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Process Engineering Thesis

2021-10

EFFECT OF TYPES OF CEREALS ON

QUALITY OF ETHIOPIAN
TRADITIONAL BEVERAGE (TELLA)

MINBALE, GASHU

http://ir.bdu.edu.et/handle/123456789/14374
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 BAHIR DAR UNIVERSITY
BAHIR DAR INSTITUTE OF TECHNOLOGY
SCHOOL OF RESEARCH AND GRADUATE STUDIES
FACULTY OF CHEMICAL AND FOOD ENGINEERING

PROCESS ENGINEERING
MSc THESIS ON:
EFFECT OF TYPES OF CEREALS ON QUALITY OF ETHIOPIAN
TRADITIONAL BEVERAGE (TELLA)

BY: MINBALE GASHU

OCTOBER, 2021
BAHIR DAR, ETHIOPIA
 BAHIR DAR UNIVERSITY
BAHIR DAR INSTITUTE OF TECHNOLOGY
FACULTY OF CHEMICAL AND FOOD ENGINEERING

Effect of types of cereals on Quality of Ethiopian Traditional Beverage

(Tella)

a Thesis Submitted in Partial Fulfillment of the Requirements for the Degree

of Master of Science in Process Engineering

Advisor: Dr. Metadel Kassahun (PhD)

October, 2021
Bahir Dar, Ethiopia

©2021Minbale Gashu

ii
 Approval of Thesis for defense
I hereby certify that I have supervised, read, and evaluated this thesis titled “Effect of
types of cereals variation on Quality of Ethiopian traditional Beverage (tella)” Prepared
by Minbale Gashu under my guidance. I recommend the thesis to be submitted for oral
defense.
Dr. Metadel Kassahun
Advisor’s name
____________________
Co-Advisor’s name Signature Date

i
 Approval of Thesis for defense result
I hereby confirm that the changes required by the examiners have been carried out and
incorporated in the final thesis.

Name of Student: Minbale Gashu Signature: Date; October 10/2021

As members of the board of examiners, we examined this thesis entitled "Effect of

types of cereals Variation on Quality of Ethiopian traditional Beverage (tella)" by
Minbale Gashu. We hereby certify that the thesis is accepted for fulfilling the
requirements for the award of the degree Of Masters of Science in Process
Engineering

Board of Examiners

Name of Advisor

Dr. Metadel Kassahun

Name Of External examiner

Dr. Beteley Tekola

Name Of Internal Examiner
Dr. Nigus Gabbiye

Name Of Chairperson

Dr. Solomon Workneh

Name of Chair Holder

Mekuanint Demeke

Name of Faculty Dean

Dr. Metadel Kassahun

ii
 Acknowledgments
I would like to express my deepest gratitude to Bahir Dar University, Faculty of
Chemical and food engineering, Bahir Dar institute of technology, for creating this
opportunity to conduct research. A very special gratitude goes out to my advisor Dr.
Metadel Kassahun for his unreserved, all rounded support and enriching comments
during the course of my study.
Together with this my sincere appreciation goes to Dr.Nigus Gabbiye for his invaluable
and critical comments during the course of my study.
Then, I would like to thank you to Ethiopian conformity assessment enterprise and Bless
Agri Food Laboratory for allowing me to do my research in their laboratory. Last but not
least, I am grateful to thank my family for their encouragement throughout the study.

iii
Table of Contents
Approval of Thesis for defense ............................................................................................ i

Approval of Thesis for defense result ................................................................................. ii

Acknowledgments.............................................................................................................. iii

Declaration ........................................................................................................................ vii

List of Tables ..................................................................................................................... ix

List of Figures .................................................................................................................... ix

Abstract .............................................................................................................................. xi

Chapter one ......................................................................................................................... 1

1. Introduction ..................................................................................................................... 1

1.1. Background .............................................................................................................. 1

1.2. Statement of the problem ......................................................................................... 3

1.3. Objective .................................................................................................................. 4

1.3.1. General objective ............................................................................................... 4

1.3.2. Specific objectives ............................................................................................. 4

1.4. Significant of the study ............................................................................................ 4

1.5. Scope of the study .................................................................................................... 5

Chapter Two........................................................................................................................ 6

2. Literature Review............................................................................................................ 6

2.1. Ethiopian traditional beverage (Tella)...................................................................... 6

2.2. Tella fermentation .................................................................................................... 7

2.3. Quality of tella ........................................................................................................ 11

2.4. Comparison of Ethiopian tella with beer................................................................ 12

2.5. Ingredients of Ethiopian traditional beverage; tella ............................................... 13

2.5.1. Gesho, Rhamnus prinoides L. Herit ................................................................ 13

iv
 2.5.2. Bikil ................................................................................................................. 14

2.5.3. Grawa (Vernonia amygdalina Del.)................................................................. 15

2.5.4. Weira (Olea europaea L. sub sp cuspidate Ciferri) ......................................... 16

Chapter Three.................................................................................................................... 17

3. Materials and Methods .................................................................................................. 17

3.1. Sample Preparation ................................................................................................ 17

3.2. Chemical and reagents ........................................................................................... 18

3.3. Apparatus and instruments ..................................................................................... 19

3.4. Preparation of Ethiopian traditional beverage; tella ............................................... 19

3.5. Proximate analysis of cereals ................................................................................. 20

3.6. Determination Quality content of malting barley .................................................. 21

3.6.1. Determination of moisture of malting barley .................................................. 21

3.6.2. Determination of nitrogen and protein content of malting barley ................... 22

3.7. Effect of cereals variation on Fermentation process of Ethiopian traditional

beverage (tella) .............................................................................................................. 23

3.8. Effect of cereals variation on Quality of Ethiopian traditional beverage (tella) .... 24

3.8.1. Evaluation of pH value .................................................................................... 25

3.9. Effect of cereals variation on Shelf life of Ethiopian traditional beverage (tella) . 25

3.10. Comparison of Ethiopian traditional beverage tella against St. George beer ...... 26

3.11. Experimental Design ............................................................................................ 26

3.12. Statistical analysis ................................................................................................ 27

Chapter Four ..................................................................................................................... 28

4. Results and Discussion ................................................................................................. 28

4.1. Proximate analysis of different cereals .................................................................. 28

4.1.1. Proximate analysis of barley............................................................................ 28

v
 4.1.2. Proximate analysis of corn .............................................................................. 29

4.1.3. Proximate analysis of wheat ............................................................................ 30

4.1.4. Proximate analysis of sorghum........................................................................ 30

4.1.5. Proximate analysis of teff ................................................................................ 31

4.2. Determination of Quality of malting barley ........................................................... 33

4.3. Effect of cereals variation on fermentation process of Ethiopian traditional

beverage (tella) .............................................................................................................. 33

4.3.1. Determination of Original wort extract (OE), Real extract (RE) and apparent
extract ........................................................................................................................ 33

4.3.2. Determination of real degree of fermentation ................................................. 35

4.4. Effect of cereal variation on Quality of Ethiopian traditional beverage (tella) ...... 37

4.4.1. Determination of Ethanol ................................................................................ 37

4.4.2. Determination of pH value .............................................................................. 41

4.4.3. Determination of Relative density ................................................................... 43

4.5. Effect of cereals variation on Shelf life of Ethiopian traditional beverage (tella) . 44

4.5.1. Determination of Real degree of fermentation ................................................ 44

4.5.2. Determination of Ethanol ................................................................................ 46

4.5.3. Determination of pH value .............................................................................. 48

4.6. Comparison of traditional beverage tella against beer ........................................... 50

4.6.1. Determination of real degree of fermentation ................................................. 50

4.6.2. Determination of Ethanol ................................................................................ 51

4.6.3. Determination of pH ........................................................................................ 52

Chapter Five ...................................................................................................................... 53

5.1Conclusion ................................................................................................................... 53

5.2 Recommendation ........................................................................................................ 53

References ......................................................................................................................... 54

vi
Declaration
This is to certify that the thesis entitled “Effect of types of cereals on Quality of Ethiopian
Traditional Beverage (tella)”, submitted in partial fulfillment of the requirements for the
degree of Master of Science in Process engineering under Faculty chemical and food
engineering, Bahir Dar Institute of Technology, is a record of original work carried out
by me and has never been submitted to this or any other institution to get any other
degree or certificates. The assistance and help I received during the course of this
investigation have been duly acknowledged.

vii
Abbreviations

%: Percent ......................................................................................................................... 22
°C: Degree centigrade ....................................................................................................... 22
AE:Apparent extract ......................................................................................................... 23
BC:Before century .............................................................................................................. 1
CES: Conformity Ethiopian standards .............................................................................. 26
CFU: Colony forming units .............................................................................................. 11
CuSO4: Copper sulphate ............................................................................................. 18, 22
ECAE: Ethiopian conformity assesment enterprise .......................................................... 40
H2SO4: Sulphuric acid ....................................................................................................... 22
K2SO4: Potassium sulphate ......................................................................................... 18, 22
L:Liter ................................................................................................................................. 2
ml: Mililiter ....................................................................................................................... 11
NaOH: Sodium hydroxide .......................................................................................... 18, 22
OE: Original wort extract.................................................................................................. 23
RE; Real extract ................................................................................................................ 23
V/V: Volume by volume ................................................................................................... 12
WHO: World health organization ....................................................................................... 2

viii
List of Tables
Table 2.1 Changes in total aerobic count, moisture content and pH occurring during
fermentation of tella. ......................................................................................................... 11
Table 2.2 Quality report variation of Ethiopian traditional beverage tella in the precious
studies ............................................................................................................................... 12
Table 3.1 Experimental design of fermentation process and quality of Ethiopian
traditional beverage ........................................................................................................... 26
Table 3.2 Experimental design of shelf life of Ethiopian traditional beverage ................ 27
Table 4.1 Proximate analysis result of barley ................................................................... 28
Table 4.2 Proximate analysis result of corn ...................................................................... 29
Table 4.3 Proximate analysis result of wheat ................................................................... 30
Table 4.4 Proximate analysis result of sorghum ............................................................... 31
Table 4.5 Proximate analysis result of teff ....................................................................... 32
Table 4.6 Quality analysis result of malting barley .......................................................... 33
Table 4.7 Original wort extract, real and apparent extract results of Ethiopian traditional
beverage tella .................................................................................................................... 34
Table 4.8 Real degree of fermentation (%) of Ethiopian traditional beverage tella ......... 35
Table 4.9 Analysis result of real degree of fermentation using one way ANOVA .......... 36
Table 4.10 Comparison result of real degree of fermentation of tella .............................. 37
Table 4.11 Ethanol content of Ethiopian traditional beverage tella.................................. 38
Table 4.12 Analysis results of Ethanol using one way ANOVA ...................................... 40
Table 4.13 Comparison results of Ethanol content of tella ............................................... 40
Table 4.14 pH result of Ethiopian traditional beverage tella ............................................ 41
Table 4.15Analysis result of pH value using one way ANOVA ...................................... 42
Table 4.16 Comparison result of pH value of tella ........................................................... 43
Table 4.17 Relative density of Ethiopian traditional beverage tella ................................. 44
Table 4.18 Real degree fermentation results of Ethiopian traditional beverage tella ....... 44
Table 4.19 Analysis result of real degree of fermentation using Factorial ANOVA ....... 46
Table 4.20 Ethanol content of Ethiopian traditional beverage tella.................................. 46
Table 4.21Analysis result of Ethanol using Factorial ANOVA ....................................... 48
Table 4.22 pH value of Ethiopian traditional beverage tella ............................................ 48

ix
Table 4.23 Analysis result of pH value using Factorial ANOVA .................................... 50
Table 4.24 Comparison of RDF against St.George beer .................................................. 50
Table 4.25 Comparison ethanol result of against St.George beer..................................... 51
Table 4.26 Comparison of pH value against St.George beer ............................................ 52

x
List of Figures
Figure 2.1 Flow chart of traditional preparation of tella .................................................... 8
Fig.2.2 From the left, powdered leaves, leaves, and stem shreds of gesho ....................... 9
Figure 2.3 Tella made with toasted barley flour after 1 week of fermentation ................ 10
Figure 3.1Ttraditionaly roasted different cereals .............................................................. 17
Figure 3.2 Sun light drying of barley malt (bikil) ............................................................. 17
Figure 3.3 Sun drying of gesho leaves and stem shreds ................................................... 18
Figure 3.4 Preparations of Ethiopian traditional beverage tella using different cereals. .. 20
Figure 3.5 Testing pH value of Ethiopian traditional beverage tella ................................ 25
Figure 4.1 Comparison of crude fat result of cereals ........................................................ 34
Figure 4.2 Comparison of original wort extract................................................................ 36
Figure 4.3 Comparison of carbohydrate contents for cereals and their RDF ................... 39
Figure 4.4 Comparison of crude protein for various cereals ............................................ 42
Figure 4.5 Effect of cereals variation and fermentation time on degree of fermentation . 45
Figure 4.6 Effect of cereal variation and fermentation time on ethanol ........................... 47
Figure 4.7 Effect of cereals variation and fermentation time on pH ................................ 49

xi
Abstract
Previous studies have presented the existence of degree of fermentation, quality and shelf
life problems upon Ethiopian traditional beverage tella, and adversely affect the
industrialization of tella in the beverage industry. In this study, Ethiopian traditional
beverage tella was brewed in Ethiopian conformity assessment enterprise laboratory
considering variation of cereals such as barley, corn, wheat, sorghum and teff to find the
best cereals with respect to degree of fermentation, quality and shelf life. The study
revealed tella prepared from teff demonstrated optimum degree of fermentation (%
60.32) and then produced higher ethanol content (%V/V 5.43) relative to others.
Analyses of ethanol and pH values of the beverage were carried out after 20 days of
storage with the intention of finding the optimum cereal regarding on shelf life.
Therefore, traditional beverage prepared from wheat withstands the declining of pH value
(3.43) within 20 days of storage compared with the others. Hence, tella prepared from
wheat is the optimum cereal regarding on shelf life of the beverage. In the study, cereal
variations significantly affect (P ≤ 0.05) RDF (%), ethanol (%V/V) and pH value of
Ethiopian traditional beverage tella.

Key words: Cereal, bikil, gesho, traditional beverage (tella), RDF (%), Ethanol (% V/V)
and pH value.

xii
 Chapter one

1. Introduction

1.1. Background
Alcoholic beverages are part of human dietary culture and have an inseparable
relationship with the life of mankind in history. The making and drinking of alcoholic
beverages are ways of enhancing the nutritional significance as well as social
relationships for human beings. Exactly when mankind started to produce and consume
alcoholic beverages is not known but beer is known to have been produced by the
Sumerians before 7,000 BC (Dufour J, Verstrepen K and Derdelinckx G, 2003.), while
wine has an unequivocally recorded history stretching back nearly 6,000 years, with the
earliest evidence dating between 5,400 and 5,000 BC (Soleas GJ, Diamandis EP and
Goldberg DM 1997). Almost all countries and regions all over the world have traditional
alcoholic beverages, which utilize indigenous agricultural products. Like in other parts of
the world, alcohol has been used in Africa for centuries and played an important role in
the culture and local economy. Given the abundance of raw materials, varieties of
fermented drinks are produced and consumed for various reasons including component of
standard diet, a means to settle disputes, facilitating marriages, to hold festivities after a
successful hunt or harvest, and for hygienic or medicinal reasons (Platt, 1955). In Africa,
fermented alcoholic beverages are consumed in occasions such as marriage, naming and
rain making ceremonies (Zvauyaet al., 1997), at festivals and social gatherings, at burial
ceremonies and settling disputes (Steinkraus, 1983). They are also used as medicines for
fever and other ailments by adding barks or stems of certain plants (Okafor, 1972).
Fermented beverages produced from cereals usually referred to as beers while those
produced from fruits are classified as wines (Pederson, 1979). Traditional recipes are
handed down through generation and are still used for food processing in many
developing countries (Kebede et al., 2002). The traditionally fermented beverages are
low-cost product in all aspect as they are usually manufactured using only rudimentary
equipment. Because of their cheapness, low income groups mostly consume them. Home-
brewed beverages have several names mostly reflecting the areas where they are

1
produced but most of them are produced in almost similar ways by distillation and
fermentation of grain cereals, fruits and/or vegetables (Shale et al., 2014).

Some of the most culturally important traditional beverages in Africa are: `tella` in
Ethiopia (Shale et al., 2014) ` thobwa` in Malawi (Matumba et al., 2011), `burukutu` in
Nigeria (Sawadogo-Lingani et al., 2010) `tonto` in Uganda (Mwesigye et al., 1995) and
`muratina` in Kenya (Aka et al., 2014). These drinks are very popular, perhaps the most
widely consumed beverage types because of the nutritional, therapeutic, social and
religious values attached to them (Solange, 2014). Indeed, studies also confirmed that
fermented products in general can improve sensory qualities and nutritional values by
enriching the product with essential proteins and vitamins (Steinkraus, 1986).

Furthermore, home brewed beverage production and local trade have also provided
livelihood for many urban and rural poor, particularly women, for whom it is often the
primary means of supporting their families (Adelek an et al., 2008).

Ethiopia is a country rich in history, ethnicity, and cultural and crop genetic diversity.
Traditional alcohol beverages are widely consumed among different ethnic groups of the
country as a prominent part of the local traditions of major social events including public
holidays, wedding, funerals and other forms of festivities. The WHO global status report
on alcohol and health released in 2014 showed that in Ethiopia the volume of unrecorded
alcohol consumption is estimated to be 3.5 L of pure alcohol per capita indicating the
highest consumption rate compared to other African countries such as Nigeria (1
L),Uganda (1.5 L) and Angola (1.6 L) (WHO, 2014). Its production depends on the
naturally present micro flora in the substrates, utensils, and equipment used, and the
environment of the households (Ashenafi M 2002).
However, Ethiopian traditional fermented products are not sufficiently studied. There are
quite few works on Ethiopian traditional fermented beverages including data on fuel oil
and alcohol content of Tej (Bekele, 2001). The ethanol level, pH and sensory, evaluation
of Tej, Areki and Tella were also reported by (Yohannes et al. 2013).
Alcoholic beverages can be classified into three main categories: wines, beers, and
spirits. This classification is based on production methods: (Dufour J, Verstrepen K and
Derdelinckx G. 2003) by mono-fermentation; (Soleas GJ, Diamandis EP and Goldberg

2
DM 1997) by malting and fermentation; and (Tamang JP.; 2010) by distillation after
fermentation. Yeasts can produce ethanol primarily through metabolism of the low-
molecular weight sugars that can be transported into the cell cytoplasm. However, in
production processes utilizing cereals or tubers, fermentations must be preceded by
depolymerization of storage polysaccharides and proteins yielding the sugars and amino
acids that can be utilized by the microorganisms (Paterson A, Swanston JS and Piggott
JR. 2003).

In Ethiopia, however, traditional beverage tella is consumed widely; there

industrialization is critically limited in the country. Few studies upon tella reported that
there is low degree of alcoholic fermentation in the traditional fermentation process
(Mooha Lee a *. M., 2015). (Tadele Yohannes, 2013). In the study, low quality of tella
was stated and varied within a given locality. In addition, short shelf life is one of the
crucial factors that influence the development of tella in the country (Weber et al., 2008).

1.2. Statement of the problem

Though, Ethiopia is well known for brewing tella for a long time, tella is not on the
market as a commercial product, but it is produced and sold domestically. There are
essential points which need researchers’ concentration to improve Ethiopian traditional
beverage tella in the beverage industry. Some of them are low degree of alcoholic
fermentation, Low quality content and alcoholic variations from place to place as well as
short shelf life of tella is the repetitive draw backs which violently affects the
modernization of tella in the beverage industry.
Importantly, the quality of tella is studied in different times by taking samples from
vendors and the alcoholic content of the beverage is varied from 2.2 to 6.65 and also the
pH results are from 3.28 to 4.99. On top of high range variation upon the reported
alcoholic and pH contents, the study doesn’t answer which cereals generate the reported
quality result. Since researchers used tella samples from vendors in different areas the
analysis result doesn’t demonstrate the source of cereals employed during the preparation
which may be the cause of low quality content and large range of variation.
According to the study, pH and ethanol content are in the range of 4.5-4.8 and 2.8-5.0%
(v/v) respectively when tella is considered to be the most suitable for consumption (Sahle

3
and Gashe, 1991). After ten days of fermentation, tella becomes too sour to consume due
to the growth of Acetobacter spp. which convert ethanol to acetic acid under aerobic
conditions. The quality of tella may be deteriorated and affected by contaminants such as
fungi and different types of bacteria. These organisms fasten shelf life of tella, and can
produce different undesired byproducts.
This thesis entitled as “Effect of types of cereals variation on Quality of Ethiopian
traditional beverage (tella)” is designated to study the effect of cereals variation on
fermentation process, quality and shelf life of tella by preparing tella in the laboratory
from different cereals. As a result of the study, tella brewers may choose the optimum
cereal in terms of degree of fermentation, quality, and shelf life during their preparation.
On the other hand, the study may serve as a base for researchers who are interested to
study the development of Ethiopian traditional beverage tella in the brewing industry.

1.3. Objective

1.3.1. General objective

➢ To study the Effect of cereals variation on Fermentation process, Quality and
Shelf life of Ethiopian traditional beverage; tella.

1.3.2. Specific objectives

1. To study the proximate analysis of different cereals and Quality of malting

barley to be used for Ethiopian traditional beverage tella.
2. To study the effect of cereal variation on Fermentation process, Quality
and Shelf life (in terms of pH) of Ethiopian traditional beverage tella.
3. To compare traditional beverage tella against standard beer

1.4. Significant of the study

This study realized the impact of cereals variation on Fermentation process, Quality and
Shelf life of Ethiopian traditional beverage tella; and will provide information for tella
brewers to choose optimum cereals for good tella preparation. Additionally, these
findings serve as a base for researchers who are interested in improving the quality as
well as the process of traditional beverage tella from small scale to large industrial scale.

4
1.5. Scope of the study
This thesis was designated to study the effect of cereals variation on fermentation
process, quality and shelf life of Ethiopian traditional beverages; tella by employing malt
(bikil) prepared from barley as common input for all types of trials prepared from barley,
corn, wheat, sorghum and teff. The study was started by testing nutritional content of
cereals and quality of malting barley. The optimum fermentable cereal was identified by
using degree of fermentation as a key parameter. Finally, the effect of types of cereals on
quality and shelf life of Ethiopian traditional beverage tella was evaluated by employing
ethanol and pH value respectively.

5
 Chapter Two

2. Literature Review
2.1. Ethiopian traditional beverage (Tella)
Ethiopia is one of the countries where a wide variety of traditional fermented beverages
are prepared and consumed. The various traditional fermented beverages are produced on
a small scale and usually for local consumption. Indigenous Ethiopian fermented
beverages include tej (Vogel &Gobezie, 1983; Fiteet al., 1991), tella (Sahle &Gashe,
1991), borde and shamita (Ashenafi& Mehari, 1995; Bacha et al., 1998, 1999). In
everyday life people enjoy fermented beverages and particularly when having guests,
they like to treat them to traditional alcoholic beverages. Among various Ethiopian
fermented alcoholic beverages, tella has many varieties in the various regions. Depending
on the type of cereal ingredients used to make, it has different names: Amhara tella,
Oromo tella, and Gurage tella (Fite et al., 1991). Amhara tella has gesho
(Rhamnusprinoides) and concentrated. Gurage tella is delicately aromatized with a
variety of spices. Oromo tella has no gesho (R. prinoides), and it is thick and sweet
(Vogel and Gobezie, 1983).
Tella is made from different cereals. The most popularly used grains are Teff and corn,
however in some areas barely, millet or sorghum can be used (Selinus, 1971). Tella is
widely brewed and consumed in both rural and urban part of Ethiopia. It is well known as
local beer (Getahun, 1976) since it is malt based beverage like that of commercial beer
(Shale and Gashe, 1991; Kunze, 1996). Tella brewing requires largely malted barley
(Hordeumvulgars), Enkuro, gesho (Rhamnusprinoides) leaves and stems chopped to
small pieces, kita (from different grains) or enkuro and derekot (Shale and Gashe, 1991).
Addition of malt and gesho which has the same function as hops in a beer. The main
fermentation yeast is Saccharomyces cerevisiae and saccharification of cereal starch
seems to depend on malt. Fermentation of Tella like other traditionally fermented
alcoholic beverages relies on the microorganisms present in the substrates, fermentation
vats, or equipments. So, with the variable micro flora of such spontaneous fermentation
variability of the product is imminent. Ethiopian indigenous fermented beverages are

6
products of acid-alcohol type of fermentation. The preparation of many indigenous or
traditional fermented beverages is still a household art. As the fermentation proceeds and
the environment becomes more and more acidic, yeasts and lactic acid bacteria dominate
the fermentation. These two groups of microorganisms usually determine the alcohol
content and flavor of the final product.

2.2. Tella fermentation

Tella has various vernaculars in the various regions and is a malt beverage based on
substrates such as barley, wheat, maize, millet, sorghum, teff or other cereals. It is, by far,
the most commonly consumed alcoholic beverage in Ethiopia (Ashenafi 2006]).
According to Samuel Sahle and Berhanu Abegaz Gashe (1991), over 2 million hectoliters
of tella is thought to be produced annually in households and tella vending houses in
Addis Ababa.
The way of preparing tella differs between the ethnic groups and depends on tradition
and the economic situation. Although the basic processing steps are similar, every tella-
maker seems to have own recipe. The clay container (insera) is washed with water and
fresh leaves of grawa (Vernonia amygdalina) several times. The well-cleaned container
is then inverted over smoking splinters of weyra (Olea europaea) for about 10 minutes.
This will eliminate microorganisms sensitive to antimicrobial components of wood
smoke. It also contributes to the desirable flavor of the fermented product.

7
 Sleeping barley Germination or malting

Milling Kilning

Mash Mixer Fermented tella

Figure 2.1 Flow chart of traditional preparation of tella (Lemi, 2020)

To make bikil (malt), grains of barley or wheat are moistened while in a container and
left to germinate for about three days. And this is finally sun-dried. Bikil is the source of
amylase for the fermenting cereals used in tella preparation. The gesho plant
(Rhamnusprinoides), which is different from hop (Humulus lupulus) is widely cultivated
in Ethiopia and is available dried in the local market. Although gesho may have
antibacterial effect against some groups of bacteria, its main purpose in the process is to
impart the typical bitter taste to tella. Gesho is responsible for the bitter taste of tella. It is
also thought to be the source of various chemicals (Sahle and Gashe, 1991; Kebede,
1994). It is assumed that gesho maintains acidic pH during tella fermentation to modify
the nature of the mash and impedes the growth of unwanted microorganism (Kebede,
1994).

8
The fermentable grains for tella preparation are usually prepared in two forms. Flours of
millet, barely or teff (dark variety) are toasted, milled, mixed in water and baked on a
wide metal pan into kita (unleavened bread). The kita is broken into small pieces. Barley
flour is separately toasted on a metal pan sprinkling water on it during toasting until it
turns dark brown. This is called enkuro. The color of tella, which may vary from light
yellow to dark brown, is determined by the extent of baking the kita or toasting the
enkuro.
Samuel Sahle and Berhanu Abegaz Gashe (1991) described the processes and
microbiology of tella fermentation. The fermentation is divided into four phases. During
the first phase, powdered leaves of gesho are mixed with water in a small earthen pot and
allowed to ferment for four to five days. The fermenting material is commonly called
tinsis. This is transferred to a large earthen pot and the second stage begins by mixing it
with barley malt, pounded stems of gesho, pieces of kita and water. This is left to ferment
for two more days.

Fig.2.2 From the left, powdered leaves, leaves, and stem shreds of gesho (Mooha Lee a *.
M. 2015)
During the third stage, chopped pounded stems of gesho, bikil, enkuro and water are
added to the container and the contents are mixed into thick slurry called difdif. This is
also allowed to ferment for two more days. At the final stage, the container is filled with
water to the brim and the contents are again mixed thoroughly. The container is then
sealed to create anaerobic conditions and left to ferment for two days or more.

9
Figure 2.3 Tella made with toasted barley flour after 1 week of fermentation (Mooha Lee
a *. M.2015)
Tella is consumed directly or after filtration. At the end of the fermentation, most
suspended materials settle to the bottom of the container. The clear liquid is tella. In
general, about 1 kg of gesho (leaves and pounded stems), 0.5 kg of bikil, 15 kg of grains,
in the form of kita (5 kg) and enkuro (10 kg) are mixed with 30 liters of water to prepare
tella.
When the clear tella is completely decanted from the sediment, fresh water is added to
the sediment and mixed well. This is left to ferment. The resulting beverage is known as
kirari and is weaker than the regular tella. It is most often used for family consumption,
and sometimes is given to children. The better quality is often kept for guests.
Sometimes, at the end of the third stage, a smaller volume of water is mixed with the
difdif and a more concentrated tella is obtained by filtering the difdif through a cotton
cloth and keeping it in a closed container. Such tella is known as filtered tella. Samuel
Sahle and Berhanu Abegaz Gashe (1991) reported that the first phase was important to
extract the components of gesho. The liquid at this stage was very dark in color with a
strong bitter taste. The microbial count increased markedly towards the end of the phase
and reduction in content of total carbohydrate and reducing sugar occurred. The
microbial flora consisted of molds, Lactobacillus spp. and other bacteria. Molds
disappeared, however, towards the end of the phase.

10
Table 2.1 Changes in total aerobic count, moisture content and pH occurring during
fermentation of tella (Tafere, 2015).

Phase count Fermentation pH Moisture Total aerobic

CFU*/ml
I (0-4 days) 0 5.29 5.6 2*103
1 5.29 5.6 4*103
2 5.19 5.6 1*104
3 5.09 4.8 2*106
II (4-6 days) 4 4.79 4.5 3*106
5 4.68 3.1 3*107
III (6-8 days) 6 4.88 4.8 7*107
IV (8-12 days) 8 4.67 4.0 1*108
10 4.59 3.4 1*108
12 3.99 6.6 9*107

The fermenting organisms were composed of Saccharomyces spp., (mostly S.

cerevisieae) and Lactobacillus spp. (mostly Lactobacillus pastorianumi). The yeasts
dominated the fermenting flora after the end of the first stage till the completion of
fermentation. Increase in alcohol content was accompanied by yeast growth and decrease
in reducing sugars and total carbohydrates. After ten days of fermentation, tella becomes
too sour to consume due to the growth of Acetobacter spp. which convert ethanol to
acetic acid under aerobic conditions.

2.3. Quality of tella

The biochemical changes, the microorganisms involved in the fermentation and those
which bring about necessary and unwanted changes in the process of tella making are
described (Shale and Gashe, 1991). According to the report, the fermentation process of
tella is divided into four phases. The first occurs in the original mixtures of ingredients,
and the second and third phases occur after successive additions of more carbohydrate
materials. The three main carbohydrate materials are mentioned to be bikil, kita and
enkuro. The latter phase is where acidification takes place, which is not desirable.

11
Maximum ethanol production occurs during the third phase and at the beginning of the
fourth phase.
Shale and Gashe (1991) reported that the extent of heat treatment the asharo (roasted
barley) receives and the degree of steaming the enkuro (roasted barley steamed after
grinding) is subjected to have the direct bearing on the color of tella, which is determined
by the housewife preparing the tella. Tella is a beverage of variable viscosity and having
a variety of colors (grayish-white to dark brown). The fermenting organisms of tella are
composed of S. cerevisiae and Lactobacillus spp. Increase in ethanol content is directly
associated with growth in the population of yeasts and decrease in reducing sugar and
total carbohydrate (Debebe, 2006). In general, good quality of tella has a final ethanol
content of 2-8% (v/v) and the pH is 4-5 (Samuel Sahle and Berhanu Abegaz Gashe,
1991).
Table 2.2 Quality report variation of Ethiopian traditional beverage tella in the precious
studies

Author Ethanol % (V/V) pH value

Samuel Sahle and Berhanu Abegaz Gashe, 1991 2.8-5.0 4.5-4.8
Debebe, 2006 2.2 - 5 4.5 - 4.8
Haftu G. Alemayehu, 2018 4.80 3.28
Tadele Yohannes ,2013 3.98-6.48 4.00-4.99
Berhanu, A,2013 - 4.02
Alemu Fite et al, 1991 3.6 -
Belachew Desta, 1977, 5.65-6.56 -
(Mooha Lee a *. M., 2015) 2-6

2.4. Comparison of Ethiopian tella with beer

Tella is called Ethiopian traditional beer. Its production process is like beer making in
that the grain starch is converted into sugars by malting. However, there is no yeast
inoculation stage for fermentation, but it utilizes the natural yeast present on the cereals.
The dominant microorganism after the end of the first stage until the completion of tella
fermentation was reported to be Saccharomyces cerevisiae and Lactobacillus

12
pastorianumi (Sahle &Gashe 1991). Saccharomyces cerevisiae is known to be the top
fermentation yeast for ale production, while S. pastorianusor S. carlsbergensisare the
bottom fermentation yeasts used for lager beer production (Boulton C and Quain
D.2001). Alcohol is a by-product of yeast metabolism and is toxic to the yeast; typical
brewing yeast cannot survive at alcohol concentrations above 12% by volume (De
Kenkeleire D.2000).
Beer ranges from less than 3% alcohol by volume to around 14%. Tastes of tella, which
the panel described were bitterness, sweet, and sour (Yohannes 2013), are similar to the
description of the typical beer taste. The bitterness of tella is from gesho, while that of
beer is from hops (De Kenkeleire D 2000 Berhanu 2014). Leaves of gesho are like hops
in terms of its role in brewing; giving a bitter flavor and antibacterial effects for the shelf-
life (Berhanu 2014). The mouth feel of beer is from the presence of protein, polyphenols,
chloride, dextrins, and beta-glucan, etc. (Langstaff SA and Lewis MJ 1993), which are
mainly from malt and hops. The taste of tella is different depending on the different
cereals. Commercial beer is mostly made from malted barley and adjunct like corn, rice
or wheat provide the carbohydrate substrates for ethanol production by Saccharomyces
carlsbergensisor Saccharomyces cerevisiae (Kunze, 1996). Regarding the substrate, there
is no as such basic difference between tella and beer. The only difference is that the
physical environment for fermentation process, which is not as such controlled as
commercial beer and the techniques used in brewing to standard quantify of gradients and
aseptic technique and use of regular art and craft for tella brewing like that of beer
production.

2.5. Ingredients of Ethiopian traditional beverage; tella

2.5.1. Gesho, Rhamnus prinoides L. Herit

Gesho, Rhamnus prinoides, is a dense shrub or sometimes appears as small tree. It is
spineless and evergreen. It is one of the two representative species with Rhamnus staddo
of the family Rhamnacaea that are only found in Africa (Phillips, 1994). Gesho is
commonly used for preparation of traditional alcoholic beverages, tella and in Ethiopia.
Rhamnus prinoides is commonly found in eastern, central and western Africa. It is found
in different parts of Ethiopia, both in the wild and as a cultivated plant. It grows at

13
elevations of 1,400 to 3,200m above sea level, in the Montana and riverine forest on the
edges of clearings. The closely related species R.staddo A. Rich (Vernacular name, tsedo)
occurs only in the wild at the edges of Montana forest in wooded and scrub land in many
parts of Ethiopia. R.prinoides is cultivated on a large scale as a field crop. It is also grown
in home gardens. The leafy branches are sold fresh, or dried. (Fullas, 2003) 13 From a
gesho plant leaf, stem and fruits, 20 secondary metabolites have been identified (Abegaz
et al., 1999; Batterham et al., 1996, Dwivedi et al., 1998, Helen and Juliet, 1987; and Yen
et al., 1998). The identified metabolites include nine anthracene derivatives, eight
flavonoids and three naphthalenic compounds (Abegaz and Kebede, 1995; Abegaz et al.,
1999; Abegaz and Ngadjui, 1999 and Nindi et al., 1999). All the metabolites are already
identified from other plants except one metabolite geshoidin unique to gesho plant. It is
identified as a compound with bittering character and assumed to give bitter test to the
alcoholic beverages tella and tej (Abegaz and Kebede, 1995). The other metabolites from
gesho are identified as antimicrobial agents (Paneitz and Westendorf, 1999) as
pharmaceutical agents (Kartsova and Ganzaha, 2006).

2.5.2. Bikil
Bikil is prepared from malted cereals which may be wheat or barley. The cereal is
steeped in water for about two days. The second day the water is drained off and the
cereal is left packed in a container for one day to allow germination. The germinated
grain is transferred in to a wide tray to allow the germinated grain to lock with each other.
Then grain is smoked, sun dried and grounded to coarse powder (Nigusie, 2008).
Germination of cereal grains is initiated by water and its successful completion is
signaled by the emergence of the developing roots and shoots. Following the uptake of
water, hormonal signals, probably released from the embryo are believed to result in the
synthesis of hydrolytic and other enzymes by the aleurone cells and scutellum (Egwim
and Oloyede, 2006). Malted cereal grains have combination of enzymes, M-amylase and
N-amylase, dextrinase, protease, phosphatase, peptidase and others enzymes commonly
called in combination malt diastase. Malt diastase is derived from scutellum, aleuron
layers and the starchy endosperm of germinated seed to hydrolyze starch molecules into
simple sugar to nourish the growing seedling (Egwim and Oloyede, 2006; 14 Johns and
Chen, 1976 and Geoffrey and Fincher, 1989). During germination and early seedling

14
development, hydrolytic enzymes; M-amylase, N-amylase limit dextrinase and M
glucosidase enzymes convert insoluble starch granules of the endosperm into glucose
(Charalampopoulos et al., 2003).
Alpha amylase hydrolyzes alpha linkages in the two types of starch forms, M- amylose
and amylopectin, to yield a mixture of glucose and free maltose (Barber et al., 1986).
Alpha amylose consists of long chains in which all the glucose units are bound in M-(1-
4) linkages and the amylopectin is highly branched with back bone linkage of glycosidic
M-(1-4) and the branch points are M-(1-6) glycosidic linkage.
Beta amylase attacks the non reducing ends of the two starch forms at M-(1-4) glycosidic
linkage. This cleaves away successive maltose units (Barber et al., 1986). However,
neither M-amylase nor N-amylases can hydrolyze the M-(1-6) linkages in amylopectin
branch points (Berne, 2005). This hydrolysis is brought about by limit dextrinase. Due to
this reason the end product of amylase action on amylopectin is a large highly branched
molecule called dextrin. Conversion of starch enzymatically into simple sugars in malted
grains is important for preparation of fermented alcoholic beverages both in large and
small scales. There are yeast species that can only utilize simple sugars during
fermentation process for growth and survival. They cannot utilize starch as carbon source
since they do not have any enzyme to hydrolyze it.

2.5.3. Grawa (Vernonia amygdalina Del.)

Grawa is popularly known as bitter leaf. It belongs to the Compositae family. It is widely
distributed in most African countries and utilized for various purposes. Different parts of
the plant are consumed as a raw vegetable after abating bitterness by boiling and soaking
in several water changes (Okafor, 1983 and Abosi and Raroseroka, 2003). It is also used
as animals’ feed in most parts of Africa (Onwuka et al., 1989). In some parts of Africa
grawa is regarded as a medicinal plant in Africa to treat fever, kidney problems and
stomach discomfort (Hamowia and Saffaf, 1994). Additional studies also showed
bactericidal effect (Erasto et al., 2006), anti fungal property (Iwalokun et al., 2006) and
strong parasiticidal properties (Ogboli et al., 2000).
In Ethiopia grawa is usually used as ethno medicinal plant for stomach disorders and as a
cleansing agent for containers of tella fermentation. Fermentation containers are washed
repeatedly using grawa leaves and water till foams appear. The effect of grawa leaves in

15
relation to tella preparation is not well studied yet. However, the leaves are reputed to
contain disinfecting agents (Sahle and Gashe, 1991).

2.5.4. Weira (Olea europaea L. sub sp cuspidate Ciferri)

Weira is a sub-species of olive. It is distributed throughout Africa, India and China
(Forest et al., 2003). The fruits, leaves and stem of weira consist of series 12 of
compounds that represent multi chemical mechanism of defense against microbe and
insect attack (Kubo et al., 1985).
Weira is commonly used in Ethiopia for smoking fermentation containers of traditional
beverages such as tella, and tej, containers of water, containers of milk and milk products.
The smoke is reported to remove and to cause sluggish growth of coli forms in smoked
milk and its products (Ashenafi, 2002). At the same time it is also observed to delay milk
spoilage. Weira smoke give smoky flavor to water, tella, tej and milk. It has been shown
that weira smoke contains compounds which have the ability to kill or delay the rate of
growth of a number of human pathogenic bacteria and fungi (Ashenafi, 2002). However,
the effect of weira smoke in relation to tella fermentation is not yet studied.

16
 Chapter Three
3. Materials and Methods
3.1. Sample Preparation
Cereals; Different types of cereals namely barley (Asela origin), corn (Gojjam origin),
wheat (Jiru origin), sorghum (Lemi origin) and teff (Gojjam origin) were collected from
milling house randomly, and it was stripped and winnowed thoroughly to remove
unwanted materials from the grains. Then, the cleaned grains were roasted, and milled to
prepare kita and enkuro for preparation of tella, accordingly the given proportion.

Figure 3.1Ttraditionaly roasted different cereals

Malting barley; for the aim of conversion of starch enzymatically into simple sugars in
alcoholic fermentation, barley grain was brought from milling house, stripped and then
grains of barley were moistened while in a container and left to germinate for three days.
Then, germinated barley was finally sun-dried.

Figure 3.2 Sun light drying of barley malt (bikil)

17
Gesho (Rhamnus prinoides L. Herit); Gesho which role is giving a bitter flavor and
antibacterial effects for the shelf life of tella (Berhanu, 2014) was collected from the
market randomly, and then the leaf and stem part of gesho was separated clearly, dried,
and then pounded to be used tella brewing.

Figure 3.3 Sun drying of gesho leaves and stem shreds

Grawa (Vernonia amygdalina Del.); Fresh leaves of grawa (Vernonia amygdalina)
contain disinfecting agents (Sahle and Gashe, 1991).were collected from the market for
washing of containers with fresh water cleanly (Sahle and Gashe 1991).
Weira (Olea europaea L. sub sp cuspidate Ciferri); Splinting weira were brought from
the market to fumigate the well-cleaned container intending to eliminate microorganisms
sensitive to antimicrobial components of wood smoke. Weira smoke give smoky flavor to
tella and also contributes to the desirable flavor of the fermented product (Sahle and
Gashe 1991).
St. George beer; St. George beer which was brewed closely on the time of Ethiopian
traditional beverage tella preparation was brought from the company to compare quality
contents against Ethiopian traditional beverage tella.

3.2. Chemical and reagents

98% of H2SO4, K2SO4 and CuSO4 were used in ECAE laboratory to test quality of
malting barley. In addition, 4% boric acid, 40% of NaOH and 0.5N HCl (37%) was
prepared cautiously in the laboratory. Indicator was prepared by mixing 100 ml of 0.1 %
methyl red (in 95% ethanol) with 200 ml of 0.2% bromocresol green (in 95% ethanol).
Deionized water was collected using distiller in the laboratory.

18
3.3. Apparatus and instruments
25 liters of plastic containers was purchased from the market to brew Ethiopian
traditional beverage tella. NIRS™ DS2500 Feed and Forage analyzer was used to test the
proximate analysis of cereals in bless Agri food laboratory service PLC. NUVE
(FN500P) Thermostat Oven and Desiccator were also served to analyze moisture result of
malting barley and cooling purpose respectively in National tobacco enterprise
laboratory.
DMA5000M Alcolyzer was used to test original wort extract, degree of fermentation and
alcohol content of the beverage in ECAE laboratory. In the mean time, HANNA
(HI2550) was employed to test pH contents in the beverage. Besides, K-138 Kjeldahl
digestion tube, Kjeldahl distillation apparatus, W4000 Water distiller, 250ml conical
flask, 250ml Beaker, 250ml Erlemenary flask, digital balance and filter paper were used
in ECAE laboratory.

3.4. Preparation of Ethiopian traditional beverage; tella

Due to the absence of organized national standard of tella brewing in Ethiopia,
traditionally fermented alcoholic beverage of tella varies from one region to the other and
even within a specific locality. However, the general procedure of mixing the ingredients,
time required for fermentation as well as the sequences of processing’s mainly similar.
The various studies (Samuel Sahle and Berhanu Abegaz Gashe, 1991; Belachew Desta,
1977) and a general review published on Ethiopian indigenous fermented foods and
beverages (Mogessie Ashenafi, 2006) reported detailed descriptions of the preparation
steps, microbiological studies and alcoholic contents of the home made food and drinks
in Ethiopia.
For the purpose of this study, the course of Ethiopian traditional beverage tella brewing
was conducted accordingly Samuel Sahle and Berhanu Abegaz Gashe (1991) tella
fermentation description which is practically adopted in the northern part of the country,
Gondar, specifically. Therefore, tella preparation was performed based on 0.33 kg of
gesho (leaves and pounded stems), 0.17 kg of bikil, 5 kg of grains in the form of kita
(1.67 kg) and enkuro (3.33 kg) and then mixed with 10 liters of water. Proportionally,
Gesho, bikil, kita and enkuro could be presented as 6:3.09:30.36:60.54 respectively.

19
Before the start of brewing the fermentation containers were well washed repeatedly
using grawa leaves and water till foams appear. Then, the well-cleaned container was
inverted over smoking splinters of weyra (Olea europaea) for about 10 minutes. After
wards, the fermentation process was conducted into the following phases.

Phase I - Powdered leaves of gesho were mixed with water into the containers and
allowed to ferment to five days. The fermenting material is commonly called tinsis. Then,
mixing of barley malt (bikil), pounded stems of gesho, pieces of kita and water to the
containers. This was left to ferment for two days.
Phase II-Chopped pounded stems of gesho, bikil, enkuro and water were added to the
containers and the contents are mixed into thick slurry called difdif. This was also
allowed to ferment for two more days.
Phase III - At the final stage, the containers was filled with water to the brim and the
contents were again mixed thoroughly. The containers were then sealed to create
anaerobic conditions and left to ferment for two days. Accordingly to Samuel Sahle and
Berhanu Abegaz Gashe (1991) Ethiopian traditional beverage fermentation process has
taken11 days. Quality evaluation of the prepared beverages has been started from the 12th
days (288hours) of fermentation.

Figure 3.4 Preparations of Ethiopian traditional beverage tella using different cereals.

3.5. Proximate analysis of cereals

The composition of cereals such as moisture, crude fat, crude protein, crude ash, crude
fiber, carbohydrate, energy and starch content of grains (barley, corn, wheat, sorghum &

20
teff) were determined using NIRS™ DS2500 Feed and Forage analyzer. The analyzer is a
multi parameter analyzer, which able to measure nutritional content of grinded samples
and provide the results collectively.

Primarily, the prepared samples were grinded based <1 micrometer sieve size> to match
the optical performance across the full wavelength range (400 - 2500 nm) of the analyzer.
The finely grounded particles (samples) were transferred to the sample cap. Then, select
cereal product calibration mode of the analyzer. Finally, the analyzer measures the
nutritional composition of samples quantitatively.
The carbohydrate and energy composition of cereals were calculated based on the
following formula (Gebreyes, 2020).

a. (Carbohydrate = 100 − (Moisture + Crude protein + Crude fat +

Crude ash + Crude fiber))
b. (Energy = 9(crude fat) + 4(crude Protein) + 4(carbohydrate))

3.6. Determination Quality content of malting barley

Malt is the key ingredient in beer that provides the starch and enzymes necessary to
produce the fermentable sugar which yeasts then turn to alcohol. Malt also provides the
color and flavor compounds which contribute to the final character of beer. The quality of
malting barley was characterized by employing moisture, nitrogen and protein (affects
the chemical composition and enzyme levels of the finished malt) content of the barley as
a key parameter.

3.6.1. Determination of moisture of malting barley

Moisture content of malting barley was analyzed by using thermostat NUVE (FN500P)
oven drying method. Five grams (5 g) of well-homogenized malting barley samples were
transferred to the dried and weighted containers. Sample containing containers were
placed in the drying oven and dried for 1 hr at 110°C until the constant weight of the
sample was maintained. Then, the dried sample was removed from the drying oven and
then cooled in a desiccator at room temperature and reweighted. Moisture content of
malting barley was calculated based on the following formula (ASEAN, 2011).

21
Moisture (g/100g) = (W2-W3)*100
(W2-W1)
Where; W1=weight of container or empty dish (g)

W2=weight of container + sample before drying (g)

W2-W1=weight of sample (g)

W3= weight of container + sample after drying (g).

W2-W3= Loss of Weight (g)

Total solid (%) = 100- % Moisture (W/W)

3.6.2. Determination of nitrogen and protein content of malting barley

Digestion; 0.6 g of grinded sample was measured accurately and placed in Kjeldahl
digestion tube. 4.95 g of catalyst (K2SO4 and CuSO4), and 15 ml of concentrated H2SO4
were added. Covered the tube with exhaust manifold and placed in the preheated digester
and digest at 400°C for two hours. Then the tube was removed and stands until the
sample was cooled.
Distillation; Switch on distillation apparatus and pre-washed for 10 minutes. 15 ml of
4% boric acid was dispensed into a 250 ml conical flask and placed the flask under the
condenser. Connect the digestion tube containing the digested sample to the distillation
apparatus. 15 ml of 40% NaOH was poured carefully into digested sample and then
immediately turned on the steam supply valve to initiate the distillation. 50ml of distillate
was collected. Finally 1 ml indicator was added and titrates with standard 0.5N
hydrochloric acid until the solution change from green to pinkish.
Calculation; the percentage of nitrogen, on dry basis, was calculated as follows (ES669,
2001).

14
𝑁=𝑇∗
𝑀 ∗ 𝐷. 𝑀

Where,

N is the percentage of nitrogen in dry barley.

22
T is the acid titration value, with the blank subtracted, and corrected to extract tenth
normality

14 atomic weight of nitrogen

M is the mass, in grains, of barley sample taken for test, and

D.M is the percentage of dry matter

The percentage of protein content of malting barley in a dry basis was calculated as
follows (ES669, 2001).

% protein = 𝑁 ∗ 6.25

Where 6.25 is protein conversion factor

3.7. Effect of cereals variation on Fermentation process of Ethiopian traditional

beverage; tella
To study the effect of cereal variations on Fermentation process of Ethiopian traditional
beverage tella, fermentation process parameters such as original wort extract (OE), real
extract (RE), apparent extract (AE) and real degree of fermentation (RDF) of tella were
tested on the fermentation process.

Therefore, fermented samples were taken from traditional beverage tella which were
brewed in Ethiopian conformity assessment enterprise laboratory, and, filtered into 250ml
Erlemenary flask to remove suspended and residual part of the samples. Then, 50ml of
tella was measured from filtered sample and analyzed by deionized DMA5000M
Alcolyzer. The Alcolyzer which is preliminary calibrated using distilled water and able to
analyze multi parameters collectively, provide original wort extract (OE, % W/W), real
extract (RE, % W/W), apparent extract (AE, % W/W) Alcohol weight by weight (Aw/w)
and real degree of fermentation of filtered sample quantitatively.
The Alcolyzer calculated Original wort extract of the sample respecting of balling
equation. Balling’s equation was based on an implied functional relationship between
alcohol (Aw/w) and unfermented solids (real extract RE), the mass lost to carbon dioxide
and yeast, and the solids content of the original wort (OE) (Anthony J. Cutaia1, 2009):

23
OE = 100 * (2.0665 * Aw/w + RE)
(100 + 1.0665* Aw/w)

Where; OE: original wort extract

Aw/w: alcohol weight by weight
RE: Real extract
2.0665: is conversion coefficient of the grams of fermentable material necessary to
produce one gram of ethanol.
1.0665; is coefficient of the mass in grams of fermentable material lost to carbon dioxide
and yeast.
The Alcolyzer calculated real degree of fermentation (% RDF) of the sample considering
the following equation (Anthony J. Cutaia1, 2009).
RDF = [100 * (OE – RE) / OE)] *[1 / (1 – .005161 * RE)]
Where; RDF real degree of fermentation
OE; original wort extract
RE; Real extract
The correction term [1 / (1 – .005161 *RE)] is mass lost by CO2 and yeast uptake during
fermentation.

3.8. Effect of cereals variation on Quality of Ethiopian traditional beverage; tella

To study the effect of cereal variations on quality of Ethiopian traditional beverage tella,
alcoholic parameters such as Alcohol (AV/V), pH value and relative density of tella were
tested on the 12th day (288hours) of fermentation process.

Therefore, certain amount of samples were taken from traditional beverage tella which
were brewed in Ethiopian conformity assessment enterprise laboratory, and, filtered into
250ml Erlemenary flask to remove suspended and residual part of the samples. Then,
50ml of tella was measured from filtered sample and analyzed by deionized DMA5000M
Alcolyzer. The Alcolyzer which is preliminary calibrated using distilled water and able to
analyze multi parameters collectively provide Alcohol (AV/V) and density of filtered
samples quantitatively.

Relative density of tella was calculated by using density of tella and density of distilled
water analyzed by the Alcolyzer.
24
Relative density of tella = Density of tella

Density of distilled water

3.8.1. Evaluation of pH value

50ml of sample was taken from filtered Ethiopian traditional beverage; tella and then
inserted calibrated HANNA (HI2550) pH meter electrode and temperature sensor into the
sample to read the pH value of tella within the temperature. Thus, the pH meter displayed
sourness of tella within 25C temperatures.

Figure 3.5 Testing pH value of Ethiopian traditional beverage tella

3.9. Effect of cereals variation on Shelf life of Ethiopian traditional beverage; tella
The qualities of Ethiopian traditional beverages tella were evaluated again after 20 days
of storage which was kept under room temperature (23-25 C) in the laboratory to study
shelf life Ethiopian traditional beverage tella by comparing the change of Ethanol and pH
value from the previous result.

Therefore, certain amount of sample was taken from the container and filtered using
250ml Erlemenary flask and then read using deionized DMA5000M Alcolyzer. On the
other hand, pH value of the beverage was analyzed using HANNA (HI2550) pH meter,
and then the optimum cereal was identified from the analysis result.

25
3.10. Comparison of Ethiopian traditional beverage tella against St. George beer
Conformity of Ethiopian standards (CES) classified beers into three (light, strong and
extra strong beer) categories according to their quality requirements. St. George beer is
labeled in the range of strong beer. Therefore, George beer was analyzed in the laboratory
to compare the result of degree of fermentation, ethanol content and pH value against
Ethiopian traditional beverages tella prepared from various cereals.

3.11. Experimental Design

Effect of cereals variation on fermentation process and quality of Ethiopian traditional
beverage tella were evaluated by employing single- factor experiment. The main factor
considered here were cereals variations which had five levels (Barley, corn, sorghum,
wheat and teff) with 2 replicates. The main response variables considered to evaluate
fermentation process were real degree fermentation of tella. On the other hand, the main
response variables considered to evaluate quality of tella were ethanol, pH value and
relative density of tella. Then, the optimum cereal which provides the optimum degree of
fermentation and alcohol were determined in the experiment.
Table 3.1 Experimental design of fermentation process and quality of Ethiopian
traditional beverage

Cereal variations 2 Replicate

Barley 1 2

Corn 3 4
levels

Wheat 5 6

Sorghum 7 8

Teff 9 10

Effects of cereals variation on shelf life of Ethiopian traditional beverage tella were
evaluated by employing Two-Factor Factorial Design. The main factors considered here
were cereals variations which had five levels (Barley, corn, sorghum, wheat and teff) and
time designed into two (288h &640h) levels with four replications. The main response
variables considered here were degree of fermentation, ethanol and pH value of tella.

26
Then, the optimum cereal which withstands the declining of pH through 20 days of
storage was determined in the experiment.
Fermentation of tella was carried out under room temperature (23 - 25 C) in Ethiopian
conformity assessment enterprise laboratory, starting from May 31 up to July 2, 2021.
Table 3.2 Experimental design of shelf life of Ethiopian traditional beverage

Levels

Cereal variations 288h (12th days) 640h (32th days)

2 Replicate 2 Replicate

Barley 1 2 11 12
levels

Corn 3 4 13 14

Wheat 5 6 15 16

Sorghum 7 8 17 18

Teff 9 10 19 20

3.12. Statistical analysis

Data entry and detail analysis were performed using SPSS (Statistical Package for social
sciences statistical software version) 20. The significant terms in the models were
identified by analysis of variance (ANOVA) and factorial ANOVA for each response. In
addition, Microsoft excel 2007 was used for further analysis.

Significance was judge if the probability levels of the F-statistic calculate from the data
was less than 0.05.

27
 Chapter Four

4. Results and Discussion

4.1. Proximate analysis of different cereals
Proximate analysis of cereals was conducted for different grains to test their nutrition
contents, and provided information about the degree of nutrients like moisture, crude
protein, crude fat, crude fiber, crude ash, carbohydrate, and energy and starch values of
the cereals.

4.1.1. Proximate analysis of barley

Nutritional content of barley grain was tested with two replications and the result is
presented with comparison of literature review reports in table 4.1.

Table 4.1 Proximate analysis result of barley

Cereal type; Barley

S/N Parameters Unit
Literature
Results review Reference
(Yadesa Abeshu,
1 Moisture % (g/100g) 12.05±0.03 11.35-13.07 2017)
(Yadesa Abeshu,
2 Crude protein % (g/100g) 9.07±0.03 10.07-14.10 2017)
(Yadesa Abeshu,
3 Crude fat % (g/100g) 2.77±0.04 2.20-6.40 2017)
(Yadesa Abeshu,
4 Crude fiber % (g/100g) 4.00±0.06 9.65-15.88 2017)
(Yadesa Abeshu,
5 Crude ash % (g/100g) 2.56±0.04 1.43-2.34 2017)
(Yadesa Abeshu,
6 Carbohydrate % (g/100g) 69.55±0.13 45.69-58.45 2017)
(Yadesa Abeshu,
7 Energy % (g/100g) 339.41±0.16 271.61-324.3 2017)

8 Starch % (g/100g) 50.82±0.14 63-65 (Assefa, 2021)

As it is mentioned in the table, moisture, crude fat and crude ash analysis result of barley
grain are within the range of the previous reports (Yadesa Abeshu, 2017). However,

28
starch (Assefa, 2021), crude protein and crude fiber results are lower, and then higher
carbohydrate contents are recorded compared with the previous studies (Yadesa Abeshu,
2017).

4.1.2. Proximate analysis of corn

Nutritional composition of corn grain was evaluated with two replications and the result
is summarized with comparison of literature review reports in table 4.2.

Table 4.2 Proximate analysis result of corn

Cereal type; Corn

S/N Parameters Unit
Literature
Results review Reference
(Girmay Tsegay*,
1 Moisture % (g/100g) 12.52±0.06 8.52-12.86 2019)

2 Crude protein % (g/100g) 7.22±0.04 7.48-11.6 (Baye, 2014)

(Girmay Tsegay*,
3 Crude fat % (g/100g) 4.31±0.08 4.01-5.99 2019)
(Girmay Tsegay*,
4 Crude fiber % (g/100g) 2.34±0.06 1.39-2.05 2019)
(Girmay Tsegay*,
5 Crude ash % (g/100g) 1.42±0.08 0.9-1.53 2019)
(Girmay Tsegay*,
6 Carbohydrate % (g/100g) 72.19±0.31 67.64-74.88 2019)

7 Energy % (g/100g) 356.43±0.34 375 (Baye, 2014)

8 Starch % (g/100g) 63.36±0.17 72 (Baye, 2014)

Importantly, moisture, crude protein, crude fat, crude ash and carbohydrate analysis
results of corn grain are in the range of the previous studies (Girmay Tsegay*, 2019).
However, there is a slight increment regarding on crude fiber measurement, (Girmay
Tsegay*, 2019). On the other hand, starch content of corn grain is lower from the
previous records (Baye, 2014).

29
4.1.3. Proximate analysis of wheat
Proximate analysis was carried out to test the nutritional content of wheat grain, with two
replications based and the analysis result is listed in table 4.3.

Table 4.3 Proximate analysis result of wheat

Cereal type: wheat

S/N Parameters Unit
Literature
Mean review Reference
(Kassegn1,
1 Moisture % (g/100g) 12.24±0.04 10.76 2017)

2 Crude protein % (g/100g) 9.84±0.08 11.7 (Baye, 2014)

2.00
3 Crude fat % (g/100g) 2.05±0.06 (Baye, 2014)

4 Crude fiber % (g/100g) 2.64±0.08 2.00 (Baye, 2014)

1.60
5 Crude ash % (g/100g) 1.59±0.08 (Baye, 2014)
(Kassegn1,
6 Carbohydrate % (g/100g) 71.64±0.10 64.85 2017)

7 Energy % (g/100g) 344.37±0.45 359 (Baye, 2014)

8 Starch % (g/100g) 60.44±0.34 71 (Baye, 2014)

Moisture (Kassegn1, 2017), crude protein, crude fat, crude fiber, and crude ash (Baye,
2014) analysis results of wheat grain are proximately close with the previous studies, in
the mean time larger content of carbohydrate (Kassegn1, 2017) are obtained compared
with the previous results. However, a starch content of wheat grain is lower from the
previous records.

4.1.4. Proximate analysis of sorghum

Nutritional composition of sorghum grain was evaluated with two replications like other
grains, and the results are briefly presented with comparison of the literature review
reports in table 4.4.

30
 Table 4.4 Proximate analysis result of sorghum

Cereal type; Sorghum

S/N Parameters Unit
Literature
Results review Reference

1 Moisture % (g/100g) 12.35±0.16 9.66-12.94 (Gebreyes, 2020)

2 Crude protein % (g/100g) 8.50±0.28 8.20-16.48 (Gebreyes, 2020)

3 Crude fat % (g/100g) 2.69±0.06 2.48-4.60 (Gebreyes, 2020)

4 Crude fiber % (g/100g) 2.28±0.03 2.17-8.59 (Gebreyes, 2020)

5 Crude ash % (g/100g) 1.93±0.04 1.12-2.29 (Gebreyes, 2020)

6 Carbohydrate % (g/100g) 72.25±0.06 67.56 -76.42 (Gebreyes, 2020)

7 Energy % (g/100g) 347.21±0.40 370 (Baye, 2014)

8 Starch % (g/100g) 60.17±0.16 63 (Baye, 2014)

Surprisingly, the analysis result of moisture, crude protein, crude fat, crude fiber, crude
ash, and carbohydrate content of sorghum are within the range of the previous studies
reported by (Gebreyes, 2020). In addition, starch content of sorghum is approximately
close to the prvious result presented by (Baye, 2014).

4.1.5. Proximate analysis of teff

Like other cereals, proximate analysis of teff was carried out to determine the nutritional
content of grains with two replication based, and the analysis result is listed out with
comparision of litrature review reports in table 4.5.

31
 Table 4.5 Proximate analysis result of teff

Cereal type; Teff

S/N Parameters Unit
Literature
Results review Reference
(Lamesgen
1 Moisture % (g/100g) 11.90±0.06 9.92-10.90 Yegrem, 2020)

2 Crude protein % (g/100g) 3.45±0.06 11 (Baye, 2014)

3 Crude fat % (g/100g) 3.28±0.04 2.5 (Baye, 2014)

4 Crude fiber % (g/100g) 2.37±0.06 3.00 (Baye, 2014)

5 Crude ash % (g/100g) 2.31±0.04 2.8 (Baye, 2014)

(Lamesgen
6 Carbohydrate % (g/100g) 76.69±0.08 68.39-71.46 Yegrem, 2020)

7 Energy % (g/100g) 350.08±0.49 357 (Baye, 2014)

8 Starch % (g/100g) 61.25±0.28 73 (Baye, 2014)

Moisture (Lamesgen Yegrem, 2020), crude fat, crude fiber and crude ash content of the
grain are approximately close to the previous study reported by (Baye, 2014). On the
other hand, higher content of carbohydrate result is recorded compared with the literature
result (Lamesgen Yegrem, 2020). However, crude protein and starch content of teff is
much lower from the previous result (Baye, 2014).
From proximate analysis results, starch content of corn is higher than the result of other
cereals, whereas barley has low content. On the other hand, teff has higher carbohydrate
content from others, whereas barley contains low content of carbohydrates. In general,
starch content of cereals can be ordered barley, sorghum, wheat, teff and corn as 50.82,
60.17, 60.44, 61.25 and 63.36 from low to high results respectively. Likewise,
carbohydrate contents of cereals are ordered as barley, wheat, corn, sorghum and teff
69.55, 71.64, 72.19, 72.25 and 76.69 from low to high results respectively. In addition,
crude protein content of cereals can be ordered from low to high teff, corn, sorghum,
barley and wheat 3.45, 7.22, 8.50, 9.07 and 9.84 respectively.

32
4.2. Determination of Quality of malting barley
The protein content of the barley grain affects the chemical composition and enzyme
levels of the finished malt. If the protein is too high, this limits the starch content and
amount of extract available to the brewer. On the other hand, if the protein is too low
there may be insufficient enzymatic activity to modify the barley kernel and break down
starch for brewing (E.WWrigley, 2010). Therefore, for this study, moisture, Nitrogen and
protein content of malting barley were tested and the results are presented in table 4.6.

Table 4.6 Quality analysis result of malting barley

S/N Parameter Replication Mean (%) Literature References

review
1 Moisture 2 12.34±0.06 Max 12 Fox et al. 2003
2 Nitrogen 2 1.73±0.07 Max1.7 Beaven's 1902, 8, 542
3 Protein 2 10.78±0.04 9.0-11.5 Fox et al. 2003

The analysis result of malting barley are adequately met the quality requirement of
moisture (Fox et al. 2003), Nitrogen (Beaven's 1902, 8, 542), and Protein content (Fox et
al. 2003) of malting barley to be malted for brewing.

4.3. Effect of cereals variation on fermentation process of Ethiopian traditional

beverage; tella

4.3.1. Determination of Original wort extract (OE), Real extract (RE) and apparent
extract
Original extract is a measure of the solids content originally in the wort, before alcoholic
fermentation has commenced to produce the beer (Anthony J. Cutaia1, 2009). The
apparent extract (AP) refers to the density of an equivalent sucrose solution, assuming no
alcohol is present (Cutaia, 2009). The real extract (RE) represents the actual (real) solid
content of a beer (Cutaia, 2009). Original extract is one of the major measurements used
by brewers to determine the future alcohol content of a beer fermented from particular
wort. Therefore, original wort extract, real and apparent extract of traditional beverages

33
prepared from various cereals has been conducted and the results are consolidated in
table 4.7.

Table 4.7 Original wort extract, real and apparent extract results of Ethiopian traditional
beverage tella

AP
S/N Type of tella Replication OE (% W/W) RE (%W/W) (%W/W)

1 Barley 2 12.75±0.01 7.41±0.00 6.12±0.00

2 Corn 2 12.69±0.03 6.81±0.02 5.39±0.02

3 Wheat 2 13.35±0.08 7.92±0.08 6.61±0.09

4 Sorghum 2 12.84±0.05 6.82±0.05 5.37±0.05

5 Teff 2 13.85±0.03 5.75±0.01 3.81±0.01

From the analysis results, teff tella has higher amount of original wort extract and low
apparent extract compared with the other beverages, whereas corn has low original wort
extract which owned high amount of crude fat contrarily with the other cereals (See Fig
4.1).

Crude fat (g/100g)

5 4.31
Crude fat content

4 3.28
2.77 2.69
3 2.05
2 Crude fat
1
0
Barley Corn Wheat Sorghum Teff
Cereal type

Figure 4.1 Comparison of crude fat result of cereals

34
Wheat tella has high amount of apparent extract compared with the other. Original wort
extract results can be ordered from high to low, teff, wheat, sorghum, barley and corn
13.85, 13.35, 12.84, 12.75 and 12.69 % respectively.

4.3.2. Determination of real degree of fermentation

Real degree of fermentation (RDF) is a measure of expressing the degree to which the
available extract was fermented and reported as a percentage calculated from the ethanol
content and gravity of the remaining extract (James Bruner 1, 2021). In Ethiopian
traditional beverage tella, the degree to which sugar in wort has been converted into
alcohol through fermentation, in beer defined as “attenuation” has been analyzed and the
results are collected in table 4.8.
Table 4.8 Real degree of fermentation (%) of Ethiopian traditional beverage tella

Type of tella Replication Mean (%)

Barley 2 43.52±0.02
Corn 2 48.08±0.04
Wheat 2 42.41±0.31
Sorghum 2 48.62±0.18
Teff 2 60.32±0.14
Average 2 48.59±6.70

The analysis result of traditional beverage prepared from teff produced a higher value of
degree of fermentation which generated higher original wort extract (see fig 4.2)
compared with the other beverages and provided higher carbohydrate content in
proximate analysis.

35
 Original wort extract (% plato)

13.85

Original wort extract

14.00
13.35
13.50
12.75 12.84
13.00 12.69
% Plato
12.50
12.00
Barley Corn Wheat Sorgham Teff
Cereal variations

Figure 4.2 Comparison of original wort extract

Whereas, a beverage prepared from wheat has low degree of fermentation, though,
original wort extract content of the beverage was larger than sorghum, barley and corn
(see fig 4.2). Comparatively, degree of fermentation for teff, sorghum, corn, barley and
wheat are 60.32, 48.62, 48.08, 43.52 and 42.41 respectively. Corn, which had lower
content of original wort extract generated higher percentage of degree of fermentation
compared with wheat and barley beverages.

Table 4.9 Analysis result of real degree of fermentation using one way ANOVA

Sum of Squares df Mean Square F Sig.

Between
403.614 4 100.904 3312.656 .000000
Groups
Within Groups .152 5 .030
Total 403.766 9

Cereal variations are significant (P ≤ 0.05) for real degree of fermentation of tella. Hence,
variation of cereals during the preparation of traditional beverage tella has a significant
effect on real degree of fermentation which directly defines the conversion of sugar to
alcohol through yeast.

36
 Table 4.10 Comparison result of real degree of fermentation of tella

Cereal variations Cereal variations Sig.

Corn .000
Wheat .008
Barley
Sorghum .000
Teff .000
Barley .000
Wheat .000
Corn
Sorghum .119
Teff .000
Barley .008
Corn .000
Wheat
Sorghum .000
Teff .000
Barley .000
Corn .119
Sorghum
Wheat .000
Teff .000
Barley .000
Corn .000
Teff
Wheat .000
Sorghum .000

4.4. Effect of cereal variation on Quality of Ethiopian traditional beverage; tella

4.4.1. Determination of Ethanol

Ethanol is one of the most commonly used recreational drugs (causing the characteristic
effects of alcohol intoxication) worldwide and it is often ingested as a component of beer

37
(Tatjana Rakcejeva, 2013). Therefore, Ethanol content of traditional beverage tella (local
beer) was evaluated and the results are presented in table 4.11.
Table 4.11 Ethanol content of Ethiopian traditional beverage tella

Type of tella Replication Mean (%V/V)

Barley 2 3.58±0.00
Corn 2 3.94±0.01
Wheat 2 3.66±0.00
Sorghum 2 4.03±0.00
Teff 2 5.43±0.02
Average 2 4.13±0.71

Tella which was brewed from teff produced higher ethanol results compared with other
beverages. In this analysis, ethanol content of the beverage shows proportional relation
with the carbohydrate value of cereals, and their degree of fermentation except beverages
prepared from wheat grains (see fig 4.3).

Even though, the carbohydrate and starch content of wheat was greater than barley, there
is no much difference of ethanol content due to low degree of fermentation. Generally,
alcoholic contents of beverages increased from barley to wheat and then corn, sorghum
and teff. Their ethanol result is 3.58, 3.66, 3.94, 4.03 and 5.43 respectively.

38
 Carbohydrate and RDF

100 72.19 72.25 76.69

69.55 71.64
60.32
Results 43.52 48.08 42.41 48.62
50

0 Carbohydrate
Barley Corn Wheat Sorgham Teff %(g/100g)
Cereal variations
% RDF

Figure 4.3 Comparison of carbohydrate contents for cereals and their RDF
In the previous studies, minimum ethanol content of Ethiopian traditional beverage tella
was reported 2.00 and 2.2 %V/V by (Sahle and Gashe, 1991) and (Debebe, 2006)
respectively. Importantly, the minimum result of ethanol content in this study which was
recorded from barley (3.58%V/V) was above the minimum result of the previous report,
and the average ethanol content of tella prepared from various cereals is 4.13 %V/V. On
the other hand, maximum ethanol content was reported 6.65 %V/V by (Tadele Yohannes,
2013) and 5.65 - 6.56 % (Belachew Desta, 1977) by V/V. However, in this study
maximum ethanol content was recorded 5.43%V/V from teff tella.

According to the study of (Sahle and Gashe, 1991) good quality of tella has a final
ethanol content of 2-8% (v/v). Thus, the result of this study is aligned in the range of the
previous conclusion.

According to the classification of Ethiopian conformity standards (CES) strong beer

should contain 4.1 %V/V minimum ethanol content and 5.25 % V/V for extra strong beer
(CES52, 2021). In this study, tella prepared from teff (V/V 5.43) produced higher content
of the CES requirement.

39
 Table 4.12 Analysis results of Ethanol using one way ANOVA

Sum of Squares df Mean Square F Sig.

Between
4.497 4 1.124 11241.850 .000000
Groups
Within Groups .001 5 .000
Total 4.497 9

Cereal variations are significant (P ≤ 0.05) for ethanol content of Ethiopian traditional
beverage tella. Hence, a cereal variation on the Ethiopian traditional beverage tella has a
significant effect on ethanol content of the beverage.

Table 4.13 Comparison results of Ethanol content of tella

Cereal Variations Cereal Variations Sig.

Corn .000
Wheat .003
Barley
Sorghum .000
Teff .000
Barley .000
Wheat .000
Corn
Sorghum .001
Teff .000
Barley .003
Corn .000
Wheat
Sorghum .000
Teff .000
Barley .000
Corn .001
Sorghum
Wheat .000
Teff .000
Barley .000
Corn .000
Teff
Wheat .000
Sorghum .000

40
4.4.2. Determination of pH value
Like ethanol pH values of different traditionally fermented beverages tella were analyzed
and the results were incorporated in table 4.14.

Table 4.14 pH result of Ethiopian traditional beverage tella

Type of tella Replication Mean

Barley 2 4.21±0.02
Corn 2 4.16±0.00
Wheat 2 3.94±0.00
Sorghum 2 4.20±0.00
Teff 2 4.30±0.00
Total 10 4.16±0.13

The pH value of wheat tella is lower compared with the other beverages, which implies
higher sourness was occupied than other beverages, despite; it contains high content of
starch and carbohydrate values compared with barley. In addition, original wort extract of
wheat was higher from sorghum, corn and barley, and expected to generate higher result
of ethanol. However, crude protein content of wheat is high and limits starch content to
be extracted during fermentation and became too sour compared with the other beverages
(See fig 4.4). Besides, apparent extract of the beverage is higher from the others.

In the previous studies, minimum pH value of Ethiopian traditional beverage tella was
reported as 3.28 by (Haftu G. Alemayehu, 2018). Importantly, in this study, the minimum
result of pH value which is 3.94 recorded from wheat is above the minimum pH value of
the previous report which implies low acidity recorded in the product. The average pH
value of tella prepared from various cereals is 4.16. On the other hand, maximum pH
value was recorded 4.99 by (Tadele Yohannes, 2013) which are higher from this study
obtained 4.30 from teff tella. From a sensory stand point of beer, optimum pH is reported
from 4.0 to 4.4 (Tatjana Rakcejeva, 2013). The result of this study is approximately in the
range of the specification (3.94 to 4.30).

41
 Crude protein
9.84
9.07
10 8.5
7.22
Results 8
6
3.45
4 Crude protein (%)
2
0
Barley Corn Wheat sorghum Teff
Cereal variations

Figure 4.4 Comparison of crude protein for various cereals

According to the study of (Sahle and Gashe, 1991) good quality of tella has a final pH
value of 4-5. The result of this study is aligned the range of the previous study.
Generally, pH value of wheat, corn, sorghum, barley and teff are ordered 3.94, 4.16, 4.20,
4.21 and 4.30 respectively. Traditional beverage tella prepared from teff has higher
amount of pH value which reveals lower content of sourness.

Conformity of Ethiopian standards (CES) levels the pH value of beers from 3.6 to 4.8
(CES52, 2021). Beneficially, in this study the pH value of Ethiopian traditional beverage
is in the range of the specification.

Table 4.15Analysis result of pH value using one way ANOVA

Sum of Squares df Mean Square F Sig.

Between
.143 4 .036 397.889 .000002
Groups
Within Groups .000 5 .000
Total .144 9

Cereal variations are significant (P ≤ 0.05) for pH value of Ethiopian traditional beverage
tella. Hence, variation of cereals has a significant effect on pH value of Ethiopian
traditional beverages tella as of ethanol.

42
 Table 4.16 Comparison result of pH value of tella

Cereal variations Cereal variations Sig.

Corn .026
Wheat .000
Barley
Sorghum .980
Teff .001
Barley .026
Wheat .000
Corn
Sorghum .041
Teff .000
Barley .000
Corn .000
Wheat
Sorghum .000
Teff .000
Barley .980
Corn .041
Sorghum
Wheat .000
Teff .001
Barley .001
Corn .000
Teff
Wheat .000
Sorghum .001

4.4.3. Determination of Relative density

Relative density of Ethiopian traditional beverages brewed from different cereals has
been tested like ethanol and pH values, and the results are presented in table 4.17.

43
 Table 4.17 Relative density of Ethiopian traditional beverage tella

S/N Type of tella Replication Mean

1 Barley 2 1.0239±0.00

2 Corn 2 1.0209±0.00

3 Wheat 2 1.0258±0.00

4 Sorghum 2 1.0208±0.00

5 Teff 2 1.0146±0.00

Accordingly the analysis result, relative density of beverages decreased proportionally

with increasing of ethanol content except beverages prepared from wheat. Therefore,
relative density of teff is lower compared with other beverages. On the contrary, wheat
contains higher relative density from barley, though; ethanol content of wheat is larger
than barley. The result shows that the original wort extract of wheat was not fermented
adequately as of barley.

4.5. Effect of cereals variation on Shelf life of Ethiopian traditional beverage; tella

4. 5. 1. Determination of Real degree of fermentation

On the 2nd phase of analysis (after 20 days of storage from first phase analysis) the results
of real degree fermentation for Ethiopian traditional beverages tella was increased
proportionally with increasing storage time.

Table 4.18 Real degree fermentation results of Ethiopian traditional beverage tella

Cereal variations Time (hrs) Replication Mean (%)

288 2 43.52±0.02
640 2 51.84±0.17
Barley Average 2 47.68±4.81
288 2 48.08±0.04
640 2 58.47±0.06
Corn Average 2 53.27±6.00
288 2 42.41±0.31
640 2 52.25±0.03
Wheat Average 2 47.33±5.68
Sorghum 288 2 48.62±0.18

44
 640 2 58.02±0.03
Average 2 53.32±5.43
288 2 60.32±0.14
640 2 63.15±0.13
Teff Average 2 61.73±1.63
288 10 48.59±6.70
640 10 56.75±4.47
Total Average 10 52.67±6.94
Note; Time was counted from starting preparation period of traditional beverage tella

In this phase of analysis, some cereals particularly corn, wheat and sorghum performed
higher degree of fermentation with increasing of fermentation time compared with the
other beverages.

Figure 4.5 Effect of cereals variation and fermentation time on degree of fermentation
Their degree of fermentation difference from the previous result demonstrated to study
the optimum fermentation time to produce desirable ethanol from cereals. There is no
more change degree of fermentation regarding on teff tella when storage time increases,
which provided the desired content of ethanol within a short fermentation time.

Interaction of cereal variations and time are significant (P ≤ 0.05) for the real degree
fermentation of tella. Hence, the interaction of cereals variation and fermentation time
has a significant effect on the real degree of fermentation upon Ethiopian traditional
beverage tella.

45
 Table 4.19 Analysis result of real degree of fermentation using Factorial ANOVA

Source Type III Sum df Mean Square F Sig.

of Squares
Corrected Model 915.948a 9 101.772 4990.046 .000000
Intercept 55475.204 1 55475.204 2720039.443 .000000
CV 545.420 4 136.355 6685.708 .000000
FT 332.683 1 332.683 16312.000 .000000
CV * FT 37.845 4 9.461 463.896 .000000
Error .204 10 .020
Total 56391.356 20
Corrected Total 916.152 19

4.5.2. Determination of Ethanol

Ethanol content of the beverage has been tested after 20 days of additional fermentation
from the first phase of analysis, and the result is proportionally increased compared with
the previous phase of analysis. Hence, ethanol content of traditional beverage tella
increase directly proportional with fermentation time in the bounded time frame.
Accordingly the findings, tella prepared from teff produced higher ethanol content
whereas barley is lower likewise the first phase of the analysis result.

Table 4.20 Ethanol content of Ethiopian traditional beverage tella

Cereal Replication Time (hrs) Mean (%V/V)

variations
2 288 3.58±0.00
2 640 4.40±0.01
Barley 2 Average 3.99±0.47
2 288 3.94±0.01
2 640 5.17±0.06
Corn 2 Average 4.55±0.71
2 288 3.66±0.00
2 640 4.76±0.01
Wheat 2 Average 4.21±0.64
2 288 4.03±0.00
2 640 5.21±0.01
Sorghum 2 Average 4.62±0.68
2 288 5.43±0.02
Teff 2 640 5.77±0.02

46
 2 Average 5.60±0.20
10 288 4.13±0.71
10 640 5.06±0.48
Average 10 Average 4.59±0.76
Note; Fermentation time is counted from starting preparation period of traditional beverage tella.

During the 2nd phase of analysis the conversion rate of ethanol for corn, wheat and
sorghum beverages were very high compared with the other. Their result demonstrates
the necessity of additional fermentation time to convert sugar into ethanol effectively.

Maximum ethanol results were presented by (Tadele Yohannes, 2013) 6.48%V/V,

(Belachew Desta, 1977) 5.65-6.56%V/V, and (Sahle and Gashe, 1991) 8.00 % V/V in the
previous studies. In this study, however, maximum ethanol content was found from teff
tella 5.77 %V/V within 640 hrs fermentation which is close to the minimum result of
(Belachew Desta, 1977) 5.65-6.56%V/V. In average 4.59 %V/V ethanol was found from
tella prepared from various cereals within 640 hrs of fermentation.

Figure 4.6 Effect of cereal variation and fermentation time on ethanol

Interaction of cereal variations and fermentation time are significant (P ≤ 0.05) for
ethanol content of tella. Hence, the interaction of cereals variation time has a significant
effect on ethanol content Ethiopian traditional beverage tella.

47
 Table 4.21Analysis result of Ethanol using Factorial ANOVA

Source Type III Sum of df Mean Square F Sig.

Squares
Corrected Model 10.970a 9 1.219 2649.763 .000000
Intercept 421.913 1 421.913 917202.130 .000000
CV 6.066 4 1.517 3296.832 .000000
FT 4.362 1 4.362 9482.130 .000000
CV * FT .542 4 .136 294.603 .000000
Error .005 10 .000
Total 432.888 20
Corrected Total 10.975 19

4.5.3. Determination of pH value

One of the main concerns of brewers is to preserve the organoleptic stability of beer
during ageing. A key factor influencing beer ageing and its stability is pH (Tatjana
Rakcejeva, 2013). The pH values of traditional beverages were tested after 20 days of
storage and the results were decreased proportionally compared from the previous phase
analysis. The analyses result were clearly demonstrated the occupation of sourness in the
beverage with time increment.
Table 4.22 pH value of Ethiopian traditional beverage tella

Cereal variations Replication Time (Hrs) Mean

Barley 2 288 4.21±0.02
2 640 3.57±0.01
2 Average 3.89±0.37
Corn 2 288 4.16±0.00
2 640 3.48±0.00
2 Average 3.82±0.39
Wheat 2 288 3.94±0.00
2 640 3.43±0.01
2 Average 3.68±0.30
Sorghum 2 288 4.20±0.00
2 640 3.46±0.01
2 Average 3.83±0.43
Teff 2 288 4.30±0.00

48
 2 640 3.51±0.01
2 Average 3.90±0.46
Average 10 288 4.16±0.13
10 640 3.49±0.05
10 Average 3.82±0.36
Note; Fermentation time was counted from starting preparation period of traditional beverage tella.

Particularly, the pH value of teff, sorghum and corn tella were violently declined
compared from the previous result. However, pH value of wheat tella has performed
better resistance within twenty days of storage. Hence, beverages prepared from wheat
comparatively ensured to encounter the effect of microorganisms which exposed the shelf
life of traditionally fermented beverage tella under aerobic condition.

Figure 4.7 Effect of cereals variation and fermentation time on pH

In the previous studies minimum pH result was presented by (Alemayehu, 2018) which
was 3.28. (Sahle and Gashe, 1991) stated that after ten days of fermentation, tella
becomes too sour to consume due to the growth of Acetobacterspp, which convert
ethanol to acetic acid under aerobic conditions. As of optimum pH is reported from 4.0 to
4.4 (Tatjana Rakcejeva, 2013), and the result of this study in this phase (3.46 to 3.57) is
approximately close to the previous study and not suitable for consumption.

Interaction of cereal variations and storage time are significant (P ≤ 0.05) for pH value of
tella. Hence, the interaction of cereals variation and time has a significant effect on pH
value content Ethiopian traditional beverage tella.

49
 Table 4.23 Analysis result of pH value using Factorial ANOVA

Source Type III Sum of df Mean Square F Sig.

Squares
Corrected
2.439a 9 .271 3387.333 .000000
Model
Intercept 292.460 1 292.460 3655744.000 .000000
CV .121 4 .030 378.063 .000000
FT 2.271 1 2.271 28392.250 .000000
CV * FT .047 4 .012 145.375 .000000
Error .001 10 8.000E-005
Total 294.899 20
Corrected
2.440 19
Total

4.6. Comparison of traditional beverage tella against beer

St.George beer, which is strong beer based on the classification of conformity of
Ethiopian standards (CES52, 2021) was tested to compare against the analysis result of
Ethiopian traditional beverage tella in terms of degree of fermentation, ethanol and pH
results. The analysis result enabled to compare the quality of Ethiopian traditional
beverage tella against St.George beer result.

4.6.1. Determination of real degree of fermentation

Degree of fermentation of St. George beer samples were analyzed using DMA5000M
Alcolyzer to compare against Ethiopian traditional beverage tella and the results are
consolidated in table 4.24

Table 4.24 Comparison of RDF against St.George beer

Type of tella Replication Mean

Barley 2 43.52±0.02
Corn 2 48.08±0.04
Wheat 2 42.41±0.31
Sorghum 2 48.62±0.18
Teff 2 60.32±0.14
St.George beer 2 66.96±0.04

50
Degree of fermentation for St.George beer is larger compared from fermentation range of
Ethiopian traditional beverage tella. Tella prepared from teff which has optimum degree
of fermentation is lower from St.George beer. After 640 hrs of fermentation, however,
degree of fermentation of tella prepared from teff is lower from St.George beer. The
analysis result aligned with the previous unrecorded study stated low degree fermentation
of tella by (Mooha Lee a *. M., 2015). Hence, the comparison result apparently intends a
mechanism to improve the degree of fermentation of Ethiopian traditional beverage tella
as of St. George beer.

4.6.2. Determination of Ethanol

Ethanol content of St. George samples was conducted using DMA5000M Alcolyzer to
compare against Ethiopian traditional beverage tella and the results are presented in table
4.25.

Table 4.25 Comparison ethanol result of against St.George beer

Type of tella Replication Mean

(beer)
Barley 2 3.58±0.00
Corn 2 3.94±0.01
Wheat 2 3.66±0.00
Sorghum 2 4.03±0.00
Teff 2 5.43±0.02
St.George beer 2 4.73±0.02

Importantly, ethanol content of Ethiopian traditional beverage tella prepared from teff is
larger than St. George beer, despite, real degree of fermentation is lower. However,
ethanol content of traditional beverage tella prepared from barley, corn, wheat, and
sorghum are lower from the result of St.George beer.
Ethanol results of barely, corn, wheat and sorghum tella were increased during the 2nd
phase of analysis 4.40, 5.17, 4.76, 5.21 and 5.77 respectively. In this phase of analysis
ethanol content of traditional beverage is above the result of St.George beer except
51
barley. Hence, traditional beverage tella needs additional study to find the optimum time
of fermentation for desirable alcohol and pH results.

4.6.3. Determination of pH
Like degree of fermentation and ethanol, pH value of St. George beer samples were
evaluated to compare against Ethiopian traditional beverage tella and the results are
consolidated in table 4.26.

Table 4.26 Comparison of pH value against St.George beer

Type of tella (beer) Replication Mean

Barley 2 4.21±0.02
Corn 2 4.16±0.00
Wheat 2 3.94±0.00
Sorghum 2 4.20±0.00
Teff 2 4.30±0.00
St.George beer 2 4.14±0.00

The pH values St.George beer is below from pH values of Ethiopian traditional beverage
tella prepared from barley, corn, sorghum and teff. Thus, St.George beer is more sourness
from traditionally prepared beverages during 288 hrs of fermentation. However, pH value
of tella was declining when fermentation time increases. If the declining of pH
maintained on this level Ethiopian traditional beverage tella would grow in the alcoholic
beverage industry like beers.

52
 Chapter Five

5.1 Conclusion
Cereal variations on the preparation of Ethiopian traditional beverage tella have a
significant effect on degree of fermentation, quality, and shelf life of the beverage. In this
study, traditional beverage prepared from teff fermented efficiently and produced
optimum ethanol content of tella compared from the others. Sorghum is the next cereal in
terms of producing better alcohol. In general, alcohol content of Ethiopian traditional
beverage tella prepared from different cereals can be ordered as teff, sorghum, corn,
wheat and barley.

On the other hand, Tella prepared from wheat withstand declining of pH value during 20
days of fermentation. Therefore, wheat is an optimum cereal to prepare better shelf life of
tella under aerobic condition.

Alcoholic result of Ethiopian traditional beverage prepared from barley, corn, wheat and
sorghum are under the alcohol content of St. George beer. Importantly, Alcoholic result
of traditional beverage prepared from teff is higher from St. George beer.

5.2 Recommendation
Real degree of fermentation for Ethiopian traditional beverage tella is lower from St.
George beer. Thus, further research needs to improve degree of fermentation of tella. In
addition, additional study needs to determine the optimum fermentation time of tella to
produce desirable alcohol and pH.

It is crucial to prepare national standards to improve quality variation of Ethiopian

traditional beverage tella. On the other hand, effect of tella ingredients such as weira and
gesho, and constituents like derekot should be studied to improve shelf life of Ethiopian
traditional beverage tella.

53
 References
1. Abegaz, B. a. (1995). Geshoidin: A bitter principle of Rhamnus prinoides and
other constituents of the leaves. Bulletin. Chem.Soc. Ethio. 2: 107-114 , 2: 107-
114.

2. Alemayehu, H. G. (2018). Physico-chemical characterization of commercial

Local Alcohol beverages available in South Nations, Nationalitiesand People’s
Regional State, Ethiopia. International Journal of ChemTech Research , pp 227-
231,.

3. Anthony J. Cutaia1, A.-J. R. ( 2009). Examination of the Relationships Between

Original, Real and Apparent Extracts and Alcohol in Pilot Plant and
Commercially Produced Beers. J. Inst. Brew. , 115(4), 318–327,.

4. Ashenafi, M. &. (1995). Some microbiological and nutritional properties of Borde

and Shamita. Ethiop. J. Health Dev. 9: , 105-110.

5. Ashenafi, M. (2002). The microbiology of Ethiopian foods and beverages. SINET

Eth. J. Sci , 25(1): 97-140.

6. Ashenafi, M. (2000). Traditional brewing in Ethiopia. In proceeding:Modern and

traditional brewing in Ethiopia. The chemical society of Ethiopia , pp. 1-12.

7. Berhanu, A. (2014). Microbial profile of Tella and the role of gesho (Rhamnus
prinoides) as bittering and antimicrobial agent in traditional Tella (Beer)
production. International Food Research Journal , 21(1): 357-365 .

8. Berne, L. (2005). The endogenous endoprotease inhibitors of barley and malt and
their roles in malting and brewing. J. Cereal. Sci. , 42: 4-7.

9. Debebe, G. (2006). Determination of ethanol level in beverages. M. Sc. Thesis,

Department of chemistry, Addis Ababa University, Addis Ababa .

10. Dersehilign Awegichew Teshome1, M. R.-C. (2017). Chemical compositions of

traditional alcoholic beverages and consumers’ characteristics, Ethiopia. African
Journal of Food Science , 11(7) pp. 234-245.

54
11. Desta, B. (1977). A survey of the alcoholic contents of traditional beverages.
Ethiop. Med. J. 15: , 65-68.

12. Dinesh Kumar, V. K. (2013). quality parameter requirement and standards for
malt barley-a review. agricultural research communication centre , 34 (4) : 313-
317.

13. Fite, A. T. (1991). Methanol, fusel oil and ethanol contents of some Ethiopian
traditional alcoholic beverages. SINET. Ethiop J Sci. 14 , : 19-27.

14. Fullas, F. (2003). Spice plants in Ethiopia: their Culinary and Medicinal
applications.

15. Gebreyes, M. M. (2020). Characterization of Nutritional, Antinutritional, and

Mineral Contents of Thirty-Five Sorghum Varieties Grown in Ethiopia. Hindawi
International Journal of Food Science .

16. Getachew Tafere Department of Biotechnology, M. U. (2015). A review on

Traditional Fermented Beverages of Ethiopian. Journal of Natural Sciences
Research .

17. Girmay Tsegay*, L. S. (2019). Assessment of Proximate Chemical Compositions

and Tryptophan Content of Released and Improved Ethiopian Maize Varieties.
Academic Research Journal of Agricultural Science and Research , Vol. 7(1), pp.
14-20,.

18. Helen, E. a. (1987). Antifungal and other compounds isolated from the roots of
New Zealand flax plants (the genus Phormium). J. Natural. Prodc. , 50(4): 767-
776.

19. James Bruner 1, 2. ,. (2021). Dry-Hop Creep Potential of Various Saccharomyces

Yeast Species and Strains.

20. Kebede, A. F. (2002). Indigenous processing methods and raw materials of borde,
an Ethiopian traditional fermented beverage. J. Food. Technol. Africa , 7: 59-64.

55
21. Kebede, T. (1994). The bitter constituent of Gesho (Rhamnus prinoides) leaves.
M.Sc.Thesis, Department of Chemistry, Addis Ababa University,Addis Ababa.

22. Lemi, B. W. (2020). Microbiology of Ethiopian Traditionally Fermented

Beverages and Condiments. Hindawi International Journal of Microbiology .

23. Megersa, N. (2015). Disposable residue of traditionally fermented ethiopian local

beer (tella) for quantitative uptake of pesticide pollutants from contaminated
waters. Ethiop. J. Sci. , 38(2):133-147.

24. Mooha Lee a, *. M. (2015). Uniqueness of Ethiopian traditional alcoholic

beverage of plant origin,tella. Journal of Ethnic Foods , 110-114.

25. Nigusie, N. (2008)). . Evaluation of the efficiency of starch conversion in

theprocess of tella fermentation. M. Sc. Thesis, Department of Biology
(Microbiology), Addis Ababa University, Addis Ababa.

26. Okafor, N. (1972). The microbiological basis of a method for palm-wine. J Appl.
Bacteriol. , 43:59-161.

27. Sahle, A. a. (1991). The microbiology of tella fermentation. SINET Ethiop. J. Sci ,
14:81 - 92.

28. Selinus, R. .. ((1971)). The traditional foods of the central Ethiopian highlands.
Scandinavian Institute of African Studies.

29. Steinkraus, K. ((1983) (ed).). Handbook of Indigenous Fermented Foods. New

York: Marcel Dekker, Inc.

30. Tadele Yohannes, F. M. (2013). Preparation and physicochemical analysis of

some Ethiopian traditional alcoholic beverages. African Journal of Food Science ,
pp. 399-403.

31. Tatjana Rakcejeva, V. S. ( 2013). ASLT Method for Beer Accelerated Shelf-Life
ASLT Method for Beer Accelerated Shelf-Life. World Academy of Science,

56
 Engineering and Technology International Journal of Nutrition and Food
Engineering , Vol:7, No:2,.

32. Tesfaye, G. M. (2017). Technology and microbiology of traditionally fermented

food and beverage products of Ethiopia: A review. African Journal of
Microbiology Research , (21), pp. 825-844,.

33. Vogel, S. a. (1983). Ethiopian tej. In: Handbook of Indigenous Fermented Foods.
Steinkraus, K.H. (ed). New York.: Marcel Dekker, Inc.pp 363-365.

34. Yadesa Abeshu, E. A. (1(3): 97-100). Evaluation of Proximate and Mineral

Composition Profile for Different Food Barley Varieties Grown in Central
Highlands of Ethiopia. World Journal of Food Science and Technology , 2017;.

35. Yegrem, L. (2019). Comparative analysis of proximate and mineral composition

of released Teff (Eragrostis tef (Zucc.) Trotter) varieties in Ethiopia. academic
research journal .

57