CHAPTER ONE

1.0 INTRODUCTION
Fruit juice is defined in the European Economic Community (EEC) as juice
obtained from fruit by mechanical processes, fermentable but unfermented, having
the characteristic color, odour and flavour typical of the fruit from which it comes
from. The definition has been extended to include the product obtained from a
concentrate, which must have sensor and analytical characteristics equivalent to
those of juice obtained directly from the fruit. Fruit Juices are valuable from the
nutritional point of view (Manay and Shadaksharawamy, 2001).
Fruit contain a wide range of different compounds and therefore show
considerable variations in composition and structure. Each individual fruit is
composed of living tissues that are metabolically active and it is constantly
changing in composition depending on the physiological role and stage of maturity
of the fruit (Salunhhe et al. 1991).
The nutritional value if fruit depends on their composition although fruits
play a very significance role in human nutrition. The composition of fruits is such
that it is not recommended as a sole source of nutrition. However, it can be used
advantageously to supplement deficiencies in other foods. The most important
component in fruits can be grouped as follows: water, protein, carbohydrates, fats,
minerals, fibres and vitamins (Potter, 1986; Salunkhe et al. 1991; Holland et al,
1992).
In developed countries, fruits juice commonly form part of the breakfast and
is produced in large quantities. The preparation of fruit juice is limited mostly to
home scale production. Fruit generally used for making juice are orange, pawpaw,
mango and apple etc. of late, the manufacture of squashes (consists of strained
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juice containing moderate quantities of pulp to which sugar is added for
sweetening) on a commercial scale has made some progress (Manay and
Shadaksharaswamy, 2001).
Fruit juices are best in taste, aroma, and color when freshly expressed. The
most important problem, therefore, in the fruit industry is to use such methods as
would help retain these properties to the maximum extent. The steps involved in
the processing of juice are selected based on the type of fruit being processed.
These are usually minor variation and generally do not depict any technological
novelty, nor do they involve any fixed scientific principle, but are found to be
indispensable steps if products of the possible quality are to be obtained. The
variation I process steps are invariably related to anatomical and compositional
characteristics of the different fruit (Okaka, 2009). Some fruits may contain
considerable amount of fibrous and lignified portions, which must be removed, or
they will adversely affect texture and mouth-feel. Others contain seeds which are
usually associated with destabilizing enzyme systems which if not promptly
inactivated cause product deterioration, yet others may contain small amount of
coagulable proteins which slow down filtration processes, and may cause
sedimentation of product components during storage (Okaka, 2009).
However, the generally followed steps involved in juice processing include:
selection and weighing, washing and peeling, juice extraction, juice formulation,
mixing, bottling and corking, pasteurizing, cooking and packaging (Fiiro, 2005). Due
to lack of appropriate storage facilities, up to 40-60% losses are recorded annually
during peak harvesting season. In other to minimize the losses, fruits are being
processes in large quantities in various forms and may be preserved with chemicals
to make them available even at off seasons. Fruit juice is important in human

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nutrition far beyond its use as a refreshing source of liquid. Many fruits contain a
variety of minor ingredients, particularly vitamins and minerals, as well as
carbohydrates, which are the predominant solid component. Although fruit contains
small amount of protein and fat, these are not important ingredients of juice. (Alan
and Sutherland, 1994).
Nutrients frequently consumed in sub-optimal concentrations by human are
proteins, calcium, iron, vitamin A, thiamin (Vitamin B1), riboflavin (Vitamin B2) and
ascorbic acid (Vitamin C). Some of these nutrients occur in higher concentrations in
fruit juices than is other foods. There is experimental evidence that indicates that
ascorbic acid of natural origin is apparently superior to that of synthetic origin
(Arthey and Ashurst, 1996).
It has been established that the above phenomenon is caused by the
presence of certain falconoid compounds in fruit juice that influence blood
circulation, increasing the permeability and elasticity of capillaries. This action is
known as vitamin P activity, but the falconoid showing this property are not
classified as vitamins, because there are several substances with this activity and so
serious deficiency disease occurs if they are consumed. There are indications that
there falconoid have a useful protective action, in particular against some
respiratory diseases, but they are readily decomposed in the body and it is
impossible to maintain an effective concentration in the blood (Arthey and
Ashurst,1996).
Apart from the obvious benefits of fruits juice, such as being a source of
potassium, it contains other substances that have or are claimed to have useful
pharmacological activity. For example, limonin and other related limonoid
substances present in citrus fruits are believed to have a role in inhibiting certain

3
 forms of cancer. Sorbitol which occurs in many fruit juice has a laxative effect
(Arthey and Ashurst, 1996).
Whatever the nutritional interest, it should be noted that changes occur
during storage, particularly to the minor components of juices and particularly
under adverse conditions (example light, increasing temperature, time) (Arthey and
Ashurst, 1996).

1.1 Objectives
1. There are different kinds of fruit flavoured drinks that are sold in the market.
This work is therefore aimed at evaluating the quality characteristics of the fruit that
are sold within Makurdi metropolis for the purpose of ascertaining their compliance
with regulatory standards.
2. To evaluate the physiochemical properties of fruits juice products.
3. To evaluate the microbiological characteristics of the juice products.

1.2 Justification of Study

Fruit juice is obtained from fruit by mechanical process, which is consumed
for several reasons including quenching stimulant effect, health value and
enjoyment, but this work is mainly focus on evaluating the quality characteristics of
fruit juice that are sold within Makurdi, Metropolis for the purpose of ascertaining
their compliance with regulatory standards as well to evaluate the physiochemical
properties of fruit juice products and microbiological characteristics of the juice
products.

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 1.3 Scope/Limitation of the Study

Quality characteristics of some fruits based drinks sold within Makurdi

metropolis is a project researched work designed to cover the quality and
constituents of fruits juice, the importance and classification of beverage, the
physiochemical analysis of juice and microbiological characteristic of juice products.

1.4 Statement of Problem

Many fruits juice contains variety of minor ingredients which are important to
human nutrition far beyond it use as a refreshing source or liquid, this nutrient
includes proteins, calcium, iron, vitamin A, B&C. Apart from being source of
potassium fruit contain other substance that have or are claimed to have useful
pharmacological activity e.g limonin. But the most important problem therefore is
for the industry to process and retain these properties to the maximum extent and
to ensure the nutrients are maintained during storage and preservation.

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 CHAPTER TWO
2.0 LITERATURE REVIEW
2.1 Beverages
Beverages are foods that are distinguished by two principal characteristics
from other foods. First, they are liquids or are consumed in liquid state but the
relative lake of actual food value differentiates them from other like milk and milk
products. Secondly, they are either consumed for their thirst quenching properties
or for their stimulating effect (Ihekoronye and Ngoddy, 1985).
Beverage can also be said to be a drinkable liquid, consumed for variety of
reason including thirst quenching effect, health value and enjoyment. Examples of
each type are water, caffeinated drinks (cocoa coffee, tea), milk, fruit and vegetable
juice and carbonated soft drinks respectively. The essential component of any
beverage is the water that it contains other component such as stimulants, coloring
and flavouring ingredients may perform some useful functions but are not essential
to proper physiological functioning of the body (Adrian, et al, 2000).

2.2 Classification of Beverages

Beverages as food can be classified majorly into alcohol and non-alcohol
beverages. They can also be classified based on their functions in the body. Fig.2.1
shows the classification of beverages. The carbonated non-alcoholic beverages have
their major ingredients are sugar, flavourings, colures, acid, water and carbon
dioxide. Carbonation adds sparkle, vim and zest to this class of beverages. Generally,
the process of manufacturing comprise of the preparation of syrups, adding

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acidulants, flavouring and blending the mixture of ingredients (Unekoronye and
Ngoddy, 1985).
Ethanol or simply alcohol is a very simple substance containing only cardon,
hydrogen and oxygen. In the body, it acts in two quite distinct ways: it is a food
which is broken down in the body to give energy and it is a drug which affects the
central nervous system. Alcoholic drinks are judged in terms of flavour and
stimulant effect and hardly at all as a source of calories. They are made by the
process of fermentation (Ihekoronye and Ngoddy, 1985). Alcoholic beverages are
wine, beers and spirits. The beverage classifications are based on the function they
perform. A particular beverage may have more than one functions. (Bernard, 1985).

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 FOOD BEVERAGES

Non-alcoholic Alcoholic

Non-carbonated Carbonated Fermented Fermented

Fruit juice Soda (Non-distilled) (Distille)
Fruit drinks Coca-Cola
Fruit nectars ginger ale fruit Grain Miscellaneous Fruit Grain Miscellaneous
Vegetable juiceTonic water Branches
Chocolate drinks Wine champagne
Vodka Beer Whiskies Vodka

Ale Stout Lager

lllicit Gin Rum Tequila

Fig 2.1 Food beverage classification

Source: (Ihekoronye and Ndoggy, 1985)

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2.3 Quality index of Beverages
In the manufacturing of beverage, there is a concern for economic and quality
consideration. Thus the development of standards for fruit juice is incorporated into
most country’s regulatory codes. These regulation stands to maintain a high quality
and safely compliance of these beverages. These regulations includes the processing
employed, the soluble solid and acid levels, amount of added substances allowed
such as sugar, acid, water, preservatives and reasonable sanitary standards (Ashurst,
1999).
Grades may be depending on the colour, flavor, consistency and absence of
defects. Codex Alimentarius grades and standards are being promulgated and
continue to be refined to provide a useful guide for standardized national norms
and facilitating intra and international trade (FAQ, 2000 b). In the matter of a large
volume beverage (juice) trade specific standards are negotiated between major
suppliers and user (Ashurst, 1999).
Quality index of beverages consist of the compositional organolepic
specification as well as limits for microbial and chemical contaminants. Table 2.1
indicates some appropriate general specification worth developing to control the
quality index of beverage (Bollag and Liu, 1990).

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Table 2.1: Some Quality Specification for Fruit Juice
Attribute Rationale
Solute solid (brix) Define juice strength
Titrable acidity and pH Define acid balance
Colour Visual appeal
Freedom from defect-decay, insert Aesthetics, susceptibility to
or damage, mechanical injury etc spoilage and contamination
Maturity Optimum quality
Flavour Defines quality
Absence of pathogen, chemical and Defines safety
Extraneous matter
Low microbial Quality, shelf-life

Source: FAO, 2000.

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11
2.4 The Constituents of a Soft Drink
The term ‘soft drink’ applies to beverages containing flavour rings and/or fruit
juice together with other constituent of technological or nutritional value designed
to enhance the appearance and stability of the product and to ensure its
organoleptic properties remain intact during a reasonable shelf life. These factors
are taken into consideration in all development work, and in order to meet current
stringent quality and legislative controls a new beverage is subjected to extensive
trials to assess the suitability and performance of all components in its makeup. It
becomes essential to arrive at the correct ingredient formulation to achieve a
reproducible product. (Hollan et al. 1992).
Table 2.2 lists the functional constituents of soft drinks and their tropical
usage levels. Each category of ingredients, other than fruit juices are discussed in
more detail in the following sections:

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 Table 2.2 soft drinks components, General Usage and Contribution
Component typical use level

1. water (quality must meet regid Up to 98% v/v when high-intensit

Requirements) sweeteners in use.

Blend carrier for other ingredients. Provides essential hydration effect of enable
body metabolism.
2. Sugar 7-12% m/v when sole source of sweetener
Contribute sweetness, body to drink, act as synergist and give balance to
flavour.
3. Fruit juice Widely variable usage. Usually up to
10% as natural strength, although
Some specialized lines in this

Provides fruit source identity, flavour, mouth feel effect. Also contributes to
sweetness and acidity.
4 High-density sweeteners use based upon sucrose equivalence
(e.g aspartame might be employed
At 0.40-6% m/v as sole sweetener).

Provides sweetness, calorific reduction. Synergist action. Often used in

combination e.g aspartame with acesulfame K.

5. Carbon dioxide 0.30-6% m/v

Provide mouth feel and sparkle to drink (carbonate only).
6. Acids (e.g citric) 0.05-0.3% m/v
Contributes sharpness, sourness, background to flavour, increases thirst-
quenching effect.
7. Flavours Nature identical and artificial: 0.10
28% m/m. natural: up to 0.5% m/m.
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Provide flavour, character and identity to the drink.
8. Emulsion (flavour, colour, cloud etc) 0.1% m/v.
Carrier for oil-based flavours or colours. Gives cloudy effect in drink to replace
or enhance cloud from natural juice.

9. Colours (natural or synthetic) 0-70ppm.

Standardize and identify colour tone of drink.
10. preservatives statutory limits apply (e.g
Sorbic acid up to 250ppm in EU)

Restrict microbial attack and prevent destabilization of the drink.

11. Anti oxidants (e.g BHA, ascorbic acid) Less than 100ppm, subject to
User-country legislation.
Prevent oxidation, limit flavour and color deteritoration
12. Carragerenans, alginates, polysaccharides, cardoryl methyl cellulose etc.
Provide mouth feel, shelf life stability, and viscosity.
13. Vitamins/minerals AOI applies
Used in ‘healthy-living’ drink to provide nutritional requirement.

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2.4.1 Water
Water as the main component of a soft drink, usually accounts for between
85 and 95% of the product and acts as a carrier for the ingredients. Its quality must
conform to rigid requirements and not interfere with the taste, appearance,
carbonation or other properties of the drink. Subject to the location of the bottling
plant, the source of water and product specifications, it may be necessary to carry
out treatment to improve the quality of the water used in the manufacture of soft
drinks. (fellow 2002).
Most soft drinks factories will carry out their own treatments to counteract
the likelihood of a possible change in quality. This is most important in areas where
variations are introduced as a result of the use of a national grid system for water
supply. (fellow 2002).
In less developed countries, water treatment becomes an essential
prerequisite where microbial loading could provide cause for concern. It is necessary
for a full water treatment to be effective and to ensure the wholesomeness of water
supplies for boiling purpose. (Hansen 1987).

2.4.1.1 Requirements
Water should comply with the following quality requirement, it should
be free from:
 High level of elements and mineral salts;
 Objectionable tastes and color
 Organic materials.

It should also be:

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  Clear and colorless, free from dissolved oxygen, sterile, that is, free from
micro-organisms.

Ideally, a non-variable supply of water should be at all season of the

year to allow a standard manufacturing process to be established. (FIIRO
2010).

2.4.2 Acidulants

The use of acidulants is an essential part of beverage formulation, with the

acid component usually third in order of concentration. Acidulants performs a
variety of functions in addition to their primary thirst-quenching properties, which
are the result of stimulation of the flow of saliva in the mouth. Because it reduces
pH, an acidulant can act as a mild preservation and in some respects as a flavour
enhancer, depending on the other components present. (Manay and
Shadaksharaswany, 2011).

In addition, by functioning as a synergist to antioxidant such as butylated

hydroxyl anisole (BHA), butylated hydroxyl toluene (BHT) and ascorbic acid,
acidulants can indirectly prevent discoloration and rancidity (Broihier, 1999).

In carbonated beverages there is the additional effect of dissolved carbon

dioxide gas. Although it is not officially recognized as an acidulant, the inclusion of
carbon dioxide, under pressure, will certainly provide extra sparkle, mouth feel,
flavour and sharpness in a drink. The following are the most commonly encountered
acidulants (Broihier, 1999).

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2.4.2.1 Citric Acid

Citric acid is the most widely used in fruit-flavoured beverages. It has a light
fruit character that blends well with most fruit flavours, which is to be expected as it
occurs naturally in many fruit types. For example, unripe lemons contains 5-8% citric
acid, it is also the principal acidic constituents of such fruits as black currants and
cranberries and is associated with malic acid in apple, apricots, blue berries,
cherries, peaches, pears, strawberries and raspberries (Deshpande and salunkhe,
1982).

Citric acid was originally produced commercially from lemons, limes or

bergamots by pressing the fruit, concentrating the pressed juice and precipitating
citric acid as it calcium salt, from which subsequently purified. It is now produced by
the action of enzymes on glucose and other sugars (Deshpande and salunkhe, 1982).

Citric acid is a white crystalline solid and it can be purchased as a granular

powder in its anhydrous state or as the monohydrate present day soft drink
formulations usually employ. The anhydrous form, which is seen to have cost-in-use
advantage over what was the more traditionally used monohydrate (Deshpande and
salunkhe,1982).

2.4.2.2 Tartaric Acid

Tartaric acid occurs naturally in grapes. Where it is present as the acid

potassium salt, during the fermentation of grapes, tartaric acid precipitates from
solution as crystals, as its solubility decrease with the increasing alcoholic
concentration of the wine. This acid has a sharper flavour that citric and it may

17
 therefore be used at a slightly lower level to give equivalent palate acidity.
(Sivasanker 2008).

Tartaric acid can be isolated from the crude deposit of tartrates obtained
during the fermentation of wine, in a similar manner to that used for citric acid, that
is, by leaching the deposit with sulphuric acid and further purification by
crystallization. Tartrate acid (dextro-form) is a white crystalline solid with melting
point (m.p) 171-174oC. tartaric acid has a strong, tart taste and it complements
natural and synthetic fruit flavour, especially graps and guaranteed for good use.
One problem that may need to be addressed is that tartaric acid salts, particularly
the calcium and magnesium tartrates, have lower solubility than citric acid.
Consequently, there is a tendency for unsightly precipitates of insoluble tartrates to
form in hard water, and in such conditions it is preferable to use citric acid
(Sivasanker 2008).

2.4.2.3 Phosphoric Acid

Phosphoric acid is the only inorganic acid to be widely used in food

preparation as an acidulant. It does however, occur naturally in the form of
phosphates in some fruits, for example, lime and grapes. In the soft drinks industry,
its use is confined almost entirely to cola-flavoured carbonated beverages, where its
special type of astringent acidity complement the dry, sometimes balsamic
character of cola drinks. (Wu and Nofziger, 1999).

Phosphoric acid has a driver, and perhaps sharper, flavour than either citric or
tartaric acid, tasting rather of flat ‘appears’, in contrast with the sharp fruitiness of

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 citric acid. It therefore, appears to blend better with most non-fruit drinks (Wu and
Nofziger, 1999).

Pure phosphoric acid is a colorless crystalline (m.p. 42.350C) and it is usually

employed in solution as a strong syrupy liquid, miscible in all proportions with
water. It is commercially available in solution concentrations of 75, 80 and 90% and
is the result of hydrogen bonding between the phosphoric and molecules Georgre
and Pamploma-Roger, 2007)

2.4.2.4 Acetic Acid

Acetic acid has very limited use in beverage, only finding use where its
vinegary character can contribute to a suitable flavour balance in the intended
product. It is seldom used in anything except non-fruit beverages. Pure glacial acetic
acids is a colorless crystalline solid (m.p 160C) with a suffocating, pungent aroma. It
is one of the strongest of the organic acids in terms of its dissociation constant and
can displace carbonic acid from carbonates. (Salunkhu and Reddy, 1991).

2.4.2.5 Malic Acid

Occurring widely in nature, malic acid is closely associated with apples. It is

the second major acid after citric, found in citrus fruits and it is present in most
berry fruits. Malic acid is slightly stronger than citric in perceived acidity, imparting a
fuller, smoother fruity flavour. Malic acid is a crystalline white solid (m.p 1000C) and
is highly soluble in water. Being less hygroscopic than citric acid it provides good

19
 storage and shelf life properties. Unlike tartaric acid its calcium and magnesium salts
are higkly soluble and it therefore presents no problem in hard-water areas.
(Salunkhu and Reddy, 1991).

Malice acid finds use in a variety of products, mostly in fruit-flavoured

carbonates. It is the preferred acidulant in low calorie drinks and on cider and apple
drinks, enhancing flavour and stabilizing colour in carbonate and non-carbonated
fruit flavoured drinks. Malice acid may also be used to mask the off-taste of some
sugar substitutes. Blends of malice and citric acids are said to exhibits better taste
characteristic than either acidulant individually. (Salunkhu and Reddy, 1991).

2.4.2.6 Asorbic Acid

Ascorbic acid, known more as vitamin C, is used not only as an acidulant but
also as a stabilizer within the soft drinks system, and its antioxidant properties serve
to improve the shelf-life stability of flavour components. Many of the ingredients
used in flavourings are susceptible to oxidation, particulary aldehydes, keto-esters.
Ascorbic acid shields these from attack by being preferentially oxidized and lost,
leaving the flavour component unaffected (Nzeagwa and Udugwu, 2009).

It should be noted that although ascorbic acid acts well as a browning

inhibitor in unprocessed fruit juice, its effect can be destroyed should the juice be
subsequently pasteurized or heat treated. In such cases, ascorbic acid can initiate its
own chemical browning reaction (Nzeagwu, and Udugwu, 2009).

Another disadvantage of ascorbic acid is the effect on some colours in the

presence of light. In the case of azo-colours, such as carmoisine, a light-catalyzed

20
 reaction occurs, resulting in cleavage of the N-N linkage and consequent destruction
of the chromospheres. This accounts for the disappearance of colour and bleaching
of the characteristic hue associated with some soft drinks (Ndugwu, and Udugwu,
2009).

2.4.3 Flavourings

It is the flavour of a drink that provides not only a generic identity but also its
unique character. This part of the sensory profile is responsible for pleasing and
attracting the consumer. For example, having decided on a cola drink, the consumer
will be a able to differentiate between colas by virtue of the background flavouring
components, which collectively provide a reference point to which the consumer
can return, consciously or not, on future occasions, whenever a particular brand of
drink is selected. (Salunkhu and Reddy, 1991).

A flavouring consists of a mixture of aromatic substances carefully balanced

to convey the right message to the sensory receptors of the consumer. The
preparation of such a mixture is a serious matter, the soft drinks flavourist, like the
perfumer, must be well versed in the technique, be creative and be able to translate
ideals into a practical solution. (Salunkhu and Reddy, 1991).

Although the art of the flavourist depends largely on individual sensory

abilities, it is frequently necessary in present day flavour work to enlist analytical aid
at an early stage of the project. Modern instrumental analytical techniques are
capable of detection to extremely low limits, but it is still usually necessary to
prepare an extract or concentrated version of the target flavour before carrying out

21
 the analysis. This may be achieved by solvent extraction, distillation, absorption
chromatography, dialysis, headspace concentration and cryogenic or adsorpbent
trapping, among other methods. (Ndugwu, and Udugwu, 2009).

A good gas chromatography (GC/MS) system can be used to identify the

profiles of compounds and individual flavouring substances up to say, 98% of the
target flavour, thus by passing much of the time consuming preliminary work
associated with organoleptic flavour matching (Nzeagwu and Udugwu, 2009).

2.4.3.1 Flavourings and Legislation

Food ingredients have, in most instances, been well investigated in terms of

use and effect. They have been categorized under permitted list as appropriate to
local legislation. Flavourings however, by virtue of their complexity have always
existed as a separated group. A distinction is made under various systems as to
whether they are of natural origin or not. This follows adoption by the FAO/WHO
food standard programmed and the codex Alimentanus Commission of a proposal
put forward around 1975 by the international organization of the flavour industry
(IOFI) to divide flavouring substances into three categories. (Dykaar and Hiyanidis,
1996).

Natural flavour and natural flavouring substance: These are preparations and
single substance, acceptable for human consumption, obtained exclusively by
physical processes from vegetable or sometimes animals raw materials either in
their natural state or processed for human consumption. (Dykaar and Kitanidis,
1996).

Natural identical flavouring substance: These are substance chemically isolated from
aromatic raw materials or obtained synthetically. They are chemically identical to
substance present in natural intended for human consumption whether process or
not. (Manay and Shadaksraswany, 2001).

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 Artificial flavouring substance: for the purpose of the codex alimentanus, these are
the substance which have not yet been identified in natural product intended for
human consumption, whether process or not. They are hence made available only
through synthesis. (Manay and Shadaksharaswany, 2001).

2.4.4 Colours

Those of us fortunate enough to possess optical powers capable of

distinguishing a variety of colour will appreciate the influence that this particular
sensory dimension exercise on our judgment of matters important to our well being,
such as food and drinks.

The perception of colour influence a taste reception of a drink and as a

consequence there is inevitably some controversy over the use of colouring in food
and drink. (Somogyi, and Hui, 1996).

One point of view states that, colour which possess no measurable nutritional
value, can have no role in food or drink other than that of deceiving the consumer.
To a certain extent this is true, but to appreciate the full value of colour as a food
additive, or more specifically as a drink additive. It is necessary to appreciate the
synergy between the sensory responses of sight and taste. (Somogyi, Ramaswamy
and Hui, 1996).

Colour provides a means of presenting a beverage to the consumers so that

the perceived organoleptic attributes are correctly placed in an ordered sequence of
appreciation. Both quality and quantity of colour are of important, and certain
colour will provoke or perhaps complement a particular taste. Reds will flavour the
fruitiness of soft drinks, for example, blackcurrent, raspberry and strawberry.
Orange and yellow tend towards the citrus flavours. Green and blues reflect the
character of peppermit, spearmint and cool flavour, sometimes herb like and
balsamic flavours. Therefire the deceit, if ever intended, is aimed at ensuring that
consumers are able to maximize their enjoyment of the beverage. (IHEKORONYE
AND NGODDY, 1985).

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 When a soft drink is based in part on fruit juice it may be necessary to restore
the appearance of the juice concerned. If its natural colour have been destroyed by
heat processing, or to intensity these colour when the contribution from the juice is
weaker than that normally associated with the effect the compounded drink is
intended to convery. Colour adjustment may also be necessary to ensure uniformity
of product and to offset natural variations in colour tone and intensity associated
with the juice type employed in the beverage formulation. (Somogyi, and Hui, 1996).

Above all, colour is a major parameter in the assessment of quality serving at

the time of production to standardize the product. Colour deterioration can provide
useful information on any quality changes during storage due to temperature
fluctuation or microbial spoilage effect for example (salt marsh, 2000).

2.4.5 Preservatives

A preservative can be defined as any substance that is capable of inhibiting

retarding or arresting live growth of micro-organisms or any deterioration of food
due to micro-organisms or as masking the evidence of any such deterioration. The
p-hydroxy benezoates previously cited in the legislation are no longer permitted for
use in soft drinks although they are till included under certain food uses.
(Sivasanker, 2008).

As mentioned earlier, carbon dioxide, although are not added specifically as a

preservative, contributes to the inhibition of the growth of micro-organisms and
coupled with other factors (e.g pH), contributes to the stability of a drink. Carbon
dioxide is deemed to be effective at volumes over 2.5 or 3.0, and for this reason, the
incidence of spoilage in carbonated beverages is less that in non-carbonated
versions (volumes of CO2 in general terms refers to the number of times the total
volume of gas, adjusted to 760mmHg and 00C, can be divided by the volume of
liquid in which it is dissolved. (Sivasanker, 2008).

Although preservative can be used to good effect in beverages formulations,

they should never be considered infallible and there is no substitute for stringent
quality and hygiene controls at every stage of manufacture. (FOA,1992).
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 Within their own product specification, raw materials should be assigned
workable limits for microbial activity, so that there is little chance of excessive
contamination in the finished beverage product. Equally, all processing plant,
machinery and containers likely to come into contact with the product during
manufacture should undergo a thorough cleaning (sanitation) before use. (No Fziger
et, al.1999).

Certain strains of yeast, moulds and bacteria can survive in relatively low Ph
conditions and some of these can exist and grow in the presence of certain
preservations so it is important that everything is done to prevent their multiplying.
Under favourable conditions, a typical rapidly growing yeast strains can double its
numbers every 30min, and at this rate in 12hours one yeast, could become 16.7
×106 cells, provided no inhibitory factor is present. (Hugo, 1969).

2.4.5.1 Sulphur Dioxide

Because of the ease with it can be produced, gaseous SO2 was one of the first
chemical compounds manufactured and used by humans. By Roman times, it was
used as a preservative by burning sulphur before sealing wine into barrels or storage
jars.

It is one of the most versatile agents used in food preservation and is well
known for its microbiocidal effect on bacteria, moulds and yeast. (Wikipedia. 2010).

The micobiocidal effect increases as the pH falls below 4.0, and because of this,
SO2 is ideally suited for most soft drink formulations. However, its preserving action
is impaired by a tendency to react with many of the fruit components of soft drinks
to form organic sulphites, in which state the SO2 is said to be ' bound', Although the
preservative properties are due mainly to free SO2. It is necessary to analyze for
total SO2 (i.e free plus bound) as legislation for safe levels refers only to maximum
total concentrations. (Encarta, 2009).

Disadvantages associated with sulphur dioxide are that some tasters can detect
it as an unpleasant back note or taint and it has a tendency to provoke allergic

25
 reactions in some individuals. Asthma sufferers tend to be affected by gaseous
sulphur dioxide, small traces of which can promote as asthmatic attack. There is a
risk with foods containing sulphates of gas liberation upon swallowing. (
Indiaparening.com, 2007).

2.4.5.2. Benzoic and Benzoates

Benzoic acid occurs naturally in some fruits and vegetables, notably in

cranberries. It is also found in some resins, chiefly in gum benzion (from styrax
benzoia) and in coal tar. Commercially available benzoic acid is produced by
chemical synthesis ( merriam webster 1981).

Pure benzoic acid is a white powdery crystalline solid ( m.p 122oC) only sparingly
soluble in water at normal temperature. Because of this, it is added to the drink in
the soluble form of its sodium or potassium salts. It is normal practice to disperse
the benzoate completely during batch makeup before addition of acid component,
with the resulting pH reduction, to avoid localized precipitation of the ' free' benzoic
acid to its solubility having been exceeded ( The solubility of benzoic acid = 0.25%
m/v at 20oC). It is the free or un-dissociated form of benzoic that exhibits
preservative action and hence its use is only effective when low ph value are
encountered ideally below pH 3, at which point the degree of dissociation reduces
to below 10%. ( Guhr ans Lachance, 1997).

Benzoic acid is generally considered to exhibit an inhibitory effect of

microbial growth, although it is of little use for bacterial control, where the greatest
growth problem will occur at pH values above 4, outside the effective limit
mentioned above. Improved results are obtained when it is used in conjunction with
other preservatives, for example SO2 or sorbic acid, due to synergistic effects. It is
interesting to note that the current European Directive, which sets individual limits
of 300mg/1 for sorbic acid and 150mg/1 for benzoic acid in non-alcoholic flavoured
drinks, nevertheless permits a joint preservative use of up to 250mg/1 sorbic acid
with 150mg/1 benzic zcid. (Britannica encyclopedia, 2010).

26
 Allergic response to benzoic acid have been reported, particularly among
children known to be made hyperactive by other agents, for example tartrazine. The
maximum ADI for benzic acid, recommended by JECFA, is 5mg/kg body weight.
(George and Pamploma Roger, 2007).

2.4.5.3 Sorbic Acid and Sorbates

Sorbic acid is found naturally in a number of fruits and vegetables, notably in

the juice of unripe mountain ash be berries (from sorbus aucuparia), where it
occurs together with malic acid. sorbic acid and its salts are among the most widely
used preservative in the world. In soft drinks the most commonly used form is
potassium sorbate, because like benzoic acid, there are problem in preparing its
solution (the solubility of sorbic acid = 0.16% m/v at 20oC), in common with benzoic
acid, as a microbial inhibitor, sorbic acid and its sorbates show reduced
effectiveness with increased pH. Although activity is greatest at low pH
value.(Friedman, 1991).

In addition to being less toxic than benzoates, sorbates seems to be less

obtrusive in terms of taste detection by certain individuals all allergenic reactions.
Overall, sorbates are considered to be one of the safest food preservative in use,
and WHO has set the sorbates at 25mg/kg body weight. (Friedman, 1991).

2.5 Micro-Organisms and Beverages

Contamination of fruit juices by the bacteria may occur when the organism
enters the processing plant or on the surface of the fruit having originated from soil,
untreated surface water, dust and decomposing fruit. The degree of contamination
varies depending upon haw the fruit was handled from field and in the processing
plant (Kader, 1992).

Although there is little evidence of the formulation of toxic fermentation

products in beverages, the problem of spoiling frequently arises. Because of their
utilization of sugars, yeasts are of most immediate concern. (kader, 1992).
27
 Yeasts are classified with the fungi and are unicellular for most of their life
cycle. Together with molds and bacteria, they can bring about deterioration in
flavour, producing taints, off-notes; differences in mouth feel and so on. Most yeast
can grow with or without oxygen, whereas most bacteria cannot survive in it. The
majority of yeasts thrive in temperatures between 25 and 270C, some can survive at
temperature over 700C and others can exist apparently quite comfortably at 0-10oC.
Bacteria exhibit some similar diversity in their characteristics, with an optional
growth temperature at around 37oC. (Alan and Sutherland, 1994).

Soft drinks provide an idea growth substrate for many micro-organisms

providing them with adequate supplies of the nutrients they require. Apart from
water, the environmental necessity, typical requirements are source of carbon
(Carbohydrates), nitrogen (amino acid), phosphorus (phosphate), potassium,
calcium (mineral salt) and traces of their minerals, for example, sulphur, iron and
even vitamins. Because of its obvious link with protein formation during cell growth,
the presence of combined nitrogen is of particular importance. Also, when it is
introduced to beverages via fruit pulp or caramel (colouring), there will be a greater
susceptibility to spoilage by certain micro-organisms. (Hasler, 1998).

2.6 Safety Preservation of Fruits Juices

Fruit juice and fruit-based beverages is mildly acidic product usually

containing fermentable sugars, organic acids, vitamins and trance element, and are
subject to contamination by and growth of a variety of spoilage organisms, notably
yeast and moulds. One to some outbreak of illness due to consumption of fruits or
juice contaminated with pathogenic micro-organism, the USFDA has issued a
guidance document to minimize microbial food safety hazards in fresh and

28
minimally processed fruit and vegetables and mandated a Hazard Analysis and
Critical Control Point (HACCP) program to achieve an s-log reduction of pathogens
(Encarta, 2010).

In order to achieve the desired hygienic condition and shelf-life, a hazard

analysis and critical control point (HACCP) program has to be involved to bring out
the good manufacturing practice (amp). The HACCP program is a systematic,
proactive and preventive approach to food safety assurance that involved
identifying and assessing the microbiological, chemical and physical hazards from a
particular food production process or practice (Hazard Analysis) and minimizing the
risk by controlling or eliminating the hazard at the point of production process
where a failure would likely result in a food hazard being introduced or allowed to
persist (critical control point) (Encarta, 2010).

The HACCP system has been effectively used to control microbial hazard in
low acid foods. Before developing an HACCP plan, the juice processor must have an
implemented amp requirement and sanitary standard operation procedure (SSOPs)
(Pearson, 1991).

In the processing of fruits, microbial are reduced by the use of preservatives.

Among them, potassium sorbate, citric acids, sodium benzoate. All these
preservative performing different function on microbes, potassium sorbate has little
effect in reducing E. Coli 0157:H7 (Encarta, 2010). Sodium benzoate 0.2% increase
the z-value from 6 to 26%0C. induction of acid resistance can also have wide ranging
effects on the ability of bacteria to resist other stresses such as heating,
antimicrobials and exposure to UV light. (Adams and Moss, 1999).

29
2.7 Other Functional Ingredients

The functionality of the ingredients already discussed tends to be self-evident

as contributing to the main identity of a drink. Except for the preservative, these
ingredients are used primarily for taste and colour. These are other whose
contribution is to improve performance and further characterize the drink. (Fellow,
2000).

2.7.1 Stabilisers

They are used in soft drinks formulation to impart stability to natural clouds.
For example, dispersions of fruit solids and to improve mouth feel characteristic by
increasing the viscosity of the drink. (Holland et, al. 1992).

In EU legislation on more than 10 are used on a regular basis in soft drink

formulations. These include the alginates, carrageens, vegetable gums, pectin,
acacia, guar, tragacanth, xanthan and carboxy methyl cellulose. (Holland et,
al.1992).

2.7.2 Antioxidants

Perhaps the most common problem encountered during storage of a

beverage relates to the oxidation effects involving certain ingredients. Both flavour
and colour components can be subject to deterioration in the presence of dissolved
oxygen, to the detriment of the product. Antioxidants are therefore included in
those formulations containing ingredients most vulnerable to oxidation. (Beom and
Myung, 1998).

30
 Oxidation can frequently be attributed to the oxygen permeability of the
plastic materials used in container manufacture, but it is essential that the oxidation
process should not start at the production stage of the drink or any of its
ingredients. (Hanseen, 1987).

Citrus-flavoured drinks, notably lemon drinks, are frequently susceptible to

oxidation and so antioxidants may feature in their formulation.

Oil-based, water-dispersible flavour (emulsions) are protected by the addition

of oil-solute antioxidants such as butylated hydroxyl anisole (BHA) and butylated
hydroxyl toluene (BHT) to the oil phase before the emulsification process, 1000mg/I
is the typical usage level in essential oils. Since the flavour emulsion will be used at
the rate of about 0.1% the level of antioxidant in the finished beverage will be of the
order of 1mg/I, who will safety, comply with an ADI of 5mg/kg body weight for
either additive (FIIRO, 2010).

Increasing use in being made of natural and nature-identical antioxidants

because in many countries use of BHT continues to be restricted on health grounds,
ascorbic palmitate (6-0-palmitoyl-L-ascorbic acid) and its sodium and calcium salts,
natural extracts rich in tocopherols (vacuum-distilled) and synthetic a-,y-,
tocopherols are used to good effect in preventing oxidation deterioration in oil-
based system, in combination, ascorbyl palmitate and tocopherol (vitamin E)
synergise to exhibit enhanced anitioxidant properties (FIRO, 2005).

31
2.7.3 Calcium Disodium Edta

This mixed salt of ethylene diamine tetraacetic acid (EDTA) is prepared by

reacting, the acid with a mixture of calcium and soldium hydroxides. It acts as
sequestrants, its binding action removing traces of metal irons present in raw
materials or process water. These metals, for example, iron, can destabilize a
beverage by a tendency of catalyse degradation of flavouring by components,
causing oxidation and off-note. Their removal serves to maintain stability of the
products during storage and to increase shelf life. (Agricultural Research Service,
2000).

2.7.4 Carbohydrates

Carbohydrate still feature as important components of many non-carbonated

beverages e.g funman, chivita and they are particulary important in the
manufacture of dilutable drinks. Some carbohydrates are mentioned below.

2.7.4.1 Sucrose

The preferred carbohydrate for most manufactures is still sucrose. Sucrose is

ready available as a bulk dry solid or as a 67oBrix syrup and it is in this latter form
that most manufacturers will use it. (Ashurst, 1999).

32
2.7.4.2 Invert Sugar

Invert sugar, sometimes referred to as partially refiner’s syrup, is produced by

acid or enzymic hydrolysis of the disaccharide sucrose into its 2004 component
parts of fructose and dextrose (glucose). Invert syrups usually contain a mixture of
sucrose, fructose and dextrose. (Plant food for Human Nutrition, Vol 46, no: 2)

The main advantage of such a syrup is to reduced likehood of crystallization

and an increase in osmolality, which may be useful in reducing spoilage risk, sugar
has numerous functions which makes it important ingredient in fruit juices, apart
from sweetness, textures preservation, fermentation, appearance, freezing point
and antioxidant activity. (Remeo, 1998).

2.7.4.3 Glucose Syrups

Glucose syrups are a group of industrial syrups manufactured from starch-

usually corn starch (Maize). The starch may be hydrolysed by either acid or enzymic
hydrolysis, or more. Usually, a combination of the two (Arthey and Ashurst, 1996).

2.7.5 Vitamin c

Vitamin C is an important antioxidant and helps to protect against cancer,

heart disease and stress. It is part of the cellular chemistry that provides energy,
essential for sperm production, for making collagen, involved in the building and
health of cartilage joint, skin and blood vessels. And it is assimilated with the use of

33
 mineral eg. Calcium, phosphorous, potassium, sodium etc. it is vital for overall mental
and physical well being (Walker, 1974).

2.7.6 Intense Sweeterners

-Saccharin: intense sweeteners have been widely used for many years. The
oldest, saccharin, was used as a sugar substitute during the Second World War
and for many years was used together with sucrose as a mainstay of beverage
sweetener. Despite commercial advantages, saccharin is little used now because
of its bitter after taste. (Shacklady, 1969).

-Aspartame: this is a widely used intense sweetener that has excellent taste
characteristics. It is a peptide made from two amino acids, phenylalamine and
aspartic acid, but will in an acidic beverage medium slowly hydrolyse to its
components which brings about loss of sweetness. (Sizer and Balasubramanian,
1999).

-Sucralose: this is a mostly recently permitted artificial sweetener. It is a

chemically modified sugar but has a very high sweetness factor, comparable with
that of saccharin, but without the unpleasant after taste. It’s sweetness profile is
claimed to be excellent. (Ross and perlmutter, 1985).

34
 CHAPTER THREE

3.0 MATERIAL AND METHODS

3.1 Materials

Fruit drinks like five alive, chivita premium, funman and chi exotic were
bought from modern market, Makurdi.

3.2 Physio-Chemical Analysis

The physio-chemical parameters analysed include: pH, percentage titratable

acidity, vitamin C, viscosity, specific gravity and degree of brix of the juice.

3.2.1 pH Determination

The pH of a fruit bas drinks was determined using the pH meter (AOAC, 2000.)
Were I immersed the glass electrode previously in water for several hours. Then I
rinse well with water, and blot the water gently with a piece of filter paper. When
the pH meter is adjusted at one pH value, I rotated the temperature compensation
dial to set to the temperature of the pH standard solution, I immersed the detecting
unit of the pH standard solution which have a pH near that of the sample solution,
and after more than 2 minutes, adjust the zero point adjustment dial so the reading
of the pH meter is set to the pH of the pH standard solution then I inserted the rod
to each of my sample and took measurement of each sample, component which
bring about loss of sweetness. (Sizer And Balasubramanian, 1999).

35
 -Sucralose: This is the mostly recently permitted artificial sweetener. It is a
chemically modified sugar but has a very high sweetness factor, comparable with
that of saccharin. But without an unpleasant after taste, It’s sweetness profile is
claimed to be excellent. (Ross And Perlmutter, 1985).

3.2.2 Percentage Titratable Acidity

Titratble acidity was determined by titrating with 0.1m NaOH to end point
and expressed as percent citric acid. Ten milliliters of each fruit based drink was
pourd into a conical flask and three drops of bromothymol blue indicator was added
added and titrated with 0.1m sodium hydroxide (NaOH) till colour changes (AOAC,
2000).

Total acidic content of juice = titre valvue x standard x 100

volume of sample taken

Where standard for juice = 0.07

3.2.3 Determination of Juice Viscosity

The viscosity of each juice was determined using a Brookfield viscometer (model LV-
3) with spindle number 2 and at speed 60rpm(AOAC,2000). The viscometer was put
on and the spindle dipped into the various fruit juice and allow for a while before
taking the readings.

3.2.4 Determination of Specific Gravity

This was done using the specific gravity bottle as described by AOAC (2005)
The bottle was washed, clean and dried in an oven, the cooled inside a desicator. It

36
 was then weighed (w1) using a weighing balance and filled with the fruit drinks, (w2),
and recorded the specific gravity bottle was then filled with distilled water and
weighed (w3). Specific gravity was ten calculated using the equation.

SG= weight of sample

weight of equal of volume of water
=w2 -w1

w3 - w1

3.2.5 Determination of Degree brix (0brix) (total sugar)

The degree brix was measured using a refractometer at 200c weight (Brix)
(Pearson, 1991).

3.2.6 Determination of Vitamin C

The vitamin C content was estimated by the indophenols method. This

procedure involve 50:50ml of water and sample into their respective beakers
(100ml), then measure out 2 grams of oxalic acid into a volumetric flash and
measure 200ml of water into and mix. Take out 25ml from the volumetric flask and
pour into each sample in beaker and mix thoroughly, then measure 10ml of the
prepared sample into a conical flask and add 1.5ml of acetone acid into it, mix
properly and then titrate using indophenols immediately there is colour change,
stop and take readings.

Vit. C(mg/10m) is calculated as = 20(V)(C)

where C=concentration of standard=0.1
V=volume of titrate

37
3.3 Microbiological Analysis

The fruit drinks were analyzed microbiologically in terms of total viable count
(TVC) and fungal count according to Harrigan and McCance (1976).

Nine milliliter of distilled water was transferred into 24 test tubes and
covered with cotton wool, for 6 samples, 4 for each sample and sterilized in an
autoclave. Appropriate culture media nutrient agars were use for total viable count
and SDA for fungal count were used to culture micro-organisms in the fruit based
drinks.

3.4 Statistical Analyses

The data obtained was subjected to analysis of variance (ANOVA).

Where the significant differences exist; Tukey’s test was use in separating the means

as described by IWE, 2002

38
 CHAPTER FOUR
4.0 RESULT AND DISCUSSION
4.1 Physico-Chemical Analysis

Results of the physico-chemical analysis of some fruit base drinks were not
significantly different from each other. The pH was all low. The low pH of fruit juice
greatly limits the number and the types of bacteria that can survive or grow at this
low pH of fruit juice are higher than that of fruit flavoured drinks (lacasera).

4.1.2 Degree Brix

The sugar level in lacasera is the highest among the other, it contain
0
15.07 Brix because it is a fruit drink with sugar level very high which is the source of
sweetness and gives balance to flavour.

While that of salwe is the lowest, it contains 8.07. This is because it already
contains five (5) different kind of natural fruit which have natural sweetener so no
need to add much sugar for sweetness or other reasons.

4.1.3 Vitamin

The vitamin content of lacasera (sample B=1.74) is very low and this is
because it does not contain fruits juice but it is a fruit drink. And the chivita (sample
E=12.26) have the highest amount of vitamin because it is a fruit juice varieties of
fruit content in the juice. Followed by chi-exotic and funman (5.74 and 5.66)
respectively.

39
4.1.4 Viscosity

The viscosity of happy hour drink (sample C=113.3) is the highest which
indicate that it is more viscous than the other drinks and this could be as a result of
an ingredient called stabilizer in it which increases viscosity of fruit drink. While that
of the other juice were not significantly different.

40
Table 4.1: Physico-chemical properties of some fruit based products
samples
parameter A B C D E F LSD
Vitamin C 5.7340±03b 1.73±0.23e 3.97±0.06c 2.93±0.23d 12.3±0.08a 5.66±08b 0.6937
(Mg/10ml)
Specific 1.05±0.27a 1.05±0.02a 1.05±0.003a 1.05±0.003a 1.06±0.006a 1.04±0.001a 5-232
gravity
Acidity 0.08±0.007a 0.15±0.0083a 0.04±0.004a 0.19±0.002a 0.17±0.006a 0.16±0.02a 5.232
PH 5.4±0.084b 5.04±0.21c 5.4±0.23b 5.53±0.02a 5.53±0.03a 5.55±0.04a 0.239
Viscosity 46.8±6.88b 40±3.5b 111.7±1.76a 40±3.5b 36.7±3.25b 36.7±2.35b 7.029
Degree 11.49±0.03c 15.1±0.12a 10.5±0.02d 9.50±0.06e 12.1±0.12b 10±0d 0.133
of Brix
Parameters are mean ± standard Deviation (SD) of triplicate determination. Means with
different superscripts within the row are significantly different (p<0.05). LSD: least
significant difference.

Chi exotic
Lacasera
Happy hour
5alive
Chivita
Fumman

41
4.4 Microbiological Analysis
we know that fruit juice may not be totally free from contamination,
contamination of fruit juice by the bacteria may occur when the organism enter the
processing plant or on the surface of the fruit having originate from soil, untreated
surface water, dust and decomposing fruits, the degree of contamination varies
depending upon how the fruit was handle in the field and in processing plant. But
preservation measures are put in place so as to inhibit, retard, or arrest live growth of
micro organisms.
In the analysis sample E=chivita have the highest fungal count (8.8×105)this
could be because of high content of fruit in the juice, while sample B=Lacasera has the
least count(2.4×105) this is because it does not contain fruits, it is a fruit flavoured
drink. Also in the bacterial count chivita have the highest count while lacasera have
the least count.
Result of microbiological analysis of the fruit based drinks are presented
in table 4.2 and expressed in CFU/ml.

42
Table4.2: Total viable count (CFU/ml) of some fruit based drinks
parameters A B C D E F

MICROBIAL 1
fungal
count 3.1×105 2.4×105 3.5×105 4.7×105 8.8×105 4.1×105
(CFU/ml)

MICROBIAL 2
bacterial
count 3.0×105 2.4×105 4.8×105 3.5×105 2.3×106 3.0×105
(CFU/ml)

KEY:
A =Chi exotic
B= Lacesera
C=Happy hour
D=5alive
E=Chivita
F=Funman

43
4.3 Comparison of Result with Standard from NAFDAC

I compared my result with analysis carried out by the department of

biochemistry, college of medicine, university of Lagos which used NAFDAC standard
as their guide and I confirm that there was no significant differences in all the analysis
carried out (e.g. PH; sugar content, vitamin c, total viable count etc. And they were
approved by the appropriate regulatory agency; which is the national agency for food
and drug law administration and control (NAFDAC). Here are the result obtain by the
university of lagos.

Table 4.3 show the result of the physico-chemical analysis carried out by
university of Lagos using NAFDAC standard:

44
 Table 4.3: Physico-Chemical Property Of Some Fruit Juice By University Of
Lagos:

Nutritive Chivita Funman Five Alive

Composition
% TTA 0.090±0.002 0.12±.006 0.10±0.04
pH 6.00±0.04 5.90±0.05 6.02±0.03
% Sugar(OBrix) 12.75±1.61 10.12±1.46 12.56±0.76
Vitamin C 11.06±1.84 9.74±1.52 12.500.62
(Mg/10ml)

45
 CHAPTER Five
5.0 CONCLUSION AND RECOMMENDATION

5.1 Conclusion

The physico-chemical properties of the various fruits base products were not
significantly different from one another and most parameter fall within permissible
limits of regulatory bodies.

Also with vitamin C content of fruit drinks is more than in fruit flavoured drinks.

5.2 Recommendation

1. It is necessary to store fruit based product under recommendation storage

condition to prevent loss in quality.

2. it can be categorically stated that, within the limit of experiment error, the
above brand of fruit based drinks sold within makurdi metropolis are pure and free
from microbial contamination and, therefore, fit for human and animal
consumptions.

46
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 APENDIX A

Nigerian industrial standard

According to the Organization of Nigeria, data were collected from local

manufacturers and research institutes in drafting the standard. The codex
Alimentarius Commission standards for fruit juices were also referred to:

 ESSENTIAL REQUIEMENTS AND QUALITY FRUIT

The fruit used for the juice manufactured shall be sound, fresh suitably mature and
free from infestation, pesticides and their residues if present, shall be in accordance
with the NATIONAL AGENCY FOR FOOD ADMINITRATION AND CONTROL (NAFDAC)
REGULATIONS

 SOLUBLE SOLIDS:

The soluble fruit solids content of juices (exclusive and added sugars) shall not be
less than X%m/m (3.2.1) as determined by refractometer at 20oC, uncorrected for
acidity and read as Brix on the Intertaional sucrose scales.

 DETERMINATION OF SOLUBLE SOLIDS (SUGER CONTENT) BY REFRACTOMETER

Ref: 22.024 AOC (1980)

Method: Adjust instrument to read 0% sucrose with water at 20oC

Determine refractomemter reading of solution at 20oC and obtain correction by

reference to table expressed as % sucrose by weight (Brix).

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MINIMUM SOLUBLE SOLIDS OF FRUIT FUICE X% M/M

Orange Lemon Apple Pine-apple Mango

Juice Juice Juice juice Juice

X% m/m 10 6 10 10 3

 SUGARS:

One or more of the following sugars may be added. Sucrose, invert sugar, dextrose.
The total quantity of added sugars shall not exceed ‘x’ g/kg (3.4.1) expressed as
sucrose.

MAXIMUM SUGAR CONTENT OF FRUIT JUICES

Orange Lemon Apple Pine-Apple Mango

Juice Juice Juice juice Juice

X% m 50 - - 40 100

 MICROBIOLOGICAL SPOILAGE

There shall be no microbiological spoilage of the product when incubated

The incubated container shall be suspected of having undergone microbiological

spoilage if

a) There is evidence of gas built-up in the container

b) The container leaks or
c) There is evidence of bacterial or mould proliferation as judged by significant
change in Ph value or by disintegration, decomposition or dicolouration of the
product when compared with un-incubated samples

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Evidence of bacterial proliferation shall be supported by direct microscopic or
cultural evidence or both.

In addition to the above, the mould mycelium count of the un-incubated product
shall not exceed 40% of the microscopic field as determined by howard mould count
method.

 ADDITIONAL REQUIREMENT

Where the fruit juice require to be kept under conditions of refrigeration, there
should be information for keeping and if necessary thawing of the product.

 MICROBIOLOGICAL EXAMINATION
a. Incubation at 37oc – Incubate two-thirds of the sample containers for 14 days
at 37oc. Examine all these containers for microbiological examination using
appropriate media.
b. Incubation at 55oc. incubate one-third of the sample containers for 10 days at
55oc examine all these containers for microbiological examination using
appropriate media. If the total colonies plate count is greater than 10 colonies
the product shall be regarded as unsafe.

 DETERMINATION OF VITAMIN C

Titration methods (Standard Indophenol Solution) Dissolve 0.05g 2,6 –

dicholorophenolindophenol in water. Dilute to 100ml and filter. To standardize
dissolve 0.05g pure ascorbic acid (Vitamin C) in 600ml of 20% methaphosphoric acid
and dilute with water to exactly 250ml. pipette 10ml o this solution into a small
flask, and titrate with the indophenols solution until a faint pink colour persists for
15 seconds. Express the concentration as mg Ascorbic acid equivalent to 1ml of the
dye solution.

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Procedure

Pipette 50ml of unconcentrated juice into a 100ml volumetric flask, add 25ml of
20% metaphosphoric acid as a stabilizing agent and make up to the mark with
water. Pipette 10ml of the solution into small flash, add 2.5ml of acetone and titrate
with the indophenols solution until a faint pink colour persists for 15 seconds.
Calculate the vitamin C content in the sample as mg per 100ml (or 100g).

 DETERMINATION OF TITRATABLE ACIDS

Reference: AOAC (1980)

Procedure: Dilute 10g of the juice to about 250ml, with neutralized or recently
boiled water and titrate to just before and point with 0.1N alkali (NaOH) using 0.3ml
phenonlphthalein for each 100ml solution being titrated. Transfer measured volume
(2 60 3ml)- of solution into about 20ml neutralized water in a small beaker in this
extra dilution the colour of the fruit juice becomes so pale that phenolphthalein
colour is easily seen. Titrate with the NaOH solution until a pink color is obtained
which persists for 30 seconds. Repeat the titration to get average result. Express
result as ml NaOH/100ml of the sample.

55