CHAPTER ONE

1.0 INTRODUCTION

1.1 BACKGROUND OF STUDY

Syrup is a thick, sugary liquid made by boiling down or otherwise concentrating

plant sap, juice or grain extracts. It can also be defined as a thick, sweet, sticky

liquid, consisting of a sugar base, natural or artificial flavorings and water. Syrup

is obtained by the hydrolysis of starch to glucose (i.e. glucose syrup), fructose,

maple, maltose, etc. The hydrolysis of starch is achieved by using acid treatment,

enzyme treatment or a combination of acid and enzymes. Glucose syrup is an

aqueous solution of several compounds principally glucose, dextrose and

maltose, and/or a purified, concentrated solution of nutritive monosaccharide and

higher saccharine, obtained by hydrolyzing starch with specific enzymes under

conditions (OINO, 2001). At present, most of the dextrose (i.e. glucose) in

commerce is prepared in the form of pure dextrose monohydrate by a combined

acid-enzyme process and the starch used in the manufacture of glucose syrup

must be as pure as possible with a low protein content (particularly soluble

protein), (Grace,1977). According to Amandikwa, (2012), the proximate

composition of cocoyam flour ranges from; 6.55% to 13.2% for moisture, 1.2%

to 2.5% for ash, 1.5% to 2.4% crude fiber, 1.7% to 2.6% fat, 7.4% to 8.9%

protein and 72.1% to 79.3% carbohydrate. The syrup can be applied to the

manufacture of various products such as beverages, corn syrups, solids, standard

1
glucose syrup, dextrose, D-glucose, sweets and confectionaries among others.

The syrup form being a concentrated form is less bulky and suitable for

transportation. It’s relatively high solutes content and lower water activity will

contribute to its storage stability.

Glucose, an important industrial product of starch hydrolysis find application as

bulk sweetener in the food, and confectionary industry (Fox and Cameron,

1982). The production of glucose, maltose and dextrins from starch of maize

(Sutherland et al., 1986), banana (Igoe, 1989; Bello – perez et al., 2000), cassava

and sweet potato (Omemu et al., 2004) has been well documented in many parts

of the world. However, production of these important products of starch

hydrolysis in Nigeria has been largely obtained from starch of tubers such as

cassava whose cultivation is in large scale in the southern part of the country.

Glucose syrup produced from enzymatic or acidic hydrolysis of starch is a

concentrated aqueous solution of glucose, maltose and other nutritive

saccharides. Glucose or dextrose sugar is found in nature in sweet fruit such as

grapes or honey. It is less sweet than sucrose (Cane syrup). Glucose syrup is

made from the enzymatic or acid hydrolysis of starch which could be from

potatoes, wheat, barley, rice and cassava (Peter hull, 2010).

Cocoyam is a root crop and belongs to the family Aracea (Anikwe, et al., 2005).

It has two major cultivated varieties namely; Colocasia spp., and Xanthosoma

spp. Cocoyam is an ancient crop grown throughout the humid tropics for its

2
edible corms, cormels and leaves, as well as other traditional uses. The Aracea

family is made up of some hundred genera and more than fifteen hundred

species. They are mostly tropical and subtropical and grow mainly in moist or

shady habitats. Cocoyam is a well-known food plant which has a long history of

cultivate, its corms are important source of starch which is the main component

of tuber crops.

Yam belong to the genus Dioscorea in the family Dioscoreaceae. The family is

believed to be among the earliest angiosperm and probably originated in

southeast Asia (Coursey, 1976).

Starch, the raw material is required for the production of low molecular weight

products (glucose/dextrose, maltose, maltotriose and dextrin) and is widely

applied in sugar, spirits, textile as well as brewing (Selmi, 2000).Starch is found

in the endosperm of cereal grains, roots and tuber crops (Omemu, et.al. 2004).

The conversion of starch to various sweeteners is achieved through a chemical

(acid) or an enzymatic process. Starch can be used in the manufacture of

confection and biscuits because of its ability to gel firmly on cooling. Starch may

be used in baking industries. Its enzyme and acid conversion can produce

maltotriose, maltose, glucose syrup, fructose syrup (Vuilleumier, 1993). The

biggest user of starch worldwide is the sweetener industry. It can easily be

hydrolyzed to form syrup containing dextrose, maltose and other

oligosaccharides. The production of sugar syrups by enzymatic method is

3
amongst the most advanced food technologies, characterized by higher yields,

wide range of products, higher product quality and energy economy (Bindumole,

et.al., 2001).

Therefore, the study is interested in the production of sugar syrup from crude

enzymes in malted wheat, millet and sorghum using cocoyam and yam flour

1.2 Objectives
The main objective of this research project is;
 To produce sugar syrup from Cocoyam flour, Aerial yam flour and potato

starch using crude enzymes from malted cereals and Amyloglucosidase

enzyme and to evaluate and compare its sugar profile

The specific objectives of this research project are;

 To malt cereal grains such as sorghum, millet and wheat to develop their

inherent enzymes in order to serve as a source of crude enzyme for syrup

production

 To produce aerial yam flour and cocoyam flour

 To extract starch from potato white root

 To produce syrups by hydrolyzing the flours and starch using the crude

enzymes developed in the malted cereal grains.

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 To determine and compare thesugars, present in thesyrupsproduced by means

of crude/inherent enzymes from malted cereal grains and exogenous enzyme

using high performance liquid chromatography.

1.3 Justification

The success of this research will help provide a guide for the use of

crude/inherent enzymes present in the malting of cereal grains (Pennisetum

glaneum) for millet, (Triticum eastivum) for wheat and (Sorghum vulgare) for

sorghum and dry milling of tubers crops (yam, cocoyam), in the production of

high quality sugar syrup.This will lead to the industrialization ofusingcrude

starch (tuber flour) for the production of sugar syrup to a large extent. This will

encourage the use of crude enzyme gotten from malted cereal grains thereby

reducing the money spend in importing exogenous enzymes and this will

promote the use of our indigenous cereals in enzyme production. And help to

compare the various source of enzyme and know the enzyme source which is

equivalent to that of exogenous enzyme (Amyloglucosidase). sugar syrup which

is cheap, nutritious, wholesome and generally acceptable by the consumer would

be elaborated in the course of this study.

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 CHAPTER TWO

2.0 LITERATURE REVIEW

Sugar syrup is foods syrup, made from the hydrolysis of starch. Maize (corn) is

commonly used as the source of the starch in the US, in which case the syrup is

called "corn syrup", but sugar syrup is also made from other starch crops,

including potatoes, wheat, barley, rice and cassava (Hull,2010). Sugar syrup

containing over 90% glucose is used in industrial fermentation, (Dziedzic et al

1995) but syrups used in confectionery manufacture contain varying amounts of

glucose, maltose and higher oligosaccharides, depending on the grade and can

typically contain 10% to 43% glucose (Jackson 1995). Sugar syrup is used in

foods to soften texture, add volume, prevent crystallization of sugar and enhance

flavour. By converting some of the glucose in corn syrup into fructose (using an

enzymatic process), a sweeter product, high fructose corn syrup can be

produced.

2.1Types of syrups

Depending on the method used to hydrolyze the starch and on the extent to

which the hydrolysis reaction has been allowed to proceed, different grades of

sugar syrup are produced, which have different characteristics and uses. The

syrups are broadly categorized according to their dextrose equivalent (DE). The

further the hydrolysis process proceeds, the more reducing sugars are produced,

and the higher the DE. Depending on the process used, sugar syrups with

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different compositions, and hence different technical properties, can have the

same DE.

2.1.1 Confectioner's Syrup

The original sugar syrups were manufactured by acid hydrolysis of corn starch at

high temperature and pressure. The typical product had a DE of 42, but quality

was variable due to the difficulty of controlling the reaction. Higher DE syrups

made by acid hydrolysis tend to have a bitter taste and a dark colour, due to the

production of hydroxymethylfurfural and other by-products (Hull 2010). This

type of product is now manufactured using a continuous process (Norman et

al.,2011) and is still widely used due to the low cost of acid hydrolysis. The

sugar profile of a confectioner's syrup can also be mimicked by enzyme

hydrolysis (Norman et al 2011) A typical confectioner's syrup contains 19%

glucose, 14% maltose, 11% maltotriose and 56% higher molecular mass

carbohydrates.

2.1.2 High maltose corn syrup

By using β-amylase or fungal α-amylase, sugar syrups containing over 50%

maltose, or even over 70% maltose (extra-high-maltose syrup) can be produced.

This is possible because these enzymes remove two glucose units (i.e. one

maltose molecule) at a time from the end of the starch molecule. High-maltose

glucose syrup has a great advantage in the production of hard candy: at a given

moisture level and temperature, a maltose solution has a lower viscosity than a

glucose solution, but will still set to a hard product. Maltose is also less

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humectant than glucose, so candy produced with high-maltose syrup will not

become sticky as easily as candy produced with a standard glucose syrup

(Hull,2010).

2.1.3 Commercial preparation

Irrespective of the feedstock or the method used for hydrolysis, certain steps are

common to the production of glucose syrup:

Before conversion of starch to glucose can begin, the starch must be separated

from the plant material. This includes removing fibre and protein (which can be

valuable by-products, for example wheat or maize gluten (Hull, 2010). Protein

produces off-flavours and colours due to the Millard reaction, and fibre is

insoluble and has to be removed to allow the starch to become hydrated. The

plant material also needs to be ground as part of this process to expose the starch

to the water.

2.1.4 Soaking

The starch needs to be swelled to allow the enzymes or acid to act upon it. When

grain is used, sulfur dioxide is added to prevent spoilage.

2.1.5 Gelatinization

By heating the ground, cleaned feedstock, starch gelatinization takes place: the

intermolecular bonds of the starch molecules are broken down, allowing the

hydrogen bonding sites to engage more water. This irreversibly dissolves the

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starch granule, so the chains begin to separate into an amorphous form. This

prepares the starch for hydrolysis.

2.1.6 Hydrolysis

Sugar syrup can be produced by acid hydrolysis, enzyme hydrolysis, or a

combination of the two. Currently, however, a variety of options are available.

Formerly, sugar syrup was only produced by combining corn starch with dilute

hydrochloric acid, and then heating the mixture under pressure. Currently, sugar

syrup is mainly produced by first adding the enzyme α-amylase to a mixture of

corn starch and water. α-amylase is secreted by various species of the bacterium

Bacillus; the enzyme is isolated from the liquid in which the bacteria are grown.

The enzyme breaks the starch into oligosaccharides, which are then broken into

glucose molecules by adding the enzyme glucoamylase, known also as "γ-

amylase". Glucoamylase is secreted by various species of the fungus

Aspergillus; the enzyme is isolated from the liquid in which the fungus is grown.

The glucose can then be transformed into fructose by passing the glucose

through a column that is loaded with the enzyme D-xylose isomerase, an enzyme

that is isolated from the growth medium of any of several bacteria.

2.1.7 Clarification

After hydrolysis, the dilute syrup can be passed through columns to remove

impurities, improving its colour and stability.

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2.1.8 Evaporation

The dilute glucose syrup is finally evaporated under vacuum to raise the solids

concentration.

2.1.9 Uses

Its major uses in commercially prepared foods are as a thickener, sweetener, and

humectants. A humectant is an ingredient that retains moisture and thus

maintains a food's freshness (Elaine 2008). Sugar syrup is also widely used in

the manufacture of a variety of candy products. In the United States, cane sugar

quotas raise the price of sugar hence, domestically produced corn syrup and

high-fructose corn syrup (HFCS) are less expensive alternatives that are often

used in American-made processed and mass-produced foods, candies, soft drinks

and fruit drinks to help control cost (Elaine 2008)

Sugar syrup was the primary corn sweetener in the United States prior to the

expanded use of HFCS production. HFCS is a variant in which other enzymes

are used to convert some of the glucose into fructose. The resulting syrup is

sweeter and more soluble. Corn syrup is also available as a retail product. Sugar

syrup is often used as part of the mixture that goes into creating fake blood for

films.

2.2 Millet

Millet are tiny in size, round in shape and minor cereals of the small seeded

grass family (poaceae). It is characterized by their remarkable ability to survive

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in less fertile soil, drought – resistant, resistance to pest and disease, short

growing season (Devi et al., 2011)and cultivated round the year and all the

world. The world millet derived from the beginning of human civilization.

Millets are considered as the first domesticated cereal (Shahidi et al., 2013).

Millet have uniqueness because of its richness in protein, calcium, dietary fibre

and polyphenol (Devi et al., 2011). It is founded that millets contain significant

amount of Sulphur containing essential amino acids like methionine and cysteine

(Obilana et al., 2002).

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Fig 2.0 Structure of millet kernel

2.2.1 Nutritional attributes of millet

Millet contain 60 to 70% carbohydrates, 7 to 11% proteins, 1.5 to 5% fat, and

2.7% crude fibre and are also rich in vitamins and minerals. They are excellent

source of vitamin B, magnesium and antioxidants. Millet is also a good source of

other dietary millet like manganese, phosphorus and iron. Millet proteins are

good source of essential amino acid except lysine and threonine but are

relatively high in Sulphur containing amino acids methionine and cysteine

(Singh et al., 2012). Millet is an alkaline forming grain that is gluten – free

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(Moreno et al., 2014), vitamin B such as Niacin, folacin, riboflavin and thiamine

and phosphorus are present in millets that play a key role in energy synthesis in

the body.

2.2.3 Millet as probiotic and prebiotic

Prebiotics aid the existing flora or help repopulate the colon when bacteria level

are reduced by antibiotics, chemotherapy or disease. Probiotics are “living

microorganisms” which when administered in adequate amounts confer a health

benefit on the host. Fermented millet products act as a natural probiotic

treatment for diarrhea is young children (Lei et al., 2006). Prebiotics are non-

digestible food ingredients that beneficially affect the host by selectively

stimulating the growth and activity of one or a limited number of bacteria in

colon (Laminu et al., 2011). Millets’ whole grain also shows prebiotics activity.

Which helps to increase the population of friendly bacteria that plays a key role

to promote digestion. Malting induces important beneficial biochemical changes

in millet grain.

2.2.4 Health benefits of millets

Millets have many nutraceutical properties that are helpful to prevent disease,

prevention of cancer and cardiovascular disease, decreasing tumour cases etc.

other health benefits are increasing the time span of gastric emptying, provides

roughageto gastro intestine (Gupta et al., 2012). Millet is an alkaline forming

food. Alkaline based diet is often recommended to achieve optimal health,

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meaning when it combine with digestive enzymes. The soothing alkaline nature

of millet helps to maintain a healthy PH balance in the body crucial to prevent

illnesses.

2.3 Wheat

Wheat (Triticiuma estivum L) belongs to the grass family (graminea) wheat is a

type of grass grown all over the world for its highly nutritious and useful grain.

It is one of the top three most produced crops in the world, along with the corn

and rice. It is an important staple crop particular in Europe, America and

middleeast country. It is widely grown in the tropics usually on a large scale

unlike maize or rice that can be grown by small holders. It is the number one

cereal of the temperate region.

2.3.1 Origin and history of wheat

Wheat originated within India.The three major categories of wheat, hard, soft

and Durum, have slight variations in their botanical descriptions. Hard wheat

contains hard, small wheat kernels while soft wheat contains larger, softer

kernels. Durum wheat are completely different than both hard and soft wheat. Its

kernels are much larger and is has a unique shape than the other wheat varieties

(Eborn, 2008). The cultivation of wheat spread from the centre of its origin that

is India to Pakistan and china. The greatest diversity of this specie Triticum is

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found in the Ethopia. Africa produces only2.46% of the total worldproduction of

wheat. Only four countries are known in Africa forwheat productionthey include

Zimbabwe, Ethiopia, Kenyaand Sudan.

Fig 2.1 Structure of a wheat kernel

2.3.2 Nutritional composition of wheat

The nutritional value of wheat is addressed through its macronutrient and

micronutrient components. These groups consist of carbohydrate, proteins and

lipids, for macronutrients, and vitamins, minerals and phytochemicals for

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micronutrients. First to macronutrients, grains consist of approximately 75%

carbohydrate (McKevith, 2004), and therefore many believe that the importance

of carbohydrate and fiber within wheat takes precedence over their

concentrations of vitamins, minerals, and phytochemicals (Basey, 2005).

Carbohydrates are categorized into “good carbs” and “bad carbs,” the difference

between them being rates of fiber digestion. “Good carbs” are unrefined

complex carbohydrates, such as whole grains, and have a very slow rate of fiber

digestion, delivering a slow, steady rate of glucose to the blood, creating a

feeling of full-ness. Refined complex carbohydrates, such as white flour and

white pasta, are known as “bad carbs.” These have a very fast rate of fiber

digestion, which leads to a feeling of hunger soon after a meal. This is where the

idea of “low-carb” diets originated, believing that eating no carbs would help to

eliminate the amount of “bad carbs” being consumed (Basey, 2005).

The second major macronutrient within wheat is protein, the most concentrated

being gliadins and glutenins. However, these proteins are still in relatively low

amounts and therefore, essential amino acids must be supplied from another

source of the diet (McKevith, 2004). Lastly, lipids are a very minor component,

only consisting of about 1-3% within wheat (McKevith, 2004).

Unlike macronutrients, micronutrients are not as concentrated within wheat and

therefore cannot be compared within their percentages. They do, however, serve

an extremely important purpose to the human body, specifically in their roles of

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promoting health and preventing disease (Basey, 2005). The most significant

vitamins found within wheat are B, specifically thiamin, riboflavin and niacin,

and Vitamin E (McKevith, 2004). The dominant mineral found in all wheat is

potassium, however, in whole grain wheat, iron, magnesium, manganese and

zinc are also found in high concentrations (Wheat Germ 8). Lastly, the most

interesting micronutrient found within wheat are phytochemicals or plant

bioactive substances. Research has shown that these substances may have many

health promoting effects, similar to antioxidants, which are extremely beneficial

to disease prevention (McKevith, 2004). The three major categories of wheat

are hard, soft and durum, have a great variety in their nutritional aspects,

specifically in their protein levels. Hard wheat typically has high protein and

gluten levels which make it particularly useful for bread making. Bread making

requires a 12% protein level and specific varieties of hard wheat can contain up

to a 16% protein level. Soft wheatcontain a 9-11% protein level, making them

ideal for making pastries and cakes. Durum, although containing high levels of

protein, does not contain the type of protein needed to form a strong gluten and

therefore, is used to make pasta and is commonly referred to as “macaroni

wheat” (Eborn, 2008).

2.4 Sorghum

Sorghum is a genus of plants in the grass family and is an indigenous crop to

Africa and though commercial needs and uses may change over time, sorghum

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will remain a basic staple food for many rural communities. The latter is

especially true in the more drought prone areas of South Africa where this hardy

crop provides better household food security than maize. In Nigeria, sorghum is

also known as guinea corn. Sorghum, guinea corn or millet is the world’s third

important food grain being exceeded in utilization for food by wheat and rice. It

is well utilized in Africa, Asia and South America. It is prepared in a multiple of

ways for human consumption. Most commonly, it is ground or cracked, prepared

into dough and baked as flat, unleavened bread or cooked like rice. It is

frequently mixed with the cereals, legumes, crude sugar or spices for various

preparations. Sorghum grain is processed to remove the fibrous and often high

colored pericarp and testa layers and to reduce the remainder of the seeds to

flour. It consists of the endosperm, the main starchy portion and the embryo or

germ, which are separated by their scutellum.

2.4.1 Origin of Sorghum

Sorghum originated in North-Eastern Africa with domestication having taken

place there around 5000-8000 years ago. The largest diversity of cultivated and

wild sorghum is also found in this part of Africa. The secondary center of origin

of sorghum is the Indian subcontinent, with evidence for early cereal cultivation

dating back about 4500years. Sorghum is the world’s fifth major cereal in terms

of production and acreage (International Crops Research Institute for the Semi-

Arid Tropics, ICRISAT, 2004). It is a staple food crop for millions of the poorest

18
and most food-insecure people in the semi-arid tropics of Africa, Asia and

Central America. The crop is genetically suited to hot and dry agro-ecologies

where it is difficult to grow other food grains.

Fig. 2.2: Structure of the sorghum kernel (caryopsis), (Adewole and Sanni,

2005).

2.4.2 Production and Cultivation of Sorghum

Sweet sorghum is a warm-season crop that matures earlier under high

temperatures and short days. It tolerates drought and high-temperature stress

better than many crops, but it does not grow well under low temperatures. It can

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be grown on soils ranging from heavy clay to light sand. Loam and sandy loam

soils generally allow the best syrup production. Sweet sorghum is ideally shown

during June, coinciding with the South-West monsoon. Rainfall of 500-600mm

distributed ideally across growing period is the best, unless the soil can hold

much water (deep). The crop does not prefer high rainfall as high soils moisture

or continuous heavy rain after flowering may hamper sugar increase. If irrigation

is available, sowing can be done before June so that the crop does not face heavy

rains after flowering and more so during the last half of grain malting period.

Sowing during rabi or summer season may result in low biomass and sugar

yield. For the seed rate, a good crop may have 1,000,000 to 120,000 plants/ha.

This can be achieved with 8kgseed/ha. Sowing can be done on ridges or in

furrows at a spacing of 60cm between rows and 15cm within rows. Three to four

seeds are dibbled in each hill/planting hole and the seedlings are to be eventually

thinned to one per hill. If a planter is used, then the existing seed rate will be

reduced.

2.4.3 Nutritional Attributes of Sorghum

Grain sorghum is mostly used as a cereal grain energy source and is a good feed

stuff for poultry, pigs and ruminants. Its composition is roughly similar to that of

maize and it is particularly rich in starch (more than 70% of the dry matter).

Crude protein content in sorghum grain ranges from 9 to 13% DM and is slightly

higher than that of maize, though much more variable depending on growing

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conditions. Like maize, it has low lysine content and its utilization may require

amino acid supplementation. Fat content is also slightly lower in sorghum grain

than in maize. Sorghum grain is devoid of Xanthophylls and 70% of its

phosphorous is bound in phytate (Sauvant et al., 2004).

2.5 Cocoyam

2.5.1 Origin and History of Cocoyam

The cocoyam are aroids, grown primarily for edible corms, belonging to the

family Araceae, which has 110 genera of which Colocasia, Xanthosoma,

Amorphallus, Alocasia and Cytospermaare important and about 2000 species.

Alocasia, Xanthosoma and Colocasia belong to the tribe Colocasia, it is in the

sub-tribe Colocasinae; Xanthosoma in the sub-tribe Caladinae. The family

Araceae is grown in a number of tropical and sub-tropical countries and has been

identified as a major group of under exploited root crops with an uncertain future

through limited demand that may lead to reduced production until it becomes a

minor niche crop, (O’Hair, 1984).

The terms ‘edible aroids’ or ‘cocoyam’ are used for both Colocasia and

Xanthosoma, referred to as taro and tannia respectively, when differentiated in

post-harvest characteristics. Other edible aroids, of lesser economic importance

include; Alocasia, Cyrtosperma and Amorphophallus, cultivated locally to form

important food crops in parts of India, Southeast Asia and Pacific Islands,

(O’Hair, 1984; Purseglove, et a.l., 1979;Rubatzkyet al., 1997). Cocoyams are

21
less important than other tropical root crops such as yam, cassava and sweet

potato but form subsistence and emergency crops in many countries and are

stable crops in parts of Ghana, Japan, Nigeria and Hawaii. In South Pacific

Island countries, edible aroids, principally taro, form a high proportion of the

root crops, however in Caribbean and West Africa, tannia dominates, (Opara,

2000).

Tannia is a crop of South (tropical) American origin domesticated by the tropical

Amerindians and people of the Caribbean and is called ‘malanga’ in Cuba and

‘yautia’ in the Dominican Republic and Puerto Rico. During the slave trade it

was taken to Africa and since the nineteenth century it has been cultivated in

Pacific Islands and Asia because of its resistance to pests and diseases. Other

names include mankani, tanier, belembe, maduma, yautia de Anglo-Saxon,

macabo, gualusa, tannie, Chou caraibe and mangarito. (O’Hair, 1984;

Purseglove, et al.,1979; Rubatzky, et al., 1997).

Taro (Colocasia esculenta) or taro eddoe, dasheen gabi, keladi, taru, arvi, kolkas,

dalo or sato-imo has been the focus of cocoyam research and its agronomic and

phenotypic properties have been more studied.

2.5.2 Cocoyam Characterization

Cocoyams are characterized morphologically by subterranean stems, corms,

enclosed by dry scale-like leaves. The cocoyams are differentiated in leaf

attachment; tannia has a stalk attached to the leaf edge while in taro it emerges

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near centre. In tannia, secondary corms (cormels) are used for human

consumption while the corms are used for vegetative propagation. Taro is a

versatile crop grown in both low land and upland conditions whereas tannia

cannot stand water-logging and is a partial shade-loving annual crop that prefers

deep well-drained soils of pH 5.5-6.5 and tolerates saline soils, (O’Hair, 1984).

Biophysical and socio-economic factors accounting for the lower yields include

low yielding cultivars, pest and disease, poor husbandry practices, deteriorating

soil fertility, lack of processing and innovation of products. Cocoyam is

susceptible to attack by pests and diseases. A typical pest is the slugs that wound

corms, providing entry points for spoilage microorganisms. Losses can reach

60% of corms. Prevention is by use of disease-free planting materials, weeding

and hilling and treatment with copper-based pesticides, (Opara, 2000).

2.5.3 Cocoyam Composition

Tagodoe and Nip (1994) concluded that taro flours are rich in starch and total

dietary fibers are low in fat, protein and ash. Moisture content are 69.1% and

67.1% respectively for taro and tannia; energy values of 4800 and 5210kj/kg; fat

contents 0.10% and 0.11%, sugar contents 1.01% and 0.42%, ash contents

0.97% and 1.01%, (Bradbury, et al., 1988). Taro corms contain pigment

anthocyanins such as cyaniding-3-glucoside, pelargonidin-3-glucoside and

cyaniding-3-rhamnoside and anthocyanogens.

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2.5.4 Cocoyam Anti-Nutrient Components

Food and feed usage of cocoyam is restricted because of the acrid nature of the

corms that irritate upon ingestion and lowers palatability, (Gohl, 1981). This has

reduced possibilities for processing. The acridity is such that if eaten raw, corms

cause swelling of the lips, mouth and throat as well as bitterness, astringent taste

and scratchiness in the mouth and throat. Anti-nutritional and off-taste problems

have been related to content of needle-like raphides of calcium oxalate crystals

and other acidic and proteinous factors, (Bradbury, et.al., 1988; Bradbury, et.al.,

1995; Sakai, 1979). Bradbury and Nixon (1998) have explained the acridity is

due to the mechanical sharp raphides in puncturing the soft skin and irritant

proteases and other compounds, (Holloway, et.al., 1989). Paullet.al.,(1999) have

proposed the presence of one or more chemical irritants on raphide surfaces.

Content of calcium oxalate raphides has been reported to decrease from outer to

the centre of the corm, (Sunell, et.al., 1979) and be more abundant in distal

sections than mid or apical, (Sefa-Dede, et.al., 2002). Effects of cocoyam anti-

nutritional factors range from reduction of food and feed intake, with depression

or weight gain, to pancreatic hypertrophy in experimental animals, (Fertuga,

et.al., 1976; Gerpacia, et.al., 1975; Moy, et.al., 1979; Ravindarn, et.al, 1996;

Samarasinghe, et.al., 1992) and (Tang, et.al., 1983). Other specific

antinutritional factors have been reported such as trypsin inhibitors, (Liener,

24
et.al., 1980; Sumathi, et.al., 1979), alpha-amylase inhibitors, (Krishnasharma,

et.al., 1980) and Sapotoxins, (Watt, et.al., 1962).

Philipy, et.al.,(2003), concluded levels of phytate in taro at 0.169% were higher

than cassava (0.133%). Bradbury, et.al., (1995), reported contents of cyanide

present in the leaves (0-30mg HCN/kg fresh weight) and in the stems (0-3mg

HCN/kg) of taro and tannia were only about 1%-5% that of cassava leaves and

tubers and are thus not a cause for concern for human nutrition.

2.5.5 Cocoyam Utilization

Cocoyam usage can be similar to that of potato in the Western world and corms

can be converted into several specific food and feed products and also for

industrial purposes. Processes for stabilizing and adding value by conversion to

semi-finished and end products include boiling, roasting, baking, frying in oil,

and pasting, milling and pounding. Arnaud-vinas and Lorenz, (1999), have also

considered the possibility of production of pasta from blends of wheat and taro

flours. It is also used in soup thickeners and baking flours, in beverages, as

porridge and in producing foods for people with gastrointestinal disorders,

(Ihekoronye, et.al., 1985; Onwueme, 1978). Subhadhiraskul, et.al., (2001),

reported that taro starch can effectively replace maize as a binding agent in tablet

manufacture. (Lawal, 2004) has suggested cocoyam starches could be modified

as for other industrial starches.

25
The high digestibility of cocoyam starches and the small size of taro granules

form a good basis for processed baby foods. In parts of West Africa, boiled

corms are mashed to form a weaning diet. Onwulata and Konstance, (2002),

have reports on the process of formulation of weaning food with taro flour

extruded with whey protein concentrate, whey protein isolate and latalbumin.

Mature aroids are processed into flour for ‘fufu’, commonly eaten in Nigeria

with stew. In South eastern parts of Nigeria in particular, tannia is used in small

quantities as a soup thickener after boiling and pounding to obtain a consistent

paste, (Obiechina, et.al., 1987; Onwueme, et.al., 1978. Taro flours have unique

properties from small starch granules (greater 1.5 micrometer) and high

mucilage (gum) content, suggesting a replacement for corm on wheat starch in

weaning foods. The use of cocoyam as a raw material for brewing has been

reported by Onwuakaet.al., (1996). The final beer, though slightly bitter, was

acceptable to local assessors

2.6 Yam

2.6.1 The origin and history of yam

The yam (Dioscorea sp.) is a monocotyledous tuber bearing plant, belonging to

the family Dioscoreacea within the genus Dioscorea. The Dioscorea species is a

climbling, vigorously twining herbaceous plant that coils swiftly around the

stake. They are perennial through root system but are grown as annual crops

(Udensi et al 2008; IITA, 2006). Having over 600 species but in which a few are

26
cultivated for food and medicines. The most cultivated species in Nigeria are the

white yam (Dioscorea rotundata), yellow yam (Dioscorea cayenesis), water yam

(Dioscorea alata) and trifoliate yam (dioscorea dumetorum) (Amusa et al 2003).

2.6.2 Dioscorea bulbifera

Dioscorea bulbifera is available in two varieties, the edible and non-edible. The

edible varietiesare cultivated and widely distributed in west Africa, west indies,

south pacific and south east Asia. (FAO, 1985). It is an aerial yam also known

with common names as potato yam, cheeky yam, bulbils-bearing yam. In south-

eastern states in Nigeria it is known as “Adu” while in south-south Nigeria it is

called “Odu”. (Sanful et al., 2013). Though it possesses a distinctive flavor and

comparable in nutritional content to most preferred yams, it does not have the

same appeal compared to Dioscorea alata and Dioscorea rotundata and so it is

less studied and has high rate of postharvest losses. In Nigeria, aerial yam is

almost going into extinction and it is counted among the underutilized plant

species. (Ojinaka et al, 2016).

2.6.3 Dioscorea rotundata,

The "white yam", and Dioscorea cayenensis, the "yellow yam", are native to

Africa. They are the most important cultivated yams. In the past they were

considered two separate species but most taxonomists now regard them as the

same species. There are over 200 cultivated varieties between them.

27
White yam's tuber is roughly cylindrical in shape, the skin is smooth and brown

and the flesh usually white and firm. Yellow yam is named after its yellow flesh,

a colour caused by the presence of carotenoids. It looks similar to the white yam

in outer appearance; its tuber skin is usually a bit firmer and less extensively

grooved. The yellow yam has a longer period of vegetation and a shorter

dormancy than white yam.

The Kokoro variety is important in making dried yam chips (Dumont, 2000).

They are large plants; the vines can be as long as 10 to 12 meters (33 to 39 ft.).

The tubers most often weigh about 2.5 to 5 kilograms (5.5 to 11.0 lb.) each but

can weigh as much as 25 kilograms (55 lb.). After 7 to 12 months’ growth the

tubers are harvested. In Africa most are pounded into a paste to make the

traditional dish of "pounded yam" (Kay, 1987).

2.6.4 Dioscorea alata

Dioscorea alata, commonly called water yam or tenth month yam is the most

widespread yam species and more important as food in the west Africa and the

Caribbean than in Asia and America where it originated and has been competing

with the most important native species Dioscorea rotundata. It is an excellent

source of starch which provides calorific energy and protein three times more

superior than the one of cassava and sweet potato. (Coursey. 1973).It has low

sugar content necessary for diabetic patients (Udensi, et al., 2010). They are also

known for their high nutritional content, with crude protein content of 7.4%

starch content of 75- 84% and vitamin C content ranging from 13.0 to

28
24.7mg/100g. (Osagie, 1992). Due to the high starch content of Dioscorea alata

tubers they provide a good source of dietary carbohydrates in the tropics and

subtropical regions (Osagie, 1992).

2.6.5 Dioscorea dumetorum

The bitter yam, is popular as a vegetable in parts of West Africa; one reason

being that their cultivation requires less labour than other yams. The wild forms

are very toxic and are sometimes used to poison animals when mixed with bait.

It is said that they have also been used for criminal purposes (Kay, 1987)

There are many cultivars of yam throughout the humid tropics. The most

economically important are discussed below:

2.6.6 Health benefits of yams

 Yam is a good source of energy; 100 g provides 118 calories. Its crunchy

edible part chiefly composed of complex carbohydrates and soluble dietary

fibre.

 Dietary fibre helps reduce constipation, decrease bad (LDL) cholesterol

levels by binding to it in the intestines and lower colon cancer risk by

preventing toxic compounds in the food from adhering to the colon mucosa.

29
2.7 Cassava

2.7.1 The origin and history of cassava

Cassava has its genetic, geographical and agricultural origin in Latin America.

Its domestication began 5000-7000 years BC in the Amazon, Brazil (Allen,

2002) and it was distributed by the Europeans to the rest of the world (Henry and

Hershey, 2002). Cassava was taken from Brazil to the west coast of Africa by

Portuguese navigators in the 16th century (Nweke, 1994). It was introduced to

most of Asia and the Pacific in the late 18th and early 19th centuries.

Cassava as known in English, is "manioc" in French, "yuca" in Spanish, and

"mandioca" in Portuguese. Cassava belongs to the class Dicotyledoneae, family

Euphoribiaceae, tribe Manihoteae, genera Manihot tournefort and species

Manihot esculenta Crantz.

2.7.2 Chemical and Nutritional Composition of Cassava Roots

Cassava roots and leaves are used for human consumption and animal feed

(Dahniya, 1994) cassava roots are rich in digestible carbohydrates, mainly

starch. Cassava root starch consists of both amylose (20 percent) and

amylopectin (70 percent). Generally, cassava roots have less than 1percent free

sugars (Bradbury and Holloway, 1988).

Cassava roots are low in protein and fat and as a result should be eaten along

with other crops rich in essential amino acids to supplement the deficit, such as

vegetables, cereals, fish and meat (Okigbo, 1980; FAO, 1990). Cassava has a

high content of dietary fibre, magnesium, sodium, riboflavin, thiamine, nicotinic

30
and citrate (Bradbury and Holloway, 1988). The major constraint on cassava

roots as human food is the presence of toxic cyanogenicglucosides compounds in

the tissues.

2.7.3 The problems of cyanogenic glycosides in cassava

Cyanogenicglucosides are phytotoxins that occur in about 2000 plant species of

which a number are used as crops (Conn, 1980). Cassava roots and leaves

contain cyanide in two different forms;

 The glycosides; linamarin and lotaustraline which are considered "bound"

and;

 The non-glycosides; hydrogen cyanide (HCN) and cyanohydride which

are considered "free".

Free cyanide comprises of 8-12 percent of the total tuber cyanide. This

cyanide can under some circumstances lead to human toxicity problems and

cassava for food use has to be processed to remove cyanide containing

substances. There is a great variation in toxicity between cultivars. A

distinction is usually made between "sweet cultivars" with relatively low

content of cyanogenic glycosides (below 10mg/100g of fresh weight) and

"bitter cultivars" with high cyanogenic glycoside content (above 20mg/100g

fresh weight) although many intermediate forms exist. Traditionally the sweet

cultivars were considered non-toxic while the bitter ones were considered

toxic. Cyanide levels in the range 6 to 370mg/kg have been found depending

on the particular cultivar, growing conditions (soil type, humidity,

31
 temperature) and the age of the plant. The highest proportion of HCN is

found in the peels (Hahn, 1984; Onwueme, 1978) it is for this reason the

cassava root is always peeled before being processed or consumed.

2.8 Potato

The cultivation of potatoes for starch mainly takes place in Germany, the

Netherlands, China, Japan, France, Denmark, and Poland, but also in Sweden,

Finland, Austria, the Czech Republic, Ukraine, Canada, and India. Many types

of potatoes are grown; for the production of potato starch, potato varieties with

high starch content and high starch yields are selected. Potato starch contains

typical large oval spherical granules ranging in size between 5 and 100 μm.

Potato starch is a very refined starch, containing minimal protein or fat. This

gives the powder a clear white colour, and the cooked starch typical

characteristics of neutral taste, good clarity, high binding strength, long texture

and a minimal tendency to foaming or yellowing of the solution. Potato starch

contains approximately 800 ppm phosphate bound to the starch; this increases

the viscosity and gives the solution a slightly anionic character, a low

gelatinization temperature of approximately 60 °C (140 °F),and high swelling

power. These typical properties are used in food and technical applications.

(BeMiller, et al., 2009).

32
2.9 Physico-Chemical Properties of Sugar Syrup

Akinola and Ayanleye (2004) recorded that the glucose syrup obtained from acid

and enzyme hydrolysis of cassava starch had a pH of 5.9, specific gravity of

1.0033, total reducing sugars of 30.4%, 80% total solids and 38% Dextrose

Equivalent. All glucose syrup is palatable, the low conversion being tasteless

(around 30 DE) Allikonis standard. The sweetness of glucose syrup has been

attributed principally to fructose, dextrose, maltose and the saccharide

composition and distribution of glucose syrups vary with degree and method of

hydrolysis. As the DE decreases, the saccharide content of higher molecular

weight might increase while the lower molecular weight saccharides increase

with increase in DE.

2.9.1 Amylose and Amylopectin

Plant starches come in two different forms; amylase (20-30%) and amylopectin

(70-80%), each with their own physical and chemical properties.

Amylopectin is a soluble polysaccharide and highly branched polymer of

glucose found in plants. It is one of the two components of starch, the other

being amylose. Glucose units are linked in a linear way with alpha-(1-4)

glycosidic bonds. Plants store starch within specialized organelles called

amyloplasts. When energy is need for cell work, the plant hydrolyzes the starch

releasing the glucose subunits. Humans and other animals that eat plant foods

also use amylase, an enzyme that assists in breaking down amylopectin.

33
Starch is made up of about 70% amylopectin by weight, though it varies

depending on the source (higher in medium-grain rice to 100% in glutinous rice,

waxy potato starch and waxy corn and lower in long-grain rice, amylomaize, and

russes potatoes for example.). Amylopectin is highly branched, being formed of

2000 to 200000 glucose units. Its inner chains are formed of 20-24 glucose

subunits. Dissolved amylopectin starch has a lower tendency of retro gradation

(gelling) during storage and cooling. For this main reason, the waxy starches are

used in different applications mainly as a thickening agent or stabilizer.

2.9.2 Dextrose Equivalent

This is the measure of the amount of reducing sugars present in a sugar product,

relative to glucose, expressed as a percentage on a dry basis. The degree of

hydrolysis of starcheswhether by acid or enzyme or a combination of the two

governs the composition of the final product. Complete or 100% hydrolysis of

starch gives dextrose, and starch and dextrose are thus the two extremes of the

range. The DE scale is therefore related to these two products. Starch has

undergone 0% hydrolysis and is thus given a nominal value of 0 DE while

dextrose the ultimate hydrolysis product, is given a value of 100 DE.

Intermediate values represent the gradual degradation or breakdown of the starch

first into large glucose polymers of 20-30 units in size then into much smaller

products of 20 units and below. Depending on its DE, glucose syrup will

therefore contain a greater or lesser percentage of high and low molecular weight

34
glucose polymers and it is the percentage of the individual saccharides in syrup

which gives that syrup its unique properties and which differentiates syrup from

another.

Unfortunately, it is not simply a matter of determining the DE of a glucose syrup

to deduce its properties, as it is possible using different hydrolysis techniques to

produce two (or even three) syrups of the same DE with different carbohydrate

composition and thus different properties. While, the scale of DE covers the

range from 0 to 100, not all these products are considered as glucose syrups

industrially. From 3-20 DE, the products are described as maltodextrins, from 20

to about 75 DE as glucose syrups, and above about 75 DE as hydrolysates.

2.9.3 Viscosity of Syrup

Viscosity of a fluid is a measure of its resistance to gradual deformation by shear

stress of tensile stress. For liquids, it corresponds to the informal concept of

‘thicknesses’. For example, honey has a much higher viscosity than water.

Viscosity is a property arising from collisions between neighboring particles in a

fluid that are moving at different velocities. Viscosity is a measure of the ratio of

shearing stress to rate of shear.

Shear stress (dynes) = Poises rate of shear (cm/sec).

2.9.4 Colour of Syrup

Colour scales: to be useful, a color scale should relate to how we see color, be

simple to understand, be linear throughout color space and be able to quantify

35
color differences. The color scales that are most widely used by the food

industry are the Hunter L, a, b and CIE (Commission Internationale d’ Eclairage)

L*, a*, b* scales. These are 3-dimensional scales. Both are based on the

opponent colors theory that states that red, green and the human eye cone

responses are re-mixed into black-white, red-green, and yellow-blue, opponent

coders as they move up the optic nerve to the brain. The *L, *a, *b type of scales

simulates this as;

*L (lightness) axis - 0 is black, 100 is white.

*a (red-green) axis - positive values are red; negative values are green and 0 is

neutral.

*b (yellow-blue) axis - positive values are yellow, negative values are blue and 0

is neutral.

36
 CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Raw Material Collection

The Cocoyam (Colocasia esculenta) tuber known as xanthosoma mafafa, yam

(Dioscorea bulbifera), commonly known as Bulb yam, (Adu in igbo land) and

potato (Ipomoea batatas L),white colored sorghum (Sorghum vulgare) with

varietal name fara-fara, millet (Pennisetum glaneum) known as Ex –Borno and

wheat (Triticum eastivum)know as Ex – Jos used in this work were purchased

from the Department of Crop Science of the Research Institute of Michael

Okpara University of Agriculture Umudike, Umuahia, Abia State, Nigeria. All

solvent used in HPLC analysis were of HPLC grade and obtained from E-Merck

(Darmstadt, Germany). Other reagents were purchased from Sigma Aldrich

(Steinheim, Germany). The equipment used in carrying out this work were

gotten from Federal University of Technology Owerri, Imo State, Nigeria and

International Institute of Tropical Agriculture, Ibadan, Oyo State, Nigeria.

3.2 Methods

3.2.1 Preparation of Arial yam and cocoyam tuber to yam flour

The cocoyam (Colocasia esculenta) and yam (Dioscorea bulbifera) were

harvested. These freshly harvested tubers of yam and cocoyam were washed

separately, manually peeled, washed, sliced and oven dried at 400C for 60

37
minutes and afterwards was milled to get the flour (Pancoast and Junk, 1980;

Novo 1999) and (Nwanekezi et al., 2004).

Cocoyam and yam tuber

↓
Washing
↓
Peeling
↓
Washing
↓
Slicing
↓
Oven dried at 400C for 60 minutes
↓
Milling
↓
Packaging

Fig 3.1 Flow diagram of production of yam and cocoyam flour

38
3.2.2Preparation of potato roots to starch

The method described by Osuji and Anih, (2011) was used. Starch

extraction was carried out from the fresh potato roots. The roots were

washed, peeled and milled into slurry. The slurry was properly stirred

and allowed to settle for about 6 hours. After settling of the potato

slurry, a heterogeneous mixture was observed, the top part o of the

mixture was a transparent liquid and the bottom part was a white thick

liquid which is starch. The supernatant was decanted and the sediment

which contains the starch was filtered with muslin cloth and oven-

dried at 45-55˚C for 30 minutes to produce the dry starch, which was

later milled to produce a fine powder

39
 Potato roots

Washing
↓

Peeling
↓

Grating/rasping
↓

Mixing with water

↓

Filtering/screening
↓

Settling
↓

Starch washing/decanting
↓

Settling/dewatering in a clean bag by pressing

↓

Drying
↓

Milling
↓

Potato starch
Fig 3.3 Flow diagram for potato starch production

40
3.2.3 Production of Malted Sorghum

Sorghum variety of guinea corn Farafara (sorghum vulgare) was used as one of

the sources of enzymes for the hydrolysis of starch. The sorghum was washed

and soaked/steeped in portable water for 24hours and the water was changed

every 6hours interval to undergo air rest. It was spread on a jute bag in an air

tight room for the commencement and completion of sprouting. Water was

sprinkled every 6hours interval on the grains to enhance sprouting. It was

germinated for 24hours before the emergence of sorghum rootlets. The properly

sprouted grains were sun dried for 3days and were milled to get the malted

sorghum.

41
 Sorghum grains
↓
Sorting
↓
Washing
↓
Steeping/germination (24hours)
↓
Sun drying
↓
Milling
↓
Packaging
↓
Malted sorghum

Fig 3.3 Flow diagram of production of malted sorghum

42
3.2.4 Production of Malted Wheat

Wheat variety Ex –Jos (Triticum eastivum) was used as one of the sources of

enzymes for the hydrolysis of starch. The wheat was washed and soaked/steeped in

portable water for 24hours and the water was changed every 6hours interval to

undergo air rest. It was spread on a jute bag in an air tight room for the

commencement and completion of sprouting. Water was sprinkled every 6hours

interval on the grains to enhance sprouting. It was germinated for 24hours before

the emergence of sorghum rootlets. The properly sprouted grains were sun dried

for 3days and were milled to get the malted wheat flour.

43
 Wheat grain

Sorting

Washing

Steeping (24hours)

Sprouting/germination (24hours)

Sun drying

Milling

Packaging

Malted wheat

Fig 3.4 flow diagram for production of malted wheat

44
3.2.5 Production of Malted Millet

The millet variety Ex –Borno (Pennisetum glaneum) was used as one of the

sources of enzymes for the hydrolysis of starch. The millet was washed and

soaked/steeped in portable water for 12hours and the water was changed every

6hours interval to undergo air rest. It was spread on a jute bag in an air tight room

for the commencement and completion of sprouting. Water was sprinkled every

6hours interval on the grains to enhance sprouting. It was germinated for 24hours

before the emergence of sorghum rootlets. The properly sprouted grains were sun

dried for 3days and were milled to get the malted millet flour.

45
 Millet grain

Sorting

Washing

Steeping (12hours)

Sprouting/germination (24hours)

Sun drying

Milling

Packaging

Malted millet

Fig 3.5 flow diagram for production of malted millet

46
3.3 PROCEDURE FOR SYRUP PRODUCTION USING EXOGENOUS

ENZYME APPLICATION (AMYLOGLUCOSIDASE)

The method described by Osuji and Anih (2011) was used. The mash water to be

used for the syrup production was prepared to a pH of 11 with aid of calcium

hydroxide. 20grams of the starch samples were weighed respectively into clean

pots. Slurry was made by adding 250ml of the mash water respectively into the

weighed starches. The temperature of the slurries were raised to 45˚C, after which

20grams of the amylogucosidase enzyme was added to each of the slurries, The

slurries were stirred and maintained at that temperature for 20 minutes.

Temperature was raised to 55 , they were stirred and allowed to rest for 10

minutes. Iodine tests were carried out by adding 2 drops of iodine to a few drops of

the samples on a ceramic tile. The temperature of the slurries were raised to 65˚C

and maintained for 1 hour. Another iodine tests were carried out. Temperatures

were further raised to 90-93˚C and maintained for another 1 hour. The slurries

were then boiled for 5 minutes, after which iodine tests were carried out. The

samples were then cooled to 60˚C by placing them in an ice water bath. The pH of

the samples was checked. After hydrolysis, the liquors were boiled for 10 minutes

to denature enzymes. The converted slurries were then filtered across a double-

layered muslin cloth. The samples were then evaporated and concentrated through

evaporation using a water bath, and then packaged.

47
3.3.1 PROCEDURE FOR SYRUP PRODUCTION USING CRUDE

ENZYME APPLICATION FROM MALTED CEREALS

The method described by Okafor et al (2017) was used. The mash water to be used

for the syrup production was prepared to a pH of 11 with aid of calcium hydroxide.

20grams of the starch samples were weighed respectively into clean pots. Slurry

was made by adding 250ml of the mash water respectively into the starches. The

temperature of the slurries were raised to 45˚C, after which 3grams of the malted

grains respectively was added to each of the slurries, The slurries were stirred and

maintained at that temperature for 20 minutes. Temperature was raised to 55 ,

they were stirred and allowed to rest for 10 minutes. Iodine tests were carried out

by adding 2 drops of iodine to a few drops of the samples on a ceramic tile. The

temperature of the slurries were raised to 65˚C and maintained for 1 hour. Another

set of iodine tests were carried out. Temperatures were further raised to 90-93˚C

and maintained for another 1 hour. The slurries were then boiled for 5 minutes,

after which iodine tests were carried out. The samples were then cooled to 60˚C by

placing them in an ice water bath. The pH of the samples was checked, after

hydrolysis, the liquors were boiled for 10 minutes to denature enzymes. The

converted slurries were then filtered across a double-layered muslin cloth. The

samples were then evaporated and concentrated through evaporation using a water

48
bath, and then packaged. This same procedure was repeated for 6, 9, 12, 15 and 20

gram of the malted cereals respectively on each of the three starches.

3.3.2 PROCEDURE FOR BRIX DETERMINATION

The apparent ˚Brix was determined using a portable digital handheld refract meter

(VBR32T Bellingham and Stanley UK Brix/ATC 0-32%).The digital refractometer

was cleaned with a clean wiper and standardized with distilled water at 20˚C until

the brix value reads zero. Two drops of syrup sample at 20˚C was dropped on the

lens (sensitive surface) of the refractometer and measured. The syrups with the

highest brix level was chosen for the sugar spectra analysis using high performance

liquid chromatography because HPLC sugar analysis is very expensive. The cost is

highly exorbitant.

3.3.3 DETERMINATION OF SUGAR PROFILE WITH HIGH

PERFOMANCE LIQUID CHROMATOGRAPHY USING REFRACTIVE

INDEX DETECTOR (HPLC-RI)

Determination and quantification of specific reducing sugars: fructose, glucose, D-

xylose. D-raffinose, stachyose and maltose was performed by HPLC with

Refractive Index detector (Waters e2695, USA; 2414 RI Detector)using Sugar

Analysis 300 x4.6mm column. The analysis was performed at 35°C with a flow

rate of 1 ml min-1using isocratic elution with 75% acetonitrile (AcN):25% water

49
(H2O) mixture as a mobile phase. All samples were centrifuged at 4,000 rpm for

10 min (HermleLabnetZ323) and the supernatant was filtered through a 0.22 μm

PES membrane (Sartorius StedimBiotech, Germany). Then the filtrate was diluted

10 times before direct injection into the HPLC. The sugars seen and detected were

recorded.

3.3.4 STATISTICAL ANALYSIS

Data were expressed as mean ± standard deviation (SD) from two parallel

measurements. The analysis of variance (ANOVA) and least significant difference

(LSD) were performed with the SPSS 20.0 software (SPSS Inc.) significance of

difference was defined at Ɵ =0.05.

50
 CHAPTER FOUR

4.0 RESULTS AND DISCUSSION

4.1 RESULTS

Table 4.1: Brix Reading of Different Crude Enzyme Application at Different

Concentrations for Different Starch Sources
Sources of Sources of Quantity of crude enzymes applied (Grams)
Enzymes carbohydrates 0 3 6 9 12 15 20

Millet Cocoyam flour 0.0 6.5 7.6 9.4 11.3. 12.0 12..0

Aerial yam flour 0.0 7.3 8.7 10.9 12.3 13.0 13.0

Potato white starch 0.0 10.5 11.8 13.8 15.5 18.5 18.0

Sorghum Cocoyam flour 0.0 8.5 10.7 12.3 14.5 19.0 19.0

Aerial yam flour 0.0 5.8 6.3 7.7 8.9 10.0 10.0

Potato white starch 0.0 8.5 9.8 11.5 13.7 15.0 14.8

Wheat Cocoyam flour 0.0 6.6 9.8 11.3 13.6 15.0 15.0

Aerial yam flour 0.0 8.3 10.2 12.3 13.8 15.0 15.0

Potato white starch 0.0 11.3 12.6 15.0 19.0 25.0 24.8

51
Table 4.2 Sugar Profiles of Flours and Starch Source Interaction

Starch Fructose Glucose Sucrose Maltose D-Xylose D-Raffinose D-Stachyose

source

COF 12.1599 9.7191 6.2552 14.2028 0.0247 0.0115 0.000

AYF 7.3599 8.9246 11.3104 21.1073 0.0408 0.0079 0.000

PWS 5.7823 9.6994 11.0100 26.0931 0.0082 0.1218 0.000

KEY:

COF= Cocoyam flour

AYF= Aerial yam flour

PWS= potato white starch

52
 Table 4.1.2: Sugar Profile of Enzyme Source Interaction

Enzyme Fructose Glucose Sucrose Maltose D-Xylose D-Raffinose D-Stachylose

source
AMG 14.7494 3.3299 3.0343 6.6713 0.0092 0.0158 0.000

Millet 11.3014 24.2293 24.1151 40.7690 0.0922 0.0801 0.000

Sorghum 2.5692 1.9467 1.5257 9.5760 0.0022 0.0128 0.000

Wheat 5.1161 8.2849 9.4258 24.8547 0.0042 0.0795 0.000

KEY:

AMG= Amyloglucosidase

53
 Table 4.4 Showing the Sugar Spectra of the Interactive Effect of Enzymes and Starches.

Crop Enzyme Fructose Glucose Sucrose Maltose D-xylose D-Raffinose D-Stachyose

0.000
AMG 21.7177f±0.101 5.0523b±0.133 0.5237g±0.160 0.6003d±0.007 0.0010c±0.010 0.0092g±0.001
Millet 25.7137a±0.101 31.9163a±0.133 19.0147h±0.160 41.4443d±0.007 0.0960c±0.010 0.0142i±0.001 0.000
COF 0.000
Sorghum 0.0106h±0.101 1.0203k±0.133 1.2997j±0.160 0.9023f±0.007 0.0010f±0.010 0.0122i±0.001
Wheat 1.1977b±0.101 0.8873j±0.133 4.1827i±0.160 13.8643j±0.007 0.0010e±0.010 0.0102g±0.001 0.000

AMG 22.3137c±0.101 4.1213d±0.133 7.7797b±0.160 16.2453e±0.007 0.0080b±0.010 0.0052h±0.001 0.000

Millet 3.1757g±0.101 23.7643c±0.133 32.8997a±0.160 33.9003g±0.007 0.1760a±0.010 0.0012e±0.001 0.000
AYF Sorghum 0.2857f±0.101 0.9573i±0.133 2.06067l±0.160 21.6203i±0.007 0.0010e±0.010 0.0222f±0.001 0.000
Wheat 3.6647f±0.101 6.8553h±0.133 2.50167k±0.160 12.6633h±0.007 0.0070d±0.010 0.0032d±0.001 0.000

AMG 0.2170e±0.003 0.8160e±0.751 0.7997e±0.004 3.1683b±0.006 0.0187d±0.006 0.0330c±0.036 0.000

PWS Millet 5.0150g±0.265 17.0073f±0.472 20.4310f±0.527 46.9623d±0.006 0.0047f±0.006 0.2250a±0.036 0.000
Sorghum 7.4113g±0.751 3.8623f±0.635 1.2167d±0.930 6.2053c±0.006 0.0047f±0.006 0.0040a±0.360 0.000
Wheat 10.4860d±0.854 17.1120g±0.390 21.5930c±0.536 48.0363a±0.006 0.0047e±0.006 0.2250b±0.360 0.000
LSD 0.584 0.205 0.379 0.387 0.006 0.001 0.000
The values are means of triplicate determination±SD
Values within the same column, followed by the same letter is not significantly different at 0.05level.
KEY: COF= Cocoyam flour, AYF= Aerial yam flour, PWS= Potato white starch, AMG= amyloglucosidase, LSD= least significant different.

54
4.2 DISCUSSION

The result showed that there was an interaction between the starches and enzymes

and that had effect on the sugars identified

4.2.1 SUGAR PROFILE OF WHEN ONLY FLOURS AND STARCH

SOURCE INTERACTION IS CONSIDERED

For fructose, interaction between the flours and starch showed that, cocoyam flour

(COF) gave the highest mean yield of fructose (12.16g/l) followed by Aerial yam

flour (7.36g/l) while potato white starch gave the least fructose yield (5.78g/l)

irrespective of the enzyme used. For glucose, interaction between the flours and

starch showed that, cocoyam flour (COF) gave the highest mean yield of glucose

(9.72g/l) followed by potato white starch (9.70g/l) while Aerial yam flour (AYF)

gave the least glucose yield (8.92g/l) irrespective of the applied. For sucrose, the

interaction of flours and starch showed that, potato white starch (PWS) liberated

the highest sucrose portion (11.31g/l and 11.01g/l) while cocoyam flour being the

least yield of 6.36g/l notwithstanding the enzyme used. For maltose, interaction

between flours and starch showed that, potato white starch gave the highest mean

yield of maltose (26.09) followed by Aerial yam flour (21.11g/l) while cocoyam

flour (COF) gave the least mean yield (14.20g/l) irrespective of enzyme used. For

55
D-Xylose, interaction between the carbohydrate sources showed that Aerial yam

flour gave the highest mean yield of D-Xylose (0.05g/l) as against cocoyam flour

(0.02g/l) while potato starch gave the least (0.01g/l) irrespective of enzyme used.

For D-Raffinose, interaction between the carbohydrate source showed that PWS

gave the highest mean yield (0.12g/l) followed by COF and AYF (0.01g/l) and

trace of D-Stachyose was not found. In fact Ding, et al., (1997) have worked on

fruit juices and revealed the existence of significant differences among the sugar

content. Among the sugars identified include glucose, Xylose.

4.2.2 SUGAR PROFILE WHEN ENZYME INTERACTION ONLY IS

CONSIDERED.

When the enzymes interaction only is considered without paying attention to the

starch source, AMG enzyme gave the highest total yield of fructose with mean (14

75g/l), followed by Millet enzyme (11.30g/l), Wheat enzyme (5.12g/l) and

sorghum enzyme gave the lowest yield (2.57g/l). Millet enzyme gave the highest

total glucose yield (24,23g/l), followed by wheat enzyme (8.29g/l), AMG enzyme

(3.33g/l) and sorghum enzyme being the least (1.95g/l). Millet enzyme gave the

highest total yield of sucrose (24.11g/l), followed by wheat enzyme (9.43g/l),

AMG enzyme (3.03g/l) and sorghum enzyme (1.52g/l). Millet enzyme gave the

highest total maltose yield (40.77g/l), followed by wheat enzyme (24.85g/l),

56
sorghum enzyme (9.36g/l), and AMG enzyme (6.67g/l). Millet enzyme gave the

highest total yield of D-Xylose(0.10) whereas AMG, Sorghum and Wheat yielded

0.00 of D-Xylose. Millet enzyme and Wheat enzyme gave the highest total yield of

D-Raffinose (0.10g/l) each followed by AMG enzyme (0.02g/l) and Sorghum

being the least (0.01g/l)

4.2.4: SUGAR PROFILE AS SHOWN BY THE INTERACTION OF BOTH

ENZYME AND CARBOHYDRATE SOURCES

When the interaction of both enzyme and starch sources are considered together,

the sugars spectra are affected. Cocoyam flour and millet enzyme gave the highest

mean yield of fructose (25.71g/l), followed by the interactive effect of Aerial yam

flour and AMG with yield of 22.31g/l while potato white starch and wheat gave

the lowest yield (10.49g/l). The interactive effect between Cocoyam flour and

millet enzyme gave the highest mean yield of glucose (31.92g/l), followed by the

interaction effect of Aerial yam flour and millet enzyme with yield of 23.76g/l,

with Potato white starch and wheat given the lowest glucose yield of 17.11g/l. The

interaction effect between Aerial yam and millet enzyme gave the highest yield of

sucrose (32.90 g/l), followed by the interaction effect of Potato white starch and

wheat enzyme with yield of 21.22g/l, with Cocoyam flour and millet given the

lowest yield of 19.01g/l. The interaction effect between potato white starch and

57
wheat enzyme gave the highest mean yield of maltose (48.04g/l), followed by the

interaction effect between Cocoyam flour and millet with yield of 41.44g/l with

Aerial yam flour and sorghum enzyme given the lowest yield (21.62g/l). The

interaction effect between Aerial yam flour and millet enzyme gave the highest

mean yield of D-Xylose (0.18g/l), followed by the interaction of Cocoyam flour

and millet enzyme with mean yield of 0.10g/l with Potato white starch and AMG

enzyme given the lowest mean yield (0.02g/l). The interaction effect between

Potato white starch and millet enzyme gave the highest mean of D-Raffinose yield

(0.22g/l), followed by the interaction effect between Aerial yam flour and sorghum

enzyme with mean yield (0.03g/l), with Cocoyam flour and millet enzyme given

the lowest mean yield of (0.01g/l).

For fructose yield, there is no significant difference between cocoyam flour on

AMG and Aerial yam on sorghum enzyme and wheat enzyme as they has the same

superscript (f), likewise no significant difference exist between water yam starch

on wheat and Aerial yam flour on millet and potato white starch on millet as both

has the same superscript (g). Other carbohydrate sources and enzymes are

significantly different with respect to yield of fructose sugar.

For glucose yield, potato white starch on millet and potato white on sorghum are

not significantly different as both have the same superscripts (f), but all other

58
carbohydrate sources on all other enzymes are significantly different with regards

to the glucose yield.

For sucrose yield, cocoyam flour on wheat enzyme and Aerial yam flour on

sorghum enzyme are not significantly different because they have the same

superscript (i). All other carbohydrate sources on all other enzyme sources are

significantly different.

For maltose yield, cocoyam flour on AMG and cocoyam flour on millet are not

significantly as both has the same superscript (d).while other carbohydrate sources

on all enzymes are significantly different with regards to maltose yield.

For Dxylose yield, Aerial yam flour on AMG and millet enzymes are significantly

different, while other carbohydrate sources on all other enzymes sources are not

significantly different in their yield of D-Xylose.

For D-Raffinose yield, cocoyam flour on millet enzyme and sorghum enzyme are

not significantly different, cocoyam flour on AMG enzyme and wheat enzyme are

not significant and potato white flour on sorghum enzyme and millet enzyme are

not significantly different while significant different exist between other

carbohydrate source on all enzymes sources with regards to D-Raffinose sources.

My explanations are in agreement with the results of the study conducted by other

researchers. In fact Osuji and Okafor (2013) have worked on Soymilk and revealed
59
the existence of significant differences among the sugar content of soymilk after

different enzyme treatment using HPLC. Among the sugars identified include

glucose, fructose, maltose, Raffinose, Xylose and stachyose. The presence of

stachyose as identified in their work can be attributed to the action of the cell wall

degrading enzyme used as opposed to this research where stachyose was not

identified because the hydrolysis was mainly as a result of Oligosaccharide

degrading enzymes.

60
 CHAPTER FIVE

5.0 CONCLUSION AND RECOMMENDATIONS

5.1 Conclusion

From the results obtained from this research, Cereal Grains such as millet,

sorghum, and wheat could be used as a source of enzyme by steeping, germinating

and subsequently drying/kilning the germinated cereal grains which helps to

develop the enzyme (a process otherwise known as malting). it is evident that

enzymic hydrolysis by means of endogenous enzyme can hydrolyze starches to

produce syrups and also sources of the starches hydrolyzed determines to a great

extent the type of sugars to be produced in a hydrolysis, as the constituents

enzymes developed in a particular malted grain and the cell structure of the starch

source must be compatible to the intended end product (sugar) in that particular

hydrolysis. With the data and information obtained from this research work, this

paper concludes that syrup producers in the country and private investors should

utilize the opportunity of producing syrups from crude enzymes and not only that

but make choices as to what type of sugar they want to dominate the syrups. This

can be achieved by choosing among the starches and enzymes that produce the

best result for that particular sugar. For syrup rich in fructose cocoyam flour with

malted millet as source of enzyme is the best. For maltose rich syrup, white potato

61
starch with malted wheat as source of enzyme is the best. For syrup rich in

glucose cocoyam flour with malted millet as source of enzyme is an ideal one. For

sucrose rich syrup Aerial yam flour with millet as source of enzyme is the best.

5.2 Recommendations

The production of sugar syrup from Aerial yam starch and cocoyam starch should

be done and also the physico-chemical properties (dextrose equivalent, colour,

viscosity) of these syrups should be done by another research worker. Production

of crude enzymes should be encouraged. Industrial application of crude enzymes

in food processing (example syrup production) should be embarked on by our

country Nigeria.

5.3 Contribution to Knowledge

From the novel research work conducted, it can be well understood that;

 Enzymes can be developed from cereal grains.

 A particular sugar type can dominate in particular syrup and how this can be

achieved.

 A particular sugar can be obtained in syrup at different preferred or desired

concentrations to meet consumer’s preferences.

62
 Endogenous enzyme present in malted cereal grains can be used to hydrolyze

complex carbohydrates to simple and smaller sugar units.

63
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77
APPENDIX

78
79
80
81
82
Effect of Enzymes and Crop type on concentration of Fructose Extracted

Descriptive Statistics
Dependent Variable: Fructose

Crop Enzyme Mean Std. Deviation N

AMG 21.71767 .010066 3

Millet 25.71367 .010066 3

Cocoyam Sorghum .01067 .010066 3

Wheat 1.19767 .010066 3

Total 12.15992 12.167358 12

AMG 22.31367 .010066 3
Millet 3.17567 .010066 3
Aerial Yam Sorghum .28567 .010066 3
Wheat 3.66467 .010066 3
Total 7.35992 9.117756 12
AMG .21700 .003000 3
Millet 5.01500 .264575 3
Potato White Sorghum 7.41133 .750555 3
Wheat 10.48600 .854400 3
Total 5.78233 3.951366 12
AMG 14.74944 10.902391 9

Millet 11.30144 10.839279 9

Total Sorghum 2.56922 3.652870 9

Wheat 5.11611 4.188529 9

Total 8.43406 9.226537 36

Tests of Between-Subjects Effects

Dependent Variable: Fructose

Source Type III Sum of df Mean Square F Sig.

Squares

Corrected Model 2976.786a 11 270.617 2380.527 .000

Intercept 2560.799 1 2560.799 22526.493 .000
Crop 264.809 2 132.405 1164.720 .000
Enzyme 841.600 3 280.533 2467.758 .000
Crop * Enzyme 1870.377 6 311.730 2742.181 .000
Error 2.728 24 .114

83
Total 5540.313 36
Corrected Total 2979.514 35

a. R Squared = .999 (Adjusted R Squared = .999)

Multiple Comparisons
Dependent Variable: Fructose
LSD

(I) Enzyme (J) Enzyme Mean Std. Error Sig. 95% Confidence Interval
Difference (I-J) Lower Bound Upper Bound

Millet 3.44800* .158941 .000 3.11996 3.77604

AMG Sorghum 12.18022* .158941 .000 11.85219 12.50826

Wheat 9.63333* .158941 .000 9.30530 9.96137

AMG -3.44800* .158941 .000 -3.77604 -3.11996
Millet Sorghum 8.73222* .158941 .000 8.40419 9.06026
Wheat 6.18533* .158941 .000 5.85730 6.51337
AMG -12.18022* .158941 .000 -12.50826 -11.85219
Sorghum Millet -8.73222* .158941 .000 -9.06026 -8.40419
Wheat -2.54689* .158941 .000 -2.87493 -2.21885
AMG -9.63333* .158941 .000 -9.96137 -9.30530

Wheat Millet -6.18533* .158941 .000 -6.51337 -5.85730

Sorghum 2.54689* .158941 .000 2.21885 2.87493

Based on observed means.

The error term is Mean Square(Error) = .114.
*. The mean difference is significant at the .05 level.

Multiple Comparisons
Dependent Variable: Fructose
LSD

(I) Crop (J) Crop Mean Std. Error Sig. 95% Confidence Interval
Difference (I-J) Lower Bound Upper Bound

Aerial Yam 4.80000* .137647 .000 4.51591 5.08409

cocoyam
Potato White 6.37758* .137647 .000 6.09349 6.66167
Cocoyam -4.80000* .137647 .000 -5.08409 -4.51591
Aerial Yam
Potato White 1.57758* .137647 .000 1.29349 1.86167
Cocoyam -6.37758* .137647 .000 -6.66167 -6.09349
Potato White
Aerial Yam -1.57758* .137647 .000 -1.86167 -1.29349

84
Based on observed means.
The error term is Mean Square(Error) = .114.
*. The mean difference is significant at the .05 level.

LSD=0.584

Effect of Enzymes and Crop type on concentration of Glucose Extracted

Descriptive Statistics
Dependent Variable: Glucose

Crop Enzyme Mean Std. Deviation N

AMG 5.05233 .013317 3

cocoyam Millet 31.91633 .013317 3

Sorghum 1.02033 .013317 3

85
 Wheat .88733 .013317 3

Total 9.71908 13.499141 12

AMG 4.12133 .013317 3
Millet 23.76433 .013317 3
Aerial Yam Sorghum .95733 .013317 3
Wheat 6.85533 .013317 3
Total 8.92458 9.210400 12
AMG .81600 .075080 3
Millet 17.00733 .047173 3
Potato White Sorghum 3.86233 .063540 3
Wheat 17.11200 .388884 3
Total 9.69942 7.771406 12
AMG 3.32989 1.928422 9

Millet 24.22933 6.465249 9

Total Sorghum 1.94667 1.437391 9

Wheat 8.28489 7.109494 9

Total 9.44769 10.151582 36

Tests of Between-Subjects Effects

Dependent Variable: Glucose

Source Type III Sum of df Mean Square F Sig.

Squares

Corrected Model 3606.582a 11 327.871 23910.479 .000

Intercept 3213.321 1 3213.321 234336.154 .000
Crop 4.928 2 2.464 179.688 .000
Enzyme 2821.877 3 940.626 68596.506 .000
Crop * Enzyme 779.777 6 129.963 9477.729 .000
Error .329 24 .014
Total 6820.233 36
Corrected Total 3606.911 35

a. R Squared = 1.000 (Adjusted R Squared = 1.000)

Multiple Comparisons
Dependent Variable: Glucose
LSD
(I) Enzyme (J) Enzyme Mean Std. Error Sig. 95% Confidence Interval

86
 Difference (I-J) Lower Bound Upper Bound

Millet -20.89944* .055202 .000 -21.01337 -20.78551

AMG Sorghum 1.38322* .055202 .000 1.26929 1.49715

Wheat -4.95500* .055202 .000 -5.06893 -4.84107

AMG 20.89944* .055202 .000 20.78551 21.01337
Millet Sorghum 22.28267* .055202 .000 22.16874 22.39660
Wheat 15.94444* .055202 .000 15.83051 16.05837
AMG -1.38322* .055202 .000 -1.49715 -1.26929
Sorghum Millet -22.28267* .055202 .000 -22.39660 -22.16874
Wheat -6.33822* .055202 .000 -6.45215 -6.22429
AMG 4.95500* .055202 .000 4.84107 5.06893

Wheat Millet -15.94444* .055202 .000 -16.05837 -15.83051

Sorghum 6.33822* .055202 .000 6.22429 6.45215

Based on observed means.

The error term is Mean Square(Error) = .014.
*. The mean difference is significant at the .05 level.

Multiple Comparisons
Dependent Variable: Glucose
LSD

(I) Crop (J) Crop Mean Std. Error Sig. 95% Confidence Interval
Difference (I-J) Lower Bound Upper Bound

Aerial Yam .79450* .047806 .000 .69583 .89317

cocoyam
Potato White .01967 .047806 .684 -.07900 .11833
cocoyam -.79450* .047806 .000 -.89317 -.69583
Aerial Yam
Potato White -.77483* .047806 .000 -.87350 -.67617
cocoyam -.01967 .047806 .684 -.11833 .07900
Potato White
Aerial Yam .77483* .047806 .000 .67617 .87350

Based on observed means.

The error term is Mean Square(Error) = .014.
*. The mean difference is significant at the .05 level.

LSD=0.205

87
Effect of Enzymes and Crop type on concentration of Fructose Extracted

Descriptive Statistics
Dependent Variable: Sucrose

Crop Enzyme Mean Std. Deviation N

AMG .52367 .016042 3

Millet 19.01467 .016042 3

cocoyam Sorghum 1.29967 .016042 3

Wheat 4.18267 .016042 3

Total 6.25517 7.824923 12

Aerial Yam AMG 7.77967 .016042 3

88
 Millet 32.89967 .016042 3
Sorghum 2.06067 .016042 3
Wheat 2.50167 .016042 3
Total 11.31042 13.229250 12
AMG .79967 .004041 3
Millet 20.43100 .526641 3
Potato White Sorghum 1.21667 .093029 3
Wheat 21.59300 .536224 3
Total 11.01008 10.461589 12
AMG 3.03433 3.561025 9

Millet 24.11511 6.622147 9

Total Sorghum 1.52567 .405691 9

Wheat 9.42578 9.158333 9

Total 9.52522 10.684542 36

Tests of Between-Subjects Effects

Dependent Variable: Sucrose

Source Type III Sum of df Mean Square F Sig.

Squares

Corrected Model 3994.429a 11 363.130 7570.249 .000

Intercept 3266.275 1 3266.275 68092.750 .000
Crop 193.020 2 96.510 2011.965 .000
Enzyme 2870.993 3 956.998 19950.744 .000
Crop * Enzyme 930.416 6 155.069 3232.764 .000
Error 1.151 24 .048
Total 7261.855 36
Corrected Total 3995.581 35

a. R Squared = 1.000 (Adjusted R Squared = 1.000)

Multiple Comparisons
Dependent Variable: Sucrose
LSD

(I) Enzyme (J) Enzyme Mean Std. Error Sig. 95% Confidence Interval
Difference (I-J) Lower Bound Upper Bound

Millet -21.08078* .103245 .000 -21.29387 -20.86769

AMG
Sorghum 1.50867* .103245 .000 1.29558 1.72175

89
 Wheat -6.39144* .103245 .000 -6.60453 -6.17836
AMG 21.08078* .103245 .000 20.86769 21.29387
Millet Sorghum 22.58944* .103245 .000 22.37636 22.80253
Wheat 14.68933* .103245 .000 14.47625 14.90242
AMG -1.50867* .103245 .000 -1.72175 -1.29558
Sorghum Millet -22.58944* .103245 .000 -22.80253 -22.37636
Wheat -7.90011* .103245 .000 -8.11320 -7.68702
AMG 6.39144* .103245 .000 6.17836 6.60453

Wheat Millet -14.68933* .103245 .000 -14.90242 -14.47625

Sorghum 7.90011* .103245 .000 7.68702 8.11320

Based on observed means.

The error term is Mean Square(Error) = .048.
*. The mean difference is significant at the .05 level.

Multiple Comparisons
Dependent Variable: Sucrose
LSD

(I) Crop (J) Crop Mean Std. Error Sig. 95% Confidence Interval
Difference (I-J) Lower Bound Upper Bound

Aerial Yam -5.05525* .089413 .000 -5.23979 -4.87071

cocoyam
Potato White -4.75492* .089413 .000 -4.93946 -4.57038
cocoyam 5.05525* .089413 .000 4.87071 5.23979
Aerial Yam
Potato White .30033* .089413 .003 .11579 .48487
cocoyam 4.75492* .089413 .000 4.57038 4.93946
Potato White
Aerial Yam -.30033* .089413 .003 -.48487 -.11579

Based on observed means.

The error term is Mean Square(Error) = .048.
*. The mean difference is significant at the .05 level.

LSD=0.379

90
91
Effect of Enzymes and Crop type on concentration of Sucrose Extracted

Descriptive Statistics
Dependent Variable: Sucrose

Crop Enzyme Mean Std. Deviation N

AMG .52367 .016042 3

Millet 19.01467 .016042 3

cocoyam Sorghum 1.29967 .016042 3

Wheat 4.18267 .016042 3

Total 6.25517 7.824923 12

AMG 7.77967 .016042 3
Millet 32.89967 .016042 3
Aerial Yam Sorghum 2.06067 .016042 3
Wheat 2.50167 .016042 3
Total 11.31042 13.229250 12
AMG .79967 .004041 3
Millet 20.43100 .526641 3
Potato White Sorghum 1.21667 .093029 3
Wheat 21.59300 .536224 3
Total 11.01008 10.461589 12
AMG 3.03433 3.561025 9

Millet 24.11511 6.622147 9

Total Sorghum 1.52567 .405691 9

Wheat 9.42578 9.158333 9

Total 9.52522 10.684542 36

Tests of Between-Subjects Effects

Dependent Variable: Sucrose

Source Type III Sum of df Mean Square F Sig.

Squares

Corrected Model 3994.429a 11 363.130 7570.249 .000

Intercept 3266.275 1 3266.275 68092.750 .000
Crop 193.020 2 96.510 2011.965 .000
Enzyme 2870.993 3 956.998 19950.744 .000
Crop * Enzyme 930.416 6 155.069 3232.764 .000
Error 1.151 24 .048

92
Total 7261.855 36
Corrected Total 3995.581 35

a. R Squared = 1.000 (Adjusted R Squared = 1.000)

Multiple Comparisons
Dependent Variable: Sucrose
LSD

(I) Enzyme (J) Enzyme Mean Std. Error Sig. 95% Confidence Interval
Difference (I-J) Lower Bound Upper Bound

Millet -21.08078* .103245 .000 -21.29387 -20.86769

AMG Sorghum 1.50867* .103245 .000 1.29558 1.72175

Wheat -6.39144* .103245 .000 -6.60453 -6.17836

AMG 21.08078* .103245 .000 20.86769 21.29387
Millet Sorghum 22.58944* .103245 .000 22.37636 22.80253
Wheat 14.68933* .103245 .000 14.47625 14.90242
AMG -1.50867* .103245 .000 -1.72175 -1.29558
Sorghum Millet -22.58944* .103245 .000 -22.80253 -22.37636
Wheat -7.90011* .103245 .000 -8.11320 -7.68702
AMG 6.39144* .103245 .000 6.17836 6.60453

Wheat Millet -14.68933* .103245 .000 -14.90242 -14.47625

Sorghum 7.90011* .103245 .000 7.68702 8.11320

Based on observed means.

The error term is Mean Square(Error) = .048.
*. The mean difference is significant at the .05 level.

Multiple Comparisons
Dependent Variable: Sucrose
LSD

(I) Crop (J) Crop Mean Std. Error Sig. 95% Confidence Interval
Difference (I-J) Lower Bound Upper Bound

Aerial Yam -5.05525* .089413 .000 -5.23979 -4.87071

cocoyam
Potato White -4.75492* .089413 .000 -4.93946 -4.57038
cocoyam 5.05525* .089413 .000 4.87071 5.23979
Aerial Yam
Potato White .30033* .089413 .003 .11579 .48487
cocoyam 4.75492* .089413 .000 4.57038 4.93946
Potato White
Aerial Yam -.30033* .089413 .003 -.48487 -.11579

93
Based on observed means.
The error term is Mean Square(Error) = .048.
*. The mean difference is significant at the .05 level.

LSD=0.379

Effect of Enzymes and Crop type on concentration of Maltase Extracted

Descriptive Statistics
Dependent Variable: Maltase

Crop Enzyme Mean Std. Deviation N

AMG .60033 .007371 3

Cocoyam Millet 41.44433 .007371 3

Sorghum .90233 .007371 3

94
 Wheat 13.86433 .007371 3

Total 14.20283 17.353114 12

AMG 16.24533 .007371 3
Millet 33.90033 .007371 3
Aerial Yam Sorghum 21.62033 .007371 3
Wheat 12.66333 .007371 3
Total 21.10733 8.402343 12
AMG 3.16833 .125133 3
Millet 46.96233 .125133 3
Potato White Sorghum 6.20533 .125133 3
Wheat 48.03633 .125133 3
Total 26.09308 22.389975 12
AMG 6.67133 7.266362 9

Millet 40.76900 5.678992 9

Total Sorghum 9.57600 9.320750 9

Wheat 24.85467 17.394139 9

Total 20.46775 17.286726 36

Tests of Between-Subjects Effects

Dependent Variable: Maltase

Source Type III Sum of df Mean Square F Sig.

Squares

Corrected Model 10458.955a 11 950.814 180912.183 .000

Intercept 15081.436 1 15081.436 2869557.261 .000
Crop 855.631 2 427.816 81400.851 .000
Enzyme 6663.214 3 2221.071 422605.066 .000
Crop * Enzyme 2940.110 6 490.018 93236.186 .000
Error .126 24 .005
Total 25540.518 36
Corrected Total 10459.082 35

a. R Squared = 1.000 (Adjusted R Squared = 1.000)

Multiple Comparisons
Dependent Variable: Maltase
LSD
(I) Enzyme (J) Enzyme Mean Std. Error Sig. 95% Confidence Interval

95
 Difference (I-J) Lower Bound Upper Bound

Millet -34.09767* .034175 .000 -34.16820 -34.02713

AMG Sorghum -2.90467* .034175 .000 -2.97520 -2.83413

Wheat -18.18333* .034175 .000 -18.25387 -18.11280

AMG 34.09767* .034175 .000 34.02713 34.16820
Millet Sorghum 31.19300* .034175 .000 31.12247 31.26353
Wheat 15.91433* .034175 .000 15.84380 15.98487
AMG 2.90467* .034175 .000 2.83413 2.97520
Sorghum Millet -31.19300* .034175 .000 -31.26353 -31.12247
Wheat -15.27867* .034175 .000 -15.34920 -15.20813
AMG 18.18333* .034175 .000 18.11280 18.25387

Wheat Millet -15.91433* .034175 .000 -15.98487 -15.84380

Sorghum 15.27867* .034175 .000 15.20813 15.34920

Based on observed means.

The error term is Mean Square(Error) = .005.
*. The mean difference is significant at the .05 level.

Multiple Comparisons
Dependent Variable: Maltase
LSD

(I) Crop (J) Crop Mean Std. Error Sig. 95% Confidence Interval
Difference (I-J) Lower Bound Upper Bound

Aerial Yam -6.90450* .029596 .000 -6.96558 -6.84342

Cocoyam
Potato White -11.89025* .029596 .000 -11.95133 -11.82917
cocoyam 6.90450* .029596 .000 6.84342 6.96558
Aerial Yam
Potato White -4.98575* .029596 .000 -5.04683 -4.92467
cocoyam 11.89025* .029596 .000 11.82917 11.95133
Potato White
Aerial Yam 4.98575* .029596 .000 4.92467 5.04683

Based on observed means.

The error term is Mean Square(Error) = .005.
*. The mean difference is significant at the .05 level.

LSD=0.387

96
Effect of Enzymes and Crop type on concentration of Dxylose Extracted

Descriptive Statistics
Dependent Variable: Dxylose

Crop Enzyme Mean Std. Deviation N

AMG .00100 .001000 3

Millet .09600 .001000 3

Cocoyam Sorghum .00100 .001000 3

Wheat .00100 .001000 3

Total .02475 .042974 12

97
 AMG .00800 .001000 3
Millet .17600 .001000 3
Aerial Yam Sorghum .00100 .001000 3
Wheat .00700 .001000 3
Total .04800 .077242 12
AMG .01867 .006429 3
Millet .00467 .006429 3
Potato White Sorghum .00467 .006429 3
Wheat .00467 .006429 3
Total .00817 .008376 12
AMG .00922 .008378 9

Millet .09222 .074317 9

Total Sorghum .00222 .003768 9

Wheat .00422 .004206 9

Total .02697 .052461 36

Tests of Between-Subjects Effects

Dependent Variable: Dxylose

Source Type III Sum of df Mean Square F Sig.

Squares

Corrected Model .096a 11 .009 604.059 .000

Intercept .026 1 .026 1813.156 .000
Crop .010 2 .005 332.621 .000
Enzyme .051 3 .017 1184.417 .000
Crop * Enzyme .035 6 .006 404.360 .000
Error .000 24 1.444E-005
Total .123 36
Corrected Total .096 35

a. R Squared = .996 (Adjusted R Squared = .995)

Multiple Comparisons
Dependent Variable: Dxylose
LSD
(I) Enzyme (J) Enzyme Mean Std. Error Sig. 95% Confidence Interval

98
 Difference (I-J) Lower Bound Upper Bound

Millet -.08300* .001792 .000 -.08670 -.07930

AMG Sorghum .00700* .001792 .001 .00330 .01070

Wheat .00500* .001792 .010 .00130 .00870

AMG .08300* .001792 .000 .07930 .08670
Millet Sorghum .09000* .001792 .000 .08630 .09370
Wheat .08800* .001792 .000 .08430 .09170
AMG -.00700* .001792 .001 -.01070 -.00330
Sorghum Millet -.09000* .001792 .000 -.09370 -.08630
Wheat -.00200 .001792 .275 -.00570 .00170
AMG -.00500* .001792 .010 -.00870 -.00130

Wheat Millet -.08800* .001792 .000 -.09170 -.08430

Sorghum .00200 .001792 .275 -.00170 .00570

Based on observed means.

The error term is Mean Square(Error) = 1.444E-005.
*. The mean difference is significant at the .05 level.

Multiple Comparisons
Dependent Variable: Dxylose
LSD

(I) Crop (J) Crop Mean Std. Error Sig. 95% Confidence Interval
Difference (I-J) Lower Bound Upper Bound

Aerial Yam -.02325* .001552 .000 -.02645 -.02005

Cocoyam
Potato White .01658* .001552 .000 .01338 .01979
cocoyam .02325* .001552 .000 .02005 .02645
Aerial Yam
Potato White .03983* .001552 .000 .03663 .04304
cocoyam -.01658* .001552 .000 -.01979 -.01338
Potato White
Aerial Yam -.03983* .001552 .000 -.04304 -.03663

Based on observed means.

The error term is Mean Square(Error) = 1.444E-005.
*. The mean difference is significant at the .05 level.

LSD=0.006

Multiple Comparisons
Dependent Variable: DRafinose
LSD

(I) Crop (J) Crop Mean Std. Error Sig. 95% Confidence Interval
Difference (I-J) Lower Bound Upper Bound
99
 Aerial Yam .00350* .000933 .001 .00157 .00543
Cocoyam
Potato White -.11030* .000933 .000 -.11223 -.10837
cocoyam -.00350* .000933 .001 -.00543 -.00157
Aerial Yam
Potato White -.11380* .000933 .000 -.11573 -.11187
cocoyam .11030* .000933 .000 .10837 .11223
Potato White
Aerial Yam .11380* .000933 .000 .11187 .11573

Based on observed means.

The error term is Mean Square(Error) = 5.220E-006.
*. The mean difference is significant at the .05 level.

Effect of Enzymes and Crop type on concentration of DRaffinose Extracted

100
 Descriptive Statistics
Dependent Variable: DRafinose

Crop Enzyme Mean Std. Deviation N

AMG .00920 .001153 3

Millet .01420 .001153 3

Cocoyam Sorghum .01220 .001153 3

Wheat .01020 .001153 3

Total .01145 .002234 12

AMG .00520 .001153 3
Millet .00120 .001153 3
Aerial Yam Sorghum .02220 .001153 3
Wheat .00320 .001153 3
Total .00795 .008774 12
AMG .03300 .003606 3
Millet .22500 .003606 3
Potato White Sorghum .00400 .003606 3
Wheat .22500 .003606 3
Total .12175 .108415 12
AMG .01580 .013165 9

Millet .08013 .108814 9

Total Sorghum .01280 .008138 9

Wheat .07947 .109210 9

Total .04705 .081189 36

Tests of Between-Subjects Effects

Dependent Variable: DRafinose
Source Type III Sum of df Mean Square F Sig.
Squares

Corrected Model .231a 11 .021 4015.741 .000

Intercept .080 1 .080 15266.914 .000
Crop .101 2 .050 9627.885 .000
Enzyme .039 3 .013 2468.375 .000
Crop * Enzyme .091 6 .015 2918.710 .000
Error .000 24 5.220E-006
Total .310 36
Corrected Total .231 35
a. R Squared = .999 (Adjusted R Squared = .999)
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 Multiple Comparisons
Dependent Variable: DRafinose
LSD

(I) Enzyme (J) Enzyme Mean Std. Error Sig. 95% Confidence Interval
Difference (I-J) Lower Bound Upper Bound

Millet -.06433* .001077 .000 -.06656 -.06211

AMG Sorghum .00300* .001077 .010 .00078 .00522

Wheat -.06367* .001077 .000 -.06589 -.06144

AMG .06433* .001077 .000 .06211 .06656
Millet Sorghum .06733* .001077 .000 .06511 .06956
Wheat .00067 .001077 .542 -.00156 .00289
AMG -.00300* .001077 .010 -.00522 -.00078
Sorghum Millet -.06733* .001077 .000 -.06956 -.06511
Wheat -.06667* .001077 .000 -.06889 -.06444
AMG .06367* .001077 .000 .06144 .06589

Wheat Millet -.00067 .001077 .542 -.00289 .00156

Sorghum .06667* .001077 .000 .06444 .06889

Based on observed means.

The error term is Mean Square(Error) = 5.220E-006.
*. The mean difference is significant at the .05 level.

LSD=0.001

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