2. DETERMINATION OF FATTY ACID PROFILE AND TRANS-FATTY
ACID CONTENT OF 5 DIFFERENT BRANDS OF FREQUENTLY
CONSUMED VEGETABLE OILS IN NNEWI
CHUKWUBUIKEM, IFEANYI PROSPER
2019544072
A RESEARCH PROJECT SUBMITTED TO THE DEPARTMENT OF
PURE AND INDUSTRIAL CHEMISTRY
FACULTY OF PHYSICAL SCIENCE
NNAMDI AZIKIWE UNIVERSITY, AWKA
IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE
AWARD OF DEGREE OF BACHELOR OF SCIENCE (B.sc) DEGREE
IN PURE AND INDUSTRIAL CHEMISTRY
DECEMBER, 2023.
3. CERTIFICATION PAGE
I, Chukwubuikem, Ifeanyi Prosper of 2019544072 hereby
certify that I am responsible for the work submitted in this
project and that this research project is an original work,
work which has not been previously presented nor
submitted to this University or any other institution for the
award of degree of B.sc.
______________________
___________________
Student Date
______________________
___________________
Dr. Odidika Collins. Date.
(Supervisor)
______________________
___________________
4. DEDICATION
This research project is dedicateed to God almighty for his
extravagant grace,faithfulness,and guidance throughout the
process of this work as well as my undergraduate studies
5. ACKNOWLEDGEMENT
My most gratitude to God almighty for his unquantifiable
grace and mercy for granted me the resources and very
important personnels for my academic journey as well as in
the completion of this research project.
My sincere gratitude goes to the head of department of Pure
and Industrial Chemistry department,Nnamdi Azikiwe
University, Dr. NH Okoye, My very able supervisor Dr. Odidika
Collins for his relentless effort in making sure that this project
work was a success,the entire staff of the department of Pure
and Industrial Chemistry, Nnamdi Azikiwe University Awka for
their guidance,teachings and words of encouragement,advice
throughout my journey in this University.I am forever grateful
to my guardians Mr & Mrs Emmanuel Chukwubuikem and the
entire chukwubuikem family
7. volume of oil production is expanding,as a result of
which,there is concern with regard to the authenticity of
VOs. In addition,The Market Receives Not Only Edible
Vos,but also those technologically processed for food.The
country's market is also filled with incoming imported fat
and oil products.Given the rather complex chemical
composition of VOs,there are problems with conducting a
comprehensive examination often the authenticity of all
types of VOs sold in the country's food markets[15].
A reliable way to determine the type of oil is to establish its
chemical composition.The FA profile contains stable and
informative characteristics by which the type of oil can be
identified[16].The analysis of FA profiles in food products
requires the extraction of lipids from samples,followed by
separation,
8.
9. All of these detectors use nitrogen,helium and hydrogen as the
carrier gas.However,an FID is widely used as a detector for
detecting FAS.FIDs offer more sensitivity and a higher
resolution than other detectors,and they are also reliable and
relatively easy to use.In addition,various compositions can be
detected using FIDs.These methods have been optimized for
the analysis of specific foods,such as VOs.Most studies
regarding the use of GC for the analysis of polyunsaturated
FAs,such as linoleic and linolenic acids and their isomers
present in VOS,also recommend a high polarity column of the
CPSil88 type.The main advantage is that this column can
separate the isomers of linoleic and linolenic acids well[24-
26].A method for validating the determination of FAs is the
procedure for the derivatization of FAs after the extraction of
lipids from food products. Methods of
10. derivatization include acid or alkaline catalysis.Such
methods of derivatization simplify the volatility of FAs,and
also increase the separation and sensitivity of GC
detectors.Usually,the most common way to determine FAs
is through lipid extraction,followed by the transformation
of the FAs into methyl esters(methylation).In GC
analysis,the identification of FAs is based on changing the
FAs to the corresponding fatty acid methyl
esters(FAMES),which have a higher volatility.The most
commonly used base reagents for rapidly changing FAs into
FAMES are NaOH or KOH in methanol.Other methods of
methylation used benzyl alcohol,diazomethane in a mixture
of ether,trichloride boron and sodium methoxide in
methanol.
11. All these led to carrying a full study on these commercially
sold vegetable oil to determine their authenticity by
comparing them with the standard range published by
health organizations like the NAFDAC and SON to enlighten
the public on the health implications of taking these food
products above required range.
12. 1.1. BACKGROUND OF STUDY
The background study of fatty acid profile and trans-fatty acid
content of different brands aspect in nutritional composition
and potential health risks manufacturing processes. They can b
corn and more. These oils differ in their fatty acid which directly
impacts their nutritional value and potential health effects.Fatt
essential acid has various physiological functions. They can be
saturated fatty acids (M has differentTFAs) naturally occurring
trans fats found in small amounts in animal-based products,
industrially produced TFAs has of cardiovascular inflammation,
and other health problems. Conducting a background study on
the used vegetable such as gas acid brands, researchers and
nutritionists can assess their nutritional quality and potential
health risks.
13. The determination of fatty acid profile and trans-fatty acid
content of different brands of commercially used vegetable
oil using Gas Chromatography with Flame Ionization
Detection (GC-FID) is a common analytical technique used in
food science and nutrition research.
GC-FID involves the separation of individual fatty acids
present in the vegetable oil sample based on their volatility
and affinity for the stationary phase of the chromatographic
column. The separated fatty acids are then detected and
quant IonizationBy analyzing the fatty acid profile of different
vegetable oil brands, researchers information is different fatty
acids have different health effects.For example, saturated
fatty acids are associated with increased risk of cardiovascular
diseases, while monounsaturated and polyunsaturated fatty
acids are considered healthier options.
14. Trans-fatty acids, specifically industrially produced trans fats,
are known to have negative health effects, including increasing
the risk of heart diseases. Therefore, determining the trans-
fatty acid content of vegetable oils is crucial for assessing their
potential health risks.
Overall, the determination of fatty acid profile and trans-fatty
acid content using GC-FID provides valuable information about
the nutritional quality and potential health risks associated
with different brands of commercially used vegetable oils. This
knowledge can help consumers make informed choices and
promote the use of healthier vegetable oil options that are low
in trans fats and balanced in terms of saturated,
monounsaturated, and polyunsaturated fatty acids.
15. 1.2 AIM OF STUDY
The aim of the study on the determination of trans-fatty
acid content and trans-fatty acid profile present in
commercially used vegetable oil is to assess the
nutritional quality and potential health risks associated
with different brands of vegetable oils. By analyzing the
trans-fatty acid content and profile, researchers can
determine the levels of industrially produced trans fats,
which are known to have negative health effects such as
increasing the risk of heart diseases. This information can
help consumers make informed choices and promote the
use of healthier vegetable oil options that are low in trans
fats and balanced in terms of saturated,
monounsaturated, and polyunsaturated fatty acids.
16. 1.3. OBJECTIVES OF STUDY
My main objectives based on this study is to first engage in
sample collection from the supposed commercial
outlet,after which a physiochemical analysis is carried out on
these samples.
The next step here is to make use of the GC-FID machine to
determine the fatty acid profiles present in the sample
followed by collation of result and comparing them with the
standardized values.
Finally,here we enlighten the general public on the health
impact of these food products if when consumed above the
acceptable limit needed by the human body.
17. 1.4. SCOPE OF STUDY
This study is focused on the determination of fatty acid
profile and trans-fatty acid content present in 5 different
brands of commercially sold vegetable oils by carrying out
physiochemical tests such as melting point,refractive
index,acid value,iodine value,saponification value,peroxide
value,cold test,flash point as well as making use of GC-FID (Gas
Chromatography Flame Ionization Detector) machine to
correctly determine their ranges in the different samples.
18. 1.5. SIGNIFICANCE OF STUDY
This study will be a sort of reference to related research
works in the academic world as well as expose both the
researcher and the entire public on the knowledge of the
health impact associated with the result of the research
conducted.
19. 1.6. LITERATURE REVIEW
Edible oil,as the energy source of human body nutrition,is a
very important part of human diet and an essential raw
material in food processing industry.The edible oil plays a very
important role in our daily life.The quality and safety of edible
oil can easily be affected by many factors including methods
of oil production and processing,distribution and catering
services,food additives and food production as well as
packaging materials.Edible oil serves as membrane bilayer and
sources of energy.Fatty acids are triglyceride molecules which
are composed of three fatty acid units connected to a
glycerol.The fatty acid chains attached to the glycerol units
can be identical or different depending on this According to
the joint WHO/ FAOreport,
20.
21. consumed safely and in the right amount.In Ethiopia research
related to saturated and unsaturated fatty acid reported a
study sample size(n-23)edible oil(Niger edible oil,Sun flower
edible oil,olive oil,palm oil,cotton seed oil and soya bean
edible oil)According to the report done variation in identities
and concentrations of fatty acids observed in all edible oil
samples.Depending on the type and brand of the oils,the
concentrations ranged saturated fatty acids(SFA) (9.92%-
47%),(MUFA) (24.54%-66.84%),and Poly Unsaturated Fatty
Acids(PUFA)(7.23%-59.11%)according to the study done by.
The SFA contents observed in these edible oils are slightly
higher than the previous reports.Results of SFA and PUFA of
this oil are slightly higher than previously published
data.While its MUFA contents is slightly less from the report
of(57,52,32) from this the ratio of
22. P/SThe range were(Palmoil0.18%-Sun flower oil4.4%)and
average lowest P/S index value was measured in palm edible
oils(0.18)followed by olive edible oil(0.86).These values are
closer to results reported by.In general,the SFA,MUFA and
PUFA results of this oil are within the range of Codex.
In quality control of Edible oil different parameters measured
to control the quality of the manufactured edible oil in the
factory.parameters such as iodine value(degree of
unsaturation),peroxide value(formation of primary oxidation
products),moisture content,specific gravity(purity),and acid
value(free fatty acids formation because of rancidity)are key
parameters to determine the shelf life quality and hence the
economic value of oils[16].Rancidity of vegetable oils cause
health risks including cancer and inflammation because of the
23. formation of toxic and reactive oxidation products.For
healthy consumption,unsaturated oils are better than the
saturated.Consumption of palmitic oil(highly saturated)is
associated with an increased risk of developing
cardiovascular
diseases(MukherjeeandMitra,2009).In contrast,edible
vegetable oils such as sun flower,olive,canola andNiger-seed
oils contain high levels of poly unsaturated fats which make
them susceptible for rancidity.
In one of the research study report in Ethiopia A total of
sample size(n-60)samples 30 from locally made(Niger seed at
market 14,Niger seed at production center 11,sun flower at
the market5) and 30 from imported palm oil
brands(Avena11,Hayat4,Jersey5 and Chef10).The report from
this study;Moister content were ranged(0.089%)-0.333%)
peroxide value
24. (7.05-15.09mill-equivalents of oxygen/kg),acid value(0.98-
2.43mgKOH/goil)and iodine value(115.63-
21.8g12/100goil)done by for local and imported edible
oils,respectively(Negashetal.,2019).
Previous studies found that oils which were produced using
low technology displayed a higher moisture.The higher
moisture content in the locally made oils indicated that it
undergo rancidity.This is because the presence of sufficient
amount of moisture favors microbial growth.Other studies
have also shown that fungus species such as Aspergillus niger
and Mucor species survive and reproduce when the moisture
content value is higher than 0.2%. The results highlight that
all rancidity quality parameters of the locally made oil
samples were not within the joint WHO/FAO standards while
the imported oils showed
25. greater fatty acid saturation.In this study,the mean
value(0.823)of specific gravity for locally made oils was not in
line with the WHO limit(Nigerseed0.9170.92;Sunflower0.919-
0.923).The specific gravity value which deviated from the
standards related to the poor refining and upgrading process
in the local production facilities.Insufficient refining process
lead for a higher impure oil grade. Furthermore,the specific
gravity values of the locally made oils were below the
standard ranges make them susceptible to adulteration. High
cholesterol content associated with chronic heart disease and
other heart problems.However,cholesterol also has essential
functions in the body such as providing essential components
of membrane and serving as a precursor of bile acids,steroid
hormones and vitamin D. .Consuming cholesterol in our
26. diet increases the level of Low Density Lipo proteins(LDLs).In
one of outlined study in Ethiopia a total of sample size(n-
9)nine samples of edible vegetable oils of which six are
branded and three are non-branded.From this report
Cholesterol was found in seven of the vegetable oils while for
two oil samples(Nigerseedk-16andNigerseedk-7)it was not
found(nil).Rape seed branded vegetable oil had highest
cholesterol concentration(257.1mg/L)while branded palm oil
has the least concentration(88.8mg/L)of cholesterol
According to the report done by.Niger seed had high content
of the essential fatty acid;linoleic(C18:2)acid,which has the
ability to decrease cholesterol levels,stimulate cholesterol
excretion into the intestine and inhibit biosynthesis of
cholesterol in the liver.Oils containing high level of poly
unsaturated
27. fatty acid are found to inhibit the activity of
hydroxymethylglutaryl-coenzymeA-reductase(HMG-CoA-
reductase)which is the regulatory enzyme in cholesterol
biosynthesis.
natural metal sources and environmental pollution.In
addition to these facts,the agricultural habits of the
farmers play an important role in the metal contents of
their products such as the application of fertilizers or
metal containing plant protection agents.In this study
similar with the study conducted in other study report on
the level of zinc and copper that are potentially present in
oil samples caused by environmental contamination.The
result of physico-chemical properties shows that acid
value,saponification value,iodine value,peroxide value
and the metal levels were in the acceptable range
28. compared to WHO/FAO report.
In Ethiopia limited study done on shelf life prediction of
edible oil one study report using three edible oil sample(n-
3)the qualities of edible cotton,peanut and soybean seed
oils,and predict their shelf stability using the accelerated
shelf-life testing method.According to the study report the
predominant fatty acid in edible soya bean and cotton seed
oils were linoleic acid 42.8% and 41.6%,respectively.While
oleic acid(46.3%)was the major fatty acid in edible peanut
oil.Based on this,the shelf-life of the domestic edible oils
determined at room temperature and amounts 36.9,42.1 and
37.8 is present in vegetable oils,although in small proportion
Un-weeks for soya bean,peanut and cotton seed
oils,respectively.Finding from above study supports previous
work that cholesterol like the peroxide value,
29. the physio
-chemical properties and level of essential metal revealed
that four edible oils were acceptable to human
consumption.Metals arrive in the plant via deposition as well
as bioaccumulation from the soil via the
Compared to soyabean and cotton seed oil,peanut oil has
longer shelf-life.The shelf-life of properly refined edible oils is
typically12-18 months at ambient temperature.As compared
to the literature,the shelf-life of edible oils determined in this
research was lower.Studies using soyabean oil indicate that
peroxide levels ranging from1.0 to 5.0meq/kg signify low
oxidation;5.0-10.0meq/kg signify moderate oxidation;and
10.0meq/kg and above signify highl levels of oxidation.
Industrial processing,especially catalytic hydrogenation of
vegetable oils,affects their fatty acid
30. composition.Higher hydrogenation of oil is
directly proportional to higher saturation,solidification and
presence of Trans fatty acid.Highly saturated fatty acid diets
have been found to increase the level of cholesterol.Several
studies have shown that palm oil has a hyper cholesterolemic
effect because of its high saturation to 50%palmitic
acid(C16:0).This high source of saturated fats
in the human diet thought to have detrimental effect on
human health,particularly when compared with Poly
unsaturated Fatty Acids(PUFAS).From other study in Ethiopia
report a sample size(n-16)Sixteen different types of solid and
liquid edible oils from main shops and supermarkets in Addis
Ababa examined. According to the report the solid oils have
higher iodine value,saturated,mono unsaturated,Poly
unsaturated fatty acids,
31. Cis,Trans,Omega-6fats as compared with the liquid.Palm oils
havevmoreTrans,saturated,mono unsaturated and lower
omega-6 and cis fats.In this study Report,there was 47% SFAs
and 11.57g PUFAS from100g for palm oil several studies
showed the correlations between high intakes of industrially
produced TFAs and increased risk of CHD.Imported palm oils
contain surplus amount of saturated fatty acid;and according
to the consumption,it is usually surpass the
rancidity,acidity,bitterness,soapiness and other of
flavors.WHO daily recommendation amount.
1.6.2 Microbial Safety and Quality of Edible Oil
A limited work done in Ethiopia on aflatoxin contamination of
edible oil.A study done a sample sizeof(-48)samples from this
the level of contamination for Ethiopian oils(n-21)ranged
between(0.07-145.59µg/
32. .Aflatoxin(AF) ingestion through contaminated foodstuffs
causes 250,000 deaths every year from. hepatocellular
carcinoma in China and sub-Saharan Africa.Previous studies
on oil seeds,by products,and peanuts also support the higher
rate of aflatoxins in Ethiopia.The noug cake,which is the by
product oft the Niger seed oil extraction process,was cited as
the primary source of aflatoxin detection in samples taken
from the commercial milk supply in Addis
Ababa,Ethiopia.Another study was conducted in eastern
Ethiopia with 120 groundnut samples.From
these,93(77.5%)tested positive for the total aflatoxin levels in
the positive samples,varying between 15 and
11900µg/kg.These results reveal that there is heavy
contamination in oil seeds and groundnuts in Ethiopia.
According to there port done by A total of sample
33. size(n-125)One hundred twenty five edible oil samples
examined among which the report of the result
62(48%)samples were containing a varying number of
bacteria and Moulds.The study report showed that the
aerobic plate count detected in 46(35,6%),moulds
32(24.8%),Yeasts 4(3.1%),total coliforms 6(4.5%)samples.Fecal
coliforms,E.coli and S.aureus found only in one sample.None
of the examined edible oil samples contains almonella and
shigella organisms.
The average microbial load is not higher than 105cfu/ml for
Aerobic mesophillic bacteria and 104cfu/ml for moulds.The
findings of this study shows that the mould and yeast results
are found to be in a mean count of 4.1x104+4.48x102cfu/ml
and 3.2x104:2.62x102cfu/ml respectively.In this study mould
and yeast count were
34. nearly similar with the study conducted in Nigeria by and
Gondertown,Ethiopia by with mean count of
4.43x103+1.25x104cfu/ml.In a similar study conducted in
Nigeria the presence of high load of Yeasts and Mould in
edible oil causes deterioration by secreting extracellular
lipases enzyme.This enzyme breaks down ester bonds in lipid
molecules and liberates diglycerides,monoglycerides,and
glycerols,Free fatty acids and results in increment of free fatty
acid values of edible oil leading to deterioration.Food borne
disease is critical public health
problem(Odohetal.,2017).Micro organisms are known to
cause chemical changes in edible oil that lead to deterioration
in the quality of the oil. The lipolytic activity of fungi on the
triglycerides of oils and fats used in baking formulations may
cause
reported A total of sample size(n-50)samples
35. collected from the markets and the microbial load and
physico-chemical characteristics reported.The mean Acid
Value(AV)ranged from 5.90+1.29 to
15.67+1.41mgKOH/mL.while the mean Free Fatty
Acid(FFA)Peroxide Value(PV)and lodine Value(IV)and pH value
ranged from 3.25 to 8.00%,2.00 to 26.90meq02/mL.,180.60
to 468.72g12/100mL and 6.32 to 6.80 Respectively.All the
samples had highest value as compared to WHO standard.The
result of this study is close to the finding of.
The acrobic mesophilic count of the locally produced oil
ranged from 4.545.82 to 12.131.98x103cfu/mL,the yeast and
mold count of the locally produced oil ranged from 7.74±9.64
to 16.9312.68103cfu/mL.The microbial isolates were
Staphylococcusaureus,Pseudomonasaerugenosa,Kelebsiellap
neumonie,
36. Shigellasonnei,Penicillium chrisoginum,Aspergillus
niger,Aspergillus flavus,Aspergillus fumigatus,Aspergillus
versicolor,Fusarium oxisporium,Candida albicans and Mucor
spp.The micro organisms isolated in this study are closely
similar to species reported by other studies in Nigeria.But in
this study in addition to Staphylococcus aureus,Penicillium
chrisoginum,Aspergillus flavus,A.fumigatus,Candida albicans
and Mucor spp.Pseudomonas aerogenosa,Aspergillus
niger,A.versicolor and Fusarium oxisporium also isolated and
reported.
1.6.3 Edible Oil Quality and Safety Management System The
food processing industries are suffering from quality related
problems.These problems include poor performance of
products in the export market,low quality and insufficient raw
material supply,low product
37. lconfidence due to lack of quality assurance systems and
increased pressure from high quality and competitive
products in the local market.The solution is to implement a
properly developed Quality Management System(QMS) in the
industry.
To establish and improve standards of cooking oil,there must
be apply the Edible oil quality and safety management
system.Such as HACCP as a preventive safety control system
of edible oil.From every aspect of the cultivation of raw
materials,procurement,production,storage,transportation,qua
lity control can be carried out,strictly controlling quality
security of edible oil,to eliminate the edible oil quality
problems from the source of edible oil,thereby producing
safe,healthy edible oil. Second;establish the national
standards certification of edible oil,carry out uniform
certification bodies of edible
38. oil. Thereby promote edible oil quality management system
and connect it with international standards.Third;implement
the market access system on the edible oil
market.Fourth;Cooking oil inspection and detection inspect
the production process quality control;inspect environment
and sanitation conditions;inspect product quality control.
