EFFECTS OF PROCESSING METHODS ON THE PHYSICO-

CHEMICAL PROPERTIES OF SWEET POTATO AND SORGHUM

BY

OGBONNA UJUNWA JULIET

BC/2009/287

SUBMITTED TO THE

DEPARTMENT OF BIOCHEMISTRY

FALCULTY OF NATURAL SCIENCES

CARITAS UNIVERSITY,AMORJI-NIKE ENUGU,

ENUGU STATE

A RESEARCH PROJECT PRESENTED IN PEATIAL FULFILMENT

OF THE REQUIREMENT FOR THE AWARDED OF BACHELOR

OF SCIENCE (B.Sc) DEGREE IN BIOCHEMISTRY

AUGUST, 2013
 CERTIFICATION

This is to certify that this project has been approved as meeting the

requirements of the department of biochemistry, Caritas University, Enugu

for the award of the degree of Bachelor of science (B.Sc) in Biochemistry.

HOD
DATE

SUPERVISOR
DATE

EXTERNAL SUPERVISOR

DATE
 DEDICATION

I dedicate this piece of work to God Almighty

 TABLE OF CONTENTS

TITLE PAGE

CERTIFICATION

DEDICATION

ACKNOWLEDGEMENT

ABSTRACT

TABLE OF CONTENTS

CHAPTER ONE: INTRODUCTION

CHAPTER TWO: LITERATURE REVIEW

2.1. ORIGIN AND DISTRIBUTION OF SWEET POTATO

2.1.1. DESCRIPTION OF SWEET POTATO PLANT

2.1.2. USES OF SWEET POTATO

2.1.3. NUTRITIONAL VALUE OF SWEET POTATO

2.1.4. ANTI- NUTRITIONAL FACTORS

2.1.5. NUTRIENT COMPOSITION OF SWEET POTATO

2.1.5.1. POLYPHENOLS COMPOSITION

2.1.5.2. ANTI-OXIDATIVE, ANTI-MUTAGENICITY AND ANTI-

CARCINOGENICITY

2.1.6. ANTI-DIABETES
2.1.7. ANTI-NUTRIENTS IN SWEET POTATO

2.1.8. ENZYME COMPOSITION OF SWEET POTATO

2.2. ORIGIN AND DISTRIBUTION OF SORGHUM PLANT

2.2.1. DISTRIBUTION OF SORGHUM PLANT

2.2.2. USES OF SORGHUM

2.2.3. ENZYME COMPOSITION OF SORGHUM

2.2.4. NUTRITIONAL COMPOSITION OF SORGHUM

2.2.5. ANTI-NUTRIENTS IN SORGHUM

CHAPTER THREE: MATERIALS AND METHODS

3.1. MATERIALS

3.2. METHODOLOGY

3.2.1. PROCESSING OF SWEET POTATO TUBER

3.2.2. PROCESSING OF SORGHUM GRAIN

3.3. SWEET POTATO AND SORGHUM ANALYSIS

3.3.1. PROXIMATE ANALYSIS

3.3.1.0. DETERMINATION OF FAT CONTENT

3.3.1.1. DETERMINATION OF ASH CONTENT

3.3.1.2. DETERMINATION OF CRUDE FIBRE

3.3.1.3. DETERMINATION OF MOISTURE CONTENT

3.3.1.4. DETERMINATION OF PROTEIN

3.3.1.5. DETERMINATION OF CARBOHYDRATES

3.4. ANTI-NUTRIENTS AND PHYTOCHEMICALS

3.4.1. DETERMINATION OF TANNINS

3.4.2. DETERMINATION OF HYDROCYANIC ACID

3.4.3. DETERMINATION OF ANTHOCYANNINS

3.4.4. DETERMINATION OF PHYTATE/PHYTIC ACID

3.5. DETERMINATION OF MINERAL CONTENT

3.5.1. MAGNESIUM

3.5.2. IRON

3.5.3. ZINC

3.5.4. PHOSPHOROUS

3.5.5. POTASSIUM

3.6. DETERMINATION OF PASTING PROPERTIES

3.7. DETERMINATION OF PHENOL OXIDASE

3.8. DETERMINATION OF REDUCING SUGARS; FRUCTOSE,

GLUCOSE AND SUCROSE

CHAPTER FOUR: RESULTS AND DISCUSSION

4.1. TABLE 1: PROXIMATE COMPOSITION OF THE SAMPLES

AND DISCUSSION

4.2. TABLE 2: MINERAL COMPOSITION OF SAMPLES AND

DISCUSSION

4.3. TABLE 3: PHYTOCHEMICAL COMPOSITION OF THE

SAMPLES AND DISCUSSION

4.4. TABLE 4: PHYSICO-CHEMICAL PROPERTIES OF

SAMPLES AND DISCUSSION

4.4. TABLE 5: SUGAR COMPOSITION OF THE SAMPLES

AND DISCUSSION

CHAPTER FIVE: CONCLUSION

REFERENCES
 ACKNOWLEDGEMENT

My special appreciation goes to God Almighty, for the strength and

wisdom bestowed upon me during my university years. My sincere regards

to my parents Mr. and Mrs. J.U Chinye and Mr. and Mrs. L.U Ogbonna, for

their love and financial support, to my siblings, thanks for being there for me.

I am also grateful to my HOD, Mr. Moses Ezenwali for being a loving

father from my first year in school till date, to my wonderful supervisor, Dr.

C.N Ishiwu for his fatherly love and utmost support and devoted time to go

through this work and offered relevant suggestions where necessary, to the

lecturers in the department, Dr. Yusuf, Mr. Peter, Dr. Ikpe, Mr. Ugwudike

and Mrs Chinenye, am saying a big thank you for enriching me with great

knowledge.

My greeting also goes to my fellow graduating students and every

student of the Biochemistry department.

I can’t afford to forget my room mates, Vicky, Vera, Doris and

Obianuju, as well as my friends, Ezinne, Evelyn, Adaeze and Juliet, for their

assistance. May God reward you all.

 ABSTRACT

This study evaluated "the effects of processing methods on the physico-

chemical properties of sweet potato and sorghum flour". Sweet potato

(Ipomoea batatas) is an important food crop in the tropical and sub-tropical

countries and belongs to the family convolvulaceae. Sweet potatoes are rich

in dietary fiber, minerals, vitamins, and anti oxidants such as phenolic acids,

anthocyannins, tocopherol and β-carotene. The proximate composition of

sweet potato was determined and these include moisture, lipids, ash, protein,

carbohydrates and fiber. In carrying out the analysis practically, methods

used vary according to the food material. The anti oxidants were also

determined alongside with phenol oxidase, pasting properties, minerals and

sugar contents. Sorghum is a tropical plant belonging to the family of

poaceae. More than 35% of sorghum is grown for human consumption. The

analyses carried out in sweet potatoes are same with sorghum with the

exclusion of phenol oxidase.

 CHAPTER ONE

INTRODUCTION

Sweet potato (Ipomoea batatas) is an important food crop in the

tropical and sub tropical countries and belongs to the family convolvulaceae.

It is cultivated in more than 100 countries. ( Woolfe, 1992). Nigeria is the

third largest producer in the world with china leading, followed by Uganda.

Sweet potato ranks seventh among the world food crops, third in value of

production and fifth in caloric contribution to human diet (Bouwkamp,

1985). Sweet potatoes are rich in dietary fibre, minerals, vitamins and anti

oxidants such as phenolic acids, anthocyanins, tocopherol and ß- carotene.

Besides acting as anti oxidants, carotenoids and phenolic compounds also

provide sweet potatoes with their distinctive flesh colours ( cream, deep

yellow, orange and purple). Sweet potato blends with rice, cowpea and

plantain in nigerian diets. It is also becoming popular as a substitute to yam

and garri. It can be reconstituted into fofoo or blended with other

carbohydrate flour sources such as wheat ( Triticum aestivum) and cassava

( Manihot esculenta) for baking bread, biscuits and other confectioneries

(Woolfe, 1992).
The leaves are rich in protein and the orange flesh varieties contain high beta

carotene and are very important in combating vitamin A deficiency

especially in children.

Sorghum (sorghum bicolor (S. bicolor) is a tropical plant belonging to

the family of poaceae, is one of the most important crops in Africa, Asia and

Latin America. More than 35% of sorghum is grown directly for human

consumption. The rest is used primarily for animal feed, alcohol production

and industrial products ( FAO, 1995). The current annual production of 60

million tons is increasing due to the introduction of improved varieties and

breeding conditions. Several improved sorghum varieties adapted to semi-

arid tropic environments are released every year by sorghum breeders.

Selection of varieties meeting specific local food and industrial requirements

from this great biodiversity is of high importance for food security. In

developing countries in general and particularly in West Africa demand for

sorghum is increasing. This is due to not only the growing population but

also to the countries policy to enhance its processing and industrial

utilization.
More than 7000 sorghum varieties have been identified, therefore there is a

need of their further characterization to the molecular level with respect to

food quality. The acquisition of good quality grain is fundamental to

produce acceptable food products from sorghum. Sorghum while playing a

crucial role in food security in Africa, it is also a source of income of

household . In West Africa, ungerminated sorghum grains are generally used

for the preparation of "to", porridge and couscous. Malted sorghum is used

in the process of local beer "dolo" (reddish, cloudy or opaque), infant

porridge and non fermented beverages. Sorghum grains like all cereals are

comprised primarily of starch.

The aim and objective of this work is to obtain diet low in sugars, with

enriched nutrients intended for diabetics.

 CHAPTER TWO

LITERATURE REVIEW

2.1. ORIGIN AND DISTRIBUTION OF SWEET POTATO

Sweet potato (Ipomoea batatas) is a member of the convolvulaceae

family (purseglove, 1972). Approximately 900 different species of

convolvulaceae in 400 genera have been identified around the world. Yen,

(1974) and Austin (1978,1988) recognized 11 species in the batatas, which

includes sweet potato. The closest relatives of the sweet potato appears to be

ipomoea trifida that is found wild in maxico, and ipomoeto tabascana.

Sweet potato has a chromosomes number for the genus ipomoea is 15, sweet

potato is considered to be a hexaploid. Most sweet potato cultivars are self-

incompatible, which means that when self pollinated, the cannot produce

viable seed. It is accepted that cultivated sweet potato originated in central

America or tropical south America. Sweet potatoes are cultivated where ever

there is enough water to support their growth: optimal annual rainfall for

growth range between 750-2000mm. sweet potato is a warm season annual,

 requiring 20-25°C average temperatures and full sunlight for optimal

development. Sweet potato thrives in well drained loamy soils with high

humus content that provides warm and moist environment to the roots.

2.1.1. DESCRIPTION OF SWEET POTATO

A. THE ROOT SYSTEM

When sweet potato is planted from stem cuttings, adventitious roots arise

from the cutting in a day or two. These roots grow rapidly and form the root

system of the plant. Research has shown the roots of sweet potato can

penetrate the soil to a depth of over 2m, the exact depth attained being

dependent on the soil condition (Onwueme, 1978 and Kays, 1985). Based on

its origin, the root system of sweet potato is divided into the adventitious

roots arising from subterranean nodes of a vine cutting and lateral roots

arising from existing roots. Kays (1985) subdivided the adventitious roots

into storage, fibrous and pencil roots. The lateral roots are subdivided into

primary, secondary and tertiary roots.

During the early ontogeny of young adventitious roots emerging from the

stem, they are often separated into two classes’ namely thick and thin roots

(Togari, 1950). According to Wilson (1982) and Kays (1985), thin roots are

typically tetrarch in the arrangement of their primary vascular tissue, i.e.

four xylem and phloem poles found within the vascular cylinder. The most

important functional differences between these root types are their capacity

for storage root initiation in a specific region of the thick roots. Several

factors such as exposure of potential storage roots to long photoperiod

(Bonsi et al, 1992), water logged soil conditions (Kays, 1985), high levels of

nitrogen supply (Chua & Kays, 1981), gibberellic acid application (McDavid

& Alamu, 1980), as well as exposing the plant to long days (McDavid &

Alamu, 1980; Du Plooy, 1989) encourage lignification and inhibit storage

root development. Alternatively high potassium supply (Isuno, 1971, Hahn

& Hozyo, 1984), the absence of light (Wilson, 1982), as well as well aerated

soil conditions, low temperature and short days been demonstrated to

encourage storage root formation (Du Plooy, 1989).

 a. Storage Roots

Storage roots arise from pentarch or hexarch thick young roots if the cells

between the protoxylem point and the central metaxylem cell do not become

lignified, or if only a slight proportion of these cells are lignified (Togari,

1950). The increase in storage root size is attributed to the activity of the

vascular cambium as well as the activity of the anomalous cambia (Wilson,

1982). The initial sign of storage root formation is the accumulation of

photosynthetic consisting predominantly of starch (Chua & Kays, 1982).

Storage root initiation is reported to occur between the periods of 35 to 60

days after planting (Agata, 1982, Wilson 1982). But the work of Du Plooy

(1989) indicated that storage root initiation might occur as early as 7 days

after planting. These conflicting results suggest the need for further research

on the storage root formation in sweet potato.

Agata (1982) reported that storage root formation started about 30 to

35 days after planting and the roots dry weight increased linearly until

harvest.
 b. Pencil Roots

Pencil roots are generally between 5 and 15mm in diameter, they are the

least well defined of the adventitious root emerging from the subterranean

node of the culting. They develop mainly from young thick adventitious

roots under conditions not conducive for the development of storage roots.

In pencil roots lignification is not total, but result in uniform thickening of

the entire root.

c. Fibrous Roots

According to Chua and Kays (1981), fibrous roots develop mainly from

tetrarch, thin adventitious roots. The fibrous roots are generally less than

5mm in diameter and are branched with lateral roots forming a dense

network throughout the root zone constituting the water and nutrient

absorbing system of the plants. Fibrous roots have heavily lignified steels

and very low levels of vascular cambial activity. High nitrogen and low

oxygen within the root zone favors their formation (Chua & Kays, 1981).

d. Lateral roots

The lateral roots of sweet potato emerge from existing roots adventitious

roots (storage, pencil and fibrous) have a profusion of lateral roots at varying
densities along their axis. The primary lateral roots emerge from

adventitious roots. Lateral emerging from the primary laterals are called

secondary lateral and those emerging from the secondary laterals are named

tertiary laterals (Kays, 1985).

B. ABOVE GROUND PLANT ORGANS

a. Vines

Sweets potato has long thin stems that trail along the soil surface and can

produce roots at the nodes. Sweet potato genotypes are classified as either

erect, bushy, intermediate, or spreading, based on the length of their vines

(Yen, 1974, Kays, 1985). Stem length varies with cultivar, and highly

variable, ranging from a few centimeters up to 10cm in length. Planting

density has pronounced effect on the internode length as well as on vine

length (Somda & Kays, 1990a). The stem is circular or slightly angular.

Stem color is predominantly green, but purplish pigmentation is often

present.

Branching is cultivar dependent (Yen, 1974) and branches vary in number

and length. Normally, sweet potato produces three types of branches,

primary, secondary and tertiary, at different periods of growth. The total

number of branches varies between 3 and 20 among cultivars. Spacing,

photoperiod, soil moisture and nutrient supply influence the branching

intensity in sweet potato plant (Kays, 1985, Somda & Kays, 1990a).

b. Leaf and Petiole

The leaves of sweet potato occur spirally on the stem. The total number of

leaves per plant varies from 60 to 300 (Somda et al, 1991).

The number of leaves per plant increases with decreasing plant density

(Somda & Kay’s, 1990b), increasing irrigation (Indira & Kabeerathumma,

1990). Peptide length varies widely with genotype and may range from

approximately 9 to 33cm (Yen, 1974). The petiole retains the ability to grow

in a curved or twisted manner so as to expose the lamina to maximum light.

The petiole is swollen at its junction with the stem, and it bears two small

nectarines at the junction. The lamina is extremely variable in size and shape,

even for leaves on the same plant. The lamina is green in color, sometimes

with purple coloration.

c. Flower

The flowers of sweet potato are born solitarily or on cymose inflorescence

that grow vertically upward from the leaf axis (Purseglove, 1972, Onwueme,
1978). Each flower has five united sepals, and fire petals joined together to

form a funnel-shaped corolla tube. The stamens are five in number and are

attached to the base of the style.

The filament is white and hairy; the anther is also white and contains one

locule. Each locule contains two ovules, so that there is a maximum of four

ovules in each ovary (Onwueme, 1978).

The physiology of the sweet potato flower is complex. Firstly, the

formation of the flower is subject to environmental control, especially

photoperiodic control. Secondly, the flower is open and receptive for an

extremely short period of time. Thirdly, incompatibility complexes exist.

Fourthly, the existence of variation in stamen length with respect to the style

introduces a further morphological complication into the pollination

mechanism. All these features make seed production difficult (Onwueme,

1978).

d. Fruit and Seed

The sweet potato fruit is a capsule 5 to 8mm in diameter. A false septrum,

formed during fruit development, may divide each of the two locules into
two, thereby creating four chambers in the mature fruit. Each chamber may

contain a seed, but usually only one or two chambers in each fruits contain a

seed, but usually only one or two chambers in each fruit contain any seed.

The seed is black and about 3mm long. It is flat on one side and convex on

the other. The micropyle is located in a hollow on the flattened side.

Endosperm is present in the seed in addition to cotyledons. The testa is very

hard and almost impermeable to water or oxygen. For this reason, the seeds

germinate with difficulty. Germination can be improved by scarifying the

seed either by mechanically clipping the testa, or by treating it with

concentrated sulfuric acid for about 45minutes.

Germination of scarified seed occurs in 1 to 2days. The radical is first

to elongate, and develops into the primary root system. Germination is

epigeal, since the cotyledons are carried above the soil level. After

emergence, the bi-lobal cotyledons expand, develop chlorophyll, and

become photosynthetic (Onwueme, 1978).

2.1.2. USES OF SWEET POTATO

Sweet potato is an important crop in many parts of the world. The storage

roots of sweet potato serve as staple food, animals feed (Posas, 1989) and to
a limited extent as a raw material for industrial purposes as a starch source

and for alcohol production (Collins, 1984). Sweet potato starch is used for

the manufacture of adhesives, textile and paper sizing and in the

confectionery and baking industries.

In most parts of the tropics, sweet potato is consumed boiled, baked, roasted

or fried.

2.1.3. NUTRITIONAL PROFILE/VALUE

The orange-flesh sweet potatoes are exceedingly rich in beta-carotene. The

purple-flesh varieties are outstanding sources of anthocyanins, especially

peonidins and cyanidins. Both types of sweet potatoes are rich in unique

phytonutrients, including polysaccharides related molecules called batatins

and batatosides. Sweet potatoes also include storage proteins called

sporamins that have unique antioxidant properties. Sweet potatoes are an

excellent source of vitamin A (In the form of beta-carotene). They are also a

very good source of vitamin C and manganese.

In addition, sweet potatoes are a good source of copper, dietary fiber, niacin,

vitamin B5 and potassium. The sweet potato is rich in vitamin A, B and C

and potassium. But in sub Saharan Africa it is estimated that 32% of the
population suffers from vitamin A deficiency which can lead, amongst other

illnesses, to blindness, (Collins, 1984). The sweet potato can therefore be a

value added food staple particularly for children and pregnant women who

are more often exposed to deficiencies (Posas, 1989).

2.1.4. ANTI-NUTRITIONAL FACTORS

Raw sweet potato tubers contain medium levels of trypsin inhibitors that are

still sufficient to decrease protein digestibility in diets. Trypsin inhibitor

content is very variable between varieties (2.6 to 32.0 Tul/g). Moist heat

treatments (MHT) at temperature above 80°C are affective in eliminating

trypsin inhibitor activity in sweet potatoes . Raw sweet potatoes also contain

oxalate up to 1.2g/kg DM.

2..1.5. NUTRITIONAL COMPOSITION OF SWEET POTATO

Recent studies show that sweet potato contains such functional

components as polyphenols, anthocyaninins and dietary fiber, which are

important for human health.

Sweet potatoes are a good source of carbohydrates, while sweet potatoes

tops (leaves and stems) contain additional nutritional components in much

higher concentrations than in many other commercial vegetables in many

parts of the world. They are rich in vitamins B, β-Carotene, iron, calcium,

zinc and proteins and the crop is more tolerant of diseases, pest and high

moisture than many other leafy vegetables grown in the tropics. Because

sweet potato tops can be harvested several times a year, their annual yield is

much higher than many other green vegetables.

Sweet potato is one of the most important summer food crops in the

southern united state. It is a versatile plant. For example, it is used as food,

as livestock feed and for starch and alcohol production. Several researchers

report that sweet potato leaves are an excellent source of antioxidative

polyphenolics, among them anthocyanins and phonelics and are superior to

other commercial vegetables. The nutritional value of sweet potato leaves is

gaining recognition, as the understanding between diet and health increases.

Sweet potato leaves with their high nutritive value and antioxidants may

become an excellent leafy vegetable.

Depending on varieties and growing conditions, sweet potato leaves are

comparable to spinach in nutrient content. The average mineral and vitamin

content in a recently developed cultivar, suioh, is 117mg calcium, 1.8mg

iron, 3.5mg carotene, 7.2mg vitamin C, 1.6 mg vitamin E and 0.56mg

vitamin K. levels of iron, calcium and carotene rank among the top,

compared with other major vegetables. Sweet potato leaves are also rich in

vitamin B, β-carotene, iron, calcium, zinc, and protein. Studies have shown

that sweet potato leaves contain as many vitamins, minerals and other

nutrients as spinach. As a crop, sweet potato is more tolerant of diseases,

pests and high moisture. Sweet potato leaves are an excellent source of

antioxidative polyphenolics, among them anthocyanins and phenolic acids

such as caffeic, monocaffenylquinic (chlorogenic), dicaffenylquinine and

tricaffeoylquinic acids, and are superior in this regard to other commercial

vegetables (Ishiguro et al, 2004).

2.1.5.1.POLYPHENOLS COMPOSITIONS

Sweet potato leaves represent at least 15 anthocyanin and 6 polyphenolic

compounds. These biologically active compounds posses multifaceted action,

including antioxidation, antimutageneicity, anti-inflammation and

anticarcinogenesis.
2.1.5.2. ANTIOXIATIVE, ANTIMUTAGENICITY AND

ANTICARCINOGENICITY

Cancers occur through such processes as initiation, promotion, and

progression in body cells. Initiation is a kind of mutation that occurs in

cancer and anti cancer genes.

Thus, controlling the gene mutation brought about by the carcinogens leads

to cancer prevention. Sweet potato leaves are a good supplementary resource

of anti oxidants and antimutagenic compounds.

2.1.5.3.ANTI-DIABETES

The latter stage of noninsulin-dependent diabetes mellitus (NIDDM), one

of the major diseases of adults, is caused by the secretion of adults, is caused

by the decrease in the secretion of insulin by the pancreatic langerhans cells.

Diabetes contributes to the death of more than 213,000 Americans each

year and is also a leading cause of heart diseases, blindness and kidney

failure. Foods with anti-diabetes effect are desired for diet therapy. Several

researchers report that sweet potato leaves have anti-diabetic compounds

that reduce the blood glucose content significantly in model rats.

2.1.6. ANTI NUTRIENTS IN SWEET POTATO

The anti-nutritional factors have been reviewed (McLean, 1970).

However, the biochemical effects of the anti-nutritional factors will be

briefly highlighted. Cyanogenic glucoside on hydrolyses yields toxic

hydrocyanic acid (HCN). The cyanide ions inhibit several enzyme systems;

depress growth through interference with certain essential amino acids and

utilization of associated nutrients. They also cause acute toxicity, neuropathy

and death.

Alkaloids cause gastrointestinal and neurological disorders. The

glycoalkaloids, solanine and chaconine present in potato and solanum spp.

(Saito et al, 1990) are haemolytically active and toxic to fungi and humans.

Some of the toxicological manifestations of potato glycoalkaloids disorders.

Saponins cause hypochollesterolaemia by binding cholesterol making it

unavailable for absorption (Johnson et al, 1986). Trypsin (protese inhibitor)

causes pancreatic enlargement and growth depression.

Haemagluttinins are proteins known for agglutinating red blood cells.

Phytates bind makes them unavailable. Oxalates, like phytates, bind

minerals like calcium and magnesium and interfere with their metabolism.

They also cause gastrointestinal tract irritation, blockage of the renal tubules

by calcium oxalate crystals, development of urinary calculi and

hypocalcaemia (Oke, 1969) .

2.1.7. ENZYME COMPOSITION OF SWEET POTATO

β-Amylase is an exoenzyme that releases successive maltose units

from the non reducing end of a polysaccharide chain by hydrolysis of α-1,4-

glucan linkages. The shortest normal saccharide attacked is maltotetraose

(Myrback and Neumuller 1950). Since it is unable to bypass branch linkages

in branched polysaccharides such as glycogen or amylopectin, the hydrolysis

is incomplete and a macromolecular limit dextrin remains. Β- amylase is

found primarily in the seeds of higher plants and sweet potatoes. It yields a

single product: maltose.

The tuberous root of sweet potato is unusually rich in the enzyme,

accounting for approximately 5% of the total soluble proteins. In contrast,

other tuberous roots contain trace amounts of β-amylase activity.

2.2. ORIGIN AND DISTRIBUTION OF SORGHUM

Sorghum is the 5th most important grain crop after wheat, maize,

rice and barley. It is indigenous to Africa. Globally, it produces

approximately 70 million tons of grain from about 50 million ha of land. It is

the dietary staple of more than 500 million people in more than 30 countries.

For all that, however, sorghum now receives merely a fraction of attention of

what it could. Not only is it inadequately supported for the world’s fifth

major grain crop, it is under supported considering its vast untapped

potential. Sorghum could contribute more to food supplies than at present,

especially to those regions and peoples in greatest need.

2.2.1. DESCRIPTION OF SORGHUM PLANT

MORPHOLOGY, GROWTH AND DEVELOPMENT OF THE

SORGHUM PLANT.

Sorghum belongs to the grass family, Graminea. It is essential that

producers know the crop they are cultivating in order to develop the most

effective production practices.

ROOT SYSTEM

The roots of the sorghum plant can be divided into a primary and

secondary root system. The primary roots are those which appear first from

the germinating seed. The primary roots provide the seedling with water and

nutrients from the soil. Primary roots have a limited growth and their

functions are soon taken over by the secondary roots. Secondary roots

develop from nodes below the soil surface. The permanent root system

branches freely, both literally and downwards into the soil. If no soil

impediments occur, roots can reach a lateral distribution of 1m and a depth

of up to 2m early in the life of the plant.

The roots are finer and branch approximately twice as much as roots from

maize plants.

LEAVES

Sorghum leaves are typically green, grasslike and flat and not as broad as

maize leaves. Sorghum plants have a leaf area smaller than that of maize.

The leaf blade is long, narrow and pointed. The leaf blades of young leaves

are upright, however, the blades tend to bend downwards as the leaves

mature. Stomata occur on both surfaces of the leaf. A unique characteristic

of sorghum leaves is the rows of motor cells along the mid rib on the upper

surface of the leaf. These cells can roll up leaves rapidly during moisture

stress. Leaves are covered by a thin wax layer and develop opposite one

another on either side of the stem. Environmental conditions determine the

number of leaves, which may vary from 8-22 leaves per plant.

STEM

The stem of the plant is solid and dry, succulent and sweet, under

favourable conditions more internodes develop, together with leaves,

producing a longer stem.

The stem consists of internodes and nodes. A cross section of the stem

appears oval or round.

The internodes are covered by a thick waxy layer reduces transpiration and

increases the drought tolerance of the plants.

SEED

The ripe seed (grain) of sorghum is usually partially enclosed by glumes,

which are removed during threshing and / or harvesting. The shape of the

seed is oval to round and the colour may be red, white, yellow, brown or

shades thereof. If only the pericarp is coloured, the seed is usually yellow or
red. Pigment in both the pericarp and testa results in a dark brown or red

brown colour. The sorghum grain consists of the testa, embryo and

endosperm.

SEED COAT

The seed coat consists of the pericarp and testa

PERICARP

This is the outermost layer of the seed and consists of the epicarp,

hypodermis, mesocarp and endocarp.

TESTA

The testa is situated directly below the endocarp and encloses the

endosperm. Apart from the role of the testa in the colouring of the seed, it

contains a tannin like substance with a bitter taste. The presence thereof

results in less bird damage significantly increases. The bitter taste of

sorghum with a testa however, makes it less acceptable as food for humans

and animals.

EMBRYO

The embryo contains those parts which give rise to the new seedling. The

new plant, which is already a complete unit, depends on the right moisture

and temperature conditions to start developing.

ENDOSPERM

The endosperm consists of hard and soft endosperm. The endosperm

supplies the seedling with nutrients until it can take up its own nutrients.

2.2.2. USES OF SORGHUM

In many parts of the world, sorghum has been used traditionally in

food products and various food items, porridge, unleavened bread, cookies,

cakes, couscous and malted beverages are made from this versatile grain.

Traditional food preparation of sorghum is quite varied. Boiled sorghums are

one of the most basic uses and small, corneous grains are normally desired

for this type of food product. The whole grain maybe ground into flour or

decorticated before grinding to produce either a fine particle product or flour,

which is then used in various traditional foods. The seed is used as food, in

brewing beer, sorghum malt and meal. It is fermented to make "Lleting" ( a

sour mash), the pith is eaten and the sweet culm chewed.

Porridge and muffins can be made using sorghum meal. Parched seeds are

used as coffee substituents or adulterants.

2.2.3. ENZYME COMPOSITION OF SORGHUM

SORGHUM PHENOLIC ENZYMES BIOCHEMISTRY

Role of phenylalanine ammonia lyase in the biosynthesis of phenolic

compounds;

The biosynthesis of phenolic compounds in plant is initiated by the shikimic

acid pathway ( Tomas Barberan and Heldth, 2005). This pathway continues

with the production of phenylalanine, which is subsequently deaminated by

the enzyme phenylalanine ammonia lyase (EC 4.3.1.5, PAL).

PAL can deaminate both L- phenylalanine and L- tyrosine into cinnamate

derivatives ( Heldt 2005).

PAL is inhibited by its products e.g trans-cinnamates ( Heldt, 2005). PAL

activity has been detected in the green shoots and leaves ( Staffford, 1969)

of sorghum.

In sorghum, the infection of the plant with pathogen involved a very rapid

accumulation of PAL mRNA (Cui et al., 1996). The presence of PAL

activity in sorghum grain and its activation upon germination was assessed .
 2.2.4. NUTRITIONAL COMPOSITION OF SORGHUM

Sorghum has nutritional composition similar to or better than rice

and wheat in some aspects. The grains contain high fiber and non starchy

polysaccharides and starch with some unique characteristics. There is a

considerable variation in sorghum for levels of proteins, lysine, lipids,

carbohydrate, fiber, calcium, phosphorous, iron, thiamine and niacin .

Protein quality and essential amino profile of sorghum is better than many of

the cereals and millets. Sorghum in general is rich source of fiber and B-

complex vitamins.

Grain sorghum is rich in fiber and minerals apart from having a sufficient

quantity of carbohydrates (72%), proteins (11.6%) and fat (1.9%).

Starch is the main constituent of the grain. There is a need to popularize

sorghum foods as sorghum with its high mineral and fiber content and with

low or slow starch digestibility makes an ideal food for diabetic and obese

population in the urban as well as rural society.

2.2.5. ANTI NUTIENTS IN SORGHUM

CYANOGENIC GLYCOSIDES

Cyanogenic glycosides are mainly present in germinating seeds,

sprouts and the leaves of immature sorghum plants. Traore et al.

(2004) showed that malted red sorghum that had been dried

contained an average 320ppm cyanogens. The most abundant

cyanogens is dhurrrin, which may comprise three to four percent of

the leaves of germinating seeds (Waniska and Rooney, 2000).

Stressors such as drought, frost, heavy insect infestation, or

overgrazing can result in increased levels of these compounds, which

along with tannins, are part of the plants’ defence mechanisms. The

use of potassium nitrate fertilizer was also shown to increase

cyanogens production in sorghum (Busk and Moller, 2002). In the

stomach of livestock, cyanogenic glycosides may be converted into

hydrogen cyanide, which is toxic, and at low level chronic exposure

may result in poor growth or reduced milk production.

Although sprouted sorghum can contain high levels of cyanogens,

typical methods of processing sprouted sorghum grain for human

consumption, such as manual degermination (removal of roots and

shoots) removes most of the toxin (Traore et al., Dada and Dendy,

1987).

TANNINS

Early literature identified tannic acid as an anti nutritional factor

in sorghum grain. However, more recent research indicates that

tannic acids is not a sorghum component (Dykes and Rooney, 2006).

Some but not all sorghum varieties have pigmented testa containing

condensed tannins, polyphenolic compounds that possibly give the

seed a bitter taste and have been known to reduce intake,

digestibility (particularly protein), growth and feed efficiency of

livestock (Gilani et al., 2005; Waniska and Rooney, 2000) tannins

act as a plant defence against consumption by birds and also provide

some resistance to mold.

PHYTIC ACID

Like all grain species, sorghum contains phytic acid which binds

minerals and reduces their availability to the consumer. Since

sorghum grain is usually low in mineral content and the presence of

phytic acid likely rendering its low mineral content unavailable,

supplementation with other mineral sources is necessary where

sorghum is a major component of the diet.

 CHAPTER THREE

3.0. MATERIALS AND METHODS

3.1. MATERIALS

Sweet potato, drying oven, grater, sorghum, jute bag, filter bag

METHODS

Sweet potato

Peel

Wash

Slice

Blanch in hot water at 90⁰C for 4 minutes

↓
 Mop the excess water

Dry at 55⁰C until dried

Mill

Sieve

Sweet potato flour

Package

Fig. 3.2.1: flow chart showing the method of processing sweet potato tuber

The sweet potato tuber was shared into two parts, the first half was peeled

and then washed. It was sliced and then blanched in hot water at 90⁰C for 4

minutes. A filter bag was used to mop the excess water out of the sweet

potato and then dried at 55⁰C until it is completely dried. It was then milled

and sieved to remove the chaff that might be present. The sweet potato flour

was then packaged in air tight container.

 The second half of the sweet potato was peeled and washed. It was

then sliced and grated to obtain the pulp. The sweet potato was steeped in

excess water for 8 hours inside the fridge. After 8 hours, the water from the

sweet potato was decanted, it was then steeped again in fresh water for 8

hours in the fridge, the water was decanted again and put in a filter bag to

press out the water. The sample was dried at 55⁰C, milled, sieved and the

refined flour was then packaged.

Sweet potato tuber

Peel

Wash

Slice

Grate to obtain the pulp

Steep in excess water for 8 hours inside the

Fridge
 ↓

Decant the water

Put in a filter bag and press out water

Dry at 55⁰C

Mill

Sieve

Package (refined flour)

Fig.3.2.1. flow chart showing the method of processing sweet potato tuber
 Sorghum

Sort / clean

Mill

Sieve

Sorghum flour

Package

Fig 3.2. flow chart showing the method of processing sorghum grains

Sorghum grains were cleaned and milled to obtain a powdered sorghum

(flour). It was then sieved to remove chaff and packaged.

The second half of the sorghum grains were cleaned and soaked in water

for 10 hours. It was spread on a wet jute bag and kept in a room for 4-5 days

to germinate, after 4 days, it was dried in an electric oven at 55⁰C. the

sprouts were removed manually and milled, it was then sieved to obtain

malted sorghum flour and packaged.

Sorghum grain

Sort/ clean

Soak in water for 10 hours

Spread on a wet jute bag and keep in a room

For 4-5 days to germinate

Dry at 55⁰C

Manually remove the sprouts

Mill

↓
 Sieve

Malted sorghum flour

Package

Fig. 3.2.2. Flow chart showing the method of processing sorghum grains

3.3. SWEET POTATO AND SORGHUM ANALYSIS

3.3.1. PROXIMATE ANALYSIS

Proximate analysis of food is the determination of the major

components of food, which include: moisture, lipid (fats), ash (mineral,

protein, carbohydrates and fiber. In carrying out the analysis practically,

methods used vary according to the food material being studied and also in

details of evaluation procedure.

3.3.1. 0. DETERMINATION OF FAT CONTENT

Apparatus and reagents,soxhlet extractor, round bottom flask, beaker,

desiccators.
The fat content of the determined using the standard AOAC (2010) method.

A soxlet extractor with a reflux condenser and a 500ml round bottom flask

was set up. About 300ml of petroleum ether was poured into the the round

bottom flask. The sample (2gm) was weighed into labeled thimble and

sealed with cotton wool, then fitted into the extraction tube of the soxhlet

extractor. The soxhlets extractor after assembly was allowed to reflux for

about 6 hours, after which the thimble was removed with care and the

petroleum ether collected on top and drained into a container for re-use. The

flask is now free of ether, and was then removed and dried in dessicators and

weighed.

W2-w1

Fat (%) content = × 100

Where

W= weight of sample used

W1 = weight of empty extracting flask

W2= weight of flask and extracted oil

3.3.1.1. DETERMINATION OF ASH CONTENT

Apparatus and reagent:

Fume, cupboard, muffle furnace, crucibles, dessicators

The ash content of the samples were determined according to the

standards of AOAC (2010). A preheated and cooled crucible was weighed.
The sample was charred on a Bunsen flame inside a fume cupboard. The
charred sample was placed in a muffle furnace set at 550⁰C for 2 hours until
a white or light grey ash is obtained. The sample was removed, cooled in
desiccators and weighed.

Ash content = w3- w1 × 100

W2- w1

Where w1= weight of empty crucible

W2= weight of crucible + weight of sample

W3= weight of crucible + weight of sample after ashing

3.3.1.2. DETERMINATION OF CRUDE FIBER

This was done according to AOAC (2010) method:

Apparatus and reagents:

Sulphuric acid, sodium hydroxide, alcohol.

The sample was oven dried at 105⁰C. Powdered dried sample(2g) was
placed in a 500ml beaker and 200ml of boiling 1.25% H2so4 was added. The
beaker was placed on a hot plate and boiled for 3minutes with occasional
rotation of the beaker. The beaker was cooled and filtered by suction
through a Buchner funnel. The beaker was rinsed with two 50ml positions
of boiling water. The residue was carefully transferred into a beaker and
200ml of 1.25% NaOH was added. It was boiled for 30 minutes, cooled and
filtered and washed twice with 50ml boiling water.

Finally, the sample was washed with 25ml 95% alcohol. The residue
was oven dried for 2hours at 130⁰C and cooled in a desiccators and
weighed. The sample was heated for 30 minutes at 600⁰C and cooled in a
desiccator and weighed.

Crude fibre = weight of residue × 100

Weight of sample

3.3.1.3. DETERMINATION OF MOISTURE CONTENT

Apparatus and reagents:

Desiccators, crucibles, weighing balance

The moisture content of the sample was determined according to the

standard of AOAC (2010). The crucibles were washed and dried in an oven

at 100⁰C for 1 hour. The weight was noted as w1. 2g of each sample was

separately weighed into the crucibles and their weights were taken and noted

down as (w2) before and during drying at 100⁰C to constant weight (w3).

Moisture content= w2 –w3 × 100 = weight of moisture ×

100

W weight of sample

Where

W1 = weight of empty crucible

W2 =weight of crucible and sample before drying

W3= weight of sample after drying to a constant weight

3.3.1.4. PROTEIN DETERMINATION

The sample was digested in concentrated sulphuric acid

Apparatus and reagents:

Kjeldahl distillation unit, concentrated sulphuric acid, anhydrous sodium

sulphate, boric acid solution, hydrochloric acid, boric acid indicator,

volumetric flask, conical flask, sodium hydroxide

The protein content of the samples were determined according to the

standard methods of AOAC (2010) using kjeldahl’s method.

Digestion of the sample

2g of sample was weighed into kjeldahl’s flask and anhydrous sodium

sulphate of 5g was added. 25ml of concentration H2so4 was added with

few boiling chips. The content of the flask was heated in the fume

chamber until clear solution is obtained. The solution was cooled and

transferred into 250ml volumetric flask and made up to the level with

distilled water.

The distillation unit was carried out using a well cleaned kjedahl

apparatus. 100ml conical flask containing 5ml of 2% boric and 2 drops

of methyl red indicator was placed under the condenser. Pipette 5ml of

the digest into the apparatus through the small funnel on the distillation

unit. The digest was washed down with distilled water followed by

addition of 5ml of 60% sodium hydroxide solution, it was distilled until

the volume of the distillate (ammonium sulphate) reaches 100ml. the

 solution in the flask was then titrated with 0.04 Hcl until the first

permanent pink colour appeared. The blank was titrated in the same

way and the time value for the samples were obtained.

% nitrogen=vs. vb × Naci × 0.04 × 100

Where vs = volume (ml) of acid required to titrate sample

Vb = volume (ml) of acid required to titrate the blank

Nacid = normality of acid (0.1N)

W= weight of sample in gram (gm)

Note: most proteins contain about 16% nitrogen, so that 16mg nitrogen

=100mg protein.

1mg Nitrogen = 6.25

Therefore protein (%) = N × 6.25 (conversion factor for protein)

3.3.1.5. DETERMINATION OF CARBOHYDRATE

The carbohydrate content was determined as illustrated by

AOAC (2010) as follows: it was estimated as the remainder after accounting

for all, crude fibre, protein and fats.

Total carbohydrate contents= 100-(% moisture +% ash + % protein + % fat).

Where

Cp = crude protein

Cf = crude fibre

Ash = ash content

Fat = fat content

Moisture = moisture content

3.4. ANTI NUTRIENTS AND PHYTOCHEMICALS

Phytochemicals are chemicals that occur naturally in plants and plants

derive food from them. The chemicals are non nutritive and appear to work

alone and sometimes the phytochemicals combine with vitamins and

nutrients in food to prevent any occurrence of diseases.

3.4.1. DETERMINATION OF TANNINS

The method of AOAC (1980) was used in this procedure;

2g residues of petroleum extracts were boiled with 300ml distilled water for

2hours, cooled and diluted to 500ml with more water and then filtered. 25ml

of the filtrates were transferred separately into 2L porcelain dish. 20ml

indigo solution and 750ml distilled water added, followed by 1ml of kMno4.
(earlier standardized with 0.1N oxalic acid) at a time until the blue solution

turned green, then few drops until solution became golden yellow. Triplicate

analyses making up to 100ml with distilled water. It was vigorously shaken

several times and allowed to stand for 15 minutes each time. Suspensions

were filtered through whatman filter paper.

Spectrophotometric measurement

5mm of each sample extract was pipette into labeled test tubes, and

5ml of oxidation solution (potassium ferrocyanide:NaOH mixture 1:9v/v)

was added. Each mixture was shaken and then allowed to standard for 1

minute. Three drops of hydrogen peroxide solution was added to each test

tube and shaken again. Absorbance of each preparation was determined at

369nm against a blank prepared in the same manner, but 5ml of water added

instead of the sample extract. Triplicate determinations were carried out on

each sample extract.

Calculation

The tannin content in mg per 100g samples was obtained as follows:

Mg vit. B1= Abs × 100* × 110 × 1000

5
Where 100* is the volume to which extract was made up to. 110 is a

conversion factor. 5 is weight of sample taken for extraction.

3.4.2. DETERMINATION OF HYDROCYANIC ACID

The alkaline titration method of AOAC (1990) was employed.

10g of each powdered sample was weighed separately into

kjedahl digestion flask and 200ml distilled water added. Flasks were

connected to the distillation unit, and let stand for 3 hours before steam

distillation commenced. 150ml of distillate was collected into 250ml

volumetric flask containing NaOH solution (0.5g in 20ml distilled water).

Distillates were made up to volume with more NaOH solution. 100ml of the

mixtures were measured out into another set of flask and 8ml 6N NH4OH

and 2ml 5% KI solutions added to each flasks. These were then titrated with

0.02 N AgNO3 to faint but permanent tubidity end point. Triplicate titrations

were carried out on each sample.

Calculation

The amount of HCN in each sample was calculated using the relationship:

1ml 0.02N AgNO3= 1.08mg HCN (AOAC, 1990)

3.4.3. DETERMINATION OF ANTHOCYANINS

This was done gravimetrically by the method of Harbone (1973). 5g

of each test sample was hydrolyzed by boiling in 100ml or 2mhcl solution

for 30 minutes. The hydrolysate was filtered using whatman filter paper. The

filtrate was transferred to a separation funnel and equal volume of ethyl

acetate was added to it, mixed well and allowed to separate into two layers.

The ethyl acetate layer (extract) was recorded while the aqueous layer was

discarded. The extract was separated to dryness in the crucible over a steam

bath. The dried extract was then treated with concentrated amyl alcohol

extract and the filterate was transferred to a weighed evaporating dish and

evaporated to dryness. It was then dried in the oven at 30⁰C for 30 minutes

and cooled in a desiccator. The weight of anthocyanin was determined and

expressed as percentage of the original sample.

3.4.4. DETERMINATION OF PHYTATE/ PHYTIC ACID

The method described by oberleas (1983) was used for the

determination of phytic acid. The description of the method is as follows:

2g of the sample was weighed into a 100ml flask and extracted with

50ml of 0.2N Hcl. 5ml of the extract was measured out into a test tube fitted

with a glass stopper. 1ml of solution prepared by dissolving 0.2g ammonia

iron (111) sulphate 12H2O in 2N Hcl and made up to 100ml, distilled water

was added to the extract. The tube was heated in a boiling water bath for

30minutes and cooled in the water bath for 15minutes before allowing it to

adjust to room temperature. The contents of the tube was mixed and

centrifuged for 30 minutes at 3000rpm. 1ml of the supernatant was

transferred to another test tube and 1.5ml of solution made by dissolving 10g

2,2- bipyridine and 100ml thioglycollic acid in distilled water and made up

to 1000ml was added to it. The absorbance was measured at 519rpm against

distilled water. Calibration curve was prepared by plotting the concentration

of the reference solution against their corresponding absorbance. The

absorbance of the test tube sample was then used to obtain the concentration

from the calibration curve..5.

3.5. DETERMINATION OF MINERAL CONTENT

A mineral is a naturally occurring substance that is solid and stable at

room temperature, represented by a chemical formula, usually abiogenic and

has an ordered atomic structure. There are over 4,900 known mineral species,

over 4,660of these have been approved by the International Mineralogical

Association (IMA). Minerals can be described by various physical properties

which relate to their chemical structure and composition. Common

distinguishing characteristics include crystal structure and habit, hardness,

luster, streak, colour etc

3.5.1. Magnesium (mg)

The magnesium content of the samples was determined by the

EDTA complexometric titration methods (Udo and Ojuwole, 1986)

The wet digest extract (20ml) was measured into a conical flask. A

20% KOH solution was added to maintain the PH at 12.0 to enable the

formation of magnesium, blank was also prepared to serve as control.

3.5.2. Iron (Fe)

The Atomic Absorption method(2005) was used by Rao, A.N.

Apparatus and reagents

Concentrated hydrochloric acid, magnesium nitrate solution,

hydroxylamine hydrochloride solution, acetate buffer solution, iron standard

solution, orthophenanthroline solution, spectrophotometer.

Procedure:

10ml of aliquot ash solution was pipette into 25ml volumetric flask,

and 1ml hydroxylamine hydrochloride solution was added. After 5 minutes,

5ml buffer solution and 1ml o- phenanthroline solution was added to dilute

the sample. Absorbance of solution was determined at 510nm. From

absorbance reading, Fe content present in aliquot of ash solution was

determined by referring to standard curve.

Preparation of standard curve

0.0, o0.5, 1.0, 1.5, 2.0, 3.0, and 4.0ml of Fe standard solution was

pipette into a series of 25ml volumetric flasks and exactly 0.2ml of

concentration Hcl was added to each of them. Each of them was diluted to

exactly 10ml with water and then reagents were added in the same way as

for the sample. The quantity of iron was plotted against the absorbance.

3.5.3. Zinc (zn)

Reagents

Copper sulphate solution , ammonium citrate solution, dilute

hydrochloric acid, zinc standard solution.

The standard method of AOAC(2010) was used.

To a suitable aliquot of ash solution of sample, 2 drops of methyl red

indicator and 1ml of CuSO4 solution was added and neutralized with

NH4OH. Enough Hcl was to make solution about 0.15N with respect to Hcl.

The pH of this solution was adjusted and measured with glass electrode

from 1.9 to 2.1. Stream of H2S was passed into solution until precipitation

was completed. It was then filtered through fine paper (whatman previously
washed with Hcl (1+6) followed by water). The filtrate was received in

250ml beaker, the flask was washed and filtered with 3 or 4 small portions

of water. The filtrate was gently boiled until odour of H2S can no longer be

detected. 5ml of saturated bromine- water was added and boiling continued

until Br-free. The solution was cooled, and neutralized to phenol red with

NH4OH and slight acid was diluted to definite volume. At this stage, the

solution contained 0.2 to 1.0 µg of zinc/ ml.

5ml of ammonium citrate solution, 2ml dimethylglyoxime solution

and 10ml of α nitrose- β- naphthol solution were added to 20ml aliquot of

the prepared solution and shook for 2 minutes. Solvent layer and extract with

10ml of CHcl3 was discarded. (the extraction procedure in this para

eliminates nickel and cobalt present , if any in the solution.)

3.5.4. PHOSPHOROUS (p)

Reagents

Vavadomolybdate reagent, 50mg/l phosphate (phosphorous) stock

solution
DIGESTION REAGENTS

Concentrated sulphuric acid, Na2SO4- CUSO4 digestion mixture.

The standard method of AOAC(1995) was used.

EXPERIMENTAL PROCEDURE

0.5g of dry powdered tissue was weighed in a digestion tube, 4g

of Na2SO4 was added, it was then digested until a clear, green coloured

solution was produced and cooled. When cooled, the green colour faded

from a nearly colourless to pale blue solution and the salts in the mixture

crystallized to a nearly solid mass. Purified water was used flask to a 100ml

volumetric flask. It was allowed to cool before the mixture was diluted in the

volumetric flask to the 100ml mark.

BLANK AND STANDARDS

Blank: 4g of the Na2SO4- CUSO4 mixture and 8.0ml conc H2SO4 were

added to 50ml H2o in a 100ml volumetric flask. It swirled to dissolve. It was

cooled to room tempetature,diluted to 100ml with purified water.

Standards , 10 and 20mg/ l phosphorous : 20.0ml and 40.0ml of the

50.0mg/l phosphorous stock solution was transferred to separate in 100ml

volumetric flasks. Purified water was added to bring the volume in each

flask up to 50ml. 4g of the Na2SO4-CUSO4 mixture and 8ml conc H2so4.

Were added to each flasks (note 8.oml H2SO4 is used rather than 10ml

because this is close to the amount expected to remain in a digestion flask

after digestion is completed). It is swirled to dissolve. After cooling to room

temperature, it was diluted to 100ml with purified water.

PROCEDUDURE FOR PHOSPHATE DETERMINATION

The solution was transferred to separate 50ml beakers, 5.0ml

portions of blank, standard and each digest. 20.0ml aliquot of

vanadomolybdate reagent was added to each beaker with swirling to mix. A

yellow colour, which is stable for hours, developed in less than 5 minutes.

A spectrophotometer was used to measure the absorbance of each

standard and sample by pouring each solution in a cuvette and measured its

absorbance at 400nm. A portion of blank solution in a cuvette was used to

set 0.000 absorbance.

 A standard curve was prepared by plotting absorbance as a function of

concentration. The absorbance of each sample was used to determine the

concentration of phosphorous in that digest from the standard curve.

3.5.5. POTASSIUM (K)

Reagents:

1000mg/l potassium stock solution

Digestion reagents:

Same as in phosphorous

Experimental procedure and blank are same as in phosphorous.

Standards – 5.omg/l, mg, 25.0mg/l ca, 50.0mg/l k.

Procedure:

The standard method of AOAC(1986) was used

The blanks, standards and each digested sample 25- fold were diluted in

volumetric flasks. The potassium lamp was put in the lamp holder, and set
the atomic absorption spectrophotometer to the appropriate wavelength, slit,

lamp current and photo- multiplier voltage.

The 25- fold diluted blank was aspirated and used to zero the

instrument. After the instrument has been zeroed, the absorbances of the 25-

fold diluted standards and digests were measured.

Standard curves for potassium was prepared by plotting their

absorbance as a function of its concentration.

3.6. DETERMINATION OF PASTING PROPERTIES

The method of Newport scientific(1998) was used.

Pasting properties were determined with the rapid visco Analyzer (RVA)

3.5g of the flour sample was weighed and dispensed into the test canister.

25.0ml of distilled water was dispensed into the canister (14% moisture

basis). The visco Analyzer was switched on and the pasting performance of

the flour was automatically recorded on the graduated sheet of the

instrument.
3.7. DETERMINATION OF PHENOL OXIDASE

(J. Agric food chem.)

The reactions were carried out in test tubes at 30⁰ with a phosphate

citrate (mcllvaine) buffer of pH 6.0. This pH was found to be close to the

optimum for most of the enzyme preparations tested and was high enough to

give the full blue colour of the indophenols dye adopted without permitting

appreciable autoxidation of the leuco dye. Variation in buffer strength had

little effect on the rate of reaction. The temperature was adequately

controlled by keeping all reagents in a 30⁰ bath, since no significant change

in temperature occurred during the very short reaction period required for

the colorimeter determination. Rates were measured with the lumetron

colorimeter and multiple reflection galvanometer. Galvanometer readings

plotted on a logarithmic axis were linear with time of the reaction for a

suitable period under the conditions described.

CHOICE OF DYE AND ITS EFFECT:

A large number of reversibly oxidized dyes of suitable potential were

tested from the standpoint of solubility, stability, autoxidizability, absorption

coefficient and effect on the enzyme preparations. The substituted

indophenols were found most suitable and 2,6- dichlorobenzenoneindo-31-

chlorophenol (Eastman) was the best of these. It is readily reduced by

hydrogen and palladized asbestos and is stable in the leuco form under

hydrogen for several hours. It is conveniently standardized colorimetrically

with ascorbic acid, but weighed samples of the dye gave sufficiently uniform

solutions for routine work. For many purposes, ascorbic acid reduction may

also be acceptable for leuco dye preparation, but there were some evidence

that the oxidation products of ascorbic acid affected the enzyme reaction

adversely.

Two important effects of the indophenols dyes were observed that were

further considered. First, the dyes varied widely in their effect on the

oxidation rates of enzyme preparations from certain sources. With some

dyes initial rates were low and fell off rapidly, indicating enzyme inhibition.

The 31 chloro dye was in fact, originally substituted for the more common

2,6- dichlorobenzenoneindophenol, because the former gave higher rates of

longer linear course with potato preparations. The potato enzyme was

particularly sensitive to these dye effects, but was not typical of the plant
materials investigated. A second important characteristic of the indophenols

dyes in general and the 31- chloro dye in particular was the fact that they

were oxidized directly, without phenolic mediator, by enzyme preparations,

therefore, two measurements of oxidase activity were necessary: (1) Total

phenol oxidase (with phenolic substrate or mediator) and (11) ‘dye oxidase’

(without phenolic substrate). The difference is considered a measure of true

phenol oxidase activity. This is based on two types of evidence. First,

mixtures of two enzyme preparations of widely varying proportions of ‘dye

oxidase’ and phenol oxidase gave additive rates within experimental error,

indicating no interaction of direct oxidation and dye oxidation through

phenol mediation. Second, the two varieties could be separated by a specific

inhibitor.

CHOICE OF PHENOLIC SUBSTRATE:

A variety of phenolic compounds were tested for substrate or

mediator activity in the system.

3.8. DETERMINATION OF REDUCING SUGAR

DETERMINATION OF FRUCTOSE, SUCROSE AND GLUCOSE

This method was conducted according to the AOAC(1995).

Fructose, sucrose and glucose concentration were determined by HPLC.

The HPLC system consisted of reverse phase-high pressure liquid

chromatography. All sugars were separated using supelco LC-NH2 column.

The mobile phase was Acetonitrite (80% in distilled water). The flow rate

was 1.5ml/min. Data was recorded and analyzed using chromatopack-CRT-

A. Results were reported as percentage (w/w).

 CHAPTER FOUR

4.1. RESULTS AND DISCUSSION

4.1.1. TABLE 1: PROXIMATE COMPOSITION (%) OF SAMPLES

Sample Protein Fat Ash Moisture Crude Carbohydrate

fibre
WSF 13.1±0.05 3.7±0.01 0.9±0.02 10.6±0.40 1.7±0.01 70.0±0.00

MSF 10.7±0.00 3.2±0.00 1.11±0.03 9.9±0.60 1.7±0.02 73.4±0.00

GPF 6.7±0.00 1.1±0.14 0.7±0.00 10.3±0.01 2.6±0.01 78.6±0.00

SPF 5.6±0.00 2.3±0.08 1.3±0.69 10.2±0.32 2.4±0.02 78.2±0.00

Data are means of duplicate determinations ± SD.

Data in the same column bearing different superscripts differed significantly

(p ˂ 0.05)

DISCUSSION

The proximate compositions of two laboratory prepared sorghum and sweet

potato flours are presented in table 1.

The fat content of the SPF is greater than that of the GPF because it was

steeped in water and decanted two times in an interval of 10 hours which a

reasonable amount of fat were eliminated in this process. MSF is an

important ingredient in addressing the issue dietary bulk of weaning foods

required in meeting a growing infant’s energy needs, this is a quality which

WSF does not possess. The protein and starch in sorghum grain are more

slowly digested than those from other cereals, and slower rates of

digestibility are particularly beneficial for people with diabetes.

KEY

WSF = whole sorghum flour

MSF = malted sorghum flour

GPF = grated potato flour

SPF = soaked potato flour

4.1.2. TABLE 2: MINERAL COMPOSITION OF SAMPLES

Sample Phosphorous Potassium Magnesium Iron (Fe) Zinc (zn)

(p) (K) (Mg)

WSF 319.98±1.79 413.00±00 78.32±0.12 3.68±0.04 2.85±0.04

MSF 253.04±4.6 453.50±6.36 87.28±0.18 4.31±0.00 3.45±0.01

GPF 98.56±0.56 342.00±1.41 38.63±0.04 1.22±0.23 0.45±0.04

SPF 121.66±0.05 398.00±1.41 42.35±0.00 2.15±0.42 0.67±0.00

Data are means of duplicate determinations ± SD. Data in the same column

bearing different superscripts are significantly different. (p ˂ 0.05)

DISCUSSION

This table consist of the mineral content of two samples of sweet potato

flour and two samples of sorghum flour. The minerals include potassium,

iron, zinc, magnesium and phosphorous. Phosphorous content in WSF is

greater than the phosphorous contained in MSF whereas potassium content

in MSF is greater than that in WSF.

Higher content of phosphorous in HSF indicates that the adequate intake will

result in provision of bone building nutrients. All the mineral contents in

SPF are greater than those in GPF, which means that SPF will be a good

source of nutrient.

KEY

WSF = whole sorghum flour

MSF = malted sorghum flour

GPF = grated potato flour

SPF = soaked potato flour

4.1.3. TABLE 3: PHYTOCHEMICAL COMPOSITION OF THE

SAMPLES

Sample Tannin Anthocyannin Phytate HCN

(mg/100g) (%) (mg/100g) (mg/100g)

WSF 29.48±0.10 0.88±0.02 216.11±1.33 19.80±0.11

MSF 11.38±0.06 0.57±0.00 107.45±0.30 8.76±0.01

GPF 25.09±0.08 1.18±0.78 10.61±0.21 8.46±0.21

SPF 49.92±0.02 0.65±0.36 16.16±0.07 10.60±0.09

Data are means of duplicate determinations. Data in the same column

bearing different superscripts are significantly different. (p ˂ 0.05)

DISCUSSION

The phytochemical composition of the samples are represented in the above

table with HSF having the lead role of phytate content. The two sweet potato

samples contain very low phytic acid which means that sorghum as a whole

has a very high phytate content. The anthocyannin content is very low in the
four samples which indicates that their intake cannot bring about the

nutritional role of anthocyannins, one of which is their ultimate role in the

chemoprevention of human cancer. Tannins found in red grained sorghum

contain anti oxidants that protect the body against cell damage, a major

cause of aging.

KEY

WSF = whole sorghum flour

MSF = malted sorghum flour

GPF = grated potato flour

SPF = soaked potato flour

4.1.4. TABLE 4: PHYSICO-CHEMICAL PROPERTIES OF

SAMPLE

Sample Pasting time Pasting temp Phenol oxidase

(min) (ºC)

WSF 10.73.±0.01 83.00±0.00 *

MSF 11.44±0.06 90.50±0.71 *

GPF 11.22±0.01 77.00±0.00 1975.00±0.00

SPF 8.90±0.02 70.50±0.71 2950.00±0.00

RP * * 15600.00±0.00

Data are means of duplicate determinations ± SD. Data in the same column

bearing different superscript are significantly different. ( p ˂ 0.05)

DISCUSSION

This table represents the pasting properties and phenol oxidase composition

of the samples. The pasting properties includes the pasting time and pasting

temperature of the different samples.

Phenol oxidase was analyzed in only the three samples of sweet potato, with

the SPF taking the lead role. The phenol oxidase is the enzyme that causes

browning in food samples, the browning of SPF was noticed right after

processing.

KEY

WSF = whole sorghum flour

MSF = malted sorghum flour

GPF = grated potato flour

SPF = soaked potato flour

RP = raw potato

* = not determined
4.1.5 TABLE 5: SUGAR COMPOSITION OF THE SAMPLE (mg/g)

Sample Fructose Sucrose Glucose

HSF 63.24±0.00 243.50±0.00 197.08±0.00

MSF 95.40±0.12 101.50±0.00 261.45±0.00

GPF 23.18±0.02 139.50±0.00 167.80±0.00

SPF 364.16±0.01 457.00±0.00 262.38±0.00

Data are means of duplicate determinations ± SD. Data in the same column

bearing different superscript differed significantly.

DISCUSSION

The sugar compositions of the samples are represented in the above table.

The values of fructose in the four samples are quite lower than the other

sugars except in SPF. Sucrose and glucose values are quite high and so cant

be recommended for diabetics. Considering the sugar content of all the

samples, fructose is quite considerable with the lowest value found in GPF,

which means a diabetic can eat GPF and still maintain an average sugar

composition.
High sucrose in the blood increases the sugar level and so diabetics should

avoid this.

KEY

WSF = whole sorghum flour

MSF = malted sorghum flour

GPF = grated potato flour

SPF = soaked potato flour

 CHAPTER FIVE

CONCLUSION

This study showed that the grated sweet potato is low in fat and

sugar and can be recommended for diabetics as these are the main causes of

diabetics, even though sorghum grain still has a beneficial role in the life of

diabetics, even as the protein and starch content are more slowly digested

than those from other cereals, and slower rates of digestibility are

particularly beneficial, grated sweet potato still has the lead role. Further

researches are hence encouraged using other types of food samples.

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