, as well as preparation of alcoholic and non-alcoholic beverages (Purseglove 1972).

The stalks of sweet sorghum varieties with high sugar content are used to make sugar and syrup

There is increasing interest in developing the potential of sorghum for uses in human foods and beverages in western countries, in particular as a source of gluten-free food (O'Hara et al. 2013; Norwood 2015). Human food uses in Australia are minor and include gluten-free beer, breakfast cereals, and baked products.

1.3.5 Biofuel

Biofuels are being developed to replace fossil sources of transport fuels in response to concerns about climate change. The biofuel industry produces ethanol from the sugars accumulated in the stalks of sweet sorghum varieties and from the starch in the seeds of grain sorghum (Almodares and Hadi 2009; O'Hara et al. 2013). The high starch content of sorghum grain (70% per grain weight) and the ability of sorghum to withstand hot dry cultivation conditions make it suitable as a feedstock for ethanol production (Wylie 2008; Almodares and Hadi 2009). The ethanol production process from sorghum also generates two co-products, the ‘Wet cake’ and syrup that are high-protein, high-value animal feed

1.3.6 Commercial Propagation

Sorghum is propagated by seed. While sorghum is considered to be predominantly a self-pollinated crop, high levels of outcrossing can also occur. It has even been suggested that seed crops should be separated from one another by a distance of 1000 m due to their ability to cross-pollinate. Further, an isolation distance of 400 m from Johnson grass or other weedy sorghums is recommended (Gupta 1999).

Hybrid seed lines are the predominant planted material in Australia. For the production of hybrid seed lines, the recommended isolation distances are 300 m for basic seeds and 200 m for certified seeds (OECD 2016). Major providers of sorghum seed undertake seed production in the Ord River (WA) and the Macquarie, Lachlan and Murrumbidgee valleys in NSW (North Queensland Register; FarmOnline). Sorghum seed production is also undertaken in South-East, Central, and North Qld. In addition, all major seed companies carry out various testing and breeding activities in South-east Qld, central Qld, and the Lockyer Valley.

1.3.7 Morphology of Sorghum

Sorghum is a cane-like grass with stout and erect stems (culms), 0.5 - 6 m tall. Most types used in grain production have a terminal compact or semi-compact head (Kimber 2000). Cultivated sorghum is generally treated as an annual crop, but may be maintained over several seasons under suitable conditions and has been described as annual or weakly perennial (Kimber 2000).

Figure 1: Sorghum plants showing upright stalk growth and alternate leaf pattern

The sorghum root system is highly organised and develops in two stages, the seminal roots and the adventitious crown roots. In early stages of growth, seminal roots develop from the radicle of the germinating seedling. These have a limited functional life of approximately three weeks. Adventitious crown roots emerge from the coleoptile (first) node and potentially from several leaf nodes above the coleoptile node. These roots form the extensive secondary root system which branches freely, both laterally and down into the soil. Sorghum plants have a fibrous root system, characteristic of grasses, which can reach a depth of up to 1.5 - 2.4 m (Kimber 2000). An extensive root system and the ability to become dormant during water stress contribute to the drought resistance of sorghum, making it an adaptable crop in marginal dryland farming systems (Whiteman and Wilson 1965).

The stem can be slender to very stocky, 5 - 50 mm in diameter, tapering at the upper end. It is solid with a hard cortex and softer inner pith that may be sweet or insipid, juicy or dry (House 1985). Each stem node contains root bands and above them growth rings that can produce new stems if the upper part of the stem is damaged. The lowest nodes in the stem contain buds that can give rise to axillary tillers while basal tillers will form at the first node (House 1985; Doggett 1988).

Sorghum leaves are concentrated near the base in some sorghum varieties, while in others they are evenly distributed along the stem. The leaves are broad and coarse, linear to lanceolate in shape and look like maize leaves. They are 90 - 100 cm long and approximately 10 - 12 cm wide. Leaves are usually shorter and smaller at the top, with the top leaf known as the flag leaf. The leaves alternate in two ranks on opposite sides of the stem and 14 - 18 leaves have been recorded on a plant at flowering. The leaf sheaths encircle the stem and there is a short membranous ligule at the junction of the leaf blade and the sheath (House 1985; Doggett 1988).

Under very dry conditions, leaves curl upwards and inwards, reducing transpiration and moisture loss by decreasing the surface area exposed. Irregular shaped silica deposits found in the leaves have been linked to drought tolerance and shoot-fly resistance (Doggett 1988). Silica deposited on the leaf surface acts as a physical barrier that alleviates water stress by decreasing transpiration and prevents the physical penetration of pests in plant tissues (Ma 2004)

1.3.8 Anti-nutritional Factors

The anti-nutritional factors (ANFS) may be defined as those sbstances generated in natural food stuffs by the normal metabolism of species and by different mechanisms (for example; inactivation of some nutrients, diminution of the digestive process, or metabolic utilization of feed) which exert effects contrary to optimum nutrition (Kumar, 1992). Being an ANF is not an intrinsic characteristic of a compound but depends upon the digestive process of the ingesting animal. For example, trypsin inhibitors, which are ANFs for monogastric animals, do not exert adverse effects in ruminants because they are degraded in the rumen (Cheeke and Shull, 1985). The utility of the leaves, pods and edible twigs of shrubs and trees as animal feed is limited by the presence of ANFs (Kumar, 1992).

Phytate (is also known as Inositol hexakisphosphate (InsP6) is the salt form of phytic acid, are found in plants, animals and soil. It is primarily present as a salt of the mono- and divalent cations K+, Mg2+, and Ca2+ and accumulates in the seeds during the ripening period (Muller, 2001). Phytate is regarded as the primary storage form of both phosphate and inositol in plant seeds and grains. In addition, phytate has been suggested to serve as a store of cations, of high energy phosphoryl groups, and, by chelating free iron, as a potent natural anti-oxidant (Muller, 2001). Phytate is ubiquitous among plant seeds and grains, comprising 0.5 to 5 percent (w/w) (Loewus, 2002). The phosphorus bound to phytate is not typically bio-available to any animal that is non-ruminant. Ruminant animals, such as cows and sheep, chew, swallow, and then regurgitate their food. This regurgitated food is known as cud and is chewed a second time. Due to an enzyme located in their first stomach chamber, the rumen, these animals are able to separate, and process the phosphorus in phytates. Humans and other non-ruminant animals are unable to do so (Harold, 2004). Phytate works in a broad pH-region as a highly negatively charged ion, and therefore its presence in the diet has a negative impact on the bioavailability of divalent, and trivalent mineral ions such as Zn2+, Fe2+/3+, Ca2+, Mg2+, Mn2+, and Cu2+. Whether or not high levels of consumption of phytate-containing foods will result in mineral deficiency will depend on what else is being consumed. In areas of the world where cereal proteins are a major and predominant dietary factor, the associated phytate intake is a cause for concern (Muller, 2001).

Oxalate is a salt formed from oxalic acid is known as an Oxalate: for example, Calcium oxalate, which has been found to be widely distributed in plants. Strong bonds are formed between oxalic acid, and various other minerals, such as Calcium, Magnesium, Sodium, and Potassium. This chemical combination results in the formation of oxalate salts. Some oxalate salts, such as sodium and potassium, are soluble, whereas calcium oxalate salts are basically insoluble. The insoluble calcium oxalate has the tendency to precipitate (or solidify) in the Kidneys or in the Urinary tract, thus forming sharp-edged calcium oxalate crystals when the levels are high enough. These crystals play a role to the formation of kidney stones formation in the urinary tract when the acid is excreted in the urine (Nachbar et al., 2000).Oxalate is an anti-nutrient which under normal conditions is confined to separate compartments. However, when it is processed and/or digested, it comes into contact with the nutrients in the gastrointestinal tract (Noonan and Savage, 1999). When released, oxalic acid binds with nutrients, rendering them inaccessible to the body. If food with excessive amounts of oxalic acid is consumed regularly, nutritional deficiencies are likely to occur, as well as severe irritation to the lining of the gut (Habtamu and Negussie, 2014). In ruminants, oxalic acid is of only minor significance as an anti-nutritive factor since ruminal microflora can readily metabolize soluble oxalates, and to a lesser extent even insoluble Ca oxalate. While the importance of the anti-nutritive activity of oxalic acid has been recognized for over fifty years it may be a subject of interest to nutritionists in the future (Oladimeji et al., 2000; Liebman and Al-Wahsh, 2011). Oxalic acid forms water soluble salts with Na+, K+, and NH4+ ions, it also binds with Ca2+, Fe2+, and Mg2+ rendering these minerals unavailable to animals. However Zn2+ appears to be relatively unaffected. Calcium oxalate is insoluble at a neutral or alkaline pH, but freely dissolves in acid (Liener, 2005).

A number of plant species produce hydrogen cyanide (HCN) from cyanogenic glycosides when they are consumed. These cyanogens are glycosides of a sugar, often glucose, which is combined with a cyanide containing aglycone (Habtamu and Negussie, 2014). Cyanogenic glucosides are classified as phytoanticipins. Their general function in plants is dependent on activation by b-glucosidases to release toxic volatile HCN as well as a ketones or aldehydes to fend off herbivore and pathogen attack (Golden, 2009).

CHAPTER TWO

2.0 METHODOLOGY

2.1 Materials

2.1.1 Chemicals and Equipment used

Apart from conventional glass wares used, the list of chemicals and equipment used in this research will be presented in the appendix

2.1.2 Collection of Sample

The seed of Sorghum bicolor will be collected from Aliero, Aliero LGA of Kebbi State area, Nigeria. The seed were destalked, washed with deionized water to remove the impurities and air dried at room temperature. After drying,the seed were ground into a fine powder and stored in a well labeled air-container for analysis.

2.2 Methods

2.2.1 Proximate analysis

The methods of Association of Official Analytical Chemists (AOAC, 1998) were used for the determination of moisture content, crude protein, crude lipids and crude fibre contents.

2.2.2 Determination of Moisture Content (AOAC, 1998)

Two grams (2g) of fresh leaves of Sorghum was weighed into a crucible and dried in an oven at 105°C for 24 hours. The dried sample was cooled in a desiccator for 30 minutes and weighed. The percentage lost in weight indicates the moisture content and was calculated using the equation below.

Where:

wb= mass of sample and crucible before drying

wa= mass of sample and crucible after drying

ws = mass of the sample

2.2.3 Determination of Ash Content (AOAC, 1998)

Two grams (2g) of the sample was placed into a muffle furnace and heated at 500°C for 2 hours. The percentage ash content was calculated using the formula in the equation below;

% ash = weight of ash × 100

weight of sample

2.2.4 Determination of Crude Protein (Kjeldahl, 1883)

The crude protein of the sample was determined using micro-Kjeldehl method.The method involves three major steps:-

⦁ Conversion of organic nitrogen to ammonium sulphate by digestion using NaOH solution and collection into boric acid solution in form of ammonium ion:

Sample nitrogen + H2SO4 → (NH4)2SO4

⦁ Distillation of ammonia in the digest using NaOH solution and collection into boric acid solution inform of ammonium ion:

(NH4)2SO4 + 2NaOH(aq) → Na2SO4(aq) + 2H2O (l) + 2NH3 (g)

3NH3 (g) + H3BO3 (aq) → (NH4)3BO3 (aq)

⦁ Estimation of nitrogen content by titration of the borate ion with standard acid (H2SO4 or HCl) using a suitable indicator.

2(NH4)3BO3 (aq) + 3H2SO4 (aq) → 3(NH4)2SO4 + 2H3BO3 (aq)

The concentration of H+ in moles required to reach the end point is equivalent to the amount of nitrogen in the sample. The protein content would be calculated by multiplying nitrogen content by the conversion factor (6.25) as most protein contain about 16% nitrogen.

Procedure:

⦁ Digestion: Twograms (2g) of the sample was placed into a cleaned dry Kjeldahl’s digestion flask. A half gramme (0.5g) of Kjeldahl’s catalyst tablet and 20cm3 of concentrated H2SO4 was added and heated on a heater coil for an hour, cooled, filtered into a 50cm3 volumetric flask and would be made to the mark with distilled water.

⦁ Distillation: Ten cubic centimeters (10cm3) of 40% NaOH solution was added to 10cm3 digest, into Kjeldahl’s distillation flask. The aliquot was steam distilled (to liberate ammonia) into 20cm3 of 2% boric acid indicator solution in a 100cm3 conical flask. The distillation process was allowed to continue for a few minutes until the solution turned green.

⦁ Titration: The distillate was titrated against 0.01M HCl to the end point. Thepercentage protein content was calculated using the equation below.

Crude protein= (a-b)×14×volume made×6.25×100

Volume of aliquot ×sample weight

Where;

a= titre value for the digested sample.

b= titre value for the blank.

2.2.5 Determination of crude lipid (AOAC, 1998)

Five grams (5g) of sample was be placed in a porous thimble and its mouth was plugged with cotton. The thimble was placed in an extraction chamber, which was suspended above a receiving flask of known weight containing petroleum ether (Bp 40-60°C) on a condenser. The flask was heated on heating mantle and the oil was extracted. After the extraction, the thimble was removed from the Soxhlet and the apparatus was reassembled and heated over water bath to recover the ether. The flask containing the crude lipid was disconnected and dried in an oven at 100°C for 30 minutes. After heating, the flask was cooled in a desiccator and weighed. The percentage crude lipid content was calculated using equation below:

% crude lipid= × 100

2.2.6 Determination of Crude Fibre

Principle: The principle of the method is based on loss of crude fibre when ignited after being digested with acid and base under specific conditions.

Procedure: The residue obtained after crude lipid extraction was placed into a 500cm3 beaker and 200cm3 of boiling 1.35% H2SO4 (w/v) was added. The content was boiled for 30 minutes, cooled, filtered through muslin cloth and the residue was washed three times with distilled water. The washed residue was returned into the initial beaker and further digested by boiling in 200cm3 of 1.25% NaOH for 30 minutes. The digest was filtered to obtain the residue and then washed three times with distilled water and finally with 25cm3 ethanol. The washed residue was dried in an oven at 105°C to a constant weight and cooled in a desiccator. The residue was scraped into a pre-weighed porcelain crucible, weighed, ashed at 550°C for two hours, cooled in a desiccator and was re-weighed. Crude fibre content was calculated using the equation below

% crude fibre = weight loss on ignition × 100

Weight of sample

2.2.7 Estimation of Available Carbohydrate and Energy Value

Available carbohydrate was estimated by subtracting the total of the percentages of crude protein, crude lipid, crude fibre and ash from 100% moisture free sample. The sample calorific value was estimated in (kcal) according to the formula in equation below (Asibey and Tayie, 1999).

Energy = (% Carbohydrate ×4) + (%lipid × 9) + (%crude protein)

2.3 Mineral analysis

The exchangeable K and Na is that which is freely exchanged with other cations in sorghum solution. During extraction by a suitable extraction solution, water soluble K or Na is also included. Therefore in any determination of exchangeable K or Na, amount of K or Na soluble in water should be subtracted from the amount determined in the extract. It is necessary for saline and calcareous sorghum. For other sorghum, however, it may not be necessary as its amount is fairly small.

Principles: In the flame photometer, a flame is used to atomise the particular element in question. The amount of atomization is proportional to the quantity of the element in the feeding solution/extract. The intensity is measured with a photocell in a selected wavelength range corresponding to the given element. The quantity of atomization (emisation) in the extract is compared with a known quantity of the element to be determined, i.e. a standard curve is prepared.

Reagents: 1. sorghum extract (Neutral normal ammonium acetate extract) from experiment on C.E.C. 2. Standard K stock solution- Dissolve 0.9533g of dried KCL in distilled water and dilutes the solution to 500ml in a volumetric flask, and mixes it. This solution contains 1000ppm K. From here prepare K standard solutions by pipetting suitable aliquot such that the final solutions range in concentration from 0-25ppm K. 3. Standard Na stock solution- Dissolve 2.542g of NaCl (dried at 1100C) in distilled water and dilute to 1 liter. This solution contains 1000ppm Na. Make standards by taking aliquots as for K.

N.B. It is advisable that the stock solution of K and Na be diluted to 100ppm and the standard curve made from the diluted standard solution.

2.3.1 Determination of potassium (K)

1. Set the flame photometer for K by inserting appropriate filter (usually of 768um wavelength).

2. Set the instrument to 100 percent transmittance by feeding 10ppm K solution.

3. Run all the standard solutions and prepare standard curve by plotting transmittance readings against concentration of standard K solution.

4. Run the sample.

5. Calculate the amount of K present in the onion as mill equivalent per 100g oven-dry weight of the soil by getting K concentration in the extract from the standard curve. Make sure that you have considered all dilutions in making calculations.

2.3.2 Determination of sodium (Na)

1. Set the flame photometer for Na by inserting appropriate filter (usually of 589mu wavelength).

2. Set the instrument to 100 percent transmittance by feeding 10ppm Na solution.

3. Repeat steps 3 to 5 as for K determination.

2.3.3 Determination of calcium (Ca)

1. Pipette 1ml aliquot or the sample in titration flasks and add distilled water to get a volume of about 20ml.

2. Add 3 drops each of KCN, NH2OH.HCL and triethanomine and add enough 10% NaOH(usually about 4ml) to raise the pH 12 on slightly higher. Check the pH with a pH-meter or hydration paper. Add a tip of murexide and titrate with EDTA to purple end point.

3. Pipette 1ml aliquots of the sample in to titration flasks and dilute to 20ml with distilled water.

4. Add 5ml of buffer solution and 3 drops each of KCN, NH2OH.HCL, K4Fe (CN) 6 and triethanolamine. Allow a few minutes for reaction to take place.

5. Add 3 drops of EBT indicator and titrate the solution with EDTA in the above manner to the permanent blue colour.

2.3.4 Determination of magnesium (Mg)

1. Pipette 1ml aliquots of the sample in to titration flasks and dilute 20ml with distilled water.

2. Add 5ml of buffer solution and 3 drops each of KCN, NH2OH.HCL, K4Fe (CN) 6and triethanolamine. Allow a few minutes for reaction to take place.

3. Add 3 drops of EBT indicator and titrate the solution with EDTA in the above manner to the permanent blue colour.

4. Wash down the neck of the flask with distilled water to avoid direct contact of 5nCl2 solution with the concentrated ammonium molybdate solution.

5. Dilute the content to about 48ml with distilled water. Add 1ml of the diluted stannous chloride solution, make the solution to volume and mix immediately. Read colour intensity after 10 minutes of stannous chloride addition at 660mu wavelength.

2.3.5 Determination of phosphorus (P)

1. The ash residue was dissolve with 5ml of 20% of HCL and dilute to 50ml volume with distill water.

2. Pipette 2ml aliquot of the sample in a 50ml volumetric flask. Add 2ml of the ammonium molybdate solution and mix the content well. If other aliquot size is taken, then the same amount of ammonium molybdate should be added. Make the volume with distilled water to about 48ml. Finally add 1ml of the freshly diluted stannous chloride solution, make to volume with water and mix immediately. After 5-6 minutes and before 20 minutes, measure the colour intensity using 660mu wavelength.

3. Calculate P content in the onion after considering all the dilutions.

2.3.6 Determination of vitamin C (Rutkowski and Grzegorczyk , 2007).

Principle

This was based on color reaction with periodically prepared phosphotungstate reagent and absorbance taken at 700nm.

Procedure

1ml of the analyzed liquid into the centrifugal test-tube, 1ml of the phosphotungstate (PR) was added and was mixed thoroughly and left in a room temperature for 30minutes. The tube was centrifuged (7000xg, 10 minutes) and the whole was collected from separated supernatant with a pipette. The supernatant was the test sample for spectrophotometric measurement. The standard was prepared in the same way without centrifugation. The absorbance of the test samples A and of the standard sample A was measured at 700nmagainst the mixture PR: 50m solution of oxalic acid =1:1 (v/v) was used as the reference sample.

Calculation

Cx = As x Cs

Where Cs = concentration of the standard solution =56.8μm/L

As= absorbance of sample

As= absorbance of standard

CHAPTER THREE

3.0 RESULT

3.1 Proximate Analysis

The result of the study is presented in the following tables and described as follows: It consisted of the proximate analysis

Table 3.1 Proximate parameters (S.D.M) of white and yellow sorghums seed.

Proximate contents white sorghum yellow sorghum

carbohydrate 67.16±1.07 74.52±1.15

Moisture 14.75±0.83 9.75±1.66

Crude protein 6.89±1.01 6.56±1.09

Crude lipid 10.42±0.55 9.40±0.90

Ash 1.68±0.08 1.32±0.09

Crude fibre 2.90±0.50 2.76±0.81

Table 3.2 Mineral contents in (mg/Kg) of white sorghum and yellow sorghum.

Mineral Elements White sorghum Yellow sorghum

SODIUM (Na) 24.33±0.89 26.33±1.33

POTASSIUM (P) 623.33±4.44 52.0±0.66

CALCIUM (Ca) 0.72±0.043 0.52±0.023

MAGNESIUM (Mg) 2.083±0.022 1.52±0.023

PHOSPHORUS (P) 1.63±0.015 2.122±0.707

Table 3.3 Vitamin content of white and yellow sorghum

Vitamin content White sorghum Yellow sorghum

Vit. C 26.34±0.49 18.30±0.39

CHAPTER FOUR

4.0 Discussion, Conclusion and Recommendation

4.1 Discussion

The result show that white sorghum had high moisture content than yellow sorghum this attributed the moisture content of both white sorghum and yellow sorghum was noted to be 14.14% and 9.75%. The respective ash contents of white sorghum and yellow was 1.68% and 1.32% respectively. The ash obtained was almost the same. The lipid content of white sorghum and yellow sorghum was 10.42% and 9.40% respectively. The fiber content of white sorghum and yellow sorghum was 2.90% and 2.76% respectively.The fiber obtained was almost the same. The crude protein was 6.89% and 6.56% white andyellow sorghum. The carbohydrate content of white and yellow sorghum was 67.16% and 74.52%.

The mineral elements analyzed by various methods are sodium, potassium, calcium, magnesium and phosphorus. The sodium content of white sorghum and yellow sorghum was 24.33 and 26.33 respectively as it was reported by Cook (1988). The potassium contents of white sorghum and yellow sorghum was 623.33 and 52.0 as it was reported by (Sagnaet al 2014). The calcium was 0.72 and 0.52 for white sorghum and yellow sorghum respectively as it was reported by (Sagnaet al 2014).

The magnesium white sorghum and yellow sorghum was 2.083 and 1.52 as it was reported by Sagnaet al (2014). And also phosphorus contents was 1.63 and 2.122 respectively as it was reported Cook (1988).

The vitamin C content of white sorghum was 27.34 ± 0.49 mg /100g while yellow sorghum 18.30 ±0.39, this is high compared to 26.34 and 18.96 reported by (Nwankwo Rita Ngozi 2014) in Hibiscus cannabinusand cassia tora, the value is extremely very high compared to 0.09924±0.001 reported by (Muibatolabisi Bello et al. 2014) in Ficus exasperate vahl leaves. The higher value implies that white and yellow sorghum leaves were excellent source of Vitamin C and this is in line with literature reported by (Meddison et al. 2007)that white and yellow sorghum fruits contains three times the vitamin c of orange whereas the leaves are nutritionally superior to its fruits. Vitamin C is a highly effective antioxidant and a very small daily intake of this vitamin for an adult is required to avoid deficiency disease scurvy.

Even in small amounts it can protect indispensable molecules in the body, such as proteins, lipids (fats), carbohydrates, and nucleic acids (DNA and RNA) from damage by free radicals and reactive oxygen species that can be generated during normal metabolism as well as through exposure to toxins and pollutants.

4.2 Conclusion

The result obtained in this analysis show a very close nutritional relationship between white and yellow sorghum seeds as both contain almost similar results of protein, carbohydrate, fats and oil, crude fibre and ash contents, therefore conclusion drawn is that both cereals can be used in place of another for both can serve almost the same nutritional requirement.The proximate analysis revealed that sorghum samples contains appreciable nutrient contents.

Also the results of mineral elements analysis showed that, the fresh had high contents of potassium and calcium when compared to the stored. With the above results, the study concluded that the best state to consume this onion was the fresh state.

4.3 Recommendation

Based on the findings, the following recommendations are made:

1. There is a need for public awareness campaign to enlighten the farmers and the general public on the importance and quality of White and yellow sorghums seed.

2. The government should provide proper incentives to encourage farmers to cultivated sorghums properly for future use.

3. Farmers should be encouraged to maintain a proper sanitary condition in and around their stored houses.

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Comparative Study of White and Yellow Sorghum Seed Ground in Aliero Metropolis

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