A TECHNICAL REPORT ON THE SIX MONTHS INDUSTRIAL TRAINING PROGRAMME

UNDER THE STUDENTS’ INDUSTRIAL WORK EXPERIENCE SCHEME (SIWES)

DONE AT

CENTRAL RESEARCH LABORATORIES, UNIVERSITY OF ILORIN.

BY

AROME SPLENDOUR OCHOLEYE

19/10AQ062

PERIOD OF ATTACHMENT: APRIL 2024 TO OCTOBER 2024

SUBMITTED TO

DEPARTMENT OF HOME ECONOMICS AND FOOD SCIENCE

FACULTY OF AGRICULTURE

UNIVERSITY OF ILORIN, ILORIN KWARA STATE.

IN PARTIAL FULFILLMENT OF THE REQUIREMENT FOR THE AWARD OF BACHELOR

OF SCIENCE (B.Sc) DEGREE IN FOOD SCIENCE

OCTOBER 2024.

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 CERTIFICATION

This is to certify that this report is a detailed account of the Student’s Industrial Work Experience

Scheme (SIWES) program, undertaken by AROME SPLENDOUR OCHOLEYE with

matriculation number 19/10AQ062 at Central Research Laboratories [CRL], University of Ilorin,

Ilorin, Kwara State for a duration of twenty-six {26} weeks and has been prepared in partial

fulfillment for the award of B.Sc. in Food Science of the department of Home Economics and Food

Science, Faculty of Agricultural Science, University of Ilorin, Ilorin, Kwara State, Nigeria.

AROME SPLENDOUR OCHOLEYE DATE

DR A. ARISE (SIWES Coordinator) DATE

DR FUNMILAYO ABIODUN

(Head OF Department) DATE

DR A. O. DAUDA

(Level Adviser) DATE

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 ACKNOWLEDGEMENT

Praise to the Most High God for the precious gift of life and his grace that saw me through the

completion of this SIWES program and also for making it a success.

My profound gratitude extends to my parents, Mr Alex and Mrs Godsake Arome, for their financial

assistance, words of encouragement and unlimited prayers throughout this training program. May

God continue to bless you both.

I appreciate all the staff at CRL for their instructions and valuable teachings, most especially Dr.

Ore of the Plant Tissue Culture Laboratory, Mr. Emmanuel of the Microbiology Laboratory, Mr.

Musa of the Material Testing Laboratory, Engr. Sanusi of the Metal Analyses Laboratory and Dr.

Fatima and Dr. Mrs. Abdulkareem of the Proximate Analyses Laboratory. I also thank the Director,

Prof I. A. Katibi for the opportunity to work there.

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 REPORT OVERVIEW

This report entails the history and importance of the Student Industrial Work Experience Scheme

(SIWES). More so, it highlights the knowledge acquired throughout a period of six (6) months

Industrial Training undergone at the CENTRAL RESEARCH LABORATORIES,

UNIVERSITY OF ILORIN, ILORIN, KWARA STATE.

The background of the organization including its organogram, the importance of several

departments in this establishment and the various activities carried out are detailed in this report. It

also entails the problems encountered during the course of the work experience with the

establishment and recommendations for the improvement of the scheme.

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 TABLE OF CONTENT

Title page 1

Certification 2

Acknowledgement 3

Report overview 4

Table of content 5

CHAPTER ONE: INTRODUCTION

1.2 Background

1.2 Objectives

CHAPTER TWO: DESCRIPTION OF THE ESTABLISHMENT OF ATTACHMENT

2.1 Location and brief history of establishment

2.2 Objectives of establishment

2.3 Organization structure (including organogram)

2.4 The various departments/units in the establishment and their functions

CHAPTER THREE: ACTIVITIES CARRIED OUT DURING ATTACHMENT 1

3.1 Introduction to Plant Tissue Culture

3.2 Microbiology Laboratory

3.3 Introduction to Material Testing Laboratory

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CHAPTER FOUR: ACTIVITIES CARRIED OUT DURING ATTACHMENT 2

4.1 Metal Analysis Laboratory

4.2 Elemental Analysis Laboratory

4.3 Proximate Analysis Laboratory

CHAPTER FIVE: SUMMARY, CONCLUSION AND RECOMMENDATIONS

5.1 Summary of attachment activities

5.2 Problems encountered during my SIWES program

5.3 Recommendation for improvement of the scheme

5.4 Conclusion

LIST OF FIGURES (Fig)

Figure 1: Organogram of CRL

Figure 2: A picture of CRL Unilorin

Figure 3: Picture of a prepared media

Figure 4: pH meter

Figure 5: Banana specie (Bapune)

Figure 6: Laminar flow cabinet

Figure 7: Incubator

Figure 8: Agar containers

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Figure 9: Trinocular Microscope

Figure 10: UV Double-beam Spectrophotometer

Figure 11: Hollow Cathode Lamp

Figure 12: Atomic Absorption Spectrophotometer

Figure 13: CHNS/O Analyzer

Figure 14: Kjeldahl Digestion Unit

Figure 15: Distillation Unit

Figure 16: Moisture Analyzer

Figure 17: Soxhlet Apparatus

Figure 18: Hot Air Oven

Figure 19: Muffle Furnace

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 CHAPTER ONE

INTRODUCTION

Student Industrial Work Experience Scheme (SIWES) was established by ITF in 1973 to solve the

problem of lack of adequate practical skills preparatory for employment in industries by Nigerian

graduates of tertiary institutions. The Scheme exposes students to industry based skills necessary for

a smooth transition from the classroom to the world of work. It affords students of tertiary

institutions the opportunity of being familiarized and exposed to the needed experience in handling

machinery and equipment which are usually not available in the educational institutions.

The unit started in University of Ilorin in the Faculty of Engineering and Technology in

1979 as an Industrial Coordination Unit catering for engineering students on industrial attachment.

The foundation staff comprised an Industrial Coordinator, a Secretary/Typist and an Office

Assistant until 1993 when an Assistant Industrial Coordinator was engaged. As an accreditation

requirement by the National Universities Commission (NUC), the unit was formally upgraded to a

Directorate in the Office of the Vice-Chancellor in August, 1999. Over the years, the clientele

student population has increased from 49 engineering students in 1979 to over 2400 students from

26 disciplines spread across seven faculties of the university. The staff strength has also increased.

AIM OF SIWES

• To integrate students from the theoretical aspects of their courses of study to the

practical and industrial approach.

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1.2 OBJECTIVES OF SIWES

Among the various important objectives of SIWES, the following are the few of them

I) To compliment classroom teaching and expose students to equipment and

machines which are not available in their universities.

II) It enlightens the minds of students to work experience and prepares them for

what is expected after graduation.

III) It creates an avenue whereby students can improve on their individual

character and interpersonal relationship.

IV) To provide an avenue for students in the Nigerian universities to acquire industrial skills

and experience during their course of study.

V) To provide students with an opportunity to apply their theoretical knowledge in real

work situation thereby bridging the gap between theory and practice.

VI) To allow the transition phase from school to the world of working environment easier

and facilitate students’ contact for later job placement.

VII) Enlist and strengthen employers’ involvement in the entire educational process and

prepare students for employment in Industry and Commerce.

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 CHAPTER TWO

DESCRIPTION OF THE ESTABLISHMENT OF ATTACHMENT

2.1 Location and brief history of establishment.

LOCATION

Central Research Laboratories is situated within the main campus of University of Ilorin, along

the CBT road, Opposite the General Studies Division building.

BRIEF HISTORY OF THE CENTRAL RESEARCH LABORATORIES {CRL},

UNIVERSITY OF ILORIN.

The Central Research Laboratories was established in August, 2008. It presently occupies a

dedicated building that was commissioned in October, 2014. The aim of establishing of its

establishment is to provide functional, well maintained, cost intensive, state of the art equipment

for researchers in science-based disciplines.

It is to be noted that the CRL building houses the following units/ centers: African Herbal

Research center, Plant Tissue Culture Laboratory, Cancer Research Laboratory headed by Prof.

(Mrs.) Olorundare, HLA/HBV Research Laboratory headed by Dr. Adeyemi, a Biotechnology

unit headed by Dr. Ibrahim. Other facilities include an ultramodern animal house, a reading

room/Library, conference room for up to 30 persons (with facilities for tea and coffee break).

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The former directors of the establishment include; Prof. M. A. Akanji (2008-2011), Prof. O.

Atteh (2011-2014), Prof. D. S. Ogunniyi (2014-2016), Prof. (Mrs.) A. T. Oladiji (2016-2018),

Prof. M. T. Yakubu (2018-2021), Prof. Nasiru Abdus Salam (2021-2024)

Under the current leadership of Prof. I. A. Katibi of the University of Ilorin's MB:BS

Department, the establishment employs about 20 staffs, including administrative officers and

technologists.

The Vision Statement:

To be a world class facility supporting high quality research for national development.

The Mission Statement:

To provide research facilities that will support cutting edge research and innovation thereby

contributing to making University of Ilorin a world class university.

The Motto:

Supporting research to advance humanity.

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Fig. 1 Organogram of Central Research Laboratories.

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 Fig. 2 Central Research Laboratories, Unilorin.

Divisions of Central Research Laboratories

● General Laboratories: In this laboratory, a variety of analysis are done, including

lipid extraction, digestion, centrifugation, ashing, refrigeration, oven drying, titration, and

distillation.

● Material Testing Laboratory: focuses on the several methods used to determine and

quantify phytochemicals present in various samples.

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 ● Metal Analyses Laboratory: In this laboratory, the Atomic Absorption

Spectrophotometer (AAS) is used to analyze metals and quantify them in samples.

● Chemistry & Biochemistry Laboratory: Deals with the exploration of various

chemical processes within and majorly related to living organisms.

● Microbiology Laboratory: This laboratory uses ELISA {Enzyme Linked Immuno-

Sorbent Assay} to detect antibodies, antigens, proteins and hormones inn bodily fluid samples.

Other techniques include Microscopy, Inoculation and Culture of a variety of microorganisms.

● Elemental Analyzer Laboratory: Elemental analysis is carried out using the CHNSO

Analyzer to determine and measure various constituents, namely, Carbon, Hydrogen, Nitrogen,

Sulfur and Oxygen are done in this laboratory.

● Biotechnology / Molecular Biology Laboratory: Deals with the study of cellular

molecules e.g. nucleic acids, proteins, their biological processes, functions and maintenance.

Specialized Laboratories

● Plant Tissue Culture Laboratory: Deals with the culturing of different plant tissues

outside their natural habitat and in an aseptic environment, used to propagate plants of the same

species.

● Cancer Research Laboratory: Deals with the study of cancerous cells and

development of anticancer agents and treatments.

● HLA/HBV Laboratory: Deals with the study of Hepatitis B virus and how to combat

it.

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 CHAPTER THREE

WORKDONE DURING MY ATTACHMENT (1)

At the Central Research Laboratories I was rotated across the following departments/

laboratories listed below:

 Proximate analysis laboratory

 Microbiology laboratory

 Metal analysis laboratory

 Plant tissue culture laboratory

 Elemental analyzer laboratory

 Material testing laboratory

3.1 INTRODUCTION TO PLANT TISSUE CULTURE

Plant tissue culture is a technique used to grow and propagate plants outside their natural habitat,

in a controlled and aseptic environment. It involves taking small plant tissue samples and placing

them in a nutrient rich medium to stimulate growth and development.

This method allows for the rapid production of genetically identical plants, disease free plant

propagation and preservation of rare or endangered plant species.

The four core stages include:

 Initiation stage:

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 The plant material is collected and sterilized before being placed in the culture medium to

initiate growth.

 Multiplication stage:

The plant cells or tissues undergo rapid cell division to produce a large number of

identical cells.

 Elongation stage:

Here, the cells continue to grow and differentiate into specific plant structures such as

roots and shoots.

 Rooting/ Regeneration stage:

The plantlets are transferred to a rooting medium to encourage root development and

acclimatization to soil conditions before being transferred to the field.

MEDIA COMPONENTS

 Essential Macro-nutrients: Nitrogen, Phosphorus, Potassium, Calcium, Magnesium and

Sulfur.

 Essential Micro-nutrients: Iron, Manganese, Zinc, Boron, Copper and Molybdenum.

 Vitamins: Thiamin, Nicotinic acid, Pyridoxine, Meso-inositol: though it is a

carbohydrate and not a vitamin, it has been shown to stimulate growth in certain cultures.

 Carbon and Energy source: Sucrose

 Stock solution: The use of stock solutions reduces the number of repetitive operations

involved in media preparation and, hence, the chance of human or experimental error.

Moreover direct weighing of media components (e.g., micronutrients and hormones) that are

required only in milligram or microgram quantities in the final formulation cannot be

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performed with sufficient accuracy for tissue culture work. For these components,

preparation of concentrated stock solutions and subsequent dilution into the final media is

standard procedure.

In addition, concentrated solutions of some materials are more stable and can be stored for

longer periods than more dilute solutions.

 Growth Regulators: i) Auxins: 1H-indole- 3–acetic acid (IAA), 1H–indole–3 -butyric

acid (IBA).

ii) Cytokinin: 6-benzylaminopurine (BAP) or 6-benzyadenine (BA)

iii) Gibberellin (GA3)

iv) Abscisic acid (ABA)

 Solidifying Agent: Agar: it is the most commonly used gelling agent for preparing

semi-solid and solid plant tissue media.

 Activated charcoal: It absorbs inhibitory compounds, growth regulators and it also

darkens the medium. Activated charcoal aids with plant rooting and is added to the

rooting media to stimulate cell growth

TYPES OF MEDIA

 Murashige and Skoog (MS) Medium (1962): It is one of the most widely used nutrient

media for plant tissue culture. It is suitable for a broad range of plant species including

rice, corn, tobacco and various ornamental plants.

 Gamborg’s B5 Medium (1968): It is suitable for soya-beans, carrot, sunflower, e.t.c

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 Woody Plant Medium (1981): It is a standard medium for culturing various woody

plant species. Examples include; Oak trees, pine trees, willow trees, birch trees and so on.

STERILIZATION TECHNIQUES

Sterilization is a critical step in plant tissue culture to prevent contamination by bacteria,

fungi and other microorganisms. There are several common methods used to sterilize

plant materials and culture media.

1. Autoclaving: It is the most commonly used method for sterilization in plant tissue

culture laboratory. It involves subjecting the materials or media to high pressure and

temperature inside an autoclave. The typical conditions are 1210C at 15psi for 15-20

minutes. This process effectively kills spores and vegetative cells of most

microorganisms.

2. Chemical Sterilization: Some plant materials or equipment that cannot withstand

autoclaving can be sterilized using chemical agents. Commonly used chemicals include

ethanol, hydrogen peroxide, sodium hypochlorite (bleach) and mercuric chloride.

3. Use of Glass Bead Sterilizers: They are specialized devices used for rapid sterilization

of laboratory instruments such as scalpels and forceps. They utilize heated glass beads to

achieve high temperatures, typically around 300 Degree Celsius.

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 PLANT TISSUE CULTURE TECHNIQUES

Plant tissue culture is a collection of techniques used to maintain or grow plant cells,

tissues, or organs under sterile conditions on a nutrient culture medium. This method leverages

the totipotency of plant cells, allowing them to regenerate into whole plants from various parts.

Below are the primary techniques utilized in plant tissue culture:

1. Seed Culture

This technique involves using seeds directly to grow plants. It is particularly useful for seeds

with hard outer coatings that take longer to germinate in field conditions. The resulting plants are

uniform and grow faster than those cultivated in traditional methods.

2. Meristem Culture

Meristematic tissues, found at the tips of roots and shoots, contain undifferentiated cells capable

of developing into any plant organ. This method exploits these cells to produce disease-free

plants since meristems are typically free from viruses and microorganisms.

3. Embryo Culture

In this technique, either mature or immature embryos extracted from seeds are cultured on a

suitable medium to develop into viable plants. It is especially beneficial for rescuing embryos

that may not survive under normal conditions, such as old seeds with low germination rates.

4. Callus Culture

Callus culture involves creating an unorganized mass of cells (callus) from explants taken from

any part of the plant. This method is versatile and can be applied across different species, leading

to further differentiation into shoots or roots.

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 Apparatus used:

1. pH Meter

2. Electronic Weighing Balance

3. Refrigerator

4. Autoclave

5. Laminar Air Flow Cabinet

6. Glass beads sterilizer (steripot)

7. Forceps

8. Scalpel

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9. Glass stirrer

10. Measuring cylinder

11. Beaker

12. Pipette

13. Cooker

14. Pot

15. Spatula

16. Phyta jars

INITIATION MEDIA PREPARATION: MS MEDIA – 1 Litre

 Measure 1000ml of distilled water, pour in a pot

 Weigh 1.65g of Ammonia nitrate (NH4NO3)

 1.90g of Potassium Nitrate (KNO3)

 0.44g of Calcium Chloride (CaCl2)

 30g of Sucrose

 0.37g of Magnesium Sulphate (MgSO4)

 0.17g of Potassium di-hydrogen phosphate (KH2PO4)

 Stock Solutions: Stock solutions 1 to 4 – 2ml, Stock solution 5 – 5ml, Stock

solution 6 – 10ml

 0.1g of Meso-inositol

 4mg of BAP dissolved in one drop of Potassium hydroxide (KOH)

 40mg of Adenine Sulphate (AS) dissolved in one drop of Hydrogen chloride

(HCl)

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 1.5mg of IAA dissolved in one drop of KOH

 0.1g of Citric acid

 5.5g of Agar

 The pH of the mixture should be 5.8

 Measure with the pH meter and add drops of KOH to neutralize it

 Sieve with a funnel

 Place on the cooker and remove when it boils slightly

 Pour into the phyta jars and place in the laminar flow cabinet.

Fig. 3: Picture of a prepared media

Fig. 4: pH Meter

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Fig.5:Banana specie (Bapune) in various growth stages.

Fig. 6: Laminar Flow Cabinet.

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 3.2 MICROBIOLOGY LABORATORY

Microbiology is the branch of science that deals with the study of microorganisms, which are

microscopic organisms that can only be seen with a microscope.

In the microbiology laboratory I learned about Microscopy, Antibiotic Minimum Inhibitory

Concentration (MIC) Test, Aseptic Procedures, Isolation and Culturing Methods, Culture Media

Preparation, Stock Preparation, Serial Dilution, Gram Staining Methods and Biochemical

Testing.

ASEPTIC TECHNIQUES

To stop undesired microorganisms like bacteria, fungus, and other pollutants from entering a

working environment, aseptic technique is a sterile operation that is carried out. This method

prevents lab employees, cultures, and equipment from becoming contaminated. The danger of

contamination can be greatly decreased or minimized by using the appropriate aseptic approach.

Sanitization, disinfection, and sterilization are all part of the process.

Some specific aseptic techniques are;

 Cleaning and disinfection of working bench/surfaces with 70% ethanol prior to use.

 Washing of hands and then disinfecting with 70% ethanol and finally wearing of gloves.

 Switching on the UV lamp in the laminar air flow for fifteen minutes helps to sterilize the

interior and contents before usage to prevent contamination of the experiment.

 Sterilization of culture media and some equipment and apparatus to be used with the

Autoclave.

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  All work must be done close to a Bunsen burner flame where air currents are drawn

upwards, in a laminar flow cabinet.

 Always cap the bottles and flasks after use. Also petri-dishes containing culture media

after inoculation should be sealed with tape to prevent microorganisms and airborne

contaminants from gaining entry.

 Avoid talking, singing, or whistling while performing sterile procedures.

 Perform experiments as rapidly as possible to minimize contamination.

ISOLATION AND CULTURING TECHNIQUES

Culturing

It is the process of replicating microorganisms through controlled laboratory settings in a

predetermined culture media.

There are five basic culturing techniques used in culturing microorganisms.

They include;

 INOCULATION: The process of introducing microorganism (inoculum) into a media to

promote growth.

 INCUBATION: This is the process of cultivating microorganisms in a controlled

environment that allows growth, that is, the incubator.

 ISOLATION: This is a process of separating individual strains of micro-organisms from

a mixed sample.

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 INSPECTION: This is a process of examining the cultures for obvious growth

characteristics using the microscope.

 IDENTIFICATION: This is a process done to classify or establish the specific

organisms that has been isolated using differential staining and biochemical tests to

distinguish one micro-organism from another.

Three procedures are involved in the culturing of microorganisms: Inoculation,

Incubation, and Serial dilution, which is a series of repeated dilutions used to lower

microbial concentration/load in a sample.

Microbes are cultivated in media that is suitable for them and promotes their growth.

Microorganisms are cultured using a variety of growth media and techniques, primarily

the Pour Plate Method, Spread Plate Method, and Streaking Method.

Streaking Method: Involves spreading a diluted microbial sample across an agar plate to

isolate individual colonies.

Pour Plate Method: Involves mixing a diluted sample with molten agar and allowing it

to solidify in a Petri dish, where colonies grow throughout the medium.

Spread Plate Method: A diluted sample is spread over the surface of an agar plate to

allow for colony formation.

Incubation: After inoculation, plates are incubated under specific conditions

(temperature, oxygen levels) that favor the growth of the desired microorganism.

For Bacteria, the appropriate temperature is 37 Degrees Celsius for 18 to 24 hours. For

Fungi, it is 28 Degrees Celsius for 3 to 7 days.

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 Fig.7: An Incubator

PREPARATION OF CULTURE MEDIA

Culture media, sometimes referred to as agar, are particular mixtures of nutrients and

other materials that promote and facilitate the growth of microorganisms. It is a liquid

(broth) or gel-like material that gives organisms the vital nutrients and minerals they need

to flourish in a laboratory setting.

It can be broadly classified into three:

 General agar: This culture medium supports the growth of a wide variety of micro-

organisms. An example is Nutrient Agar.

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 Differential agar: It is used to differentiate between various types of micro-organisms

based on their biochemical properties. Examples are MacConkey Agar and Blood Agar.

 Selective agar: This favors the growth of specific micro-organisms while inhibiting

others. Examples include Eosin Methylene Blue (EMB) Agar and Mannitol Salt Agar.

In CRL, the preparation of culture media in the microbiology laboratory is based on

manufacturer’s specification as written on the container of the media. According to the

Standard Operating Procedure (SOP) and also depending on the analysis carried out, the

media used are MacConkey Agar (MAC), Nutrient Agar (NA), Potato Dextrose Agar

(PDA), De Man, Rogosa and Sharpe (MRS) Agar, Mueller Hinton Agar (MHA),

Sabourand Dextrose Agar (SDA), Eosin Methylene Blue (EMB) Agar, Blood Agar,

Chocolate agar etc.

General steps of preparing culture media;

●The required amount of dehydrated medium is suspended in a certain amount of

distilled water in a media jar.

●The jar is sealed to prevent contamination.

●It is then sterilized by autoclaving at 15 psi pressure at 121°C for 15 minutes.

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 Fig. 8: Various Dehydrated Agar Containers

SERIAL DILUTION

Serial dilution is a laboratory technique used to systematically reduce the concentration of micro-

organisms in a solution. This method involves a series of dilutions, where each dilution step

reduces the concentration by a consistent factor, typically by factors of ten (10-fold) or two (2-

fold).

The primary purpose of performing serial dilutions is to obtain manageable concentrations for

analysis or experimentation, such as estimating the number of microorganisms in a sample or

determining the minimum inhibitory concentration (MIC) of antimicrobial compounds.

PROCEDURE FOR SERIAL DILUTION

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1. Prepare a known number of dilution blanks in test tubes. Fill them with 9mL of distilled

water. Label them accordingly.

2. Take an empty test tube and add at least 2mL of the undiluted sample solution.

Thoroughly mix.

3. Using a pipette, draw 1mL of the undiluted solution and transfer it to the test tube labeled

1:10. Thoroughly mix the contents to ensure proper dilution

4. For each subsequent dilution, take 1mL from the previously diluted solution and transfer

it the next test tube containing 9mL of distilled water labeled 1:100.

5. Mix thoroughly after each transfer to a consistent dilution factor of 1:10 for each step

until you reach your final dilution.

GRAM STAINING

Gram staining is a technique that helps differentiate between different types of bacteria

based on their cell wall composition. This provides preliminary identification.

Bacterial are stained with crystal violet and then treated with Gram’s solution after being

decolourized with ethanol and treated with safranin and washed in water.

Those that retain the crystal violet are Gram-Positive and those that do not are Gram-

Negative.

Example of gram-negative bacteria is Escherichia coli and for gram positive bacteria is

Staphylococcus aureus.

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 PRINCIPLE OF GRAM STAINING TECHNIQUES

 The structure of the organism’s cell wall determines whether the

organism is gram-positive or gram-negative.

 When stained with a primary stain i.e. crystal violet and fixed by a

mordant, some bacteria are able to retain the primary stain.

 Some bacteria resist decolorization while others are been decolorized.

 The components of gram-positive bacteria cell wall are made up of thick

peptidoglycan layer (90%) and teichoic acid.

 While the cell wall of gram-negative bacteria consist of a thin

peptidoglycan layer (5 to 10%), outer membrane with

lipopolysaccharides.

Reagents and Materials Required in Gram Staining

Crystal Violet, Gram staining Lugol’s Iodine, 95% Ethanol, Safranin, Distilled water, Blotting

paper, Glass slides, Microbiological loops, Inoculating needles, Bunsen burner, Immersion oil.

Preparation of Smear

 Prepare your slide

 Preparation of slide is done by placing bacteria on a slide in a drop of water, allowing

them to dry then heat fixing.

 Heating the slide ensure that the bacteria won’t wash off during staining process.

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  Correct smear preparation should produce a monolayer of organisms sufficiently dense

for easy visualization but thin enough to reveal morphological characteristics. That is,

thick smear should be avoided.

Procedure Involved in Gram Staining Techniques

 Prepare a heat fixed smear from a 18-24-hour old culture.

 Stain with crystal violet solution for 60 seconds and rinse off the solution.

 Iodine solution was added for 30 seconds.

 Washed with distilled water and decolorized with alcohol.

 The smear was finally flooded with safranin to counter stain for 60 seconds.

 Wash with water and blot dry.

 View under microscope at 100X objective lens using immersion oil.

Using the microscope, Gram-positive bacteria cell appears purple, or crystal violet iodine

complex and Gram-negative cells appear red or pink.

MICROSCOPY

Microscopy is a technical field that involves the use of a microscope to view samples and objects

that cannot be seen with the naked eye. The primary purpose of microscopy is to magnify small

structures, allowing for study of their properties, behaviors, and interactions at a much finer

scale.

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I learned how to use a Trinocular Microscope to observe, identify and classify various micro-

organisms like gram positive or negative bacteria, parasites present in a sample of fish pond

water, and various fungi and bacteria cultures.

WET MOUNT STAINING

 Wet mount is a staining technique used in viewing fungi specimen for

identification. Fungal cells have both macroscopic as well as microscopic

structure.

 Wet mount preparation is the most widely used method of staining and observing

fungi and it is simple to prepare.

REAGENTS AND MATERIALS USED IN WET MOUNT STAINING

 Laminar flow

 Methylene blue or Lacto-phenol cotton blue

 Slides

 Cover slip

 Syringe or wire loop

 Specimen {cultured fungi}

PROCEDURE INVOVED IN WET MOUNT STAINING

 Place a drop of 70% ethanol on a clean microscopic glass slide.

 Add one or at most 2 drops of the lactophenol cotton blue (LPCB) or methylene blue

before the alcohol dries out.

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 Immerse the specimen in the drop of LPCB.

 Holding the coverslip between the index finger and thumb, touch one edge of the drop of

mounting with a coverslip edge and lower gently avoiding air bubbles.

 This preparation is now ready for examined.

 Switch to higher power (40X) objective for more detailed examination of spores and

other structures.

Fig. 9: A Trinocular Microscope.

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 BIOCHEMICALS TESTS

These tests assess the metabolic capabilities of microorganisms by examining their ability to

utilize certain substrates or produce specific enzymes.

Examples of biochemical tests include; Motility Test, Endospores Test, Pigmentation Test,

Catalase Test, Oxidase Test, Indole Test, Gelatin Hydrolysis Test, Methyl Red Test, VP Test,

Starch Test, Gas Production Test and so on.

CATALASE TEST:

This was used to differentiate those bacteria that produce enzyme Catalase. A little portion of the

bacterial growth is transferred with a sterilized wire loop to a drop of hydrogen peroxide on a

clean glass slide. The presence of catalase observed by bubbling indicates a positive test result

while absence of bubble indicates negative test result of bacteria.

OXIDASE TEST:

This is used to differentiate bacteria that produce cytochrome C oxidase located in their

membrane which catalyzes the transport of electrons from donor compounds to electron

acceptors (O2).

In the presence of oxygen, the enzyme oxidizes the phenylendiamine reagent (Kovac’s) resulting

in the production of indolephenol (a purple blue compound). A piece of filter paper is soaked in

Kovac’s reagent, and then a well isolated colony is selected to prepare a smear on the filter

paper. The appearance of a purple blue coloration indicates a positive result, while no color

change indicates a negative result.

MOTILITY TEST:

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Motility test is aimed at identifying motile bacteria. Motility can sometimes be referred to as the

way an organism grows on solid media, and it determines the presence or absence of flagella.

Tubes containing the motility medium are inoculated by making a fine stab with a loopful of the

culture to a depth of 1-2cm and is then incubated at 37 degrees Celsius for 48hrs. Motility is

observed by spreading of the organism outwards from the stab area.

INDOLE TEST:

This test is carried out for Indole production by test organism which is important in identifying

entero-bacteria. A little portion of each isolate is inoculated onto 5ml of sterile peptone-water

enriched with 1% tryptophan and incubated at 37 degree Celsius for 24 hours. To the culture,

0.5ml of Kovac’s reagent was added and gently stirred. A red color indicated positive result

while a yellow color indicated negative test result of bacteria.

METHYL RED (MR):

MR test is carried out to identify Entero-bacteria based on the ability to produce and maintain

stable end product from glucose fermentation. A little portion of each isolate is inoculated into

the glucose phosphate peptone water medium and is incubated at 37 degree Celsius for 48 hours.

Few drops of methyl red is added to the culture. MR positive is indicated by red color.

COAGULASE TEST:

A drop of distilled water is placed on two separate slides. A colony of the test organism

(previously checked by Gram staining) is emulsified in each of the drops to make two thick

suspensions. A loopful of plasma is added to one of the suspensions, and mixed gently.

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Clumping of the organisms within 10 seconds was observed which will indicate coagulase

positive. No plasma is added to the second suspension. This is to differentiate any granular

appearance of the organism from true coagulase clumping.

3.3 INTRODUCTION TO MATERIAL TESTING LABORATORY

Phytochemical analysis was majorly carried out in this laboratory. Phytochemicals are naturally

occurring compounds found in plants that contribute to their color, flavor and disease resistance.

They are not essential nutrients like Vitamins and Minerals, but they have beneficial effects on

human health. Phytochemicals have anti-oxidative properties, anti-inflammatory properties and

immune-boosting properties, among other health benefits.

Phytochemicals are also referred to as Secondary Metabolites because they are not directly

involved in the primary metabolic processes required for growth, development and reproduction.

Their role is more about enhancing the plant’s survival and adaptability in its environment than

supporting basic life functions.

Categories of Phytochemicals analyzed at the Central Research Laboratories include quantitative

and qualitative of Saponin, Tannin, Phenol, Flavonoid, Steroid, Tritepenoid, Glycoside and

Alkaloid. Also the presence of Protein and Reducing sugars are determined in the laboratory,

alongside quantitative analysis of Phosphorus and Sulphate in a sample.

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 QUALITATIVE ANALYSIS

This refers to the methods used to identify the presence or absence of specific phytochemicals in

a sample. This type of analysis helps to determine what phytochemicals are present, but it does

not provide information about their concentrations.

Below is a table showing the procedures for the determination of various phytochemicals

SECONDARY PROCEDURE OBSERVATIONS

METABOLITE

1. Saponin FOAM TEST Presence of foam indicates the

Take small amount of the presence of Saponin.

sample, add a bit of water, and

shake thoroughly.

2. Flavonoid ALKALI TEST Yellow coloration indicates

Mix the sample with the presence of Flavonoid.

increasing amount of NaOH.

3. Tannin and Phenol FERRIC CHLORIDE TEST Blue black color indicates the

5% Ferric Chloride is added to presence of Tannin and

the sample. Phenol.

4. Steroid and SALKWOSKI’S TEST Brick red color in the

Tritepenoid Chloroform and Concentrated chloroform layer and Green

H2SO4 is added to the sample. fluorescence in the acid layer

indicates the presence of

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 steroids.

NB: Presence of Steriod

indicates the presence of

Tritepenoid.

5. Glycoside BALJET TEST Yellow or orange to deep red

Mix a small amount of the indicates the presence of

sample in 2ml of Sodium Glycoside.

picrate solution.

6. Alkaloid DRAGENDORFF’S TEST Red precipitate coloration

Add 10% HCl to the sample, indicates the presence of

then boil in the water bath for Alkaloid.

3 to 5 minutes. Filter the

sample and add Dragendorff

reagent to the filtrate.

7. Protein LEAD-ACETATE TEST Brownish black precipitate

Add 40% NaOH and few indicates the presence of

drops of Lead-acetate to the Protein.

sample, and then boil for a

few minutes.

8. Reducing sugar FEHLING TEST The color changes from blue

Add equal amounts of to red, and this indicates the

Fehling’s A & B reagents to presence of Reducing sugar.

the sample and then boil.

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I carried out analysis on Noni (Morinda citrifolia) leaves as an assignment, using water as a

solvent for extraction to test for the presence of some phytochemicals and this was the result:

Phytochemical Result

1. Saponin Present

2. Flavonoid Present

3. Tannin and Phenol Present

4. Steroid and Tritepenoid Absent

5. Alkaloid Present

6. Protein Present

7. Reducing sugar Absent

QUANTITATIVE ANALYSIS

This involves measuring or quantifying the concentration of specific phytochemicals in a sample.

This type of analysis provides numerical data that can be used to compare the amount of

phytochemicals across various samples. Techniques used for quantitative analysis in CRL

include Spectrophotometry, that is, the use of Ultra-Violet (UV) double-beam Spectrophotometer

and the Fourier Transform Infra-Red (FTIR) Spectrophotometer.

Spectrophotometers operate on the Beer-Lambert’s Principle which states that the absorbance in

a sample is directly proportional to both the concentration of molecules present and the path

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length of light through the sample. It describes how light intensity decreases exponentially with

distance through an absorbing medium.

PHYTOCHEMICAL PROCEDURE READING WAVELENGTH

1. Sulphate Measure 10ml of the sample 420nm

in a beaker; add 39ml of

distilled water. Measure 1ml

of Barium chloride (reagent)

and add to the sample.

Leave to stand for 30 minutes.

2. Phenol Prepare 1ml of the sample, 725nm

mix it with 0.25ml of Folin-

C10 Calteu’s reagent. Add

1.25ml of 20% Na2CO3

solution.

Allow to stand for 40 minutes

at room temperature.

3. Flavonoid Prepare 1ml of the sample and 520nm

add 4 drops of concentrated

HCl with 0.5g of magnesium

turnings.

Leave to stand for a few

minutes.

4. Saponin Prepare 1ml of the sample 380nm

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 solution and add 2ml of 5%

FeCl3. Add distilled water to

top it up to 50ml.

Allow to stand for 30 minutes

for a blood red color to

develop.

5. Glycoside Measure 10ml of the sample 510nm

in beaker; add 50ml of

chloroform, shake for an hour.

Filter the mixture and 10ml

pyridine into a 100ml conical

flask, add 2ml of 2% sodium

nitropruside to the filtrate and

shake thoroughly for 10

minutes.

Add 3ml of 20% NaOH to

develop a brownish-yellow

color.

6. Tannin Measure 1ml of sample and 699nm

add 0.5ml of Follin-dennis

reagent, plus 1ml of Na2CO3

and 7.5ml of distilled water.

7. Phosphorus Add 4ml of reagent B to 5ml 699nm

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 of the sample solution. Then

add 16ml of distilled water.

NB: Prepare a standard solution (blank) following the same procedure for each of the

phytochemicals using distilled water as the sample.

Blank solutions are prepared to establish a baseline measurement. The blank typically contains

all the components of the sample solution prepared except the sample itself. This allows for

correction of any absorbance that may be due to the solvent or other components in the solution,

ensuring that the measurements taken for the actual sample reflects only the absorbance due to

the compounds of interest.

PROCEDURE FOR THE USE OF THE UV SPECTROPHOTOMETER.

 Turn on the spectrophotometer and select the appropriate wavelength for the analysis.

 Insert the blank solution into one of the sample holders (the reference beam).

 Close the sample compartment and zero the instrument using the blank. This sets the

baseline for the measurements.

 Insert the sample solution into the other sample holder (the sample beam).

 Close the compartment and record the absorbance reading.

 Use the predetermined calibration curve to determine the concentration of

phytochemicals in the sample based on its absorbance.

 After measurements, clean the cuvettes and turn off the instrument.

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I carried out quantitative analysis on Noni leaves extract. The results are as follows:

PHYTOCHEMICAL AVERAGE ABSORBANCE

1. Phenol 4.3261

2. flavonoid 0.8674

3. Saponin 1.8656

4. Glycoside 2.3397

5. Tannin 4.2333

6. Phosphorus 1.8874

7. Sulphate 2.8810

Fig. 10: UV Double beam Spectrophotometer.

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 CHAPTER FOUR

ACTIVITIES CARRIED OUT DURING ATTACHMENT 2

4.1 METAL ANALYSIS LABORATORY

Metal analysis involves various processes used to identify and quantify the elements present in a

metal sample. It is a critical process that involves the examination and quantification of metal

elements in various samples, which can include environmental, biological and industrial

materials.

This analysis is essential for understanding the composition and concentration of metals, which

can have significant implications for health, safety, and environmental sustainability.

Techniques used for metal analysis include Atomic Absorption Spectroscopy, X-ray

Fluorescence and Flame Photometry. The technique most employed in Central Research

Laboratories is the use of the Atomic Absorption Spectrophotometer.

ATOMIC ABSORPTION SPECTROPHOTOMETER

An atomic absorption spectrophotometer (AAS) is an analytical instrument used to determine the

concentration of specific elements in a sample. The working principle of an AAS involves

several key steps:

 Sample Introduction: The sample, typically in liquid form, is introduced into the AAS
instrument. This can be done through the nebulizer, which converts the liquid sample into
an aerosol mist. The aerosol is then carried into a flame where it will be atomized.
 Atomization: In the flame of the instrument, the aerosol is mixed with a fuel (like
acetylene) and an oxidant (like air or nitrous oxide) and introduced into a flame. The high

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 temperature of the flame causes the solvent to vaporize and atomizes the metal ions,
producing free atoms in the gaseous phase.
 Light Source: A hollow cathode lamp specific to the element being analyzed emits light
at characteristic wavelengths. For example, if analyzing lead (Pb), a hollow cathode lamp
designed for lead will be used. The light source produces monochromatic light that passes
through the atomized sample.
 Absorption of Light: As the light passes through the cloud of free atoms in the atomized
sample, certain wavelengths of light are absorbed by the atoms. Each element has unique
electronic transitions that correspond to specific wavelengths; thus, only light at these
wavelengths will be absorbed.

According to Beer-Lambert Law:

The amount of light absorbed by a medium is directly proportional to the concentration of the
element in the sample provided the path length is constant.

 Detection: After passing through the atomized sample, the remaining light is detected by
a photo-detector (such as a photomultiplier tube). The detector measures the intensity of
light that has not been absorbed by the sample.
 Data Processing: The signal from the photo-detector is processed to determine the
absorbance. This value can then be compared to a calibration curve generated from
standards with known concentrations of the target element. By comparing the absorbance
values, the concentration of the element in the unknown sample can be quantified.
 Output: The results are displayed, often as a concentration value

In summary, an atomic absorption spectrophotometer works by atomizing a sample, passing light

through it, and measuring the amount of light absorbed by the atoms in the sample to determine

the concentration of specific elements.

CALIBRATION CURVE

A calibration curve is constructed by running several samples of known element in question

concentration called standard. The amount of radiation absorbed is then compared with absorbed
standard to give the concentration of the element in question.

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The concentration of the standards is then plotted on a graph which must give a straight line
graph with no y-intercept; this signifies a direct proportionality.

THE LIGHT SOURCE

The common source of light is usually a hollow cathode lamp. This contains a tungsten anode
and a cylindrical hollow cathode made of the element to be determined. These are sealed in a
glass tube with an inert gas like neon or argon.

Fig. 11: Hollow Cathode Lamp.

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Fig. 12: Atomic Absorption Spectrophotometer

4.2 ELEMENTAL ANALYSIS LABORATORY

Elemental analysis is a technique used to determine the composition of organic and inorganic

compounds in a substance. This involves quantifying the individual elements present in a

sample.

The instrument used for elemental analysis in CRL is the CHNS/O Analyzer which measures the

amounts of Carbon (C), Hydrogen (H), Nitrogen (N), Sulfur (S) and Oxygen (O) present in a

sample.

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 OPERATING PRINCIPLE OF THE CHNS/O ANALYZER.

The CHNS/O Analyzer operates on the principle of Combustion for the analysis of Carbon,

Hydrogen, Nitrogen and Sulfur and for Oxygen analysis, it operates on Pyrolysis.

COMBUSTION ANALYSIS

It involves the complete combustion of a sample in an oxygen rich environment. This process

breaks down the organic molecules into their fundamental elements: CO2, H2O, N2 (g) and SO2.

The produced gases are then separated and detected using thermal conductivity.

PYROLYSIS ANALYSIS

Pyrolysis is a thermal decomposition process that occurs in the absence of oxygen, where

organic materials are broken down into simpler molecules through the application of heat.

Pyrolysis plays a crucial role in the analysis of oxygen content. During the pyrolysis process, the

sample is heated to high temperatures, causing it to decompose. The volatile components

released during pyrolysis can be collected and analyzed. In a CHNSO analyzer, the oxygen

present in the sample is typically converted into gaseous products, which can then be quantified.

The operating principles of a CHNSO analyzer involve several key steps to determine the

concentrations of carbon (C), hydrogen (H), nitrogen (N), sulfur (S), and oxygen (O) in a sample.

Here’s a brief overview of the process:

1. Sample Preparation: A small, accurately weighed sample of the material to be analyzed is

prepared, often in a solid form.

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2. Combustion: The sample is placed in a high-temperature furnace where it is combusted in the

presence of excess oxygen. This combustion process converts the elements in the sample into

gaseous products. For example, carbon is converted to carbon dioxide (CO2), hydrogen to water

(H2O), nitrogen to nitrogen oxides (NOx), sulfur to sulfur dioxide (SO2), and any oxygen

present is also converted to CO2 or H2O.

3. Gas Collection: The gases produced from the combustion are collected and passed through a

series of traps and filters to remove any unwanted byproducts or moisture.

4. Detection and Quantification: The resulting gases are then analyzed using various detection

methods, which are Thermal Conductivity Detectors (TCD) and Gas Chromatography (GC)

5. Data Analysis: The amounts of each element are calculated based on the detected

concentrations of the combustion products. The results are then reported, often in terms of

weight percent.

Fig. 13: CHNS/O Analyzer

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 4.3 Proximate Analysis Laboratory

Proximate analysis is a systematic method used to determine the major components of biological

materials, particularly in food and feedstuffs. This analysis provides essential information about

the nutritional value of a sample by quantifying its primary constituents. The proximate analysis

carried out at CRL typically includes the following components:

 Moisture Content: This measures the amount of water present in the sample. It is

crucial because high moisture levels can lead to spoilage and affect the stability of

nutrients.

 Crude Protein: This component estimates the total nitrogen content in the sample

using methods such as Kjeldahl’s method, which involves digesting the sample

with sulfuric acid and measuring nitrogen levels. The crude protein percentage is

calculated by multiplying the nitrogen content by a factor (commonly 6.25), based

on the assumption that proteins contain approximately 16% nitrogen.

 Crude Lipids (Fats): This measures the total fat content in a sample, typically

extracted using solvents like petroleum ether or di-ethyl ether. The lipid content is

important for understanding energy contributions and overall nutritional quality.

 Crude Fiber: This component assesses the indigestible portion of plant materials,

which includes cellulose, hemicellulose, and lignin. It is determined after treating

the sample with dilute acid and alkali solutions.

 Total Ash Content: Ash represents the inorganic mineral content remaining after

combustion of organic material at high temperatures (usually around 550°C). It

provides insight into the mineral composition of the sample.

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  Carbohydrate content or Nitrogen-Free Extract (NFE): NFE is calculated by

subtracting the percentages of moisture, crude protein, crude lipids, crude fiber,

and total ash from 100%. It primarily represents digestible carbohydrates like

sugars and starches.

The results from proximate analysis are often expressed as percentages of each

component relative to the total weight of the sample.

PROCEDURE FOR THE DETERMINATION OF PROTEIN CONTENT

1. DIGESTON OF SAMPLES.

Digesting sample is important in the starting point of protein content determination. It is the

process of decomposing a solid sample into a liquid state by dissolving it using reagents such as

strong acids, alkalis. The sample is first digested in strong sulfuric acid in the presence of a

catalyst, which helps in the conversion of the amino nitrogen to ammonium ions.

PROCEDURE

 Measure 2g of the sample on the weighing balance using filter paper as a container, then

pour into a digestion flask.

 Add 5g of Sodium Sulphate.

 Then add 0.5g of Copper Sulphate.

 Add a pinch of selenium to act as catalyst.

 Add 25ml of conc. Sulfuric acid.

 Heat in a Kjeldahl digestion apparatus until the color changes preferably to a sticky green

(light green) precipitate.

 Add little distilled water to liquify or dissolve the content.

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  Filter and then transfer liquefied mixture into 250ml volumetric flask, top up with

distilled water till it reaches the mark level.

 Pour the digest into a bottle (usually a clean table water bottle) and label appropriately,

including the name of the sample and the date.

Fig. 14: Kjeldahl Digestion Unit.

2. DISTILATION

PROCEDURE

 Measure 5ml of the digest into a digestion flask and add 5ml of 40% NaOH.

 Measure 5ml of boric acid into a conical flask

 Place both in the distillation unit

 The analyte in the digestion flask is separated by steam distillation. At first, it is

evaporated and then it condenses into the receiving flask (Conical flask) through the

receiving outlet. Once the distillation is completed and it reaches 50ml, the residue left in

the sample tube is aspirated and removed.

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 Fig. 15: Distillation Unit.

3. TITRATION

The nitrogen content is then estimated by titration of the ammonium borate formed with standard

(0.01m) hydrochloric acid, using a slight colour change to determine the end-point of the

reaction. Record the Titre value.

Calculations:

%N

Where:

Titre Value = Titre of Sample, 0.01 HCl = Molarity of Acid Used, 0.014 = Nitrogen

Constant per 1000ml, 250 = Volumetric Flask used and 5 = sample.

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 The formula above is used for determining the nitrogen content in the sample. Once the
nitrogen content has been determined it is converted to a protein content using the
appropriate conversion factor: %Protein = conversion factor (6.25) X nitrogen content
previously determined.

MOISTURE CONTENT DETERMINATION

In Central Research Laboratories, moisture analysis is carried out using a Moisture Analyzer
which is a relatively faster way to carry out this analysis compared to the Hot-Air Oven Method.

PROCEDURE

1. Sample Preparation: A representative sample of the material to be tested is prepared. This may
involve grinding or homogenizing the sample to ensure uniformity.

2. Weighing the Sample: The initial weight of the sample is measured using the moisture
analyzer's built-in scale. The standard weight is 2 grams.

3. Heating the Sample: The moisture analyzer applies heat to the sample, often using a halogen
lamp. The heat causes the moisture in the sample to evaporate.

4. Monitoring Weight Loss: As the moisture evaporates, the moisture analyzer continuously
measures the weight of the sample. The decrease in weight is monitored until it stabilizes,
indicating that all moisture has been removed.

5. Results Interpretation: The moisture analyzer provides a read-out of the moisture content
percentage (%M), percentage dryness (%D), ratio of the moisture to dryness in percentage (%R),
final weight (in grams) and the time taken (in seconds) to carry out the analysis.

Fig. 16: Moisture Analyzer.

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 CRUDE LIPID EXTRACTION USING THE SOXHLET APPARATUS.

Soxhlet apparatus is the equipment used for the determination of the crude lipid present in a
sample. It consist of 4 major components

1. An extractor: comprising the thimble which holds the sample.

2. Condenser: for cooling and condensing the ether vapor.

3. 250ml flask.

4. Heating mantle.

PROCEDURE

1. Weigh the filter paper to be used for weighing on the weight balance and record it as

W1

2. Weigh the desired amount of the sample (noting the weight down)

3. Weight of filter paper + weight of sample, record it as W2

4. Wrap the sample and staple with pin, then record as W3 (you can name the sample for

easy recognition after extraction).

5. Rinse all the glass apparatus of the Soxhlet apparatus with petroleum ether and dry

them in the oven.

6. Load W3 into the thimble and place the thimble in the Soxhlet extractor.

7. Fill a clean round bottom flask with 90ml of petroleum ether and place the whole

setting on a heating mantle, allowing the petroleum ether to boil.

8. The process goes on for about six hours, after then the condensing unit is removed

from the extraction set up, thereby removing the lipid.

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 9. Collect all the solvent after extraction.

The sample is dried up using oven and then placed inside a desiccator and finally

weighed (W4).

Calculations:

% Lipid:

Fig. 17: Soxhlet Apparatus.

CRUDE FIBER ANALYSIS

This is basically sequentially treating the sample with acid and alkali to remove the soluble
components like sugar and starch, leaving behind the indigestible fiber fraction. The residue is
then filtered, dried and weighed to determine the crude fiber content. This analysis is carried out
on the defatted sample from the crude lipid analysis.

PROCEDURE
1. Weigh 2g of the sample into a beaker labeled W1.

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 2. Then add 20ml of 1.25% Sulfuric acid.
3. Warm the solution of the acid‐sample for about 20-25mins in a water bath.
4. Allow to cool before filtering using filter paper and funnel.
5. Wash away with distilled water any traces of the Sulfuric Acid until it is colorless.
6. Allow to drain off completely, and then add 20ml of NaOH.
7. Wash away again any traces of NaOH with distilled water.
8. Scrape carefully the left-over content in the filter paper into the crucible and label it
W2.
9. Remove the crucibles and determine the weight after drying it up in the oven.

Fig. 18: Hot Air Oven.

DETERMINATION OF TOTAL ASH CONTENT.

The total ash content represents the inorganic matter in a given sample. It is the residue obtained
after incineration of the dry material at high temperatures in a muffle furnace which in turn
appears as grey-white colored powder.

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 Fig. 19: Muffle Furnace.

PROCEDURE

1. Heat a platinum crucible to 600°C in a muffle furnace for 1 hour, cool in a desiccator and
weigh (W₁).

2. Weigh accurately 2 g of the dried sample (W₂) in to a crucible.

3. Place the crucible inside the previously set muffle furnace and heat for 6 to 8 hours to greyish
white ash.

4. Cool the crucible in a desiccator and weigh (W3).

5. Heat the crucible again for further 30 minutes to confirm completion of ashing, cool and
weigh.

Calculation:

% of Ash content

Where; W1= Weight of crucible.

W2= Weight of dry matter and crucible.

W3= Weight of crucible with ash.

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 CHAPTER FIVE

SUMMARY, CONCLUSION AND RECOMMENDATIONS

5.1 SUMMARY OF ATTACHMENT ACTIVITIES

My Industrial Training took place at Central Research Laboratories, University of Ilorin and it
started on the 2nd of April, 2024. My colleagues and I were welcomed warmly and introduced to
various departments in the organization; General Laboratories, Metal Analysis Laboratory,
Chemistry & Biochemistry Laboratory, Microbiology Laboratory, Material Testing Laboratory,
Plant Tissue Culture Laboratory, and Elemental Analysis Laboratory.

I was first assigned to the Plant Tissue Culture Laboratory, where I was introduced to the
concept of plant tissue culture, media preparation, sub-culturing of plantlets and transplanting
plantlets into a net house. I spent a total of 5 weeks there and was moved to the Microbiology
laboratory where I learnt agar preparation, gram staining technique, serial dilution and
microscopy. I also spent 5 weeks there.

The next assignment was both the Material Testing Laboratory and the Elemental Analysis
Laboratory, where I spent 4 weeks. In the Material Testing Laboratory, quantitative and
qualitative analysis of phytochemicals was carried out. While in the Elemental Analysis
Laboratory, Carbon, Hydrogen, Nitrogen, Sulfur and Oxygen analysis was carried out using the
CHNS/O Analyzer.

In the Metal Analysis Laboratory, I learnt how to use the Atomic Absorption Spectrophotometer
(AAS) to carry out metal analysis on various samples. There I spent 5 weeks. At the General
Laboratory, I was introduced to proximate analysis in great details.

5.2 PROBLEMS ENCOUNTERED DURING THE SIWES PROGRAM

1. Restriction of student to some equipment unless strictly monitored by a technologist.

2. Experimental works cannot be carried out in the laboratory except there are real samples
on ground.

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 5.3 RECOMMENDATIONS
The Students’ Industrial Work Experience Scheme is essential for students because it gives the
opportunity to expose students to tools, facilities, and equipment related to their course of study
that may not be available in their respective institutions. It also gives them a feel on how a work
place operates. Therefore, I suggest that:

1. The Institution should also help in securing more placements for students.
2. Regular monthly allowances for students on attachment should be paid promptly in order
to motivate the students and contribute to the progress of the establishment.
3. Institutions should improve their relationship with available industries to allow students
have easy access to all necessary practical experiences for which the SIWES program
established.
4. The SIWES Unit should endeavor to pay the students impromptu visits to ensure that the
students are being trained as expected.
5. Industrial training should be made compulsory for all students.

5.4 CONCLUSION

My 26 weeks industrial training program at Central Research Laboratories, University of Ilorin

was highly productive and engaging. It provided an avenue for me to understand practical
aspects of the theoretical knowledge already acquired in some of the courses I have offered so far
as an undergraduate of Food Science department. Thanks to the training, there was an
opportunity to see more equipment and instruments used for food analysis. The training has
exposed me to professional work methods and due to this, I am confident to build my future
career as a Food Scientist.

All of these priceless experiences and insights were obtained not just from working directly but
also from seeing and conversing with colleagues, supervisors, and other people associated with
my field of study.

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