Journal of Food Composition and Analysis 115 (2023) 105027

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Journal of Food Composition and Analysis

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

Shade and postharvest processing effects on arabica coffee quality and

biochemical composition in lowland and midland coffee-growing areas of
southwestern Ethiopia
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: Hitherto, studies on shade and postharvest processing (PHP) effects on coffee quality across elevation ranges (ER)
Coffee quality are limited. The effects of shade, PHP and their interaction on quality and caffeine, sucrose and chlorogenic acids
Acidity (CGAs) contents of arabica coffee beans in lowland (1100–1550 m asl) and midland (1550–1900 m asl) were
Caffeine
evaluated. The results showed that physical, total preliminary and total specialty qualities were higher for beans
Chlorogenic acids
Shade
grown in midland than those grown in lowland, but CGAs (57.5 g kg-1 dw) were higher for beans grown in
Lowland lowland. Dry-processed midland coffee had higher physical and total preliminary qualities with lower 3,5diCQA
Midland (6.9 g kg-1 dw) and FQA (4.4 g kg-1 dw) than wet-processed lowland and midland coffees. Conversely, wet-
processed lowland coffee had lower physical and total preliminary qualities with higher 3,5diCQA (8.7 g kg-1
dw) and FQA (4.7 g kg-1 dw) than dry-processed lowland and midland coffees. Coffee beans grown with shade in
lowland had lower acidity than those grown without shade in lowland and midland. But, coffee beans grown
without shade in lowland had lower caffeine (13.8 kg-1 dw) than those grown with shade in lowland and
midland. Physical and total preliminary qualities were negatively correlated with caffeine, 4-CQA, 3,5diCQA,
FQA and total CGAs. This study is the first to show the relationship between ER and PHP in coffee quality and
biochemical composition of green arabica coffee beans.

1. Introduction bioactive properties, such as antioxidant, α-glucosidase inhibitory and

Quality of the green coffee beans, which includes both bean physical
and cup qualities, is the main criterion that determines market price and
consumer preference of coffee. Biochemical composition of green coffee
beans is decisive for cup quality through roasting-induced chemical
reactions. For example, high trigonelline and 3,4-dicaffeoylquinic acid
levels in green 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 Maillard
reactions (Flament, 2002). Conversely, high levels of caffeoylquinic
acids, feruloylquinic acids and their oxidation products are associated
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 (CGAs) and trigonelline, also have useful
antimicrobial activities (Bhattacherjee et al., 2014; Bhattacherjee and
Datta, 2015; Zhou et al., 2012), whereas other biochemicals, e.g.,
caffeine and diterpenes, may have harmful effects by affecting endo­
thelial function and raising a fraction of blood lipids, respectively
(Godos et al., 2014). On the other hand, consumption of caffeine could
reduce the risk of Parkinson’s disease development (Higdon and Frei,
2006). Biochemical composition can also be associated with bean
physical quality (Ramalakshmi et al., 2007).
Several factors, such as genetics, growing environment, agricultural
practice and postharvest processing, determine quality and biochemical
composition of green coffee beans (Ahmed et al., 2021; Amalia et al.,
2021; Hameed et al., 2018; Getachew et al., 2022; Koutouleas et al.,
2022; Worku et al., 2018). Elevation (mainly affecting the temperature
of the area) and shade (mainly affecting the microclimate at coffee farms
and surrounding areas) are among the growing conditions that

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

https://doi.org/10.1016/j.jfca.2022.105027
Received 10 September 2022; Received in revised form 28 October 2022; Accepted 5 November 2022
Available online 7 November 2022
0889-1575/© 2022 Elsevier Inc. All rights reserved.
M. Worku et al. Journal of Food Composition and Analysis 115 (2023) 105027

considerably influence these attributes of coffee beans (Ahmed et al., the fungal population occurring during postharvest processing and
2021; Koutouleas et al., 2022; Tolessa et al., 2017; Worku et al., 2018). storage negatively affects coffee quality (Waters et al., 2017).
Particularly, higher elevations and shade at lower elevations are found However, except few recent studies (e.g., Worku et al., 2018, 2022),
to improve physical and cup quality attributes of coffee beans, which empirical data on individual and interaction effects of shade and post­
have been primarily attributed to cool climates (Bertrand et al., 2012; harvest processing on quality and biochemical composition of green
Decazy et al., 2003; Muschler, 2001). Lower temperatures have been coffee beans in a range of elevations (agro-ecological zones) are scant.
suggested to lengthen the maturation period of coffee berries, which in Information on the association between different quality attributes and
turn leads to higher accumulation of aroma precursors (Vaast et al., biochemical compositions of Ethiopian coffee is also scarce. Therefore,
2006). In line with this, Getachew et al. (2022) reported a negative the objectives of the present study were to determine (1) the quality
correlation between soil temperatures and green bean quality and be­ attributes and biochemical compositions, (2) the individual and inter­
tween soil temperatures and biochemical compositions (e.g., caffeine, action effects of shade and postharvest processing on quality attributes
trigonelline and CGAs contents), and Joët et al. (2010) demonstrated a and biochemical compositions, and (3) the links between quality attri­
direct impact of mean temperatures during seed development on the butes and biochemical compositions of green arabica coffee beans
time-window of CGAs biosynthesis and final CGA isomer composition growing in two agro-ecological zones (lowland: 1100–1550 m asl and
through subtle transcriptional regulations. Shade also leads to a signif­ midland: 1550–1900 m asl) of southwestern Ethiopia.
icant reduction in sucrose content, but an increase in reducing sugars
(glucose and fructose contents), sucrose synthase, sucrose-phosphate 2. Materials and methods
synthase activities and CaSUS2 gene transcripts levels in developing
beans (Geromel et al., 2008). 2.1. Study area description
Studies also showed the interaction effect of elevation and shade
level on microclimate (e.g., soil and air temperature), coffee leaf traits Samples of green arabica coffee beans were collected from ten sites,
(e.g., specific leaf area, water use efficiency, N content and C:N ratio) located at two elevation ranges (lowland: 1100–1550 m asl and
(Getachew et al., 2021), coffee quality attributes and biochemical midland: 1550–1900 m asl) and in eight districts of southwestern
compositions (Tolessa et al., 2017; Worku et al., 2018), and coffee Ethiopia. Majority of the coffee-growing areas in southwestern Ethiopia
productivity (Anhar et al., 2021). Shade has a positive effect on overall are located at these elevations. Southwestern Ethiopia is the center of
quality and productivity of coffee at lower elevations (Koutouleas et al., origin and diversity for Coffea arabica and one of the four major coffee-
2022; Anhar et al., 2021). However, elevation and shade effects on growing regions of Ethiopia. It is generally characterized by hot and
quality and biochemical composition of coffee beans can be highly site humid tropical rainforest climate with unimodal rainfall and dominated
specific or specific to the coffee variety and the growing conditions other by moderately acidic nitosols (Dewitte et al., 2013; Worku, 2019). Other
than elevation and shade; e.g., precipitation, soil, growing season, or soil groups in the region include vertisols, leptosols, regosols, cambisols,
management practice (Worku et al., 2018, 2022). Moreover, this may be alisols and acrisols (Dewitte et al., 2013). In the region, both
reinforced by postharvest processing of coffee cherries, as reported in wet-processed (washed) and dry-processed (unwashed) coffees, with a
some recent studies (Taveira, 2014; Tolessa et al., 2017; Worku et al., higher share for the latter, are produced.
2018, 2022). The specific sampling sites were: (1) Bebeka (1150 m asl, South
Various studies also showed the impact of postharvest processing on Bench District), (2) Yayu (1440 m asl, Yayu District), (3) Godere (three
quality and biochemical composition of green coffee beans (Amalia sites located at three different elevations; i.e., 1352, 1564, 1745 m asl,
et al., 2021; Tassew et al., 2021). Germination and fermentation of the Godere District), (4) Gomma II (1545 m asl, Gomma District), (5) Gore
beans are the two bioprocesses that take place during postharvest (1783 m asl, Ale District), (6) Limmu-Kossa (1802 m asl, Limmu Kossa
treatment and may lead to significant modifications of these coffee at­ District), (7) Jimma (1818 m asl, Manna District), and (8) Lem-Kaffa
tributes. The germination process is initiated while the beans are still (1820 m asl, Gewata District). Geographical and environmental details
inside the cherry, and the evolution of germination depends on how the of each sampling location are shown in Table 1.
beans are processed. A range of metabolic reactions takes place during Nearly all these sampling sites were from large commercial farms
germination and can influence the quality attributes and biochemical and samples were collected from adult coffee shrubs; i.e., 7–15 years old
compositions of beans. Moreover, the microbiota associated with coffee planted trees or 2–10 years old coppiced trees. These farms use
cherries and beans during postharvest processing (e.g., bacterial, yeast improved varieties, row planting (2 m inter-row and 1.5–1.8 m intra-
and fungal species) affects coffee processing from cherries to beans and row spacing), fertilizers and herbicides. Shade trees in the sampling
play a role in the degradation of pulp or mucilage, and their metabolism sites are a mixture of locally preferred coffee shade tree species; mainly
can affect the sensory attributes of coffee (Waters et al., 2017). Conse­ from old forest stands with an average density of 64–114 trees ha− 1 and
quently, sensory scores (profiles) (Ferreira et al., 2013; Taveira, 2014)
and biochemical compositions (e.g., free amino acids, CGAs, trigonelline Table 1
and sugar contents) (Arruda et al., 2012; Duarte et al., 2010; Knopp Geographical and environmental data of sampling locations: latitude (Lat),
et al., 2006) of green coffee beans were observed to considerably vary longitude (Long), elevation ranges (Elev), mean annual rainfall (Rf), mean
with postharvest processing method. annual maximum temperature (Tmax) and mean annual minimum temperature
The effect of postharvest processing on quality and biochemical (Tmin); adapted from Worku et al. (2018).
composition of green coffee beans may also depend on the characteris­ Location Lat Long Elev (m asl) Rf Tmax Tmin
tics of fresh coffee cherries (e.g., mucilage and moisture contents) that (mm) (◦ C) (◦ C)
can vary with genetics and growing environments of coffee (Hameed Bebeka 6◦ 52′ N 35◦ 52′ E 850–1200 1760 31.0 15.0
et al., 2018; Tolessa et al., 2019; Worku et al., 2022). This is because the Yayu 8◦ 26′ N 36◦ 03′ E 1200–2000 2100 26.1 12.7
characteristics of coffee cherries can affect the extent of germination and Godere 7◦ 08′ N 35◦ 20′ E 1200–1900 1737 27.0 12.0
fermentation processes as well as the proper processing of beans Gomma II 7◦ 57′ N 36◦ 37′ E 1450–1750 1540 29.0 13.0
Gorea 8◦ 09′ N 35◦ 31′ E 1770–2085 – – –
differently when coffee cherries are processed by different ways; e.g., Jimma 7◦ 40′ N 36◦ 50′ E 1780–2100 1580 26.3 14.5
wet and dry methods. Proper processing of large volumes of coffee Limmu- 7 57′ N
◦
36 53′ E
◦
1600–1900 1920 27.0 12.0
cherries with high mucilage and moisture contents, which are the Kossa
characteristics of some coffee varieties or coffees grown under shade by Lem-Kaffa 7◦ 24′ N 36◦ 13′ E 1450–2370 1723 26.4 11.9
using a dry processing method, is difficult, particularly in areas having a a
Climate data are not available, but it is generally known that this site is the
moist climate during harvesting. This leads to fungal development and wettest and coolest area in the region.

2
M. Worku et al.
a canopy cover of 30–40 % (Worku et al., 2015).
2.2. Research design and sample collection
We sampled ten sites at two elevation ranges (lowland: 1100–1550 m
asl and midland: 1550–1900 m asl), including two shade levels (with
and without shade over the canopy of the coffee plants) and two post­
harvest processing methods (wet and dry) per site, and eight locations
(Bebeka, Yayu, Gomma II, Godere, Gore, Jimma, Limmu-Kossa and Lem-
Kaffa). The sampling sites in the first four locations and two sites at
Godere are located between 1150 and 1550 m asl (at lowland) while the
remaining ones are located between 1550 and 1820 m asl (at midland)
(see the above section). The ten sampling sites were one from each of the
seven locations, and three from the eighth location (i.e., Godere). Per
site, twelve samples of ripe red coffee cherries (each about 6–7 kg)
consisting of six samples grown with shade (30–40 % shade level) and
six samples grown without shade were randomly collected from about
15 ha coffee farm per site and by selective hand picking of ripe red
cherries at peak harvesting time. The six samples were divided into a dry
or a wet processing treatment (3 each). The 6–7 kg fresh red cherries
results in around 1 kg clean dry green beans (11.5 % moisture).
2.3. Sample processing
According to locally recommended practices, each sample was har­
vested in the morning and transported to the processing station for
processing in the afternoon. From the six samples of ripe-red coffee
cherries collected from shade and no shade per site, three samples were
subjected to wet processing. The cherries were de-pulped using a coffee
pulper (Aagaard pregrader, McKinnon, Brazil), followed by 36 h
fermentation and mucilage removal with water. Consequently, the
washed-parchment-beans were sun-dried on raised drying-beds until a
moisture content of ca. 11.5 %. Finally, parchments were removed by
using a coffee hulling machine (Coffee huller, Pinhalense, Brazil) at
TechnoServe, Jimma branch. The other three samples were subjected to
dry processing by direct sun-drying on raised drying-beds until they
attained moisture content of ca. 11.5 %. Subsequently, the dried-coffee
cherries were de-husked using a coffee hulling machine (Coffee huller,
McKinnon, Scotland) at Jimma University College of Agriculture and
Veterinary Medicine. Then, clean beans of each sample were packed in 1
kg plastic bags and stored at room temperature until quality and
chemical analyses were carried out.
2.4. Quality analysis
Quality was determined at the Ethiopia Commodity Exchange (ECX)
cupping laboratory in Jimma, Ethiopia following a coffee quality
assessment procedure of ECX (2011) that has been adopted from the
cupping protocols of the Specialty Coffee Association of America
(SCAA). We carried out both preliminary and specialty quality assess­
ments, which are the main coffee quality assessments at ECX. The pre­
liminary quality assessment includes 4 bean physical quality attributes
(i.e., primary defect, secondary defect, odor and color of green beans)
(out of 40 scores), 4 cup quality attributes (i.e., acidity, body, cup
cleanness and flavor of a brew, each at scale of 0–15 scores) (out of 60
scores), and total preliminary (overall) quality (the sum of physical
quality and cup quality scores) (out of 100 scores). 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. Specialty
quality assessment includes 10 cup quality attributes (i.e., aroma, body,
acidity, flavor, cup cleanness, balance, sweetness, uniformity, aftertaste
and overall cup preference of a brew, each at scale of 0–10 scores) and
total specialty quality (the sum of the scores of the ten quality attributes
(out of 100 scores) of coffee samples that received at least 75 scores out
of 100 scores in the preliminary quality assessment.
3
Journal of Food Composition and Analysis 115 (2023) 105027
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
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) (100/100) per sample
were calculated (Worku et al., 2016, 2018). Specialty quality assessment
was made for 10 cup quality attributes (each at scale of 0–10 scores) (see
these cup quality attributes above) following the procedures described
above and then, total specialty quality (the sum of the scores of the ten
quality attributes) (100/100) per sample was calculated (Worku et al.,
2016).
Data on bean physical quality (the sum of the 4 bean physical quality
attributes), acidity, body, flavor, cup quality (the sum of the 4 cup
quality attributes, see above), total preliminary quality and total spe­
cialty quality were used for statistical analysis.
2.5. Chemical analysis
The contents of caffeine, sucrose, the seven different positional iso­
mers of CGAs [3-caffeoylquinic acid (3-CQA), 4-caffeoylquinic acid (4-
CQA), 5-caffeoylquinic acid (5-CQA), 3,5dicaffeoylquinic acid
(3,5diCQA), 4,5dicaffeoylquinic (4,5diCQA), feruloylquinic acid (FQA)
and feruloyl-caffeoylquinic acid (FCQA)] and total CGAs (TCGAs) in
each 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 CGAs; i.e., 3-CQA, 4-CQA, 5-CQA, 3,5diCQA, 4,5diCQA, FQA and
FCQA.
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
the analyzes of CGAs and caffeine contents, the standard stock solutions
of caffeine and CGAs at a concentration of 1 mg mL-1 were prepared by
dissolving CGAs standard (≥ 98.0 %, Sigma-Aldrich, Steinheim, Ger­
many) and caffeine standard (≥ 99.8;%, AZ Chem., Thun-der Bay, ON,
Canada) in methanol (≥ 99.9 %, Sigma-Aldrich, Steinheim, Germany)
and stored at 4 ◦ C in darkness. A 100 mg powder of coffee beans per
sample was subjected to direct solvent extraction with 10 mL of meth­
anol (≥ 99.9 %, Sigma-Aldrich, Steinheim, Germany): water (LabChem
Inc, Zelienople, PA, USA): acetic acid (≥ 99.8 %, Sigma-Aldrich, Stein­
heim, Germany) (12:27:1, v-v:v) containing 2 mg mL-1 ascorbic acid (≥
99 %, Sigma-Aldrich, Steinheim, Germany) in an ultrasonic bath for 15
min. Ascorbic acid enhances the extractions of CGAs and caffeine. Then,
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 measure­
ment took 70 min. The detection was done using a photodiode array
(PDA) detector at 280 nm for caffeine and at 320 nm for CGAs. The
mobile phase was a stepwise gradient composed of 0.2% acetic acid (≥
99.8 %, Sigma-Aldrich, Steinheim, Germany) in water (v/v) and HPLC
grade methanol (≥ 99.9 %, Sigma-Aldrich, Steinheim, Germany)
M. Worku et al. Journal of Food Composition and Analysis 115 (2023) 105027

ranging from 10 % to 70 %, with a flow rate of 1 mL min-1 and an in­ sample. Quantification was performed by external standardization using
jection volume of 50 µL (representative chromatograms are in Supple­ analytical curves, which were obtained by plotting peak area versus
mentary Fig. 1). Calibration was carried out by injecting concentration amount injected and comprised of 6 concentrations of sucrose. LOD was
series of CGAs and caffeine standards, which were prepared from the 0.05 mg g-1 and LOQ was 0.1 mg g-1. Each sample was analyzed in
standard stock solutions by appropriate dilution processes using meth­ triplicate.
anol, every 20 samples using an adapted response factor for different To express the biochemical composition data on dry weight basis, the
CGAs. Quantification was performed by reporting the measured inte­ dry matter of each sample of green coffee beans was determined by
gration areas in the calibration equation of the standard, which exhibits taking the weight of the same amount of the grind subsamples of each
a similar pattern of UV spectra (Alonso-Salces et al., 2009). Thus, sample used for chemical analysis (i.e., 100 mg for caffeine and CGAs
chromatographic peaks that present UV spectra with a maximum at analyses and 0.5 g for sucrose analysis) after 24 h oven drying at a
324–328 nm and a shoulder at 295–305 nm were reported to the seven temperature of 105 ◦ C and air-cooling down to a temperature of ca.
different CGAs (3-CQA, 5-CQA, 4-CQA, 3,5-diCQA, 4,5-diCQA, FQA and 21 ◦ C.
FCQA) and at 271–273 nm to caffeine. Limits of detection (LOD) and
quantification (LOQ) respectively were 0.8 and 1.6 µg g-1 of sample for
2.6. Statistical methods
CGAs, and 1.5 and 3.0 µg g-1 of sample for caffeine. Peak identification
was based on retention time and confirmed using an already established
The main effects of (1) Elevation range (2 levels: lowland and
liquid chromatography time of flight mass spectrometry (LC-TOF-MS)
midland), (2) Shade (2 levels: shade and no shade) and (3) Postharvest
method (Alonso-Salces et al., 2009; Perrone et al., 2008) on selected
processing (2 levels: dry and wet), plus two-way interaction effects be­
samples (deviation from theoretical mass < 1.6 ppm). 3-CQA, 5-CQA
tween shade and postharvest processing on quality attributes (bean
and 3,5-diCQA were identified by the dehydrated base peaks in MS2 at
physical quality, acidity, body, flavor, cup quality, total preliminary
m/z 191 and in MS3 at m/z 85; 4-CQA was identified by the base peaks in
quality and total specialty quality) and biochemicals (caffeine, sucrose,
MS2 at m/z 173 and in MS3 at m/z 93; 4,5-diCQA was identified by the
3-CQA, 4-CQA, 5-CQA, 3,5diCQA, 4,5diCQA, FQA, FCQA and TCGAs) of
base peaks in MS2 at m/z 353 and in MS3 at m/z 173; FQA was identified
green arabica coffee beans were examined using a nested-crossed
by the base peaks in MS2 at m/z 193, 173 and 191 and in MS3 at m/z 134,
design. A factor is nested in another factor when the levels used in the
93 and 85, and FCQA was identified by the base peaks in MS2 at m/z 367
other factor are similar but not identical, and a factor is crossed with
and in MS3 at m/z 193 and 173 (Alonso-Salces et al., 2009; Deshpande,
another factor when the levels used in the other factor are identical
2014). Caffeine was detected by the base peak in MS2 at m/z 138
(Montgomery, 2020). Shade and postharvest processing are crossed, and
(Alonso-Salces et al., 2009; Perrone et al., 2008). Electrospray ionization
both are nested within elevation range. The components of the model
source (ESI) of TOF-MS was operated in positive mode with the nebu­
are shown in the source of variation columns of Table 2 and Table 3. The
lizer gas (N2) flow set at 3 L min-1 and desolvation temperature at
relationship between quality attributes and biochemicals was deter­
300 ◦ C. Data were acquired by ChromQuest software (Thermo Fisher
mined using Pearson’s correlation coefficient. The analysis was con­
Scientific, Waltham, MA, USA), Version 4.1 SP2. Each sample was
ducted using the Mixed procedure of SAS 9.4 (SAS, 2014). For each
analyzed in triplicate.
response variable, the validity of model assumptions was verified by
For analysis of sucrose content, first, an internal standard solution
examining the residuals as described in Montgomery (2020). For sig­
was prepared using 600 mg fenyl-beta D-glucopyranoside (≥ 95 %,
nificant (p < 0.05) effects, multiple means comparison was conducted
Sigma-Aldrich, Steinheim, Germany) and 100 mL distilled water. Next,
using the least square means (lsmeans) statement of Proc Mixed at the 5
0.5 g powder of coffee beans from each sample was mixed with 25 mL
% level of significance.
distilled water containing 30 mg of fenyl-beta D-glucopyranoside (≥ 95
%, Sigma-Aldrich, Steinheim, Germany), as internal standard in a hot
3. Results
water bath (60 ◦ C, 30 min). Carrez I (15 % K4Fe(CN)6 (≥ 99 %, Sigma-
Aldrich, Steinheim, Germany) in water, 15 g L-1) and carrez II (30 %
ANOVA p-values that show the significance of the main effect of
ZnSO4 (≥ 99 %, Sigma-Aldrich, Steinheim, Germany) in water, 30 g L-1)
elevation range, and the main and interaction effects of shade and
(5 mL each) were added to the mixture and distilled water was then
postharvest processing, all nested in elevation range, on quality attri­
added to obtain a total volume of 50 mL. After mixing, the extract was
butes and biochemical contents of green arabica coffee beans are pre­
filtered through a 0.45 µm PTFE filter, and 0.5 mL of the filtrate was
sented in Table 2 and Table 3. Elevation significantly affected bean
transferred to a gas chromatograph-vial and dried using nitrogen gas. To
physical quality, total preliminary quality, total specialty quality and all
the dried extract, 0.5 mL of STOX-reagent (hydroxylamine hydrochlo­
studied CGAs, except for 5-CQA (Tables 2 and 3). Shade, nested in
ride in dry pyridine, 25 g L-1) (≥ 99 %, Fluka, Buchs, Switzerland) was
elevation range, significantly affected acidity and caffeine (Table 2).
added and the vial was placed in an oven at 60 ◦ C for 30 min. After
Postharvest processing significantly affected bean physical quality, total
cooling to room temperature, 0.5 mL of hexamethyldisilazane (HMDS)
preliminary quality, caffeine and all studied CGAs, barring 4-CQA and
(≥ 99 %, Sigma-Aldrich, Steinheim, Germany) and 0.05 mL of tri­
4,5diCQA. However, the interaction between shade and postharvest
fluoroacetic acid (TFA) (≥ 99 %, Sigma-Aldrich, Steinheim, Germany)
processing, each nested in elevation range, did not significantly affect
were added and then kept for 60 min to allow phase separation. Finally
any of the studied quality and biochemical attributes (Tables 2 and 3). In
for analysis, 1 µL was taken from the upper layer and injected into a gas
addition, none of the effects on body, flavor, cup quality and sucrose was
chromatograph (GC-3380, Varian, USA) equipped with a flame-
significant (Table 2). The overall means of body, flavor, cup quality and
ionization detector (Varian Instrument Group, Walnut Creek, CA)
sucrose content were 9.9, 10.3 and 46.3 scores and 46.03 g kg-1 dry
using a 1079 automatic injector. The chromatographic parameters were:
weight (dw), respectively.
stationary phase - (5 %-phenyl)-methylpolysiloxane (≥ 99 %, Sigma-
Aldrich, Steinheim, Germany), film thickness 0.25 µm, 30 m × 0.32
mm inside diameter (i.d.) (Agilent Technologies, Palo Alto, CA, USA); 3.1. Elevation effect
and mobile phase - He at 1 mL min-1, split 1/40, injector temperature =
250 ◦ C; detector temperature = 340 ◦ C; temperature program = 180 ◦ C As indicated in Table 4, mean scores of bean physical quality, total
for 1 min, ramp at 15 ◦ C min-1 to 290 ◦ C. The flame ionization detector preliminary quality and total specialty quality were significantly higher
was operated with hydrogen and air at 30 and 300 mL min-1, respec­ for midland coffee than for lowland coffee. But, mean contents of 3-CQA,
tively and helium at 20 mL min-1 as makeup gas. Identification was 4-CQA, 3,5diCQA, 4,5diCQA, FQA, FCQA and TCGAs were significantly
carried out according to retention times and addition of standards to the higher for lowland coffee than for midland coffee.

4
M. Worku et al. Journal of Food Composition and Analysis 115 (2023) 105027

Table 2
ANOVA p-values that show the significance of the main effect of elevation range (ER), and the main and interaction effects of shade and postharvest processing (PHP),
all nested in ER, on quality attributes, and caffeine and sugar contents 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 TSQ Caffeine Sucrose

ER 0.002 0.075 0.374 0.165 0.103 0.001 0.041 0.512 0.887

Shade(ER) 0.835 0.015 0.668 0.554 0.186 0.596 0.276 0.026 0.167
PHP(ER) 0.001 0.275 0.762 0.199 0.274 0.001 0.079 0.032 0.564
Shade(ER) * PHP(ER) 0.112 0.275 0.992 0.751 0.942 0.161 0.626 0.764 0.699

BPQ = bean physical quality, CQ = cup quality, TPQ = total preliminary quality, TSQ = total specialty quality

Table 3
ANOVA p-values that show the significance of the main effect of elevation range (ER), and the main and interaction effects of shade and postharvest processing (PHP),
all nested in ER, on CGAs contents of green arabica coffee beans. Significant effects that require multiple means comparison are shown in bold.
Source of variation 3-CQA 4-CQA 5-CQA 3,5diCQA 4,5diCQA FQA FCQA TCGAs

ER 0.001 0.001 0.744 0.001 0.001 0.001 0.001 0.001

Shade(ER) 0.313 0.337 0.416 0.752 0.130 0.641 0.174 0.630
PHP(ER) 0.001 0.110 0.005 0.001 0.163 0.024 0.001 0.048
Shade(ER) * PHP(ER) 0.864 0.740 0.211 0.907 0.498 0.580 0.876 0.479

Table 4
Mean values of bean physical quality (BPQ) (0–40 scoring), total preliminary quality (TPQ) (0–100 scoring), total specialty quality (TSQ) (0–100 scoring), and CGAs
contents (g kg-1 dw) obtained from the two elevation ranges (ER) (lowland and midland).
ER BPQ TPQ TSQ 3-CQA 4-CQA 3,5diCQA 4,5diCQA FQA FCQA TCGAs

Lowland 33.4 b 79.3 b 79.9 b 3.93 a 6.02 a 7.78 a 3.12 a 4.57 a 3.69 a 57.47 a
Midland 36.0 a 83.0 a 81.4 a 3.34 b 5.57 b 6.56 b 2.12 b 4.17 b 2.86 b 53.01 b

Within each column, means sharing the same letter are not significantly different.

3.2. Shade effect in midland had also much higher total preliminary quality than dry-
processed coffee in lowland and wet-processed coffees in lowland and
Mean value of acidity score was considerably lower for coffee beans midland, but there was no significant difference between dry-processed
grown with shade in lowland than those grown without shade in low­ coffee in lowland and wet-processed coffee in midland. Bean physical
land and midland and with shade in midland. Coffee beans grown with quality and total preliminary quality of wet-processed coffee in lowland
shade both in lowland and midland had a higher mean value of caffeine were lower than that of dry-processed coffees both in lowland and
content than those grown without shade in lowland (Table 5). But, midland and wet-processed coffee in midland.
coffee beans grown without shade in lowland and midland and those Caffeine contents of wet-processed coffee in lowland and midland
grown with shade in midland did not significantly differ in acidity. were considerably higher than that of dry-processed coffee in lowland.
Similarly, there was no significant difference between coffee beans But, there was no significant difference between dry-processed coffees in
grown without shade in lowland and midland and between those grown lowland and midland, and among dry-processed coffee in lowland and
with shade in lowland and midland and without shade in midland in wet-processed coffees in lowland and midland (Table 6).
caffeine content (Table 5). The 3-CQA content of dry-processed coffee in lowland was much
higher than that of dry- and wet-processed coffees in midland. The 3-
CQA contents of dry-processed coffee in midland and wet-processed
3.3. Postharvest processing effect
coffee in lowland were significantly higher than that of wet-processed
coffee in midland. The 5-CQA contents of wet-processed coffees in
Means (based on the main effect of postharvest processing nested in
lowland and midland were much more than that of dry-processed cof­
elevation range) of bean physical quality, total preliminary quality and
fees in lowland and midland (Table 6). The 3,5diCQA content of wet-
contents of caffeine and CGAs (3-CQA, 5-CQA, 3,5diCQA, FQA, FCQA
processed coffee in lowland was significantly higher than that of dry-
and TCGAs) are presented in Table 6. Bean physical quality was much
processed coffees in lowland and midland and wet-processed coffee in
higher for dry-processed coffees in lowland and midland than for wet-
midland. The 3,5diCQA content of wet-processed coffee in midland was
processed coffees in lowland and midland and for wet-processed cof­
much higher compared to that of dry-processed coffees in lowland and
fee in midland than for the same coffee in lowland. Dry-processed coffee
midland and that of dry-processed coffee in lowland compared to that in
midland (Table 6). The FQA content of wet-processed coffee in lowland
Table 5
was significantly higher than that of dry-processed coffees in lowland
Mean values of acidity (0–15 scoring) and caffeine (g kg-1 dw) obtained from the
and midland and wet-processed coffee in midland. The FQA content of
two shades (no shade and shade) within each of the two elevation ranges
(lowland and midland). dry-processed coffee in lowland was significantly higher than that in
midland. However, the FCQA content decreased across the two pro­
Shade(ER) Acidity Caffeine
cessing methods and elevation ranges. The TCGAs contents of dry- and
No Shade(Lowland) 11.5 a 13.80 b wet-processed coffees in lowland were much higher than that of dry- and
No Shade(Midland) 11.4 a 13.94 ab
wet-processed coffees in midland, but it did not considerably differ be­
Shade(Lowland) 10.4 b 14.31 a
Shade(Midland) 11.5 a 14.40 a tween dry-processed coffees and between wet-processed coffees in
lowland and midland (Table 6).
Within each column, means sharing the same letter are not significantly
different.

5
M. Worku et al. Journal of Food Composition and Analysis 115 (2023) 105027

Table 6
Mean values of bean physical quality (BPQ) (out of 40 scores), total preliminary quality (TPQ) (out of 100 scores), caffeine (g kg-1 dw), individual chlorogenic acids (g
kg-1 dw) and TCGAs (g kg-1 dw) obtained from the two postharvest processing (PHP; dry and wet) within each of the two elevation ranges (lowland and midland).
PHP(ER) BPQ TPQ Caffeine 3-CQA 5-CQA 3,5diCQA FQA FCQA TCGAs

Dry(Lowland) 36.1 a 81.2 b 13.74 b 4.08 a 27.82 b 6.88 c 4.44 b 4.51 a 56.94 a
Dry(Midland) 38.2 a 84.9 a 14.08 ab 3.63 b 27.72 b 5.38 d 4.05 c 3.32 b 52.33 b
Wet(Lowland) 30.8 c 77.3 c 14.38 a 3.78 ab 28.85 a 8.69 a 4.70 a 2.87 c 58.00 a
Wet(Midland) 33.9 b 81.1 b 14.26 a 3.06 c 28.75 a 7.74 b 4.29 bc 2.39 d 53.68 b

Within each column, means sharing the same letter are not significantly different.

3.4. Correlation between quality and biochemical attributes
Results of the correlation analysis between quality and biochemical
attributes are presented in Table 7. Bean physical quality and total
preliminary quality were negatively correlated with 4-CQA, 3,5diCQA,
FQA and TCGAs. Flavor and total specialty quality was also negatively
correlated with FCQA, and FCQA and TCGAs, respectively. However,
there was a positive correlation between bean physical quality and
FCQA and between cup quality and sucrose. On other hand, acidity and
body were not considerably correlated with either of the studied bio­
chemicals, and 3-CQA, 5-CQA, 4,5diCQA and FQA with any of the
quality traits.
Regardless of the statistical significance, all studied quality attributes
showed a negative correlation with caffeine, FQA and TCGAs and most
quality attributes with all the studied CGAs, excluding 5-CQA. However,
most of the studied quality attributes showed a positive correlation with
sucrose and 5-CQA (Table 7).
4. Discussion
The findings of this study indicate the importance of (1) elevation
range on physical quality, overall quality (total preliminary and spe­
cialty qualities) and CGAs contents, (2) shade, nested in elevation range,
on acidity and caffeine content, and (3) postharvest processing, nested
in elevation range, on physical quality, overall quality, and caffeine and
CGAs contents of green arabica coffee beans in southwestern Ethiopia
(Tables 2 and 3). Nonetheless, the differences observed for 3-CQA, 4-
CQA and FQA contents (≤ 0.6 g kg-1 dw) due to elevation effect (Table 4)
and for acidity (≈ 1 score) and caffeine content (≤ 0.6 g kg-1 dw) due to
shade effect (Table 5) were very minor. Besides, the two-way interaction
effect between shade and postharvest processing, each nested in eleva­
tion range, on quality and biochemical composition was not significant
(Tables 2 and 3). This indicates that coffee beans grown both with and
without shade in lowland and midland can be processed either by a dry
or a wet method without affecting its quality and biochemical content.
Similar to the findings of previous studies (Leonel and Philippe, 2007;
Tolessa et al., 2017; Worku et al., 2018, 2022), the results of this study
also showed a dominant elevation effect on coffee quality and
biochemical composition compared to the other studied factors. The
effect of postharvest processing was also dominant over that of shade.
However, there is no wicked interaction effect between micro-climate
driven by elevation and shade and bean processing on coffee quality
and biochemical composition.
The significant effects of elevation on CGAs and shade on acidity in
this study agree with previous studies (Bertrand et al., 2006; Borém
et al., 2014; Tolessa, 2017; Tolessa et al., 2017; Worku et al., 2018).
However, this and previous studies (Avelino et al., 2007; Vaast, Cilas,
et al., 2004; Tolessa et al., 2017; Worku et al., 2018) disagree for the
elevation effect on flavor, caffeine and sucrose and shade and post­
harvest processing effect on sucrose. Contrary to the present study
(Table 2), previous studies (e.g., Tolessa et al., 2017; Worku et al., 2018)
reported significant effects of elevation on flavor, caffeine and sucrose,
and shade and postharvest processing on sucrose. Similarly, there was
no interaction effect between shade and postharvest processing, each
nested in elevation range, in this study (Tables 2 and 3). Worku et al.
(2022) reported a similar finding on all studied physical quality attri­
butes and defects of green coffee beans, except for the proportion of
large beans. Yet, in Worku et al. (2018), a substantial interaction effect
between shade and postharvest processing on bean physical quality was
observed. In this study, shade considerably affected caffeine content and
postharvest processing physical quality, total preliminary quality, and
caffeine and total CGAs contents of green beans (Tables 2 and 3) while
these effects were not observed in Worku et al. (2018).
There is also a high inconsistency between different studies that re­
ported significant effects of elevation, shade, postharvest processing and
their two-way interactions on quality and biochemical composition of
green coffee beans. For example, some studies (e.g., Tolessa et al., 2017;
Worku et al., 2018) reported a negative effect of elevation on caffeine
content, while others (Avelino et al., 2007; Vaast, Cilas, et al., 2004;
Tolessa, 2017) reported a positive effect. On the other hand, the eleva­
tion effect on CGAs content observed in this study is in accordance with
these former studies, but disagrees with other ones (Bertrand et al.,
2006; Borém et al., 2014) that compared coffee varieties at different
elevations. Similarly, Somporn et al. (2012) found higher CGAs content
in shade than in full-sun grown coffee beans, whereas Vaast et al. (2006)
observed the reverse for CGAs, sucrose and trigonelline contents. These
contradictory reports indicate that the effects of these factors on coffee
quality and biochemical composition are highly site and growing season

Table 7
Pearson’s correlation coefficients between quality attributes and biochemical composition of green arabica coffee beans. The values shown in bracket are the p-values
for testing the null hypothesis that there is no correlation. Significant correlation coefficients at 5 % level are shown in bold.
Biochemical Quality attribute

BPQ Acidity Body Flavor CQ TPQ TSQ

Caffeine -0.19(0.08) -0.07(0.49) -0.14(0.17) -0.10(0.35) -0.11(0.30) -0.26(0.01) -0.13(0.21)

Sucrose -0.13(0.22) 0.06(0.60) 0.19(0.07) 0.19(0.06) 0.22(0.04) -0.01(0.95) 0.10(0.35)
3-CQA -0.10(0.33) -0.01(0.92) 0.10(0.33) -0.15(0.14) -0.04(0.74) -0.12(0.27) -0.14(0.19)
4-CQA -0.28(0.01) -0.06(0.59) 0.11(0.29) -0.09(0.40) -0.01(0.91) -0.26(0.01) -0.08(0.47)
5-CQA -0.14(0.18) 0.01(0.99) -0.07(0.52) 0.03(0.80) 0.01(0.99) -0.14(0.17) 0.04(0.71)
4,5diCQA -0.19(0.07) -0.01(0.96) 0.07(0.45) -0.14(0.18) -0.02(0.83) -0.20(0.06) -0.17(0.11)
3,5diCQA -0.30(0.01) 0.06(0.56) -0.15(0.14) 0.01(0.92) -0.01(0.97) -0.28(0.01) -0.02(0.82)
FQA -0.51(0.01) -0.16(0.12) -0.12(0.26) -0.06(0.56) -0.14(0.20) -0.52(0.01) -0.17(0.11)
FCQA 0.26(0.01) -0.05(0.61) -0.06(0.60) -0.25(0.01) -0.17(0.11) 0.11(0.29) -0.26(0.01)
TCGAs -0.29(0.01) -0.07(0.51) -0.11(0.30) -0.18(0.08) -0.14(0.20) -0.36(0.01) -0.21(0.05)

BPQ = bean physical quality, CQ = cup quality, TPQ = total preliminary quality, TSQ = total specialty quality.

6
M. Worku et al.
specific or depend on coffee variety, microclimate, soil properties,
agronomic practice, or experimental setup of the study.
The effect of elevation on coffee bean quality and biochemical
composition can mainly be linked to the relationship between elevation
and microclimate, particularly temperature of the growing area
(Getachew et al., 2022; Muschler, 2001) and partly to other factors such
as soil moisture (Carr, 2001; Moat et al., 2017) and coffee diseases and
pests (Worku, 2019). It is often noted that as elevation increases, tem­
perature decreases, but moisture (rainfall) increases. Lower tempera­
tures have been suggested to extend the maturation period of coffee
berries that in turn leads to a better bean filling (Leonel and Philippe,
2007; Vaast et al., 2006). In addition, the negative effects of drought
(Moat et al., 2017) and some coffee diseases and pests in Ethiopia (e.g.,
coffee leaf rust, coffee berry borer and Antestia bug) on coffee decrease
with elevation (Daba et al., 2019; Garbaba and Garedew, 2019; Garedew
et al., 2019). It is also indicated that coffee plants growing at higher
elevations with light shade level can assimilate more CO2 with minimum
evaporative water loss due to their higher water use efficiency under
these conditions than at lower elevations with light to intermediate
shade levels (< 35–65 %) (Getachew et al., 2021). Consequently, coffee
berries grown at higher elevations mature slowly with less growth
constraints (e.g., moisture, disease- and pest-induced stresses) and
supply more assimilates to the developing beans, which can lead to well
matured, dense and pure beans than those grown at lower elevations.
Conversely, warmer climatic conditions at lower elevations give more
rapid bean maturation and climatic- and biotic-related stresses for coffee
plants, resulting in more immature and defective coffee beans. A link
between immature and defective coffee beans and higher caffeine and
CGAs contents as well as poor cup quality has been observed earlier
(Farah et al., 2006; Link et al., 2014; Joët et al., 2010). Our results agree
with these findings and indicate that lowland coffee beans have lower
scores of bean physical quality and overall quality with more CGAs
contents compared to the midland ones (Table 4). Moreover, the
opposite trend of quality scores and CGAs contents between lowland and
midland coffees (Table 4) is in line with the negative relationships be­
tween quality attributes and CGAs (Table 7). Based on the mean values
of the total quality scores (Table 4), lowland coffee (79.9 scores out of
100) had also a lower specialty quality grade (i.e., Commercial Grade or
Grade 3 with a score range of 71-< 80) than midland coffee (81.4 scores
out of 100) (i.e. Specialty 2 or Q2 with a score range of 80-< 85). But,
lowland and midland coffees (79.3 and 83.0 scores out of 100, respec­
tively) had a similar preliminary quality grade; i.e., Grade 2 (75–84
scores out of 100). This quality grade classification is as per ECX coffee
cupping and quality grading guidelines (ECX, 2011).
The findings of this study are inconsistent with those of previous
studies (Muschler, 2001; Tolessa et al., 2017; Vaast, Van Kanten, et al.,
2004) regarding the increase in acidity of beans grown under shade, and
the change in body with elevation and shade. Similarly, the findings of
this study did not agree with those of previous studies (Tolessa et al.,
2017; Worku et al., 2018) in terms of shade effect on caffeine content.
Unlike in this study (Table 5), no shade effect on caffeine content
(Tolessa et al., 2017; Worku et al., 2018), but shade-induced improve­
ments in acidity and body in the low-elevation, sub-optimal coffee-zones
(Muschler, 2001; Vaast, Van Kanten, et al., 2004) and in a medium to
high elevation range (955–1250 m asl) where fertilization levels are
high (Leonel and Philippe, 2007) were reported in previous studies. But,
at higher elevations, shade, especially a dense shade, has a negative
effect both on acidity and body (Bosselmann et al., 2009; Tolessa et al.,
2017). Moreover, the absence of significant differences between beans
grown with and without shade in midland and in lowland and midland,
respectively (Table 5) contradicts with those reported by Avelino et al.
(2005). These authors postulated that a better exposure to morning
sunlight, due to east-facing slopes, is a cause for acidity increase at two
sites of different elevation in Costa Rica. Regardless of these in­
consistencies, some studies (Bosselmann et al., 2009; Tolessa et al.,
2017; Vaast, Cilas, et al., 2004) obtained higher acidity for coffees
7
Journal of Food Composition and Analysis 115 (2023) 105027
produced at higher than at lower elevations with light to medium shade
levels. In the same manner with that of higher elevations, shade allows
more time for bean filling (Vaast, Van Kanten, et al., 2004) and possibly
for accumulation of acidity precursors (Leonel and Philippe, 2007).
These observations do not corroborate with our findings, as acidity of
coffee beans grown under shade in midland was lower than that of beans
grown without shade in lowland and midland and under shade in low­
land (Table 5). Besides, lower caffeine content for beans grown under no
shade in lowland than under shade in lowland and midland observed in
this study (Table 5) is unexpected. It is generally thought that the coffee
plant can use caffeine as one of its defense mechanisms against some
stresses (e.g., pest attacks) that are often higher for coffees grown
without shade in lowland than with shade in lowland and with and
without shade in highland. Nevertheless, data on specific biochemical
compounds controlling each cup quality attribute including acidity, as
well as how environmental (and biotic) factors interplay on metabolic
pathways of each biochemical are generally scarce. To our knowledge,
the only study that provides comparable data has been executed in
Nicaragua by Leonel and Philippe (2007). These authors found that
elevation determined coffee quality to a greater extent than shade, and
fat accumulation, favored by elevation, enhanced the intensity of
various organoleptic characteristics (e.g., aroma, body, acidity and fla­
vor) of coffee. However, this requires further investigation using
repeated experiments over several years and locations.
The differences observed between coffee beans grown in lowland and
midland in physical quality, total specialty quality, and caffeine and
CGAs contents due to postharvest processing (Table 6) clearly confirm
the findings of the previous studies (Taveira, 2014; Tolessa, 2017;
Worku et al., 2018). These studies reported the existence of interaction
between growing environment and postharvest processing for quality
attributes and biochemical contents of green arabica coffee beans. Tol­
essa (2017) and Worku et al. (2018) showed an interaction effect of
elevation and postharvest processing on sucrose and caffeine contents
and sucrose content, respectively. Taveira (2014) also found a link be­
tween elevation and postharvest processing to differentiate metabolites
of Acaiá genotype samples. However, the findings of this study did not
agree with those of Tolessa (2017) in postharvest processing effect on
caffeine and total CGAs contents at different elevation ranges. This study
showed higher caffeine and total CGAs contents for wet-processed
lowland and midland coffees than for dry-processed lowland coffee
and for dry- and wet-processed lowland coffees than for others,
respectively (Table 6). But, Tolessa (2017) showed higher caffeine
content for semi-wet and wet-processed midland coffees than for
dry-processed midland coffees and for dry-, semi-wet- and
wet-processed lowland coffees, and a statistically similar total CGAs
content across elevation ranges and postharvest processing methods. In
addition, in this study, we observed inconsistency between the contents
of individual isomers of CGAs for coffee beans grown in lowland and
midland and processed by dry and wet method (Table 6). This can
suggest the existence of different responses of metabolic processes of
individual CGAs isomer for postharvest processing across elevation
ranges.
Regarding elevation, shade or processing vs. coffee quality, it is
generally thought that higher elevation, shade or wet processing pro­
duces good quality coffee. Unlike this thought, this study indicates
higher bean physical quality scores for dry-processed lowland and
midland coffees than wet-processed ones and total preliminary quality
scores for dry-processed midland coffee than others (Table 6). Similarly,
Worku et al. (2018) showed higher scores of physical quality and pro­
portion of Grade 1 quality coffee for beans grown without shade and
processed by a dry method and for beans grown without shade at higher
elevations (1550–1820 m asl) or for those gown at higher elevations
(1550–1820 m asl) and processed by the dry method, respectively than
all other samples. Lower proportion of defected beans and higher scores
of bean physical quality and total preliminary quality, respectively have
also been reported for dry-processed highland coffee than others (Worku
M. Worku et al.
et al. 2022) and for dry-processed than for wet- and semi-wet-processed
beans (Tolessa, 2017). These findings exhibit that dry processing can
produce quality coffee that is comparable to or higher than that is pro­
duced by wet processing. However, these results may not be consistent
with the actual coffee production situation, particularly for large farms,
due to larger volumes of coffee cherry in dry processing (pulp + bean)
than in wet processing (bean only). The pulp of the fresh coffee cherries,
which represents 43.2 % and 28.7 % of their fresh and dry weights,
respectively (Bressani, 1979), consists of a large amount of water, which
affect proper drying in dry processing. Also, the climate in the study area
is very moist, which often makes proper drying difficult and leads to
fungal development. Therefore, it is vital to know that in practice, dry
processing of larger volumes of fresh cherries, containing a high amount
of water under a humid climate, may lead to lower bean quality
compared to wet processing. Nonetheless, to make a reliable conclusion,
further research using repeated experiments over several years and lo­
cations, and samples collected not only from fields, but also from bulk
productions is recommended.
Overall, the results observed in the present and previous studies
indeed confirm the initial hypothesis of this study; i.e., the effects of
shade and postharvest processing on quality and biochemical composi­
tion of coffee can vary with growing conditions (e.g., elevation, micro­
climate, season, soil, topography or aspect). Nevertheless, scientific
evidence as to how the postharvest processing method influences quality
attributes and biochemical compositions of coffee beans grown in
different environments (e.g., elevation and shade condition) is scant. To
our knowledge, except for shade effect on microclimate (mainly tem­
perature) and that of temperature on bean development, there are also
no previous studies on how shade influences these attributes of coffee
beans grown under different growing environments. So, this calls for a
comprehensive study (that includes multi-season and multi-location) to
determine the actual reason for these results.
The negative relationship between some quality attributes and bio­
chemicals observed in this study (Table 7) agrees with that reported in
the literature (Farah et al., 2006; Farah, 2012). In these studies, high
levels of CQAs, FQAs and their oxidation products are associated with
poor cup quality and off-flavor. However, a strong positive correction
between high trigonelline and 3,4diCQA levels and high quality is re­
ported in Farah et al. (2006) and between CGAs and some quality at­
tributes such as acidity, body, flavor and overall cup quality in Sualeh
et al. (2020). The negative association between bean physical quality
and 4-CQA, 3,5diCQA, FQA, FCQA and total CGAs (Table 7) is also in
line with the findings of Ramalakshmi et al. (2007). These authors found
lower contents of caffeine, protein, lipid, sucrose and total polyphenols
in defected than in normal (graded) coffee beans.
5. Conclusion
Elevation of the coffee-growing area was found to be the dominant
factor affecting green arabica coffee bean quality and biochemical
composition. This is followed by postharvest processing. Overall, better
quality coffee with lower contents of chlorogenic acids was associated
with midland (1550–1900 m asl) coffee-growing areas. Better quality
coffee with lower 3,5diCQA and FQA contents was also associated with
midland coffee-growing areas and dry processing. Conversely, lower
quality coffee with higher 3,5diCQA and FQA contents was associated
with lowland coffee-growing areas and wet processing. Wet processing
also produced beans with higher caffeine and 5-CQA contents. Growing
with shade in lowland and midland also produced beans with higher
caffeine content than growing without shade in lowland. In contrast,
growing without shade in lowland and midland produced beans with
higher acidity than growing with shade in lowland. Both physical and
overall qualities were negatively associated with caffeine, 4-CQA,
3,5diCQA, FQA and total CGAs. But, bean physical quality and FCQA,
and cup quality and sucrose were positively correlated. This is the first
report showing the characteristics of quality and biochemicals of green
8
Journal of Food Composition and Analysis 115 (2023) 105027
arabica coffee beans obtained from different elevation ranges, shading
and postharvest processing methods, and the physical quality correla­
tion with biochemicals. However, this study considered two elevation
ranges (lowland: 1100–1550 m asl and midland: 1550–1900 m asl),
modern plantation coffee, one growing season and samples directly
collected from coffee trees. Hence, to arrive at a more comprehensive
conclusion, future studies need to consider repeated experiments over
several years and multi-location elevation ranges (lowland to highland),
and samples collected not only from coffee trees in modern plantations,
but also in other production systems (e.g., semi-forest/plantation and
garden coffees) and bulk productions.
Declaration of Conflict of Interest
Authors have no conflict of interests to declare.
Funding
This work was funded by VLIR-UOS Institutional University Coop­
eration with Jimma University, Ethiopia.
Data Availability
The authors do not have permission to share data.
Acknowledgement
The Ethiopia Commodity Exchange in Addis Ababa and Jimma, all
cuppers and lab technicians who were involved in this study and all
farmers who provided us with coffee samples and sample processing
services are acknowledged.
Appendix A. Supporting information
Supplementary data associated with this article can be found in the
online version at doi:10.1016/j.jfca.2022.105027.
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