Peanut Paste

KWAME NKRUMAH UNIVERSITY OF SCIENCE AND TECHNOLOGY,
KUMASI
COLLEGE OF SCIENCE
DEPARTMENT OF FOOD SCIENCE AND TECHNOLOGY
EVALUATION OF PEANUT PASTE IN SELECTED MARKETS IN
NORTHERN GHANA
BY
YUSSIF ABUBAKARI
(BSc Community Nutrition)
A Thesis submitted to the Department of Food Science and Technology, College of
Science in partial fulfilment of the requirements for the degree of
Masters of Science in Food Science and Technology
© 2015, Department of Food Science and Technology
JULY 2016
DECLARATION
I hereby declare that the experimental work described in this thesis was performed by
me at the Food Science and Technology Laboratory of the Kwame Nkrumah
University of Science and Technology, Kumasi, under the supervision of Prof Ibok
Oduro and Prof William Otoo Ellis towards the MSc (Food Science and Technology)
degree. I certify that, to the best of my knowledge, this work has not been submitted
for the award of any other degree of the University, except where due
acknowledgement has been made in text
Yussif Abubakari ……………………… …………………
(Student) Signature Date
Certified by:
Prof. William O. ELLIS ……………………… …………………
(Supervisor) Signature Date
Prof. Mrs Ibok ODURO ……………………… …………………
(Head of Department, Supervisor) Signature Date
i
DEDICATION
I dedicate this entire work to my family and friends, my supervisors, the Ghana PMIL
(PEANUT AND MYCOTOXIN INNOVATION LABORATORY PROJECT), staff
of the CSIR-SARI and the entire 2014-2015 MSc Food Science and Technology class
of the KNUST-Kumasi
ii
ACKNOWLEDGEMENT
I thank the almighty God, the most merciful and the most beneficent for guiding me
throughout my entire MSc Food Science and Technology program. This work would
not have been successful without the help of my supervisors Prof Ibok Oduro and Prof
Williams Otoo Ellis, The Peanut and Mycotoxin Laboratory Project Team, the Food
Science and Technology Faculty and all friends of the MSc Food Science and
Technology 2014-2015 class, I thank you all and may God bless you.
iii
TABLE OF CONTENTS
DECLARATION........................................................................................................... i
DEDICATION..............................................................................................................ii
ACKNOWLEDGEMENT ..........................................................................................iii
TABLE OF CONTENTS ........................................................................................... iv
LIST OF TABLES .....................................................................................................vii
LIST OF FIGURES ..................................................................................................viii
LIST OF PLATE......................................................................................................... ix
ABSTRACT .................................................................................................................. x
CHAPTER ONE .......................................................................................................... 1

INTRODUCTION........................................................................................................ 1
1.0 BACKGROUND OF STUDY ............................................................................. 1
1.1 PROBLEM STATEMENT .................................................................................. 3
1.3 JUSTIFICATION ................................................................................................. 3
1.4 MAIN OBJECTIVE ............................................................................................. 3
1.5 SPECIFIC OBJECTIVES .................................................................................... 3
CHAPTER TWO ......................................................................................................... 4

LITERATURE REVIEW ........................................................................................... 4
2.0 PEANUT .............................................................................................................. 4
2.1 PEANUT PRODUCTS ........................................................................................ 5
2.1.1 PEANUT BISCUIT ....................................................................................... 5
2.1.2 PEANUT PASTE .......................................................................................... 6
2.2 CONSUMPTION RATE OF PEANUT PASTE IN NORTHERN GHANA ...... 7
2.3 PRODUCTION OF PEANUT PASTE ................................................................ 8
2.3 CHALLENGES WITH PEANUT PRODUCTION ............................................. 9
2.3.1 MICROBIAL CONTAMINATION OF PEANUT PASTE .......................... 9
2.3.2 AFLATOXIN CONTAMINATION ........................................................... 10
CHAPTER THREE ................................................................................................... 13

MATERIALS AND METHODS .............................................................................. 13
3.0 THE MARKET SURVEY ................................................................................. 13
iv
3.1 PREPARATION OF PEANUT PASTE CONTROL SAMPLE ....................... 13
3.1.1 SOURCE OF RAW MATERIAL (NKATE-SARI)...................................... 13
3.1.2 CONTROL SAMPLE PREPARATION OF PEANUT PASTE ................. 14
3.1.3 PEANUT BUTTER PRODUCTION .......................................................... 14
3.2 EXPERIMENTAL ANALYSIS ........................................................................ 15
3.2.1 PROXIMATE ANALYSIS ......................................................................... 15
3.2.2 MOISTURE DETERMINATION ON PEANUT BUTTER
(AOAC, 2003)............................................................................................ 15
3.2.3 CRUDE PROTEIN DETERMINATION (AOAC, 2003) ........................... 15
3.2.4 CRUDE FIBER (AOAC, 2003) .................................................................. 16
3.2.5 CRUDE FAT DETERMINATION (AOAC, 2003) .................................... 16
3.2.6 ASHING (AOAC, 2003) ............................................................................. 17
3.2.7 TOTAL CARBOHYDRATE (AOAC, 2003) ............................................. 17
3.3 MICROBIAL LOAD ......................................................................................... 17
3.3.1 EQUIPMENT AND MATERIALS STERILIZATION .............................. 17
3.3.2 SAMPLE PREPARATION ......................................................................... 18
3.3.3 TOTAL AEROBIC COUNT ....................................................................... 18
3.3.4 TOTAL COLIFORMS ................................................................................ 18
3.3.5 FUNGAL ENUMERATION ....................................................................... 19
3.3.6 MOULD IDENTIFICATION...................................................................... 19
3.4 AFLATOXIN DETERMINATION (AOAC, 2008) .......................................... 19
3.5 STATISTICAL ANALYSIS .............................................................................. 20
CHAPTER FOUR ...................................................................................................... 21

RESULTS AND DISCUSIONS ................................................................................ 21
4.0 SURVEY RESULTS ......................................................................................... 21
4.1 PROXIMATE COMPOSITION ........................................................................ 22
4.1.1 MOISTURE ................................................................................................. 22
4.1.2 CRUDE PROTEIN ...................................................................................... 23
4.1.3 FAT .............................................................................................................. 24
4.1.4 FIBER .......................................................................................................... 25
4.1.5 ASH ............................................................................................................. 25
4.1.6 CARBOHYDRATE .................................................................................... 26
4.2 MICROBIAL LOAD ......................................................................................... 26
v
4.2.1 TOTAL AEROBIC COUNT ....................................................................... 26
4.2.2 TOTAL COLIFORMS ................................................................................ 28
4.2.3 FUNGAL ENUMERATION ....................................................................... 29
4.3 AFLATOXIN ..................................................................................................... 32
CHAPTER FIVE ....................................................................................................... 35

CONCLUSION AND RECOMMENDATION ....................................................... 35
5.0 CONCLUSION .................................................................................................. 35
5.1 RECOMMENDATIONS ................................................................................... 35
REFERENCES ........................................................................................................... 36
APPENDICE .............................................................................................................. 46
vi
LIST OF TABLES
Table 2.1: Proximate composition of peanut paste ....................................................... 7

Table 2.2: USDA acceptable levels of microbes in peanut paste ................................ 10
Table 2.3: Aflatoxin levels in peanut paste at different treatment point ...................... 12
Table 3.1: Research study areas ................................................................................... 13
Table 4.1: Survey results obtained from the various peanut past producers ............... 21
Table 4.2: Proximate composition of peanut paste samples ........................................ 23
Table 4.3: Mould identified in peanut paste samples from selected markets .............. 31
vii
LIST OF FIGURES
Figure 4.1: Total aerobic counts after 48 hours incubation ......................................... 27

Figure 4.2: Total coliform count after 48 hours of incubation .................................... 28
Figure 4.3: Fungal enumeration of peanut paste samples. ........................................... 30
Figure 4.4: Aflatoxin contamination levels in peanut paste from the various
markets ..................................................................................................... 33
viii
LIST OF PLATE
Plate 2.1: Peanut paste on display at the market ............................................................ 8

Plate 3.1: De-shelled peanut into zip lock bag ............................................................. 14
Plate 3.2: Aflatoxin Analysis Procedure ...................................................................... 20
Plate 4.1: Peanut paste on display at the Bolgatanga central market ........................... 32
ix
ABSTRACT
Peanut paste is a delicacy in Ghana, but issues about its production quality in
Northern Ghana, has been a challenge in recent times. The study aimed to assess
peanut paste quality in Northern Ghana. Peanut paste samples (24) were acquired
from six major markets. A control sample, using Nkate-sari variety of peanut was
prepared in the Food Science and Technology Laboratory of the Kwame Nkrumah
University of Science and Technology. A survey was conducted using structured
questionnaires to understand the processing methods used. Proximate, aflatoxin and
microbial load of the samples were also determined. From the survey, 75% of the
producers used untreated stream water during processing. There was no sorting,
grading or blanching during processing. The traders (96%) acquire raw peanuts from
the market already de-shelled. Moisture, Crude protein and Carbohydrate content of
the samples ranged from 5.05 ± 0.07 to 6.45 ± 0.21, 23.67±0.05 to 31.56 ±0.78 and
19.44 ±1.19 to 27.65 ±0.96 respectively. Statistical analysis showed no significant
difference (p>0.05) between ash, carbohydrate and protein content. Aflatoxin analysis
of the Tamale central, Bolga central, Wa Gonomuni, Tamale Aboabu and Wa central
market samples showed concentrations of 2.89 ppb, 8.6 ppb, 55.39 ppb, 103.44 ppb
and 126.55 ppb respectively. Total aerobic count ranged from 2.5×103 cfu/g to
9.9×103 cfu/g. Coliform count were below the acceptable limit of 10 cfu/g. Fungal
enumeration was less than 101 cfu/g in all samples except for Navrongo central
market samples. Aspergilus parasiticus was isolated in Wa gonomuni, Wa central
market and Tamale Aboabu market samples respectively. Blastomyces Dermatitidis
was found in Bolga central market samples. Even though some samples had high
nutrient composition, contamination levels were significant due to poor production
practices.
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CHAPTER ONE
INTRODUCTION
1.0 BACKGROUND OF STUDY
Peanut (Arachis hypogaea) is a leguminous crop ranked as the 13th most important
food crop and 4th important oilseed crop in the world after soybean, cottonseed and
rapeseed (Li et al., 2011). Peanuts are cultivated globally, especially in developing
countries, where it constitutes about 97% of the total land area under cultivation and
94% of the global production (Craufurd et al., 2006). The major producers of peanut,
also known as groundnuts are China, India, Mali, Ghana and Nigeria (Mudenda,
2013). They are also widely consumed in Central and South America (Johanson &
Ives, 2000).
About 80 % of Ghanaians from Northern Ghana consume peanut and peanut products
out of which 32% consumes it as many as three times in a week (Yussif, 2014). The
intake of peanut and its products such as peanut butter is increasing daily in the
Northern part of Ghana (Tsigbey & Braderngurg, 2004). The production and sales of
peanut and its product (peanut paste) serves as a major source of livelihood to the
women in Northern Ghana (Millar & Yeboah, 2006).
Peanut paste is a very important product in the world, it can be used as condiment or
for the preparation of soup (Katz, 2005). It’s butter is also a good source of protein,
fat, carbohydrate, fiber and minerals and has longer shelf life, smoothness and a very
pleasant flavor (Chow, 2007).
According to Fraser et al., (1992), frequent peanut paste consumption can reduce the
incidence of coronary heart diseases by 25 to 50 %. Consumption of peanut snacks
1
daily is important to the health of the individual since it contains other nutrient
essential to the human systems development (Hu et al., 1998). Peanut paste can
therefore serve as a good product for the control of type 2 diabetes when consumed
frequently or incorporated daily into meals (Jiang et al., 2002).
Although peanut is the most important leguminous crop cultivated in Ghana (Tsibgey,
2003), with major productions in the Northern part of the country, almost all of the
peanuts grown are for domestic purposes. This may be attributed to the high
prevalence of microbial and toxic metabolite contamination. This has particularly
become dominant due to the a wide range of environmental conditions in the sub-
tropical regions that favor the growth of certain microorganisms such as Aspergillus
flavus, a major producer of aflatoxins (Narasimhulu, 2007). Aflatoxins contamination
is a major limitation to the exportation of peanut onto the global market (Emmok,
2010).
The Food and Agriculture Organization estimates that, 25% of the world’s food crops
production are lost due to aflatoxin contamination, though other approaches to
detoxification and to minimize the contaminations are being employed (Proctor et al.,
2004). Research shows that, the introduction of the new standard by the European
Union on the accept/reject aflatoxin contamination levels will cause about 64% (US$
670 million loss) of peanut coming from Africa on to the world market to be rejected
(Otsuki et al., 2001). According to Dhanasekaran & Shanmugapriya., (2011), high
levels of consumption of aflatoxin contaminated product have been linked with the
incidence of certain cancers that are very deadly in humans. Aflatoxins are noted first
class carcinogens in humans (International Agency for Research on Cancer, 2002)
2
1.1 PROBLEM STATEMENT
Northern Ghana is a leading producer of peanut products such as peanut paste, but
almost all peanut paste produced are for domestic use. This may be due to the high
risk of microbial contamination through the variable methods of production that lack
standard quality supervision. These contaminations can pose serious health problems
to consumers. In addition, there is limited data on the effects of peanut processing on
the quality of the kernels and its products
1.3 JUSTIFICATION
This research will provide information on proximate, Aflatoxin and microbial levels
in the various peanut paste in the study location, which will help in food quality and
safety decision-making. Data on proximate composition of these peanut pastes will
serve as useful information for consumers.
1.4 MAIN OBJECTIVE
To assess the quality of peanut paste in the three Northern regions of Ghana (Upper
East, Upper West and Northern Region)
1.5 SPECIFIC OBJECTIVES
1. To determine the nutrient content of peanut paste sold on Northern sector
markets
2. To assess microbial load and aflatoxin levels on the peanut paste sold on
Northern sector market
3
CHAPTER TWO
LITERATURE REVIEW
2.0 PEANUT
Peanuts, originated from South America where they grow very well under subtropical
and tropical climatic conditions. The plant grows above the ground but the pods
mature in the soil (Martinez, 2014).
Peanuts are a good source of minerals such as calcium, phosphorus, magnesium and
potassium and some vitamins such as vitamin E, K and B and contains 44 to 56% oil
and 22 to 30% protein on dry matter basis (Keenan & Savage., 1994). It also has high
amount of oleic and linoleic fatty acid profile that accounts for about 75 to 80% of the
total oil it contains (Nelson et al., 1999). Peanut contains vitamin E, C, B and niacin
which can be destroyed during extreme heating, it contains an average of 5 to 7%
moisture in freshly harvested peanut and ash of about 3% (Woodroof, 1989).
Tocopherol present in peanut oil in an amount of about 0.05 percent is a good sign of
the highest stability of the peanut oil (Aoyagi, 2015).
Varieties of peanut which are commonly grown around the world include the Virginia
type, the Valencia, the Spanish and the runner and have an average protein content of
about 25% (Ahmed & Young., 2011). Different varieties have different protein
content and are due to the differences in their gene sequence, geographical location
and season of cultivation (Nagaraj, 2009). Peanuts can simply be digested and
metabolized by humans (Rena, 2015). The digestibility index for the protein
component is 89% which makes peanut highly digestible (Woodroof, 1989). With the
several amino acids present in peanut, sixteen of them are responsible for the reaction
it undergoes during roasting which leads to browning, due to high amount of sucrose,
high temperature of roasting and high fiber content (Woodroof, 1989).
4
2.1 PEANUT PRODUCTS
Peanuts as a cheap source of protein has aroused research focus in recent times on the
utilization of oilseed proteins as edible sources of protein for human and animal
consumption. Peanuts have globally been processed for oil and the residual meal is
used either as animal feed or as fertilizers
2.1.1 PEANUT BISCUIT
The term biscuit was derived from the Latin word biscoctus, meaning twice cooked
(Macrae et al., 1993). Biscuits are popular foodstuff, consumed by a large number of
populations today, due to their pleasant taste, prolonged shelf life and easy availability
at fairly low cost (Gandhi et al., 2001). Biscuits occupy primary position, both for
production and consumption as compared to other bakery products (Gandhi et al.,
2001).
As biscuits are typically higher in fat content, it becomes difficult to prepare biscuits
by reducing fat contents in their formulation to lower the risk of diseases. To reduce
the quantity of fat in bakery products fat replacers like peanut paste are used (Sanchez
et al., 1995). According to Sadaf et al. (2013), although carbohydrate composition
decreased significantly in biscuits with the supplementation of peanut paste, There
was a significant effect of peanut paste on color of biscuits. They also concluded that,
biscuits produced through the incorporation of peanut paste to reduce the quantity of
hydrogenated vegetable shortening showed generally acceptable quality
characteristics. Hydrogenated vegetable shortening of the biscuits not only increased
the protein content but it also reduced the fat contents while imparting better flavor to
biscuits.
5
2.1.2 PEANUT PASTE
Peanut paste is prepared through the dispersion of peanut oil in peanut solids, when
roasted peanuts are ground. Peanut paste is has been proven to be a good source of
nutrients and low in fat. It is continually applied for the preparation of low calorie
improved food products (Woodroof, 1983). Fifty two percent (52 %) of the United
States peanut cultivated, are for peanut paste production, and are used as spread or
sandwich, 23% are for the production of salted peanut and 21% are used as
confectionery (Singh & Singh, 1991). In other countries such as Senegal, Brazil and
India, 70 to 100% are crushed or sold out to the world market which are used for the
production of oil and feeding animals (Lusas, 1979).
In Ghana, peanut paste can be used as sandwich, condiment or for the preparation of
soup or stew (Katz, 2005). Peanut paste is a good source of protein and has a longer
shelf life, smoothness and a very pleasant flavor (Chow, 2007). The protein
composition (amino acid content) could be compared with other high protein products
such as soya beans, bean products and animal products (Ibadullah, 2013). The paste
can be processed into different textures such as smooth without any grainy particles,
regular and having particle size of less than 0.0625 inch diameter and chunky with
particle size of more than 0.0625 (Woodroof, 1989);
Peanut paste contains very important nutrient that are needed by the body for proper
development (Ayoola & Adeyeye, 2010). Study by Özcan & Seven, (2007) and Boli
et al., (2013) shows that, moisture content of peanut paste ranged between 4.50 to
6.06 % . Crude protein with 21% to 35%, crude fat with 24.55% to 50%, ash with
1.86% to 5.5% and crude fiber with 1.00% to 6.78 as shown in Table 2.1.
6
Table 2.1: Proximate composition of peanut paste
% Moisture % Crude % Crude fat %Ash % Crude fiber

protein
4.50 - 6.06 21- 35 24.55 -50 1.8- 5.5 1.00- 6.78
Source:(Özcan & Seven, 2007)
Peanuts paste contains fat in the form of monounsaturated and polyunsaturated fats
and this can lower LDL cholesterol and triglycerides levels by promoting the increase
of HDL cholesterol (Kris-Etherton et al., 1999). Peanut paste produced from high
oleic varieties have higher stability and longer shelf life (Riveros et al, 2009).
According to Fraser et al., (1992), frequent peanut paste consumption can reduce the
incident of coronary heart diseases by 25 to 50%.
Consumption of peanut snacks daily is very important to the health of the individual
since it contains other nutrient such as calcium, fiber, protein carbohydrate and fat
essential to the human systems development (Hu et al., 1998). Peanut paste can
therefore serve as a good product for the control of type 2 diabetes (peanut butter is
low in Glycemic index and Glycemic load) when consumed frequently or
incorporated into our daily servings (Jiang et al., 2002). When peanut products is
added to meals with high glycemic load, it reduce post prenatal glycaemia (Johnston
& Buller, 2005)
2.2 CONSUMPTION RATE OF PEANUT PASTE IN NORTHERN GHANA
Peanut paste is one of the most important peanut product in the world, especially at
places where poverty is at the highest peak (Omer et al., 2001). Research by Du Toit
7
et al., (2008) shows that, median infants living in Israel between the age of 8 to 14
months consume at least 7.1g of peanut protein, which according to Yussif., (2014)
represents 8 times of peanut paste in a month. Contrary to this, Infants from the
United Kingdom takes it zero times in a month, indicating a low consumption rate in
this part of the world compared to Africa (Ghana).
In Ghana, 80% of the population living in the Northern part of Ghana consume peanut
and peanut product, and 32% percent take it as many as three times in a week (Yussif,
2014). The intake of peanut and its product such as peanut paste is increasing daily in
the Northern part of Ghana (Tsigbey & Braderngurg, 2004). The production and sales
of peanut and its product (peanut paste) serves as a major source of livelihood to the
women in Northern Ghana (Millar & Yeboah, 2006) where it is publicized on tables
for consumers to buy (Plate 2.1).
Plate 2.1: Peanut paste on display at the market

Source: (Author, 2015)
2.3 PRODUCTION OF PEANUT PASTE
The procedure in peanut paste production includes de-shelling of the dry peanut,
roasting, cooling, blanching, and removal of foreign particles (including seed coat),
grinding, cooling and packaging as shown in figure 2.1. Occasionally, salt and other
ingredient such as fat, sugar and other antioxidants are also added to increase the shelf
life and improve the quality (Woodroof, 1989). The color of peanut paste could be
8
described as being heavy roasted and some may be light roasted depending on the
type preferred by the consumer, since differences could be seen during frying
(Settaluri et al., 2012). Figure 2.1 shows the production of peanut paste from Nkate-
sari as described by Woodroof (1989)
Raw Groundnuts
Sorting and Grading
Washing
Drying (105°C at 20 min,)
Cracking using hand
Sorting and grading
Roasting (116 °C for 8 min)
Grinding
Packaging into sample containers and allow to cool at room temperature
Figure 2.1: Peanut paste production from Nkate-sari
2.3 CHALLENGES WITH PEANUT PRODUCTION
2.3.1 MICROBIAL CONTAMINATION OF PEANUT PASTE
Peanuts generally, do not pose a huge risk for food contamination (Engbe, 2007). This
largely is due to the roasting step where peanuts are reduced to the 1.25% moisture
content and <0.75 water activity (aw) (Engbe, 2007). Moisture is required for most
microorganisms to survive. The low aw inhibits growth of most bacteria and many
molds. Since peanuts are rarely eaten raw, roasting not only improves peanut aroma,
flavor, and texture, but also destroys contaminating microorganisms (Woodroof,
9
1983). Nonetheless, Very few outbreaks of foodborne with relation to peanuts were
likely due to poor handling practices after a thermal treatment step, particularly
roasting (Woodroof, 1983).
From the foodborne diseases perspective, areas of concern for peanuts primarily
revolve around from aflatoxin contamination, a toxic metabolite produced by the
mold Aspergillus, and cross contamination from sources that introduce pathogens to
peanuts after processing. Proper peanut processing and handling postharvest should
therefore, ensure a safe product for consumers. It is recommended to implement a
cleanup protocol using chemicals that are food grade and environmental friendly
(Tsitsigiannis et al., 2005).
Peanut paste should contain no levels of Salmonella, Total plate count should not
exceed 10,000 CFU/g, whiles yeast and mould count should be less than 100 CFU/g.
E.coli. In addition, total coliform should not be more than 10 CFU/g as shown in
Table 2.2 (Council, 2009)
Table 2.2: USDA acceptable levels of microbes in peanut paste
Microorganism Expected levels

Salmonella Negative
E. coli <10 CFU/g
Coliform <10 CFU/g
Aerobic plate count <10,000CFU/g
Yeast <100CFU/g
Mould <100CFU/g
(Council, 2009)
2.3.2 AFLATOXIN CONTAMINATION
Food contamination caused by Aflatoxin could occur at any point along the value
chain, it does not only cause post-harvest lost but also responsible for high prevalence
of liver diseases such as liver cancer (Bababunmi et al., 1978). According to Musingo
10
et al. (1989), contaminations that commonly occur along the value chain can be very
resilient during processes such as storage, handling and processing of food and animal
feeds.
There are several types of aflatoxin in nature, among these, aflatoxin B1 and B2, G1
and G2 are the common and the most important which caused human disease (Haydes
et al., 1991). The mechanism by which aflatoxin B1 can cause disease in humans is
by the formation of epoxides when the aflatoxin molecule is activated by the use of
cytochrome P450 reductase at the end of the furanos ring which is as a result of the
presence of proteins such as DNA, RNA (Wang & Groopman, 1999).
Major symptoms of aflatoxin infection involves immune system suppression and
acute liver cirrhosis; since the liver plays a major role in detoxification of metabolites
(Beasley, 2011). Also, severe damage and cirrhosis of the liver cells are the major
effect of ingesting very high doses of aflatoxin (Wang & Groopman, 1999).
According to Dhanasekaran & Shanmugapriya., (2011), high levels of consumption of
contaminated product have been linked with the incidence of certain cancers that are
very deadly in humans. Aflatoxins are noted first class to cause human cancers
(International Agency for Research on Cancer, 2002)
Humans or animals with low immune system are more susceptible to infection from
aflatoxin (Nigam et al., 2009). The coupling effect of some deadly infectious diseases
such as hepatitis B/C, TB, HIV and consuming high levels of contaminated food have
led to higher mortality among patients whose immune system were down due to
aflatoxin infection. Some major symptoms of aflatoxin in poultry includes poor
feeding ability, liver cirrhosis, suppression of the immune system and a comparative
11
change in weights and size of organ (McKenzie et al., 1997). aflatoxins are often
found in high concentrations in cereals, peanut, maize, and other products.
A study on the occurrence of aflatoxin in Chinese peanut paste samples (Table 2.2),
revealed that, out of 50 peanut paste samples, 41 tested positive for aflatoxin B1 at a
level of 68.51 μg/kg (Li et al, 2009). Levels of aflatoxin contamination during peanut
paste production as shown in Table 2.3 shows that, roasting at 160°C had 51%
reduction, 27% after blanching and an 11% reduction after grinding, however, most of
the contamination of peanut paste is after process of production (Siwela, 2011).
Table 2.3: Aflatoxin levels in peanut paste at different treatment point
PROCESS % AFB1 % AFB2 % AFG1 % AFG2 % TOTAL
ROASTING 37 79 48 38 51
BLANCHING 57 19 27 23 27
GRINDING 3 3 6 33 11
% TOTAL 79 99 81 94 89
REDUCTION
Source: (Siwela, 2011)
12
CHAPTER THREE
MATERIALS AND METHODS
3.0 THE MARKET SURVEY
A market survey was conducted in the three Northern regions of Ghana (Northern,
Upper West and Upper East). Structured questionnaires were administered to 24
peanut paste sellers in the sampling zones (Table 3.1) using simple random statistical
method (4 from each market and a total of 8 from each Region) to acquire knowledge
on the processing methods before buying the peanut paste from them. Quartering
sampling method was also used to determine the laboratory samples used for the
various laboratory procedures.
Table 3.1: Research study areas
Regions Sampling zone
Northern Aboabo and Tamale central market
Upper West Wa central market and Gonomuni market
Upper East Navrongo central market and Bolgatanga
3.1 PREPARATION OF PEANUT PASTE CONTROL SAMPLE
3.1.1 SOURCE OF RAW MATERIAL (NKATE-SARI)
Nkate-sari was used to produce the control peanut paste sample. Nkate-sari is an
improved variety of peanut released by the Council for Scientific and Industrial
Research of the Savannah Agricultural Research Institute at Nyankpala in the
Northern region of Ghana. Dried peanut sample (5.0kg) of Nkate-sari in storage from
13
the 2014 harvest were obtained from the Savannah Agricultural Research Institute,
bagged in a clean jute sack and transported by bus in an ice pack container in an 8h
journey to the Kwame Nkrumah University of Science and Technology, Kumasi.
Samples were stored under cold condition prior to analysis.
3.1.2 CONTROL SAMPLE PREPARATION OF PEANUT PASTE
Raw peanut samples (2.0kg) were obtained using the quartering sampling protocol.
This representative sample was used for peanut paste production. The samples were
de-shelled and apportioned into zip lock bags and labeled as shown in plate 3.1, ready
for peanut butter production.
Plate 3.1: De-shelled peanut into zip lock bag

3.1.3 PEANUT BUTTER PRODUCTION
Raw peanut samples were sorted and graded before immersion in clean water and
washed thoroughly and sun dried. They were then de-shelled, sorted/graded again
manually, and 50 g roasted in a clean frying pan at 116°C for 20 min. Roasted
samples were allowed to cool at room temperature, then grinded using the attrition
mill machine. The milled samples were immediately packaged into zip lock bags and
allowed to cool (Junior et al., 2014).
14
3.2 EXPERIMENTAL ANALYSIS
3.2.1 PROXIMATE ANALYSIS
The moisture, crude protein, crude fat, crude fiber, Carbohydrate and ash contents of
the peanut butters were determined using standard methods (AOAC, 2003).
3.2.2 MOISTURE DETERMINATION ON PEANUT BUTTER (AOAC, 2003)
Peanut paste (2g) was accurately weighed in a clean, dried petri dish of a known
weight (W1). It was quickly placed in a conventional oven at 105 °C for 6 h. The petri
dish was then placed in a desiccator for 30 min to cool. After cooling, it was weighed
again (W2). The percent moisture was then calculated.
3.2.3 CRUDE PROTEIN DETERMINATION (AOAC, 2003)
Peanut paste (2g) was weighed into a digestion tube, 5 grams of catalyst and 1 glass
bead together with 10 mL concentrated sulfuric acid was also added. Digestion tubes
were placed in the digester. Digestion commenced initially at low temperature to
prevent frothing and boiling at a temperature of 320°C until the solution was clear
which is an indicator of complete oxidation. Erlenmeyer flask (250mL) containing 50
mL of 4% boric acid with indicator was placed in a distillation unit and distillation
commenced for 10 min whiles the tip of the condenser extended below the surface of
the acid solution. 100 mL of water and 70 mL of 50% sodium hydroxide (excess) was
added to the digests during the distillation process to ensure complete release of
ammonia. 150 mL of distillate was obtained when condenser with ice cold water was
used to effectively capture all distilled ammonia and the receiver flask was lowered to
ensure that the delivery tube was properly placed for continues distillation. The
delivery tube was rinsed with water and the washing was made to drain into the flask.
Titration of the distillate was done with a standardized 0.1 N hydrochloric acid until
15
the first appearance of the pink colour was obtained. Result was recorded to the
nearest 0.05 mL volume and calculated.
3.2.4 CRUDE FIBER (AOAC, 2003)
Peanut paste (5g) was taken from the zip lock bags and defatted using the AOAC
2003 standard before subjecting to analysis. 2g of peanut samples were weighted into
a flat bottom flask and 200 mL of boiling sulphuric acid (1.25%) was added for 30
min. The resulting solution was filtered through cheesecloth using a funnel and then
washed with hot water until it was free from the acid. The residue on the cloth was
transferred into a flask and 200 mL of boiling sodium hydroxide solution (1.25%) was
added. The flask was immediately connected to the digestion apparatus and boiled for
30 min. The flask was then removed and immediately the solution was filtered and
washed thoroughly with boiling distilled water until washing was no longer basic. The
residue was rinsed with 15 ml of alcohol. It was transferred into porcelain crucibles
and dried at 105°C in an oven for 24 h. It was cooled to room temperature in a
desiccator and weighed. The crucible and its weight was incinerated in a muffle
furnace at 550 °C. It was cooled to room temperature in a desiccator and weighed.
The difference between the two weights was recorded and the percentage crude fiber
calculated.
3.2.5 CRUDE FAT DETERMINATION (AOAC, 2003)
Approximately 1 g of dried samples from the moisture analysis was weighed and
wrapped in a filter paper and placed in a fat free thimble and then placed into the
extraction tube. Clean and dried receiving beaker was weighed and filled to about
three quarter with petroleum ether and then fitted into the Soxhlet apparatus. The tap
was turned on to allow the flow of water, and heater was also turned on to start the
16
extraction process. After 6h, siphoning allowed ether to evaporate and condense back
into the receiving beaker and the process was repeated until extraction was complete.
The beaker was disconnected before the last siphoning. Extract was placed in a water
bath for the remaining ether to evaporate; the dish was then placed in an oven at
105oC for 2 hours and cooled in a desiccator. The percent crude fat was determined.
3.2.6 ASHING (AOAC, 2003)
Clean empty crucible was placed in a muffle furnace at 600 °C for an hour, it was
cooled in a desiccator and then weighed and weight noted W. 1g of sample was
weighed and transferred into the crucible and noted W2. The sample was ignited over
a burner with the help of blowpipe, until charred. Then the crucible was placed in a
muffle furnace at 600°C for 6 h for complete oxidation of all organic matter in the
sample. After the process, Crucible was cooled in the desiccator and the weight was
noted W3. Percent crude ash was calculated and recorded.
3.2.7 TOTAL CARBOHYDRATE (AOAC, 2003)
Total carbohydrate was determined by difference between 100% and the sum total of
determined proximate component
3.3 MICROBIAL LOAD
3.3.1 EQUIPMENT AND MATERIALS STERILIZATION
Sterilization of equipment was carried out before and after the analysis to prevent
cross contamination of samples and equipment. Laboratory glass wares were washed
and rinsed very well with soap and under running tap water, air dried and kept in an
oven for 3h at 160 °C to ensure glassware are clean. Media was prepared under safety
condition (fume hood). Loops and forceps for inoculation were flamed to ensure
17
sterility before and after each use. Naked flame was turned on to ensure sterilization
of the working environment and 70% alcohol was also used to clean the working
bench before and after use (Cheesbrough, 2006).
3.3.2 SAMPLE PREPARATION
Peanut paste (10g) was weighed using a sterile weighing dish and transferred into
sterile sample bottles containing 90ml (0.1%) peptone water. Sample was vortexed for
about 1 minute at moderate speed. This serves as the initial dilution from which other
dilutions to the sixth power was prepared. This dilution was then used for subsequent
analysis (Cheesbrough, 2006).
3.3.3 TOTAL AEROBIC COUNT
Plate count agar was used to determine the total aerobics present in the peanut paste
samples. Preparation of the plate count agar was done as described by Cheesbrough
(2006). After the preparations, 100µL of the serial diluted samples were then plated
using the spread plate method and the inoculated plates were then incubated at 37 °C
for 24h. Plates that showed visible colonies between 30 and 300 were identified and
manually counted. The numbers of colony forming units per gram of samples were
calculated by multiplying the number of organisms by the dilution factor
(Cheesbrough, 2006)
3.3.4 TOTAL COLIFORMS
The media used was the Violet Red Bile Lactose (VRBL) agar. This media was
chosen because of its good selective ability. The Spread plating procedure was used
for the inoculation in which 100µL of the serial diluted samples were plated and
incubated for 24 hours at 37°C (Cheesbrough, 2006).
18
3.3.5 FUNGAL ENUMERATION
Malt extract agar was used to determine the presence and the numbers of yeast and
molds in the peanut paste samples. Spread plating method was employed with a
volume of 1ml of the serial diluted peanut paste samples and incubated at 27°C for 5
days (Cheesbrough, 2006).
3.3.6 MOULD IDENTIFICATION
Mould cultures were prepared by lifting the mycelia mat of the organism with a sterile
inoculation pin into a drop of water on a slide (spreading the mat). It was then covered
with a cover slip and observed under a microscope. Different characteristic features of
the isolated organism were observed and used in their identification (Cheesbrough,
2006).
3.4 AFLATOXIN DETERMINATION (AOAC, 2008)
Peanut paste (25g) and 5g of sodium chloride were properly blended in a laboratory
blender for 2 min to obtain smooth and a homogenous mixture-using pause blending
to prevent overheating. The blended mixture was passed through a pre-folded filter
paper (Vicam fluttered) to obtain a clear filtrate. 15 ml of the filtrate was pipetted into
125 mL glass stopper Erlenmeyer flask and 30 mL of distilled water added. The flask
was closed using the stopper and mixed very well. The diluted filtrate was again
passed through a micro glass fiber filter paper for about 30 min to obtain a final and
clear filtrate. The filtrate was then passed through affinity column called the AflaTest
(Plate 3.2) which contained monoclonal antibody that are specific to aflatoxin B1, B2,
G1 and G2. aflatoxins were isolated, purified, and concentrated on the column using
70% CH3OH. 0.1ml of clear filtrate were then injected into the HPLC (Cecil HPLC)
through the chamber and were quantified by florescence measurement after reaction
19
with bromine solution as shown in plate 3.2. Individual and total aflatoxins were
quantified and recorded on the monitor. The HPLC has a column temperature of 40
degrees, mobile phase consist of 40% methanol, 60% water, 120mg of potassium
bromide and 350micrograms of nitric acid prepared in one liter. Column flow rate was
1ml/min, excitation and emission wavelengths were 360 and 435nm respectively,
whiles column types used was C18 (150mm/4.6mm/5micrometers). Limit of detection
and quantification were 0.5nanogram/gram and 1nanogram/gram respectively
Blending Filtration
Injection of peanut butter

Affinity column
sampl
Plate 3.2: Aflatoxin Analysis Procedure
3.5 STATISTICAL ANALYSIS
Statistical Package for Social Science (SPSS version 20.0) was used for Statistical
analysis. Analysis of variance (one way ANOVA) was used to determine whether the
values recorded on the various markets show any significant difference and one
sample t-test was used to compare variations between values recorded and standard
values.
20
CHAPTER FOUR
RESULTS AND DISCUSIONS
4.0 SURVEY RESULTS
The survey revealed, seventy five percent (75%) of peanut past producers in the study
were between the ages of 31 to 35 years and 25% between the ages of 20 to 25. All
the sellers interviewed, were females (Table 4.1) suggesting that, the peanut business
in the Northern parts of Ghana maybe considered, a female dominated activity with
production skills being passed down from mother to daughters. Although four percent
(4%) of the respondents acquired raw materials from their own harvest, approximately
ninety six percent (96%) of the peanut paste sellers acquired their raw materials from
the market already de-shelled, implying that peanuts may have already been exposed
to contamination even before processing. All the respondents were however, ignorant
of the peanut varieties they bought, an indication that there may be variation with
respect to organoleptic quality.
Table 4.1: Survey results obtained from the various peanut past producers
Frequency Percentage
Type of Mills Commercial 24 100.0
Sex/Gender Female 24 100.0
Age (years) 20 - 25 6 25.0
31 - 35 18 75.0
Education No Formal Education 24 100.0
Acquisition of Skills From Parent 24 100.0
Source of Peanut Own Farm 1 4.2
Market 23 95.8
Source of Water Pipe 2 8.3
Stream 18 75.0
Borehole 4 16.7
Transportation Bicycle 3 12.5
Donkey 3 12.5
Head 18 75.0
Sorting No 24 100.0
Shelflife 4 weeks and above 24 100.0
21
There was no sorting, grading or blanching before during processing, since this will
increase production cost. Seventy five percent (75%) used water from the stream to
wash their utensils during production, whiles 16.7% and 8.3% used borehole and pipe
water respectively. According to the respondents, every organism present in the water
will be killed during processing. All producers in these markets used commercial
mills to mill their products, since it is expensive to acquire a private mill. None of
them added any other ingredient during milling but water was added during milling to
help remove left overs of the paste from the mill. According to the sellers, the mills
were not cleaned between processing or batches. None of these producers had formal
education of any kind but had some knowledge on safety and hygiene. Seventy five
percent (75%) of the producers transport their product to the market on head, 12.5%
by donkey and 12.5% by bicycle respectively, this serves as their means of transport.
All the sellers packaged their product in plastic containers for display at the market.
This packaging method serves as means of advertisement.
4.1 PROXIMATE COMPOSITION
4.1.1 MOISTURE
The proximate compositions of the different peanut samples are presented on Table
4.2. The percentage moisture of the peanut paste samples generally ranged from 5.05
±0.07 to 6.45 ±0.21. This was similar to previous studies conducted by Özcan and
Seven (2007), who reported values of 4.5 to 6.06 ±0.18. These values however
contrasts reports made by Chang et al. (2013) who reported a value of 1%. The
contrast may be attributed to the varieties used as the starting material for the peanut
paste production, since different peanut varieties have different compositions of
moisture (Payman et al., 2011). Statistically, peanut paste samples from all the
sampling zones in this study, significantly varied from each other (p>0.05) except
22
samples from Navrongo central market and the Wa central market that showed no
significant difference. The variation, in addition to diversity in raw material variety
may also have resulted from the consistent addition of water during processing
(milling) to aid in the removal of the residue of samples from the mill. Peanut paste
samples from Bolgatanga central market with a moisture content of 5.05 ±0.07 may
have a longer shelf-life, due to its comparatively lower moisture content, which
implies less water for microbial activities and hence delay in food spoilage (Copetti et
al., 2011).
Table 4.2: Proximate composition of peanut paste samples
MARKETS % % CRUDE % CRUDE % CRUDE % ASH % TOTAL

MOISTURE PROTEIN FAT FIBRE CARBOHYDRTES
b a a a a
TcM 5.90±0.28 23.67±0.05 39.10±0.76 1.65±0.02 2.67±0.47 27.03±0.60a
TaM 5.25±0.21 b 30.73±5.25a 31.86±2.50 b 1.42±0.01a 3.10±0.01a 27.65±0.96a
WcM 6.45±0.21 a 31.56±0.78a 38.22±0.32a 1.13±0.01 b 3.21±0.28a 19.44±0.19a
b a a a a
WgM 5.85±0.21 30.99±0.18 39.52±0.26 1.09±0.01 2.66±0.62 19.90±0.77a
BcM 5.05±0.07 b 28.67±0.06a 36.68±0.05a 1.57±0.08 b 2.25±0.12a 25.80±0.26a
NcM 6.25±0.07a 26.16±1.15a 37.22±0.16a 1.89±0.01c 2.94±0.09a 25.55±0.27a
Control 5.45±0.35 b 26.48±0.12a 38.91±0.01a 1.27±0.01d 2.34±0.30a 25.57±0.18a
Key: TcM: Tamale central market, TaM: Tamale Aboabu Market,WcM: Wa central market,WgM: Wa
Gonomuni Market, BcM: Bolga central Market, NcM: Navrongo central Market,Control (Peanut paste
from Nkate-sari). Values are means ± standard deviation. Different Superscripts imply significant
difference at p<0.05
4.1.2 CRUDE PROTEIN
Protein content was highest in peanut paste bought from Wa central market with a
value of 31.56±0.78 and the least protein content from the Tamale central market
samples with a value of 23.67±0.05. This may imply that, peanut varieties used in the
production of peanut paste samples in Wa central market maybe a better protein
compliment that can be used to address protein deficiencies in that region. All the
samples however showed no significant difference (p<0.05). In addition, the samples
did not significantly vary from the control samples.
23
Protein content ranging from 23.67±0.05 to 31.56% ±0.78 shows that, protein values
falls within studies conducted by Boli et al., (2013) who reported values from 21 to
35% although differences in protein content may arise from differences in varieties
used in these areas or as a result of milling high protein foods such as fish or soybean
product before milling the peanut. People who require high protein may therefore use
peanut paste to compliment the inadequate proteins in their foods. Proteins available
to the body are used for the development of worn out tissues and body building
mechanism (FAO/WHO/UNU, 2007)
4.1.3 FAT
Percentage crude fat values ranged from 31.86±0.76 in the Tamale central market
(TcM) to 39.59±0.26 in the Wa gonomuni market (WgM), against the control (paste
from Nkate-sari) with a value of 38.91±0.01 (Table 4.2).
Although there were variations in the results from the different locations ranging from
31.86 ±2.50 to 39.72 ±0.26, only fat values of the Tamale aboabu market samples
showed significant difference (p<0.05) among the rest of the market samples,
including the control (butter from Nkate-sari). It was observed that, peanut paste
bought from the markets and the control were within the ranges of 24.55 to 50% as
reported by Özcan & Seven ( 2007).
The differences in some of the fat results obtained in the peanut paste samples may be
due to the varieties used (Dwivedi et al., 1993). It can therefore be recommended to
patients of higher energy requirement to use peanut paste from these sectors, since
they contain appreciable amount of fat. According to Vanessa., (2011), peanut fat
contains 9 kCal of energy per gram of fat and carbohydrate has 4 kCal per gram
carbohydrate.
24
4.1.4 FIBER
Percentage crude fiber content of the peanut paste samples shows least values
between 1.09 ±0.01 from Wa gonomuni market and highest values of 1.89 ±0.03 from
the Navrongo central market (NcM) samples. Tamale central market (TcM) samples
had the second highest fiber content of 1.65 ±0.02. Statistical analysis showed no
significant difference (p>0.05) within the Tamale aboabu market (TaM) samples,
Tamale central market (TcM) and Wa gonumuni market (WgM) samples. Also there
were no significant difference (p>0.05) between the Wa central market samples and
the Bolga central market samples but shows a significant difference (p<0.05) between
the control and the Navrongo central market samples respectively, as shown in Table
4.2.
The percentage crude fiber content of the samples obtained was relatively lower in
comparison with previous studies by Özcan & Seven., (2007), who reported values of
1.00 to 6.75% but was similar to that of the control sample. Though the fiber contents
of the samples varied from market to market, higher fiber values were obtained for
some of the peanut paste samples as shown in Table 4.2. Thus, peanut paste from
these markets may serve to contribute fiber to patients with fiber requirment (Boli et
al., 2013)
4.1.5 ASH
Percentage crude ash was determined; the analyzed data revealed that, Wa central
market (WcM) peanut paste had the highest ash content with 3.21% ±0.28. The values
recorded generally agreed with those reported by Özcan & Seven, (2007), who
recorded values of 1.86 to 5.5% Despite the fact that there were variation between
peanut paste samples bought from the various markets and the control (butter from
Nkate-sari), statistical analysis showed no significant difference (p >0.05). The
25
variations in ash values may have resulted from the wearing of the attrition mill
during milling of peanut.
High ash content in food is an indication of high minerals content, although it may
also be an indication of impurities in the samples (Ayoola et al., 2012).
4.1.6 CARBOHYDRATE
Carbohydrate content in the Tamale Abuabo (TaM) samples was higher than the rest
of the samples with a value of 27.65±0.96. Despite the fact that there were variations
in the carbohydrate values recorded in the samples as shown in table 4.2, statistical
analysis shows no significant difference (p>0.05). Carbohydrate content in some of
the samples were found to be higher than previous studies of 15 to 26% (Boli et al.,
2013). Low carbohydrate content may be due to the variety of raw peanut used for the
paste preparation (Asibuo et al., 2008). Peanut paste from these markets can be a
good source of carbohydrate for malnourished children, since carbohydrate is needed
by the body for proper cellular activity (Kunemund, 1988).
4.2 MICROBIAL LOAD
4.2.1 TOTAL AEROBIC COUNT
From Figure 4.1, Wa Gonomuni market (WgM) samples had the highest
contamination of 9.7×103 cfu/g, compared to Samples from the Tamale Aboabu
market (TaM) that had the lowest contamination level of 2.5×103 cfu/g. All the
samples were however, below the maximum acceptable limit of 104 (Council, 2009).
Bolga central market (BcM) samples and the control had no count. The absence of
these microbes in the Bolga central market suggests that the processing method, used
by the producers in that market was effective in the control of these microbes. In
26
addition Wa central market (WcM) and Wa gonomuni market samples significantly
varied (p>0.05) from the Bolga central market (BcM), Tamale Aboabu market (TaM),
Navrongo central market (NcM), and Tamale central market (TcM) samples. This
difference may be attributed to the difference in processing environments and
conditions of processing as revealed in the survey.
12
Total Aerobic Count (CFU/g x 103)
10
8
9.75e
6 9.70e
4
5.00d
2 3.95c
N.Da N.Da 2.50b

0
BcM Control Tam NcM TcM WcM WgM
Samples
Gonomuni Markt, BcM: Bolga central Market, NcM: Navrongo central Market, N.D: Non -detectable.
Different Superscripts imply significant difference at p<0.05
Figure 4.1: Total aerobic counts after 48 hours incubation
Lack of Good Manufacturing Practices (GMP) such as the addition of stream water
during processing may provide a favorable medium for microbial proliferation
(Copetti et al., 2011): the stream water used to wash the utensils during processing by
some of the producers may serve as a vehicle of contamination into the final product.
Also already de-shelled raw peanut, purchased from the market, may have already
been exposed to contamination that can survive the processing treatments (Copetti et
al., 2011).
27
4.2.2 TOTAL COLIFORMS
From figure 4.2, there were no coliforms detected in Bolga central market (BcM),
Navrongo central market (NcM), Wa central market (WcM) and the control (butter
from Nkate-sari) samples. Although Tamale central market (TcM) samples had
counts of 1.3×101, which is above the maximum acceptable limit of 101 cfu/g
(Council, 2009), Tamale Aboabo market (TaM) and Wa gonomuni market (WgM)
reported low counts below this limit. This was in agreement with Chang et al. (2013)
who also reported presence of coliforms in peanut butter samples but below the
maximum acceptable limit. Statistical analysis showed significant differences
(p<0.05) between the Tamale aboabu market samples and the Wa gonomuni market
samples, in contrast with samples from the Navrongo central market (NcM), Bolga
central market (BcM), Wa central market (WcM) and the control (paste from Nkate-
sari), which had no significant differences (p>0.05).
18
16
Total Coliform Count (cfu/g) x 101
14
12
10
8 1.35c
6 0.11c
4
2 0.50b
N.Da N.Da N.Da N.Da
0
NcM BcM WcM Control TaM WgM TcM
Samples
Gonomuni Market, BcM: Bolga central Market, NcM: Navrongo central Market, N.D: Non- detected.
Figure 4.2: Total coliform count after 48 hours of incubation
28
The differences may be attributed to variations in hygienic conditions, as total
coliform counts in food samples are indications of poor hygienic practices during and
after production on the part of the producer (Reij & Den Aantrekker, 2004).
According to the United States Drug Administration indicated that, total coliform
levels above 10 cfu/g may be a sign of high contamination which could lead to
diarrhea and other foodborne diseases (Council, 2009). The presence of coliform in
the peanut paste samples may be an indication of fecal contamination, which may also
have resulted from direct contact of the product with fecal matter, through packaging
to storage and sale at retailed markets since most of the coliforms are likely to have
been killed during processing (Smith & Stratton, 2007).
4.2.3 FUNGAL ENUMERATION
There was no fungal contamination in the Navrongo and the control samples. Even
though the remaining samples showed contaminations, they were all below the United
States Drug Administration (USDA) acceptable standard of 100cfu/g (Council, 2009).
Statistical analysis showed no significant difference (p>0.05) between the NcM,
WcM, the control, TcM and the BcM samples although there was significant
differences (p<0.05) between the WgM samples and the TaM samples as shown in
figure 4.3
29
5
4.5
Fungal Enumeration (CFU/g) x 101

4
3.5
3 4.50d
2.5
2 3.50c
1.5 2.50b 2.50b
1
0.5
N.Da N.Da N.Da
0
NcM WcM Control TcM BcM WgM TaM
Samples
Gonomuni Market, BcM: Bolga central Market, NcM: Navrongo central Market, N.D: Non -detected.
Figure 4.3: Fungal enumeration of peanut paste samples.
Lack of hazard analysis and critical control points (HACCP) programs, coupled with
inadequate understanding on Good Manufacturing Practices can lead to yeast and
moulds contaminations in food (Odu & Okonko, 2012). The mould contaminations
observed in some of the samples may have resulted from poor milling conditions,
using contaminated raw materials, lack of formal education, inadequate knowledge on
Good Manufacturing Practice and the use of stream water to wash utensils as reported
in the survey results.
4.2.3.1 MOULD IDENTIFICATION

Blastomyces Dermatitidis was identified in the peanut paste sample from Bolga
central market samples. Aspergillus parasiticus was also identified in samples from
Tamale central market, Wa central market and Wa Gonomuni market samples
respectively as presented in Table 4.3. However, the control, Tamale central market
(TcM), Navrongo central market (NcM) samples showed no contamination.
30
Table 4.3: Mould identified in peanut paste samples from selected markets
ORGANISMS ISOLATED WgM BcM TaM WcM TcM NcM CONTROL
Aspergilus parasiticus + - + + - - -
Blastomyces Dermatitidis - + - - - - -
TcM: Tamale central market, TaM: Tamale Aboabu Market,WcM: Wa central market,WgM: Wa
Gonomuni Market, BcM: Bolga central Market, NcM: Navrongo central Market, Control (paste from
Nkate-sari)
Blastomyces is a disease caused by fungus known as Blastomyces Dermatitidis
(Chapman, 2008). This disease results in chronic to acute respiratory tract infection
which usually occurs in people with low immune system (HIV infected patients) and
people with organ transplants (Chapman, 2008). Blastomyces Dermatitidis may
contaminate food products during processing, packaging and marketing (Chapman,
2008) when they are exposed to fungi. In plate 4.1, a market seller openly advertises
her product to buyers while ignorantly exposing the peanut paste to microbial
contamination.
This may be attributed to inadequate knowledge of the basic good food safety
principles which may bring about cross contamination during such exposure.
31
Plate 4.1: Peanut paste on display at the Bolgatanga central market
Source (Author, 2015)
4.3 AFLATOXIN
Figure 4.4 presents aflatoxin contamination levels of all the peanut butter samples.
Peanut paste samples from the Control, Navrongo central market (NcM ), and
Tamale aboabu market (TaM) showed aflatoxin contamination of 0 ppb, 0 ppb and
2.89 ppb respectively which was below the Codex acceptable limit of 20 ppb.
Although Bolgantaga central market (BcM) samples were below the Codex
acceptable limit of 20 ppb (Nigam et al., 2009), it risked being rejected by the
European Union as it exceeded their maximum acceptable limit of 4ppb (Holbrook et
al., 2008). All the samples, from the remaining markets were highly contaminated as
they all exceed (Figure 4.4) the maximum acceptable limit of both the United States
and European Union. The contamination was statistically significant (p<0.05), in
comparison with the non-contaminated samples. The results of the aflatoxin study was
in agreement with those reported by Siwela (2011): 51 ppb, 27 ppb, 99 ppb, 79 ppb, 3
ppb and 6 ppb. According to their report, the aflatoxin concentrations maybe
mitigated by roasting and blanching.
32
Gonomuni Market, BcM: Bolga central Market, NcM: Navrongo central Market, N.D: Non- detected
Figure 4.4: Aflatoxin contamination levels in peanut paste from the various
markets
High aflatoxin contaminations from some of the markets may be attributed to the use
of aflatoxin-contaminated raw materials and poor packaging and storage as suggested
by Ndung et al. (2013). This contamination may have come from the field during
production or during storage by the peanut paste producers (Dorner & Cole, 2002). It
was observed that, none of the producers from the markets wash their raw materials or
sorted low-grade peanuts, before processing. Contamination of peanut by Aspergilus
flavus and Aspergilus parasiticus during peanut processing maybe through improper
handling, cross contamination, use of contaminated utensils and packaging
(Diedhiou et al., 2012)
It was reported that, people living in rural areas of some West African countries are
highly exposed to appreciable levels of aflatoxin contamination through their daily
meals (Egal et al., 2005). This puts them at risk of contracting diseases like cancer
and other cardiovascular disease (Egal et al., 2005). Severe damage and cirrhosis of
33
the liver cells are some other effect of ingesting very high doses of aflatoxin (Wang &
Groopman, 1999). High levels of consumption of aflatoxin contaminated products
have been linked with the incidence of certain cancers that are very deadly to humans
(Dhanasekaran & Shanmugapriya, 2011). Major symptoms of aflatoxin health effect
involve immune system suppression and acute liver cirrhosis, (Beasley, 2011). Also,
aflatoxins are categorized as first class carcinogenic (International Agency for
Research on Cancer, 2002). aflatoxin exposure in people living with infectious
diseases such as hepatitis B/C, TB and HIV have led to high mortality rates as a result
of weakened immune systems (Turner et al.,2003). High temperatures during storage
of samples have also been noted to favor the growth of these fungi, and this is an
indication of improper storage of raw materials and the product (Lansden &
Davidson, 1983).
34
CHAPTER FIVE
CONCLUSION AND RECOMMENDATION
5.0 CONCLUSION
Producers of peanut paste in selected sampling zones of this study lack Good
Manufacturing Practices and a well-defined HACCP programs even though peanut
paste has economic value and health benefits to these inhabitants. Peanut paste from
these markets (TcM, TaM, WcM, WgM, BcM, NcM) and the control (paste from
Nkate-sari) were found to have relatively high fat, protein and ash content compared
to other studies.
Peanut paste samples from the selected areas showed some level of microbial
contamination, levels of contaminations are still within USDA acceptable standard.
Some of the peanut paste samples were however found to be unsafe, they contained
harmful moulds: Aspergilus parasiticus Blastomyces Dermatitidis. Peanut paste
samples from Wa guomani market, Tamale Aboabo market and Wa central market
were also found to contain aflatoxin, which are known to have serious health
implications upon exposure. There is the need for training programs for those in this
sector to build their capacity in food safety practices such as good manufacturing
practice and HACCP.
5.1 RECOMMENDATIONS
The varieties of peanuts used by the various producers should be investigated. Further
research should also be conducted on the different production methods in the various
research locations. Raw peanut samples from the producers may also be analyzed to
determine the base line contamination.
35
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45
APPENDICE
APPENDIX 1 QUESTIONNAIRE
KWAME NKRUMAH UNIVERSITY OF SCIENCE AND TECHNOLOGY
COLLEGE OF SCIENCE
DEPARTMENT OF FOOD SCIENCE AND TECHNOLOGY
QUESTIONNAIRE ON EVALUATION OF PEANUT PAST IN SELECTED
MARKETS IN NORTNERN GHANA
I am a student of KNUST conducting a research on the evaluation of peanut paste in
the three Northern Ghana (Northern, Upper west, Upper East) to ascertain the safety
of the product on local markets. Information received will be treated confidentially
and used for academic purpose only
46
(PEANUT PASTES SELLERS)
1. Location (market)……………………
2. Name of sample: ……………………
3. Sex of the peanut past seller
a. Male [ ] b. Female [ ]
4. Age
a. 20- 25 years [ ] b. 25-30 years [ ] c. 30-35 years [ ] d. 35 years and
above [ ]
5. 2. Level of education
a) JHS [ ] b) SHS [ ] c) Tertiary [ ] d) Nil [ ]
6. 3. Area of residence ……………………………………………….
7. Source of water
a) Pipe [ ] b) bore hole [ ] c) river [ ] d) stream [ ]
Production period
8. How long can the product stay without going bad?
a. 1-7 days [ ] b. 1-2 wks [ ] c. 3-4 wks [ ] d. above 4 wks [ ] e. I can’t
remember [ ]
Production process
9. How did you learn the method of production?
a. Formal education [ ] b. Apprentice [ ] c. From parent observation [ ]
10. Where did you acquire your raw peanut?
a. Own harvest [ ] b. Market [ ] c. Research institution
11. If from own harvest was it from
a. Storage [ ] b. Freshly harvested [ ]
47
12. How long had it been stored?
a.1mth [ ] b.2mths [ ] c.3mths [ ] d. more than 3 mths [ ]
13. Do you know the variety you use frequently?
a. Yes [ ] b. No [ ]
14. If yes what is the name of the variety? .............................
15. Do you sort or grade your peanut before cracking?
a. Yes [ ] b. 2 No [ ] c.
16. Do you wash the peanut before or after cracking?
a. Yes [ ] b. No [ ]
17. Do you sort or grade after cracking?
a. Yes [ ] b. No [ ]
18.How do you clean your utensils before using them?
a. Soap and clean tape water [ ] b. disinfectant and water [ ] c. I do not clean [ ]
19. How long after frying before you mill?
a.30 minutes [ ] b. after one hour [ ] c. 24 hours [ ] d. months [ ]
20. What type of mill do you use?
a.Commercial mills [ ] b. Personal mill [ ] c. 24 hours [ ] d. months [ ]
21. Do you clean your mills?
a.Yes [ ] b.No [ ]
22. If yes
how?.........................................................................................................
23. Apart from the peanut do you add any other ingredient during milling?
a.Yes [ ] b.No [ ]
48
24. If yes what are they?………………………………………………………….
25. How do you package the butter to the market after milling?..............................
26. How do you transport it to the market?
a. By carrying on the head [ ] b. on motor bike [ ] c. Bicycle [ ] d. car [ ]
e. Donkey [ ]
Comment
27. Have you any idea why you should produce the product under hygienic
conditions?
a. Yes [ ] b. No [ ]
28. If yes why?..........................................................................................................
29. Will your customers prefer a quality product at a higher cost?
a.Yes [ ] b. No [ ]
30. Can you differentiate between good paste and bad paste?
a.Yes [ ] b No [ ]
31. What are the signs (characteristics) of a bad peanut paste?
49
APPENDIX 2: FORMULAS USED IN CALCULATION
FORMULA USED IN MOISTURE CALCULATION
W1-W2 x100
% moisture = Weight of sample
W1 = Initial weight of crucible + Sample
W2 = Final weight of crucible + Sample
Note: Moisture free samples were used for further Analysis
(AOAC, 2003)
FORMULA USED FOR THE CALCULATION OF PERCENTAGE CRUDE
PROTEIN
1.4007 * c * (V - Vb)
%N=
Sample weight (g)
c: Concentration of the standard-acid solution: Hydrochloric acid 0.1N or c = 0.1
mol/l
Alternative: sulfuric acid 0.1N or c = 0.05 mol/l
V: Consumption of the standard acid in ml (Sample)
Vb: Consumption of the standard acid in ml (Blank Sample)
Protein (g per 100g) = % total nitrogen x appropriate nitrogen conversion
Factor (6.25) and results was reported in gram per 100 g of sample and to one decimal
place.
(AOAC, 2003)
FORMULA USED FOR THE CALCULATION OF PERCENTAGE CRUDE FIBER
% CRUDE FIBER BY WEIGHT= W1 - W2
50
Where W1 and W2 are the initial and final weight of the crucible and crucible plus
samples respectively in grams and W is the weight of the sample in grams
Test results were recorded in grams per 100 grams of sample and to one decimal
place.
(AOAC, 2003)
FORMULA USED TO CALCULATE FOR PERCENTAGE CRUDE FAT
Weight of ether extract x 100
%Crude fat = Weight of sample
(AOAC, 2003)
FORMULA USED FOR THE CALCULATION OF PERCENTAGE CRUDE ASH
%Ash= Difference in Weight of Ash x100
Weight of sample
(AOAC, 2003)
51

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KWAME NKRUMAH UNIVERSITY OF SCIENCE AND TECHNOLOGY,

DEPARTMENT OF FOOD SCIENCE AND TECHNOLOGY

EVALUATION OF PEANUT PASTE IN SELECTED MARKETS IN

(BSc Community Nutrition)

A Thesis submitted to the Department of Food Science and Technology, College of

Science in partial fulfilment of the requirements for the degree of

Masters of Science in Food Science and Technology

© 2015, Department of Food Science and Technology

me at the Food Science and Technology Laboratory of the Kwame Nkrumah

acknowledgement has been made in text

Yussif Abubakari ……………………… …………………

(Student) Signature Date

Prof. William O. ELLIS ……………………… …………………

(Supervisor) Signature Date

Prof. Mrs Ibok ODURO ……………………… …………………

(Head of Department, Supervisor) Signature Date

(PEANUT AND MYCOTOXIN INNOVATION LABORATORY PROJECT), staff

CHAPTER ONE .......................................................................................................... 1

CHAPTER TWO ......................................................................................................... 4

CHAPTER THREE ................................................................................................... 13

CHAPTER FOUR ...................................................................................................... 21

CHAPTER FIVE ....................................................................................................... 35

Table 2.1: Proximate composition of peanut paste ....................................................... 7

Figure 4.1: Total aerobic counts after 48 hours incubation ......................................... 27

Plate 2.1: Peanut paste on display at the market ............................................................ 8

University of Science and Technology. A survey was conducted using structured

questionnaires to understand the processing methods used. Proximate, aflatoxin and

19.44 ±1.19 to 27.65 ±0.96 respectively. Statistical analysis showed no significant

market samples. Aspergilus parasiticus was isolated in Wa gonomuni, Wa central

market and Tamale Aboabu market samples respectively. Blastomyces Dermatitidis

nutrient composition, contamination levels were significant due to poor production

1.0 BACKGROUND OF STUDY

women in Northern Ghana (Millar & Yeboah, 2006).

pleasant flavor (Chow, 2007).

incidence of coronary heart diseases by 25 to 50 %. Consumption of peanut snacks

frequently or incorporated daily into meals (Jiang et al., 2002).

prevalence of microbial and toxic metabolite contamination. This has particularly

flavus, a major producer of aflatoxins (Narasimhulu, 2007). Aflatoxins contamination

production are lost due to aflatoxin contamination, though other approaches to

(Otsuki et al., 2001). According to Dhanasekaran & Shanmugapriya., (2011), high

class carcinogens in humans (International Agency for Research on Cancer, 2002)

to consumers. In addition, there is limited data on the effects of peanut processing on

the quality of the kernels and its products

safety decision-making. Data on proximate composition of these peanut pastes will

serve as useful information for consumers.

1.4 MAIN OBJECTIVE

East, Upper West and Northern Region)

1.5 SPECIFIC OBJECTIVES

1. To determine the nutrient content of peanut paste sold on Northern sector

Northern sector market

mature in the soil (Martinez, 2014).

which can be destroyed during extreme heating, it contains an average of 5 to 7%

moisture in freshly harvested peanut and ash of about 3% (Woodroof, 1989).

the highest stability of the peanut oil (Aoyagi, 2015).

high temperature of roasting and high fiber content (Woodroof, 1989).

used either as animal feed or as fertilizers

2.1.1 PEANUT BISCUIT

production and consumption as compared to other bakery products (Gandhi et al.,

et al., 1995). According to Sadaf et al. (2013), although carbohydrate composition