Journal of Food Composition and Analysis 115 (2023) 105015

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Journal of Food Composition and Analysis

journal homepage: www.elsevier.com/locate/jfca

Quality and biochemical composition of Ethiopian coffee varied with

growing region and locality
Mohammed Worku a, *, Tessema Astatkie b, Pascal Boeckx c
a
Department of Horticulture and Plant Sciences, College of Agriculture and Veterinary Medicine, Jimma University, Jimma, Ethiopia
b
Faculty of Agriculture, Dalhousie University, Truro, NS, Canada
c
Isotope Bioscience Laboratory – ISOFYS, Faculty of Bioscience Engineering, Ghent University, Belgium

A R T I C L E I N F O A B S T R A C T

Keywords: Coffee quality and biochemical composition are believed to vary with geographical origin. Seed weight, quality
Arabica coffee attributes, and contents of caffeine, sucrose and chlorogenic acids of green arabica coffee beans from five coffee
100 seed weight regions (eastern or Harar, southern, southwestern, western and northwestern regions of Ethiopia) and 24 coffee
Quality attributes
localities across the five coffee regions were evaluated. Coffee samples varied with their growing region and
Caffeine
Chlorogenic acids
locality in these bean features. Coffee samples from Harar and southern regions had much higher bean physical
Sucrose quality (BPQ), acidity and total preliminary quality (TPQ) than those from southwestern region, but lower
Coffee regions in Ethiopia caffeine content than those from the remaining regions. Samples from Harar also had higher 100 seed (bean)
weight, and sucrose and total chlorogenic acids (TCGAs) contents than those from the other regions. Lem-Kaffa
coffee from southwestern region had much lower BPQ, TPQ and TCGAs than most of the other locality coffees
and significantly differed from any of the studied locality coffees in its 100 bean weight, quality and biochemical
composition. This study generally highlights the variation of Ethiopian coffee with growing region and locality,
and the presence of similarity between some locality coffees across the coffee regions in their bean weight,
quality and biochemical composition.

1. Introduction Maillard reactions (Flament, 2002). High levels of caffeoylquinic acids,

Coffee is the second most traded commodity after oil and it is a main
source of foreign currency earnings for over 80 tropical countries (Gole,
2015). In Ethiopia, the coffee industry dominates the agriculture sector
in its contribution to the national economy in general and exports in
particular, accounting for 10% of the total agriculture production, 10%
of total government revenue, 40% of total export and 25–30% of the
total export earnings of the country (Tefera et al., 2014; Worku, 2019). It
also provides an employment opportunity for about a quarter of the
population of the country (Gole, 2015; Tefera et al., 2014).
Quality is one of the important attributes of coffee that determines its
market price and consumer preference, and biochemical composition
determines the cup quality of coffee. Biochemical composition of green
coffee beans is decisive for sensory quality through roasting-induced
chemical reactions. For instance, high trigonelline and 3,4-dicaffeoyl­
quinic acid levels in coffee beans are strongly correlated with high cup
quality (Farah et al., 2006), and proteins and amino acids are essential
for the conversion of reducing sugars into aroma precursors through
feruloylquinic acids and their oxidation products are, however, associ­
ated with poor cup quality and off-flavor (Farah, 2012; Farah et al.,
2006) and caffeine is associated with bitterness and astringency of the
brew (Sualeh et al., 2020). Some of the biochemicals that contribute to
flavor, e.g., chlorogenic acids and trigonelline, have also useful bioac­
tive properties, such as antioxidant, α-glucosidase inhibitory and anti­
microbial activities (Bhattacherjee and Datta, 2015; Bhattacherjee et al.,
2014; Zhou et al., 2012), whereas others, e.g., caffeine and diterpenes,
may produce harmful effects by affecting endothelial function and
raising a fraction of blood lipids, respectively (Godos et al., 2014).
Consumptions of caffeine, on the other hand, could reduce a risk of
Parkinson’s disease development (Higdon and Frei, 2006) and rise en­
ergy availability, alertness and concentration, and learning capacity
(Farah, 2012; Jiang et al., 2014).
At farm level, various factors such as genetics (e.g., species or vari­
ety), growing environment (e.g., soil, shade, temperature and rainfall),
agricultural practice (e.g., fertilization, pruning, harvesting method)
and postharvest processing technique (e.g., dry or wet processing),

* Corresponding author.
E-mail address: mohaworku@gmail.com (M. Worku).

https://doi.org/10.1016/j.jfca.2022.105015
Received 21 June 2022; Received in revised form 25 October 2022; Accepted 30 October 2022
Available online 1 November 2022
0889-1575/© 2022 Elsevier Inc. All rights reserved.
M. Worku et al. Journal of Food Composition and Analysis 115 (2023) 105015

influence the quality and the biochemical composition of coffee (Ber­
trand et al., 2012; Duarte et al., 2010; Leonel and Philippe, 2007; Tarzia
et al., 2010; Taveira, 2014; Vaast et al., 2006). The growing environ­
ment that determines these attributes of coffee can vary with
coffee-growing region and locality across the coffee-growing regions
and within a given coffee-growing region. For example, the
coffee-growing regions in Ethiopia (i.e., eastern or Harar, southern,
southwestern, western and northwestern Ethiopia) vary in coffee genetic
diversity, agro-ecology, and/or coffee production system. The genetic
diversity of coffee and the amount and distribution of rainfall generally
decreases from the southwestern coffee region towards the eastern and
northwestern coffee regions (Moat et al., 2017a; Worku, 2019). Coffee in
the eastern (Harar), southern and northwestern coffee regions is mainly
produced under a garden system where coffee is growing with other
horticultural crops and with almost no shade. However, in the south­
western and western regions, it is largely produced under a
semi-forest/plantation coffee (managed forest coffee with a high shade
tree canopy cover) and semi-forest and garden systems, respectively.
More details on these coffee regions and production systems are avail­
able in Gole (2015) and Worku et al. (2015).
There are also several coffee-growing localities within each coffee
region of Ethiopia that can vary in their growing environment (e.g.,
elevation, temperature, rainfall, or soil) influencing coffee quality and
biochemical composition (Table 1; Worku et al., 2018). Consequently, in
each coffee region, different coffee types that are thought to vary in their
quality attributes are recognized in the local and/or world markets; e.g.,
Harar coffee in eastern coffee region; Sidamo, Yirgacheffe, Guji and
Kochere coffees in southern coffee region; Jimma, Limmu, Kaffa, Bebeka
and Tepi coffees in southwestern coffee region; Lekemt (Wellega) coffee
in western coffee region; and Wenbera and Zegie coffees in northwestern
coffee region (Boot, 2011; Worku, 2019). As per Kufa (2012), Harar,
Sidamo, Yirgacheffe, Jimma and Limmu, and Lekemt coffees have a
Table 1
Summary information of coffee samples collected from farms in 24 localities
(districts) in five coffee regions (Elev. = elevation; Lat. = latitude; Long. =
longitude; n = number of samples).
Region Sampling Elev. (m Lat. Long. n
district asl) (◦ N) (◦ E)
‘mocha’ (chocolate), spicy, floral, winy and fruity flavor, respectively.
However, according to Boot (2011), high quality Harar coffee is notable
for a fruity flavor and a creamy body; Sidamo coffee for a very wide
variety of flavors; Yirgacheffe coffee for a fruity flavor, a bright acidity
and a silky mouth feel; Limmu coffee for a mild acidity; and Lekemt
coffee for a perfume-like aftertaste. Similarly, the results of the cup of
excellence competition conducted in 2020 and 2021 in Ethiopia showed
a difference among coffee-growing regions and among localities in
cupping scores (Ethiopia, 2020; Ethiopia, 2021; Montagnon, 2020). The
results also showed a trend between genetics and geography of coffee in
cupping scores (Montagnon, 2020). Furthermore, Mehari et al. (2016a;
2016b) reported significant differences among coffee regions and coffee
types (i.e., Harar, Sidamo, Yirgacheffe, Jimma, Kaffa, Lekemt and
northwestern or Gojam coffees) in their contents of caffeine, trigonelline
and individual chlorogenic acids (CGAs); but not in the content of total
chlorogenic acids. Sualeh et al. (2020) also reported substantial varia­
tions among coffee bean samples growing in different districts in Bench
Maji and Sheka Zones, southwestern Ethiopia, in their caffeine, trig­
onelline and chlorogenic acids contents and antioxidant activity. How­
ever, data on composition of other cup quality precursors, such as
sugars, fatty acids and proteins or amino acids, of Ethiopian coffee
growing in different coffee regions and localities are limited.
Except the aforementioned general observations on quality, and
pioneer studies on composition of some biochemicals (e.g., caffeine,
trigonelline and CGAs), there is no comprehensive study on physical
attributes (e.g., weight, size or shape), quality and biochemical
composition of Ethiopian coffee across coffee-growing regions and lo­
calities. Specifically, different quality attributes and some important cup
quality precursors (e.g., sucrose) of Ethiopian coffee across coffee-
growing regions and localities have not been studied so far. Hence,
the objectives of this study were (1) to examine the variation among
Ethiopian coffees grown in different coffee regions and localities (dis­
tricts) in seed (bean) weight, quality attributes and biochemical
composition, and (2) to determine similarities among Ethiopian coffees
grown in different localities across coffee-growing regions (i.e., eastern
or Harar, southern, southwestern, western and northwestern Ethiopia)
in terms of bean weight, quality and biochemical composition.
2. Materials and Methods

Eastern (Harar) (n Bedeno 1540–2000 9.0924 41.6322 3 2.1. Description of the study regions
= 15)
Melkabelo 1500–2000 9.0833 41.3667 3
Darolebu 1873–2000 8.5942 40.3061 3 The samples of green arabica coffee beans were collected from 24
Habro 1600–2000 8.8667 40.5167 3 coffee-growing localities (districts) located in five coffee regions,
Chiro 1826–2100 9.0674 40.8657 3 namely eastern (Harar), southern, southwestern, western and north­
Southern (n = 3) Yirgacheffe 1800–1950 6.1620 38.2058 3
western Ethiopia (Fig. 1; Table 1). These coffee regions have different
Southwestern (n = Jimma 1800–2000 7.7333 36. 7333 3
33) agro-ecology and/or coffee production systems. Harar coffee region is
Limmu-Kossa 1600–2000 7.8500 36.8000 3 located in three administrative zones (East Hararge, West Hararge and
GommaIIa 1660–1820 7.9619 36.6950 3 Arsi Zones) (Fig. 1) at elevations between ca. 1200 and 2100 m asl
Yayu 1200–2000 8.3031 35.8319 3 (Table 1). It has a dryer climate than the other coffee regions and ex­
Gorea 1770–2085 8.1513 35.5357 3
Lem-Kaffaa 1700–1820 7.3793 36.2492 3
periences a bimodal rainfall, with short rains from March to May and
Bebekaa 850–1200 6.8835 35.4281 3 long rains from June to September (Abebe, 2010; Klingele, 1998). The
Tepia 1100–1300 7.1870 35.4155 3 mean annual total rainfall is about 800–1200 mm, which is low for the
Gemadiroa 1700–1800 7.4724 35.4172 3 cultivation of good coffee, and the mean temperature is about 16–20 ◦ C
Godere1a 1800–2000 7.3187 35.1269 3
(Moat et al., 2017a). Leptosols, luvisols, vertisols, cambisols and calci­
Godere2a 1350–1800 7.3187 35.1269 3
Western (n = 9) Gimbi 1700–2200 9.1927 35.7894 3 sols (Dewitte et al., 2013) are the major soil types existing in this coffee
Haru 1672–1950 8.9800 35.8253 3 region. The green coffee beans of this region, produced predominantly
Anfilo 1300–1800 8.4825 34.5847 3 under a garden system (coffee growing mostly around homesteads with
Northwest (n = Ankasha 1600–1860 10.7270 36.8042 3 other crops, e.g., fruit crops, root crops, chat, enset, vegetables and
12)
Jabitenan 1800–1917 10.6824 37.3128 3
spices, and a low level of shade tree canopy cover), are exclusively un­
Bure 2050–2091 10.7074 37.0998 3 washed and are known as Harar coffee (Boot, 2011).
Mecha 1900–2100 11.4140 37.1305 3 The southern coffee region of Ethiopia can be divided into two: Rift
Total n 72 valley and southeastern coffee zones (Moat et al., 2017b). But, the
a
GommaII, Lem-Kaffa, Gore, Bebeka, Tepi and Gemadro, and Godre1 and woina-dega agro-climatic zone (moist to sub-humid warm sub-tropical
Godre2 are sampling sites located in the Gomma, Gewata, Ale, South Bench, climate), situated between 1500 and 2300 m asl in Sidama, Gedeo,
Yeki, and Godere Districts, respectively. Guji and Bale Zones (southeastern coffee zone) (Fig. 1), is the main

2
M. Worku et al.
Fig. 1. The coffee-growing areas of Ethiopia; dark green = Eastern (Harar) coffee region, dark blue and light green = Southern coffee region (dark blue =
Southeastern coffee zone, light green = Rift coffee zone), light blue = Southwestern and western coffee regions, pink = Northwestern coffee region; adapted from
Moat et al. (2017b).
coffee producing zone in the southern coffee region (Boot, 2011). This
coffee region generally has a bimodal rainfall, with a short rainy season
from mid-February to April and a main rainy season from June to
October (Yilma, 2001). The mean annual total rainfall ranges from 1000
to 1600 mm and the mean annual temperature ranges from 16◦ to 20◦ C
(Moat et al., 2017a). In the southern region, luvisols, fluvisols, cambisols
and leptosols are the major soil types (Dewitte et al., 2013). Coffee in
this region is predominantly produced under a garden system and pro­
cessed as washed coffee. It is often designated as Sidamo and Yirgacheffe
coffee for marketing purposes (Boot, 2011).
The major coffee-growing areas in southwestern Ethiopia are located
in the Jimma, Illubabor, Kaffa, Bench Maji, Sheka and Godere Zones
(Fig. 1) and at elevations between ca. 850 and 2200 m asl (Table 1). The
climate in this coffee region varies from sub-moist tropical to hot and
humid rainforest climate with a high and reliable unimodal rainfall. The
mean annual temperature ranges from ca. 15–21 ◦ C and the mean
annual total rainfall ranges from ca. 1500–2100 mm, well distributed
over a period of 8–9 months between March and November (CPDE,
2011; ERA, 2009; Kassa, 2017; Moat et al., 2017a). The soils of the re­
gion are moderately acidic, and nitosols is the dominant soil type in the
coffee-growing areas. Other major soil types available in the highlands
3
Journal of Food Composition and Analysis 115 (2023) 105015
of the region include vertisols, leptosols, regosols, cambisols, alisols and
acrisols (Dewitte et al., 2013). The dominant coffee production system in
this region is semi-forest/plantation coffee, which is managed forest
coffee that has a high shade tree canopy. The region produces both
washed and unwashed coffee, with unwashed coffee constituting a
larger share of the total production. The coffee is designated as Limmu,
Jimma, Kaffa, Bebeka and Tepi coffee for marketing purposes (Boot,
2011).
As shown in Fig. 1, the major coffee-growing areas in western
Ethiopia are in West and Kelem Wellega Zones. The western coffee re­
gion has a similar agro-climatic and soil conditions with southwestern
coffee region. It receives a unimodal rainfall, and the rainy period ranges
from February to November. The major soil type in this coffee region is
nitosols (Dewitte et al., 2013). The coffee production systems in this
region can be categorized as semi-forest/plantation and garden systems
and the coffee produced is mainly an unwashed coffee. Washed coffee
production is recently started in the region with technical support of
TechnoServe, Ethiopia. The coffee is designated as Lekemt coffee for
marketing purposes (Boot, 2011), but locally, it is called Wellega coffee.
Northwestern region is a minor coffee producing region of Ethiopia
and the main coffee-growing area in this region is concentrated in the
M. Worku et al.
highlands (ca. 1500–2100 m asl) of three administrative zones, namely
the Metekel, Agew Awi and West Gojam Zones. This coffee area is the
smallest coffee region of Ethiopia (Fig. 1), but it covers half of the minor
coffee-growing areas that are located in various parts of the country and
accounts for ca. 2% of the total land area covered by coffee (Teketay,
1999). Furthermore, coffee production units in this region are very small
and they mainly use a garden system. The region has a unimodal rainfall
with mean annual total values ranging from ca. 1400–1800 mm,
distributed from May/June to August/September (Cheung et al., 2008;
Moat et al., 2017a). Yet, the rain in this region comes later in the year
and ends earlier, resulting in a shorter, more intense wet season and a
longer dry season, compared to the wet southeastern, southwestern and
western coffee regions (Moat et al., 2017a). In this coffee region, the
mean temperature is about 16–19 ◦ C (Fazzini et al., 2015) and the soil
types are predominantly nitosols followed by cambisols (Dewitte et al.,
2013). The coffee produced in this region is unwashed, and, until very
recently, it has been used only for local consumption.
2.2. Sample collection and preparation
Samples of natural sundried (unwashed) green arabica coffee beans
(n = 72, each about 1 kg) were collected from 24 localities or districts (3
samples from three different farms per locality or district), located in
five coffee growing regions of Ethiopia: eastern or Harar (n = 15),
southern (n = 3), southwestern (n = 33), western (n = 9) and north­
western (n = 12) (Fig. 1; Table 1). Additional 30 samples of unwashed
green arabica coffee beans [each about 1 kg that came from the 10
districts in each eastern, southern and southwestern coffee region (i.e.,
10 samples per coffee region) into Ethiopia Commodity Exchange (ECX)
warehouses at Dire Dawa (eastern Ethiopia), Awasa (southern Ethiopia)
and Jimma (southwestern Ethiopia)] were collected from ECX ware­
houses at Dire Dawa, Awasa and Jimma. The samples collected from the
ECX warehouses were used for the determination of quality attributes of
green beans (i.e., bean physical quality, acidity, body, flavor, cup quality
and total preliminary quality) and those from the farms for the deter­
mination of 100 seed weight, quality attributes (bean physical quality,
acidity, body, flavor, cup quality and total preliminary quality) and
biochemical compositions (caffeine, sucrose, total chlorogenic acids, 3-
caffeoylquinic acid, 4-caffeoylquinic acid, 5-caffeoylquinic acid,
3,5dicaffeoylquinic acid, 4,5dicaffeoylquinic, feruloylquinic acid and
feruloyl-caffeoylquinic acid) of green beans. Detailed information on
samples collected from farms is given in Table 1.
2.3. Quality analysis
The quality attributes of the green coffee beans were determined at
Jimma ECX cupping laboratory, Ethiopia, following the coffee quality
assessment procedure of ECX (Worku et al., 2016, 2018). The pre­
liminary quality assessment at ECX included bean physical quality at­
tributes (primary defect, secondary defect, odor and color of the green
beans), cup quality attributes (acidity, body, cup cleanness and flavor of
the brew) and total preliminary quality (physical quality + cup quality).
The primary defects include full black, sour, fungus attacked and insect
damaged beans as well as the presence of foreign matters. The secondary
defects include partial black, broken, insect damaged, faded and coated
beans.
The three bean physical quality attributes (primary defects, sec­
ondary defects and odor) were evaluated by using a sub-sample of 350 g.
The evaluation for primary and secondary defects was scored on a 0–15
scale and for odor on a 0–10 scale. For cup quality analysis, a sub-sample
of 150 g was roasted for ca. 8–12 min using a heated roaster (Probat, 4
Barrel Roaster, Germany) at about 250 ◦ C to obtain a medium roasted
coffee. After air-cooling, each roasted coffee sample was grounded into a
size of < 20 mesh. Then, 13.75 g of coffee powder was added into five
cups and about 250 mL of clean and odor free hot water (about 93 ◦ C)
was poured on the coffee powder. The contents of all cups were then
4
Journal of Food Composition and Analysis 115 (2023) 105015
stirred until the coffee powder was completely infused with hot water.
Subsequently, each cup was evaluated by three Q-certified cuppers for
its acidity, body, cleanness and flavor, and for each attribute, the tasting
scores were recorded on a 0–15 scale. Finally, bean physical quality (=
primary defects + secondary defects + odor) (40/100), cup quality (=
acidity + body + cleanness + flavor) (60/100) and total preliminary
quality (= bean physical quality + cup quality) (100) per sample were
calculated (Worku et al., 2016, 2018).
2.4. Chemical analysis
The contents of caffeine, sucrose, total chlorogenic acids (TCGAs)
and the seven different positional isomers of chlorogenic acids [3-
caffeoylquinic acid (3-CQA), 4-caffeoylquinic acid (4-CQA), 5-caffeoyl­
quinic acid (5-CQA), 3,5dicaffeoylquinic acid (3,5diCQA), 4,5dicaf­
feoylquinic (4,5diCQA), feruloylquinic acid (FQA) and feruloyl-
caffeoylquinic acid (FCQA)] in each coffee sample were determined at
the Department of Food Safety and Food Quality laboratory, Ghent
University, Belgium by following the procedures used in our previous
publication (Worku et al., 2018). TCGAs content was defined as the sum
of the seven identified positional isomers of chlorogenic acids; i.e.,
3-CQA, 4-CQA, 5-CQA, 3,5diCQA, 4,5diCQA, FQA and CFQA.
Before chemical analysis, 50 g of each sample was ground to a par­
ticle size smaller than 0.5 mm (M20 universal mill, IKA, Germany). For
analyses of caffeine and chlorogenic acids contents, 100 mg powder of
coffee beans per sample was subjected to direct solvent extraction with
10 mL of methanol: water: acetic acid (12:27:1, v-v:v) containing
2 mg mL− 1 ascorbic acid in an ultrasonic bath for 15 min. The crude
solvent extract was then filtered through a 0.45 µm PTFE filter and the
filtrate was injected into an HPLC system (Finnigan Surveyor, Thermo
Scientific, USA), equipped with a reversed phase analytical column
(250 ×4.6 mm i.d., 5 µm, Prevail C18, Alltech, CITY) and guard column
(10 ×3.9 mm i.d., 5 µm, Symmetry C18, Waters, Zellik, Belgium). The
system was maintained at 25 ◦ C and the total measurement took 70 min.
The detection was done using a photodiode array (PDA) detector at
280 nm for caffeine and at 320 nm for chlorogenic acids. The mobile
phase was a stepwise gradient composed of 0.2% acetic acid in water (v/
v) and HPLC grade methanol ranging from 10% to 70%, with a flow rate
of 1 mL min− 1 and an injection volume of 50 µL. Calibration was carried
out by injecting concentration series of caffeine and chlorogenic acids
every 20 samples using an adapted response factor for different
chlorogenic acid types. Limits of detection (LOD) and quantification
(LOQ) respectively were 1.5 and 3.0 µg g− 1 of sample for caffeine, and
0.8 and 1.6 µg g− 1 of sample for chlorogenic acids. Peak identification
was based on retention time and confirmed using an already established
liquid chromatography time of flight mass spectrometry (LC-TOF-MS)
method (Perrone et al., 2008) on selected samples (deviation from
theoretical mass <1.6 ppm). Electrospray ionization source (ESI) of
TOF-MS was operated in positive mode with the nebulizer gas (N2) flow
set at 3 L min− 1 and desolvation temperature at 300 ◦ C. Data were ac­
quired by ChromQuest software (Thermo Fisher Scientific, Waltham,
MA, USA), Version 4.1 SP2. Each sample was analyzed in triplicate.
For analysis of sucrose content, first, an internal standard solution
was prepared using 600 mg fenyl-beta D-glucopyranoside and 100 mL
distilled water. Next, 0.5 g powder of coffee beans from each sample was
mixed with 25 mL distilled water containing 30 mg of fenyl-beta D-
glucopyranoside, as internal standard in a hot water bath (60 ◦ C,
30 min). Carrez I (15% K4Fe(CN)6 in water, 15 g L− 1) and carrez II (30%
ZnSO4 in water, 30 g L− 1) (5 mL each) were added to the mixture and
distilled water was then added to obtain a total volume of 50 mL. After
mixing, the extract was filtered through a 0.45 µm PTFE filter, and
0.5 mL of the filtrate was transferred to a gas chromatograph-vial and
dried using nitrogen gas. To the dried extract, 0.5 mL of STOX-reagent
(hydroxylamine hydrochloride in dry pyridine, 25 g L− 1) was added
and the vial was placed in an oven at 60 ◦ C for 30 min. After cooling to
room temperature, 0.5 mL of hexamethyldisilazane (HMDS) and
M. Worku et al.
0.05 mL of trifluoroacetic acid (TFA) were added and then kept for
60 min to allow phase separation. Finally for analysis, 1 µL was taken
from the upper layer and injected into a gas chromatograph (GC-3380,
Varian, USA) equipped with a flame-ionization detector (Varian In­
strument Group, Walnut Creek, CA) using a 1079 automatic injector.
The chromatographic parameters were: stationary phase - (5%-phenyl)-
methylpolysiloxane, film thickness 0.25 µm, 30 m x 0.32 mm inside
diameter (i.d.) (Agilent Technologies, Palo Alto, CA, USA); and mobile
phase - He at 1 mL min− 1, split 1/40, injector temperature = 250 ◦ C;
detector temperature = 340 ◦ C; temperature program = 180 ◦ C for
1 min, ramp at 15 ◦ C min− 1 to 290 ◦ C. The flame ionization detector was
operated with hydrogen and air at 30 and 300 mL min− 1, respectively
and helium at 20 mL min− 1 as makeup gas. Identification was based on
retention time of known compounds. LOD was 0.05 mg g− 1 and LOQ
was 0.1 mg g− 1. Each sample was analyzed in triplicate.
To express the biochemical composition data on dry weight basis, the
dry matter of each sample of green coffee beans was determined by
taking the weight of the same amount of the grind subsamples of each
sample used for chemical analysis (i.e., 100 mg for caffeine and
chlorogenic acids analyses and 0.5 g for sucrose analysis) after 24 h
oven drying at a temperature of 105 ◦ C and air-cooling down to a
temperature of ca. 21 ◦ C.
2.5. Statistical analysis
The effect of the Major region (3 levels: Harar, Southern and
Southwestern) on quality attributes (bean physical quality, acidity,
body, flavor, cup quality and total preliminary quality) was determined
using a Completely Randomized Design (CRD). The effects of Region (5
levels: Harar, Southern, Southwestern, Western and Northwestern) and
Locality (District), nested in Region, on 100 seed weight, quality attri­
butes (bean physical quality, acidity, body, flavor, cup quality and total
preliminary quality) and biochemical compositions (caffeine, sucrose,
TCGAs, 3-CQA, 4-CQA, 5-CQA, 4,5diCQA, 3,5diCQA, FQA and CFQA)
were determined using a Nested design, with District nested in Region.
The statistical analysis was done using the Mixed procedure of SAS 9.4
(SAS, 2014). For each response variable, the validity of model assump­
tions was verified by examining the residuals as described in Mont­
gomery (2020). For significant (p < 0.05) effects, multiple means
comparison was conducted using the lsmeans statement of Proc Mixed
with Tukey adjustment at the 5% level of significance. The similarity of
the 24 districts in terms of the response variables were determined using
a Cluster analysis (multivariate analysis of complete linkage clustering),
and dendrograms were produced according to Johnson and Wichern
(2007).
3. Results
The ANOVA p-values that show the significance of the effect of re­
gion on quality of green coffee beans (i.e., ANOVA results of the samples
collected from the EXC warehouses in three main coffee regions: Harar,
Southern and Southwestern regions) are presented in Table 2. The
ANOVA p-values that show the significance of the effects of Region and
Locality (District), nested in Region, on 100 seed weight, quality and
biochemical composition of green coffee beans (i.e., ANOVA results of
the samples collected from farms in 24 districts in five regions) are
Table 2
ANOVA p-values showing the significance of the main effect of Main region on
quality attributes of green arabica coffee beans. Significant effects that require
multiple means comparison are shown in bold.
Source of variation BPQ Acidity Body Flavor CQ TPQ
Region 0.005 0.012 0.485 0.131 0.102 0.013
BPQ = Bean physical quality, CQ = Cup quality, TPQ = Total preliminary
quality.
5
Journal of Food Composition and Analysis 115 (2023) 105015
presented in Table 3 and Table 4. As indicated in Table 2, bean physical
quality, acidity and total preliminary quality in the three major coffee-
growing regions varied significantly (p < 0.05). Also, as shown in
Table 3 and Table 4, the 100 seed weight, bean physical quality,
caffeine, TCGAs, 5-CQA, 3,5diCQA, 4,5diCQA and FQA in the five
coffee-growing regions varied significantly (p < 0.05). Moreover, 100
seed weight, bean physical quality, total preliminary quality and all
studied biochemicals in the coffee-growing localities (districts) varied
significantly (p < 0.05). Cup quality marginally (p = 0.052) varied
among the coffee-growing districts (Table 3).
3.1. Variation among coffee regions
Coffees of the Harar and southern regions were significantly higher
in bean physical quality, acidity and total preliminary quality than that
of the southwestern region (Table 5). But, there were no significant
differences between coffees of Harar and southern regions in these
quality variables (Table 5) and among coffees of the three main coffee-
growing regions in terms of body, flavor and cup quality (Table 2), and
their overall means were 9.4, 9.8, and 43.6 scores, respectively. The
coffee also varied among the five coffee regions in 100 seed weight, bean
physical quality, caffeine, TCGAs, 5-CQA, 3,5diCQA, 4,5diCQA and FQA
(Tables 3 and 4).
3.2. Variation among coffee localities
The mean values of 100 seed weight, bean physical quality, total
preliminary quality, caffeine, sucrose and TCGAs obtained from 24
coffee-growing localities (districts), nested in region, are presented in
Table 6 and those of the seven positional isomers of chlorogenic acids in
Table 7.
As indicated in Table 6, Gore(Southwestern, SW) coffee had a
significantly higher 100 seed weight than the coffees of all other local­
ities, excluding Darolebu(Harar, H) and Chiro(H) coffees. Darolebu(H)
coffee had also a higher 100 seed weight compared to Tepi(SW), Anfilo
(Western, W), Ankasha(Northwestern, NW), Jabitenan(NW) and Mecha
(NW) coffees. But, the 100 seed weight of Ankasha(NW) and Jabitenan
(NW) coffees were significantly lower than those of Darolebu(E), Chiro
(E), Gore(SW) and Haru(W) coffees. The bean physical quality and total
preliminary quality of Lem-Kaffa(SW) coffee were significantly lower
than those of the coffees of all other localities, barring Ankasha(NW)
coffee in the case of the former variable, and Tepi(SW), Gemadiro(SW),
Gimbi(W) and Ankasha(NW) coffees in the case of the latter variable
(Table 6).
The caffeine content of Jabitenan(NW) coffee was significantly
higher than those of the coffees at all other localities, except for Habro
(H), Gimbi(W), Haru(W) and Mecha(NW) coffees. The caffeine content
of Haru(W) coffee was considerably higher than those of Bedeno(H),
Darolebu(E), Melkabelo(H), Chiro(H), Yirgacheffe(Southern, S), Jimma
(SW), Lem-Kaffa(SW), Yayu(SW), Bebeka(SW), Gemadiro(SW) and Bure
(NW) coffees. But, it was much lower for Jimma(SW) coffee compared to
the coffees of all other localities, other than Darolebu(H) and Chiro(H)
Table 3
ANOVA p-values showing the significance of the main effect of Region and Lo­
cality (District), nested in Region, on 100 seed weight and quality attributes of
green arabica coffee beans. Significant effects that require multiple means
comparison are shown in bold.
Source of 100SW BPQ Acidity Body Flavor CQ TPQ
variation
Region 0.001 0.041 0.772 0.782 0.243 0.819 0.163
Locality 0.001 0.001 0.193 0.285 0.083 0.052 0.001
(Region)
100SW = 100 seed weight, BPQ = Bean physical quality, CQ = Cup quality, TPQ
= Total preliminary quality.
M. Worku et al. Journal of Food Composition and Analysis 115 (2023) 105015

Table 4
ANOVA p-values showing the significance of the main effect of Region and Locality (District), nested in Region, on biochemical composition of green arabica coffee
beans. Significant effects that require multiple means comparison are shown in bold.
Source of variation Caffeine Sucrose TCGAs 3-CQA 4-CQA 5-CQA 3,5diCQA 4,5diCQA FQA CFQA

Region 0.001 0.184 0.001 0.097 0.525 0.001 0.001 0.001 0.001 0.063
Locality(Region) 0.001 0.001 0.001 0.003 0.001 0.001 0.001 0.001 0.001 0.001

TCGAs = Total chlorogenic acids.

Table 5 Table 6
Mean values of bean physical quality, acidity and total preliminary quality of Mean values of 100 seed weight (100SW), bean physical quality (BPQ), total
green arabica coffee beans obtained from ECX warehouses in the three main preliminary quality (TPQ), caffeine, sucrose and total chlorogenic acids (TCGAs)
coffee-growing regions. of green arabica coffee beans obtained from the 24 coffee-growing localities
Regiona Bean physical Acidity Total preliminary
(districts), nested in region.
quality (1–40 score) (0–15 score) quality (0–100 score) Locality 100SW BPQ TPQ Caffeine Sucrose TCGAs
(Region)a (g) (0–40 (0–100 (mg g− 1 (mg g− 1 (mg g− 1
Eastern 30.9 a 11.3 a 77.6 a
score) score) dw) dw) dw)
(Harar)
Southern 33.0 a 11.0 a 77.6 a Anfilo(W) 14.2 39.0 a 89.0 a 13.9 44.1 54.1
Southwestern 21.1 b 8. 9 b 60.6 b cde bcde abcd bcdef
a Ankasha 13.6 e 31.7 79.7 ab 13.8 53.5 abc 55.3
The number of samples per coffee region was 10 (total n = 30). Within each
(NW) ab bcde abcdef
column, means sharing the same letter are not significantly different. Bebeka 16.0 38.3 a 84.3 a 12.9 def 57.9 a 57.1
(SW) bcde abcde
coffees. In general, caffeine content is the most varied variable among Bedeno(H) 15.2 39.0 a 85.0 a 11.7 fg 43.6 55.3
bcde abcd abcdef
the localities across the five coffee regions (Table 6).
Bure(NW) 14.7 34.7 a 81.7 a 13.3 cdef 45.3 55.3
Sucrose content was considerably higher in Bebeka(SW) coffee than bcde abcd abcdef
in Jimma(SW), Lem-Kaffa(SW), Tepi(SW) and Gimbi(W) coffees. It was Chiro(H) 17.6 38.5 a 89.5 a 10.8 gh 51.3 abc 60.5 a
also higher in Yayu(SW) coffee compared to Tepi(SW) and Gimbi(W) abc
Darolebu 18.1 ab 40.0 a 83.0 a 110.0 gh 50.7 abc 53.8
coffees. But, the sucrose content of Tepi(SW) coffee was substantially
(H) defg
lower than those of Darolebu(H), Habro(H), Chiro(H), GommaII(SW), Gemadiro 15.0 34.0 a 80.0 ab 12.9 def 43.4 56.7
Yayu(SW), Bebeka(SW), Godere1(SW) and Ankasha(NW) coffees (SW) bcde abcd abcde
(Table 6). The TCGAs content was significantly higher for Chiro(H) Gimbi(W) 16.2 34.0 a 78.0 ab 14.6 abc 39.0 cd 52.6
coffee than for Darolebu(H), Yirgacheffe(S), Jimma(SW), Limmu-Kossa bcde efg
Godere1 15.4 39.0 a 82.0 a 13.9 51.1 abc 53.6
(SW), Lem-Kaffa(SW), Gore(SW), Godere1(SW), Godere2(SW), Gimbi
(SW) bcde bcde defg
(W), Haru(W) and Anfilo(W) coffees. Next to Chiro(H) coffee, it was Godere2 15.5 39.0 a 87.0 a 14.4 bcd 47.0 54.1
higher for Jabitenan(NW) coffee. Yet, it was lower for Lem-Kaffa(SW) (SW) bcde abcd cdef
coffee than for the coffees of all other localities, except for Darolebu GommaII 15.4 34.0 a 83.0 a 14.0 49.4 abc 58.3
(H), Jimma(SW), Limmu-Kossa(SW), Gore(SW), Godere1(SW), Gimbi (SW) bcde bcde abcd
Gore(SW) 20.6 a 39.0 a 88.0 a 13.9 47.5 53.2
(W) and Haru(W) coffees (Table 6). bcde abcd efg
As indicated in Table 7, the 3-CQA content of Yayu(SW) coffee was Habro(H) 16.6 37.0 a 84.0 a 14.9 abc 48.7 abc 58.7
much higher than those of Bedeno(H) and Lem-Kaffa(SW) coffees. The 4- bcde abc
CQA content of Chiro(H) and GommaII(SW) coffees was significantly Haru(W) 17.1 34.0 a 83.0 a 15.1 ab 46.9 53.4
bcd abcd efg
higher than those of Bedeno(H), Lem-Kaffa (SW), Gore(SW), Haru(W)
Jabitenan 13.9 e 40.0 a 84.0 a 16.2 a 44.9 58.8 ab
and Bure(NW) coffees. The 4-CQA content of Yayu(SW), Gemadiro(SW), (NW) abcd
Godere2(SW) and Jabitenan(NW) coffees was significantly higher Jimma 15.0 34.0 a 84.0 a 9.4 h 42.5 bcd 53.6
compared to those of Bedeno(H) and Lem-Kaffa(SW) coffees. But, it was (SW) bcde defg
much lower for Bedeno(H) coffee compared to Habro(H), Chiro(H), Lem-Kaffa 15.6 24.7 b 67.7 b 12.7 ef 40.7 bcd 49.3 g
(SW) bcde
GommaII(SW), Limmu-Kossa(SW), Yayu(SW), Gemadiro(SW), Godere2 Limmu- 16.1 38.0 a 85.0 a 13.7 45.4 51.2 fg
(SW) and Jabitenan(NW) coffees. Kossa bcde bcde abcd
The 5-CQA content of the Jabitenan(NW) coffee was much higher (SW)
than the coffees of all localities, excluding Yirgacheffe(S), GommaII Mecha 14.0 de 36.0 a 82.0 a 14.6 abc 47.9 56.4
(NW) abcd abcde
(SW), Godere1(SW), Haru(W), Anfilo(W), Ankasha(NW) and Mecha
Melkabelo 16.7 35.5 a 83.5 a 12.3 efg 45.6 58.8
(NW) coffees. Next to Jabitenan(NW) coffee, GommaII(SW) coffee had a (H) bcde abcd abc
higher 5-CQA content than the coffees of other localities, followed by Tepi (SW) 14.4 37.0 a 81.0 ab 13.9 33.3 d 57.1
Yirgacheffe(S), Haru(W) and Anfilo(W) coffees. But, Jimma(SW) and cde bcde abcde
Lem-Kaffa(SW) coffees had a lower 5-CQA content than Yirgacheffe(S), Yayu(SW) 16.6 39.0 a 84.0 a 13.0 cdef 54.6 ab 56.5
bcde abcde
GommaII(SW), Gemadiro(SW), Godere1(SW), Haru(W), Anfilo(W), Yirgacheffe 14.9 40.0 a 87.0 a 12.0 fg 45.9 54.8
Jabitenan(NW), Ankasha(NW) and Mecha(NW) coffees (Table 7). (S) bcde abcd bcdef
The 3,5diCQA content was considerably higher for Bedeno(H) and a
The number of samples per coffee locality(Region) was 3 (total n = 72).
Habro(H) coffees than for the coffees of all other localities, excluding
Means sharing the same letter within each column are not significantly different.
Melkabelo(H) and Jabitenan(NW) coffees. Next to Bedeno(H) and Habro H = Eastern or Harar, S = Southern, SW = Southwestern, W = Western, NW
(H) coffees, it was significantly higher for Melkabelo(H) coffee than for = Northwestern.
the coffees of other districts, except for Jabitenan(NW) coffee. But, it
was much lower for Limmu-Kossa(SW) coffee than for Bedeno(H),
(NW) and Mecha(NW) coffees. In general, the 3,5diCQA content is the
Melkabelo(H), Darolebu(H), Habro(H), Chiro(H), Jimma(SW), Gore
most varied variable among localities across the five coffee regions
(SW), Bebeka(SW), Tepi(SW), Jabitenan(NW), Bure(NW), Ankasha
(Table 7).

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M. Worku et al. Journal of Food Composition and Analysis 115 (2023) 105015

Table 7
1
Mean values of the seven isomers of chlorogenic acids (all in mg g− dw) of green arabica coffee beans obtained from the 24 coffee-growing localities (districts), nested
in region.
Locality (Region)a 3-CQA 4-CQA 5-CQA 3,5diCQA 4,5diCQA FQA CFQA

Anfilo(W) 3.4 ab 5.7 abcd 29.6 abcd 5.9 fghi 2.4 defg 3.8 bcdef 3.3 cd
Ankasha(NW) 3.2 ab 5.6 abcd 28.6 abcdef 7.8 cdef 2.6 cdefg 4.0 abcdef 3.4 bcd
Bebeka(SW) 4.1 ab 5.7 abcd 26.8 efgh 7.1 defgh 4.2 a 4.0 abcdef 5.2 ab
Bedeno(H) 2.8 b 4.9 d 26.7 fgh 10.7 a 2.9 cdefg 4.3 abcdef 3.1 cd
Bure(NW) 3.3 ab 5.2 bcd 27.0 efgh 8.4 bcd 3.3 abcde 3.5 ef 4.7 abc
Chiro(H) 4.3 ab 6.5 a 27.8 cdefgh 7.7 cdef 4.1 ab 4.1 abcdef 6.03 a
Darolebu(H) 4.0 ab 5.9 abcd 26.3 gh 6.8 defgh 3.2 bcde 4.4 abcde 3.3 cd
Gemadiro(SW) 3.7 ab 6.30 ab 28.5 bcdefg 6.0 efghi 2.9 cdefg 4.1 abcdef 3.9 bcd
Gimbi(W) 4.0 ab 6.1 abcd 27.7 cdefgh 4.9 hi 2.3 efg 4.3 abcdef 3.5 bcd
Godere1(SW) 3.0 ab 5.8 abcd 29.3 abcde 5.2 ghi 2.0 g 4.1 abcdef 3.5 bcd
Godere2(SW) 4.1 ab 6.4 ab 27.8 cdefgh 5. 5 ghi 2.4 defg 4.6 ab 3.4 cd
GommaII(SW) 4.4 ab 6.5 a 30.2 ab 5.8 fghi 2.9 cdefg 4.5 abc 4.1 abcd
Gore(SW) 3.3 ab 5.3 bcd 27.2 efgh 7.1 defg 2.6 cdefg 3.9 abcdef 3.9 bcd
Habro(H) 3.92 ab 6.1 abc 27.7 cdefgh 10.9 a 3.4 abc 4.4 abcd 3.8 bcd
Haru(W) 3.1 ab 5.2 bcd 29.5 abcd 6.3 efghi 2.0 g 3.9 abcdef 3.4 cd
Jabitenan(NW) 3.4 ab 6.3 ab 30.5 a 9.3 abc 2.7 cdefg 3.5 def 3.0 cd
Jimma(SW) 3.8 ab 5.6 abcd 25.9 h 7.3 cdefg 3.4 abcd 3.7 cdef 3.7 bcd
Lem-Kaffa(SW) 2.9 b 5.1 cd 26.2 h 6.4 defghi 1.9 g 4.7 a 2.3 d
Limmu-Kossa(SW) 4.1 ab 6.1 abc 27.2 efgh 4.4 i 2.3 efg 3.8 bcdef 3.3 cd
Mecha(NW) 3.5 ab 6.0 abcd 28.8 abcdef 8.0 cde 2.7 cdefg 3.9 abcdef 3.5 bcd
Melkabelo(H) 3.7 ab 5.7 abcd 26.7 fgh 10.6 ab 4.0 ab 4.1 abcdef 4.0 abcd
Tepi (SW) 3.7 ab 5.6 abcd 27.5 defgh 7.7 cdef 3.2 bcdef 4.7 a 4.8 abc
Yayu(SW) 4.5 a 6.3 ab 27.6 cdefgh 6.4 defghi 3.2 bcde 4.0 abcdef 4.6 abc
Yirgacheffe(S) 3.6 ab 6.0 abcd 29.8 abc 6.4 defghi 2.4 efg 3.4 f 3.4 cd
a
The number of samples per coffee locality(Region) was 3 (total n = 72). Means sharing the same letter within each column are not significantly different. H
= Eastern or Harar, S = Southern, SW = Southwestern, W = Western, NW = Northwestern.

The 4,5diCQA content was considerably higher for Bebeka(SW)
coffee than for the coffees of all other localities, excluding Melkabelo
(H), Habro(H), Chiro(H), Jimma(SW) and Bure(NW). Next to Bebeka
(SW) coffee, it was significantly higher for Melkabelo(H) and Chiro(H)
coffees than for the coffees of other localities, except Habro(H), Jimma
(SW) and Bure(NW) coffees. It was, however, much lower for Lem-Kaffa
(SW), Godere1(SW) and Haru(W) coffees than for Darolebu(H), Chiro
(H), Habro(H), Melkabelo(H), Jimma(SW), Yayu(SW), Bebeka(SW),
Tepi(SW) and Bure(NW) coffees (Table 7).
The FQA content of Lem-Kaffa (SW) and Tepi (SW) coffees was
significantly higher than those of Yirgacheffe(S), Jimma(SW), Limmu-
Kossa(SW), Anfilo (W), Jabitenan(NW) and Bure(NW) coffees, and
that of Godere2(SW) coffee was significantly higher than those of Yir­
gacheffe(S), Jimma(SW), Jabitenan(NW) and Bure(NW) coffees. It was,
however, much lower for Yirgacheffe(S) coffee than for Darolebu (H),
Habro(H), GommaII(SW), Lem-Kaffa(SW), Tepi(SW) and Godere2(SW)
coffees (Table 7).
The CFQA content of Chiro(H) coffee was considerably higher than
those of the coffees of all other districts, except for Melkabelo(H),
GommaII(SW), Yayu(SW), Bebeka(SW), Tepi(SW) and Bure(NW) cof­
fees. Bebeka(SW) coffee had also a higher CFQA content than Bedeno
(H), Darolebu(E), Yirgacheffe(S), Limmu-Kossa(SW), Lem-Kaffa(SW),
Godere2(SW), Haru(W), Anfilo(W) and Jabitenan(SW) coffees. Yet, the
CFQA content was significantly lower for Lem-Kaffa(SW) coffee than for

Fig. 2. Complete linkage dendrogram showing the similarity of the 24 coffee-growing localities (districts) in terms of the 100 seed weight and quality attributes of
green arabica coffee beans.

7
M. Worku et al. Journal of Food Composition and Analysis 115 (2023) 105015

all other coffees, excluding Chiro(H), Yayu(SW), Bebeka(SW), Tepi(SW)
and Bure(NW) coffees (Table 7).
3.3. Similarity among coffee localities in 100 seed weight and quality
As indicated in Fig. 2, Habro(H) and Limmu-Kossa(SW) coffees,
Bedeno(H) and Bebeka(SW) coffees, Melkabelo(H) and Haru(W) coffees,
and Bure(NW) and Mecha(NW) coffees are the most similar ones (ca. ≥
95%) in terms of 100 seed weight and quality. In these variables, the
next most similar coffees (ca. 90–95%) were Yirgacheffe(S) and Gore
(SW) coffees; Bedeno(H), Yayu(SW) and Bebeka(SW) coffees; and
Gemadiro(SW), Bure(NW) and Mecha(NW) coffees. The third most
similar coffees (ca. 80–90%) were Chiro(H) and Anfilo(W) coffees;
Habro(H), Yirgacheffe(S), Limmu-Kossa(SW) and Godere2(SW) coffees;
Melkabelo(H), Jimma(SW), GommaII(SW) and Haru(W) coffees; Bedeno
(H), Yayu(SW), Bebeka(SW) and Jabitenan(NW) coffees; and Gemadiro
(SW), Bure(NW) and Mecha(NW) coffees. The fourth most similar cof­
fees (ca. 75–80%) were Habro(H), Yirgacheffe(S), Limmu-Kossa(SW),
Gore(SW) and Godre2(SW) coffees; Melkabelo(E), Jimma(SW), Gom­
maII(SW), Haru(W) and Ankasha (NW) coffees; Bedeno(H), Darolebu
(H), Yayu(SW), Bebeka(SW), Tepi(SW), Godere1(SW) and Godere2(SW)
coffees; and Gemadiro(SW), Gimbi(W), Bure(NW) and Mecha(NW)
coffees. The fifth most similar coffees (ca. 70–75%) were Chiro(H),
Habro(H), Yirgacheffe(S), Limmu-Kossa(SW), Gore(SW), Godere2(SW)
and Anfilo(W) coffees. However, Lem-Kaffa(SW) coffee was not similar
with any of the studied locality coffees in 100 bean weight and quality;
the similarity index was nearly 0% (Fig. 2).
3.4. Similarity among coffee localities in biochemical composition
The similarity among the coffees of the 24 coffee-growing localities
(districts) in their biochemical composition is presented in Fig. 3. In
biochemical composition, the most similar coffees (ca. 90%) were Yir­
gacheffe(S) and Anfilo(W) coffees, and Gore(SW), Godere2(SW) and
Haru(W) coffees. The next most similar ones (around 85%) were Yir­
gacheffe(S), Gore(SW) and Anfilo(W) coffees; Bedeno(H) and Bure(NW)
coffees; Yayu(SW) and Ankasha(NW) coffees; and GommaII(SW) and
Mecha(NW) coffees. The third most similarity (around 80%) in
biochemical composition was observed between Habro(H) and
Melkabelo(H) coffees; between Bedeno(H), Jimma(SW) and Bure(NW)
coffees; between Gore(SW), Godere2(SW) and Limmu-Kossa(SW) cof­
fees; between Gimbi(W) and Mecha(NW) coffees; and between Darolebu
(H) and Godere1(SW) coffees. Bedeno(H), Yirgacheffe(S), Jimma(SW),
Gemadiro(SW), Anfilo(W) and Bure(NW) coffees; Darolebu(H), Yayu
(SW) Godere1(SW) and Ankasha(NW) coffees; Melkabelo(H), Habro(H)
and Jabitenan(NW) coffees; and Chiro(H), GommaII(SW) and Mecha
(NW) coffees were the fourth most similar ones (ca. 70–75%). But, no
similarity was observed among (1) Bedeno(H), Yirgacheffe(S), Jimma
(SW), Limmu-Kossa(SW), Lem-Kaffa(SW), Gore(SW), Tepi(SW), God­
ere2(SW), Gemadiro(SW), Gimbi(W), Haru(W), Anfilo(W) and Bure
(NW) coffees, and (2) Melkabelo(H) Darolebu(H), Habro(H), Chiro(H),
GommaII(SW), Yayu(SW), Bebeka(SW), Godere1(SW), Ankasha(NW),
Mecha(NW) and Jabitenan(NW) coffees in biochemical composition; the
similarity index was nearly 0% (Fig. 3).
3.5. Similarity among coffee localities in 100 seed weight, quality and
biochemicals
The similarity among the coffees of the 24 coffee growing localities
(districts) in terms of 100 seed weight, quality and biochemical
composition is presented in Fig. 4. There was a high similarity (ca.
83.5%) between Yirgacheffe(S) and Godere2(SW) coffees, between Bure
(NW) and Mecha(NW) coffees, between Habro(H) and Melkabelo(H)
coffees, and between Yayu(SW) and Bebeka(SW) coffees in 100 seed
weight, quality and biochemical composition. The next high similarity
(ca. 80%) was observed among Yirgacheffe(S), Limmu-Kossa(SW) and
Godere2(SW) coffees; among Gemadiro(SW), Bure(NW) and Mecha
(NW) coffees; between GommaII(SW) and Haru(W) coffees; and be­
tween Darolebu(H) and Godere1(SW) coffees. The third most similar
coffees (ca. 75–80%) were observed among Yirgacheffe(S), Limmu-
Kossa(SW), Gore(SW) and Godere2(SW) coffees and between Bedeno
(H) and Jabitenan(NW) coffees; and the fourth most similar ones (ca.
70–75%) among Yirgacheffe(S), Limmu-Kossa(SW), Gore(SW), Godere2
(SW) and Anfilo(W) coffees; among Melkabelo(H), Habro(H), GommaII
(SW) and Haru(W) coffees; and between Tepi(SW) and Gimbi(W) cof­
fees. However, Lem-Kaffa(SW) coffee was not similar with any of the
studied locality coffees in 100 bean weight, quality and biochemical
composition (Fig. 4).

Fig. 3. Complete linkage dendrogram showing the similarity of the 24 coffee-growing localities (districts) in terms of the biochemical compositions of green arabica
coffee beans.

8
M. Worku et al.
Fig. 4. Complete linkage dendrogram showing the similarity of the 24 coffee-growing localities (districts) in terms of 100 seed weight, quality attributes and
biochemical compositions of green arabica coffee beans.
4. Discussion
The samples collected from farms and ECX warehouses showed a
similar trend in physical quality and total preliminary quality scores. In
general, the samples of the eastern and southern regions were higher in
physical quality, acidity and total preliminary quality scores than those
of the remaining regions. Similarly, in the cup of excellence competition
conducted in 2020 and 2021 in Ethiopia, samples of the southern region
had higher cupping scores than those of the other regions (Ethiopia,
2020; Ethiopia, 2021; Montagnon, 2020). However, compared to that
collected from ECX warehouses (Table 5), the samples collected from
farms both in eastern(Harar) and southern regions had slightly higher
physical quality scores (38.2, 40.0) and total preliminary quality scores
(84.8, 87.0). This may be related to the difference between the two
sample sources in harvest and postharvest practices as well as in volume
of coffee cherries harvested and processed. For example, the coffee
cherries of our samples collected from farms were selectively harvested
(only fully ripened cherries) and properly dried on raised drying beds in
small volume (each sample with 6–7 kg) by us. However, those samples
taken from ECX warehouses were dried beans whose cherries have been
harvested and dried by famers. Famers may not be selective in har­
vesting and can dry their coffee cherries on the ground in large volume.
These conditions often influence coffee quality. Similarly, the better
quality for Harar and southern region coffees than for other ones
observed in this study and others could be related to a better coffee
management practice in these two regions compared to that in other
regions, particularly in southwestern and western regions. In Harar and
southern regions, coffee is produced under a garden system, which is
more intensive in agronomic practice and selective in harvesting than
forest and semi-forest/plantation systems, which exist mainly in south­
western and western regions (Gole, 2015; Worku, 2019).
Regardless of the statistical differences among coffee-growing lo­
calities across the five coffee regions (Table 6), Harar coffee samples
were higher in the mean 100 seed weight (16.7 g) than those samples
from the other regions, while samples from the southwestern and
western regions had equal mean 100 seed weight (15.8 g). Samples from
the southern and northwestern regions had a mean 100 seed weight of
14.9 and 14.1 g, respectively. Specifically, samples from Gore in
southwestern region had a significantly higher 100 seed weight (20.6 g)
than those samples from other localities whereas samples from Ankasha
9
Journal of Food Composition and Analysis 115 (2023) 105015
and Jabitenan in northwestern region had a much lower 100 seed weight
(ca 14.0 g) than that from Chiro(H), Darolebu(H), Gore(SW) and Haru
(W) (Table 6). These differences among coffee-growing regions and lo­
calities can be related to their differences in growing conditions, e.g.,
climate, soil and/or coffee management/production system (Gole, 2015;
Table 1; Worku, 2019). On the other hand, the equal mean 100 seed
weight for the southwestern and western coffee regions can be due to
their similar growing environment or agro-ecology (Worku, 2019).
Caffeine, sucrose and chlorogenic acids are among the compounds
found in green coffee beans that are important contributors to flavor
(Farah et al., 2006). They are found in coffee beans in small quantity,
ranging from 0.9–2.5% (i.e., ca. 9.0–25.0 g kg− 1), 6.0–9.% (i.e., ca.
60.0–90.0 g kg− 1) and 4.1–11.3% (i.e., ca. 41.0–113.0 g kg− 1) on dry
weight basis, respectively (Farah, 2012). In the present study, the mean
caffeine contents observed for southwestern, western and northwestern
region coffees (13.2, 14.6 and 14.5 mg g− 1 dw, respectively) were
slightly higher than those observed for Harar and southern region cof­
fees (12.0 mg g− 1 dw) and previously reported for green coffees of the
five regions (10.5–12.5 g kg− 1) (Mehari et al., 2016a) and worldwide
(9.0–13.0 g kg− 1) (Farah, 2012). But, the mean sucrose and TCGAs
contents of Harar coffee (47.9 and 57.1 mg g− 1 dw, respectively) were
slightly higher than those obtained from coffees of other regions (i.e.,
45.9 and 54.8, 46.6 and 53.8, 43.3 and 53.4, and 50.3 and 56.5 mg g− 1
dw from southern, southwestern, western and northwestern region
coffees, respectively). However, these values were slightly lower than
the mean values previously reported for green coffee of each coffee re­
gion in Ethiopia (65 and 60.0–64.0 g kg− 1) (Mehari et al., 2016b;
Worku, 2019) and worldwide (60.0–90.0 g kg− 1) (Farah, 2012). Yet,
those observed for TCGAs were generally within the lower range of
values previously reported for green arabica coffee worldwide
(41.0–79.0 g kg− 1) (Farah, 2012). The variations observed among the
coffee regions in their caffeine, sucrose and chlorogenic acids contents in
this study can be linked to the variation among the coffee-growing re­
gions in their climate. For example, the studied coffee regions vary in
their total amount and period of rainfall (Moat et al., 2017a). In general,
the southwestern, western and northwestern regions have a higher total
rainfall and a longer rainy period than Harar and southern regions. On
the other hand, the variations between this study and previous studies
mentioned above can be due to the difference between these studies in
their sampling sites and growing seasons. For example, a variation in
M. Worku et al.
biochemical composition of coffee beans due to specific growing locality
was observed in this and previous studies (Sualeh et al., 2020; Tables 6
and 7). However, some studies that compared the contents of caffeine
and chlorogenic acids of coffee beans from different origins (countries)
(e.g., Awwad et al., 2021; Jeszka-Skowron et al., 2016) did not find a
significant difference between coffee bean origins, except for coffee
samples originated from Brazil and Colombia in caffeine and from
Ethiopia and India in chlorogenic acids (Awwad et al., 2021).
In the present study, the mean values for 3-CQA, 4-CQA, 5-CQA,
3,5diCQA, 4,5diCQA, FQA, CFQA and TCGAs in the coffees of the five
regions varied from 3.3–3.8, 5.7–5.9, 27.0–29.8, 5.7–9.0, 2.2–3.4,
3.4–4.3, 3.4–3.9 and 53.4–57.1 mg g− 1 dw, respectively. Almost similar
values of 3-CQA, 3,5diCQA and 4,5diCQA (2.9–6.2, 5.1–8.1 and
2.1–4.9 mg g− 1 dw, respectively), but slightly higher values of 4-CQA, 5-
CQA and CFQA (5.8–8.4, 29.0–33.7 and 4.1–5.7 mg g− 1 dw, respec­
tively) were reported by Mehari et al. (2016b). In this study, the mini­
mum values of 3-CQA, 4-CQA and 5-CQA contents were observed for
northwestern, southwestern and western, and Harar coffees, respec­
tively; those of 3,5diCQA, 4,5diCQA and TCGAs contents for western
coffee; and those of FQA and CFQA contents for southern coffee and
southern and western coffees, respectively. The maximum values of
3-CQA, and 4-CQA and 5-CQA contents were observed for southwestern
and southern coffees, respectively and those of 3,5diCQA, 4,5diCQA,
FQA, CFQA and TCGAs contents for Harar coffee. Lower 4-CQA and 4,
5diCQA contents for western coffee and higher 4,5diCQA and FQA
contents for Harar coffee were also reported by Mehari et al. (2016b).
But, the results of the two studies on other chlorogenic acids did not
agree. Unlike our results, Mehari et al. (2016b) reported higher 5-CQA
and 3,5diCQA contents for southwestern region coffee, and 3-CQA and
4-CQA contents for northwestern region coffee, but lower 3-CQA,
4-CQA, 4,5diCQA and FQA contents for western region coffee. As
chemical composition of coffee beans can vary with specific growing
locality and season (Sualeh et al., 2020; Tables 6 and 7), this difference
between the two studies could be due to their variation in sampling sites
and growing season.
The difference between some coffee-growing localities in one hand
and higher similarities between some other coffee-growing localities
across the five coffee regions on the other hand observed in the present
study (Figs. 2 through 4; Tables 6 and 7) as well as in a previous study
(Sualeh et al., 2020) can show the coffee regions of Ethiopia do not
utterly differ in their coffee quality and biochemical composition de­
terminants. That is, although the coffee regions in Ethiopia are thought
to vary in coffee genetics, growing environment (e.g., rainfall, temper­
ature, shade level, soils) and/or coffee management or production sys­
tem, similarities can exist among some coffee-growing localities across
the coffee regions in these coffee quality and biochemical composition
determinants. Due to this, coffee samples collected from some localities
across the five coffee regions are highly similar in their 100 seed weight,
quality attributes and contents of caffeine, sucrose and chlorogenic acids
(Figs. 2 through 4; Tables 6 and 7). However, no similar pattern of
similarities was observed among coffee-growing localities due to their
quality attributes and biochemical compositions (Figs. 2 and 3). Owing
to a relationship between quality and biochemical composition of coffee,
this is not in line with our expectation. Furthermore, while quality at­
tributes were observed as the less varied variables across the studied
coffee-growing localities (24 districts) (Table 6), caffeine and 3,5diCQA
contents were the most varied ones (Tables 6 and 7). This may show that
the latter variables can be more sensitive to the existence of small dif­
ferences among coffee-growing areas in genetics, growing environment
and/or management of coffee than the other ones tested in this study.
Yet, to the best of our knowledge, this report on differences and
similarities among coffee-growing localities across the five coffee re­
gions in Ethiopia in seed weight, quality and biochemical composition of
green coffee beans is the first one of its kind. Nevertheless, it is from the
samples collected from fields in one growing season. So, to arrive at a
reliable conclusion on this issue and to utilize its outcome in practice,
10
Journal of Food Composition and Analysis 115 (2023) 105015
further research using repeated experiments over several years and lo­
calities, and samples collected not only from fields, but also from bulk
productions is required. As the quality and biochemical composition
determinants of coffee in each locality were not examined in this study,
including these variables in a future study is also recommended.
5. Conclusion
The results of this study generally indicate that Ethiopian coffee vary
with both growing region and locality in bean weight, quality attributes
(e.g., physical quality, acidity and total preliminary quality) and
biochemical composition (e.g., caffeine, sucrose and chlorogenic acids
contents). But, the extent of the variability depends on the source of
samples and the type of variables. For example, coffee samples from
farms did not vary with region in acidity, total preliminary quality, su­
crose, 3-CQA and 4-CQA, but that from ECX warehouses, analyzed only
for quality attributes, varied with region in the former two quality
variables. Besides, compared to other variables, quality attributes were
generally the less varied ones across the coffee-growing localities, but
caffeine and 3,5diCQA contents were the most varied ones.
This study also showed the presence of similarity between some lo­
cality coffees across the five coffee-growing regions in bean weight,
quality attributes and biochemical composition. For example, Habro and
Limmu-Kossa coffees, Bedeno and Bebeka coffees, Melkabelo and Haru
coffees, and Bure and Mecha coffees were highly similar in their bean
weight and quality; Yirgacheffe and Anfilo coffees, and Gore, Godere2
and Haru coffees in their biochemical composition; and Yirgacheffe and
Godere2 coffees, Bure and Mecha coffees, Habro and Melkabelo coffees,
and Yayu and Bebeka coffees in their bean weight, quality and
biochemical composition. However, Lem-Kaffa coffee was not similar to
any of the other locality coffees in its bean weight and quality attributes,
but it had a high similarity (around 80%) with Gimbi coffee in
biochemical composition.
This study generally elucidated the existence of variations between
coffee-growing regions and localities across the coffee regions, and
similarities between some coffee-growing localities across the coffee
regions in bean weight, quality attributes and biochemical composition.
Nevertheless, it should be confirmed by further research covering mul­
tiple years and localities, samples from fields and bulk productions, and
quality and biochemical composition determinants in each coffee-
growing locality.
Funding
The work was financially supported by VLIR-UOS Institutional Uni­
versity Cooperation with Jimma University, Ethiopia.
Declaration of Conflict of Interest
Authors have no conflict of interests to declare.
Data availability
The authors do not have permission to share data.
Acknowledgment
The Ethiopia Commodity Exchange and all farmers who provided us
with coffee samples are acknowledged.
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