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Effect of Different Fermentation Methods on Physicochemical Composition

and Sensory Quality of Coffee (Coffea arabica)

Article · June 2017

DOI: 10.9790/2402-1106023136

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IOSR Journal of Environmental Science, Toxicology and Food Technology (IOSR-JESTFT)
e-ISSN: 2319-2402,p- ISSN: 2319-2399.Volume 11, Issue 6 Ver. II (June. 2017), PP 31-36
www.iosrjournals.org

Effect of Different Fermentation Methods on Physicochemical

Composition and Sensory Quality of Coffee (Coffea arabica)
Kinyua Agnes Wamuyu1,Kipkorrir Richard1, Mungendi Beatrice1,
Kathurima Cecilia2
1
Institute of Food Bioresources and Technology, Dedan Kimathi University of Technology Nyeri, Kenya.
2
Coffee Research Foundation, Ruiru, Kenya.

Abstract: Fermentation of coffee beans is primarily done to remove mucilage and can be done using two
methods; dry and wet fermentation methods. This research aimed at determining the effect of different
fermentation methods on physicochemical composition and the sensory quality of coffee. Coffee cherries were
pulped and subjected to natural fermentation methods in different fermentation containers; plastic bucket, sack
and cement tank. After fermentation, the parchment were washed and dried. The green coffee beans were
evaluated for physicochemical composition and sensory attributes. The results showed that different
fermentation methods did not have significant variations in most of the physico-chemical parameters analysed.
However, significant variations were observed in the levels of pH with the wet fermented coffee samples
showing lower levels as compared to dry fermented samples. Sensory evaluation results showed that wet
fermented coffee samples had better colour of green beans, least silver skin discoloration and overall quality
compared to dry fermented coffee samples. There were no significant differences in the body, acidity, colour,
flavour and overall class among the coffee samples fermented using different containers. Hence different
containers used during fermentation do not affect coffee quality and processors can adopt materials that are
cheaper to reduce expenses during coffee processing.
Key Words: Arabica coffee, fermentation methods, physicochemical composition, cup quality

I. Introduction
In Kenya, coffee is largely processed by wet method. The stages included during wet processing of
coffee include; harvesting, sorting, removal of pulp, fermentation followed by washing, grading drying and
finally storage[1]. Coffee is fermented to ease the removal of a layer of mucilage surrounding the bean, after
which it is washed and dried. During fermentation the mucilage is degraded by natural enzymes in coffee
fruits and the growth of microorganisms[2]. Chemical changes that occur during fermentation are pectin
degradation by pectinase enzymes and breakdown of sugars in the mucilage[3]. Fermentation is concluded when
the mucilage is easily detached by washing. Apart from mucilage removal fermentation also has impact on
chemical composition of coffee bean and resulting cup quality. Microbial fermentation yields different flavor
materials by breaking down the carbohydrates in the coffee mucilage and thus the superior aroma characteristics
of wet-processed coffees[4].
Wet fermentation, involves water being added to submerge the coffee parchment in fermentation tank.
For dry fermentation, the coffee parchment fermented in a tank without addition of the water [5]. In Kenya,
coffee parchment fermentation is commonly done under dry or wet fermentation methods. Wet fermentation
method is commonly used when conditions are too hot and fermentation conditions need to be controlled.
Similarly coffee processors may utilize different equipment during the fermentation of coffee parchment [6].
This variations in the method and equipment used may cause variations in several parameters which could
influence the final coffee quality. However, there is limited information on the effect of different fermentation
methods and materials used on the chemical composition and the final cup quality. This research aimed at
determining the effects of different fermentation methods and materials used on physicochemical composition
and cup quality of coffee.

II. Methods And Methods

2.1Sample Collection and Treatment
Coffee cherries of Ruiru 11 cultivar was obtained from Dedan Kimathi University of Technology
coffee farm, located in Nyeri County, Kenya. The ripe cherries were harvested from coffee plants grown in the
same block by selective hand picking method to obtain uniform ripe cherries. The cherries were pulped and then
subjected to four different treatments which included dry fermentation of coffee parchment in a sack, dry
fermentation in a plastic bucket, dry fermentation in a cement tank and wet fermentation in a plastic bucket. For
dry fermentation in a plastic bucket, perforations at the bottom of the container allowed the excess water to drain

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 Effect Of Different Fermentation Methods On Physicochemical Composition And Sensory Quality Of

while for wet fermentation the parchment was submerged in water in a plastic bucket. A control sample was also
prepared by fermentation of coffee parchment in a cement tank by the dry method. The end of fermentation was
assessed by rubbing a few beans of the fermenting coffee in the hand to check whether they felt gritty and can
easily be washed. After fermentation the coffee parchment was thoroughly washed to remove the degraded
mucilage and then dried to reduce moisture content to a level of 9.5 to 10%.

2.2 Physicochemical Composition

2.2.1 Preparation of Green Beans for Chemical Analysis
The parchment were hulled and ground to obtain fine powder of particle size 0.6mm for analysis of
physicochemical components including; crude protein, titratable acidity, pH, sucrose, caffeine, chlorogenic
acids and trigonelline.
2.2.2 Determination ofCrude Protein
Crude protein was determined using Kjedhal method as described in [7]. The Protein % was calculated by
multiplying nitrogen value obtained by protein factor (6.25).
2.2.3 Determination Titratable Acidity
Total acidity in the coffee samples was determined by adding 10 g of the coffee sample in a conical flask with
75 ml 80% alcohol. Agitation was done using a shaker for 16 hours followed by filtration. Ten ml of the filtrate
was diluted to 100 ml followed by titration with 0.1N Sodium hydroxide using phenolphthalein as an indicator
[8].
2.2.4 Determination of pH
The pH of coffee samples was determined using the method as described by[9].Three grams of ground coffee
were added to 50ml hot water. The extract was cooled to room temperature and pH measured using a pH meter.

2.2.5 Determination of Sucrose Content

Sucrose was determined using High Performance Liquid Chromatography (HPLC) analysis as
described by[10]. In sample preparation, 0.5g of the sample was added to a flask containing 50ml 96 % ethanol.
The mixture was refluxed for one hour, then ultrasonicated for six minutes. This was followed by cooling and
filtration using a Whatman filter paper no.42.The filtrate obtained was evaporated to dryness using rotor vapors.
It was then reconstituted with10 ml mobile phase containing acetonitrile: water 80:20 and filtered using micro
filter (0.45 μm). A sample of 20µm was injected into HPLC fitted with a refractive index.

2.2.6 Determination ofCaffeine Content

Extraction and purification of caffeine was done as described by [11]. Five (5) grams of the ground
coffee powder was weighed in a 250 ml Erlenmeyer flask before adding 3.5 grams of magnesium oxide and 200
ml of double distilled water. The mixture was refluxed for 25 minutes and then left to cool. It was then filtered
under vacuum on celite and the filtrate recovered in a 250 ml flask topping up the volume with distilled water.
Twenty (20) ml of this preparation was then pipetted into 100 ml flask adjusting to volume with mobile phase
consisting of 35% v/v methanol, 65% v/v distilled water and 0.1% v/v glacial acetic acid. The extracts were then
filtered using 0.45 μm micro filters. A sample volume of 20 μl was injected into HPLC with pulsed diode array
(PDA) detector set at 278 nm. Peak areas were used for calculating the concentration of caffeine.

2.2.6 Determination of Chlorogenic Acid Content

The coffee samples were prepared for HPLC analysis according to the method described by [12]. Two
grams of each ground coffee sample were weighed accurately in 250 ml beakers and 100 ml distilled water was
added to each sample. The mixture was boiled for 5 minutes while stirring, and then cooled. Subsequently, the
solution was filtered using 0.45 μm filter paper. The clear filtrate with some dilution was used for the HPLC
analysis with detector, pulsed diode array set at 324 nm. A 100μl amount of the filtrate was diluted with 900 μl
of demonized water and pipetted into clean 1000 μl volumetric flasks. Peak areas were used for calculating the
concentration of chlorogenic acids.

2.2.7 Determination of Trigonelline Content

Trigonelline content was determined using HPLC analysis according to method described
by[13].Samples preparation was done by weighing 0.5g of ground coffee sample and adding it 30 ml of an
acetonitrile: water solution (5:95 v/v) at 80 °C for 10 min. Extracts were filtered and diluted in the mobile phase
before injection. Peak area were used for calculating the concentration of trigonelline.

2.3 Sensory Evaluation

The quality of the coffee samples was analyzed using the method of [14]. Three hundred grams of
green bean were used for raw bean analysis and 100g during roasting for each sample. A sensory evaluation

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 Effect Of Different Fermentation Methods On Physicochemical Composition And Sensory Quality Of

report form was used to record quality attributes for each coffee on raw and liquor quality. During raw bean
quality analysis, color and quality attributes were assessed. Liquor was assessed using acidity, body and flavor
attributes. Color of raw bean evaluated as greyish blue, greyish green with blue tinge, greyish green, greyish
green with brown tinge, brownish grey green, greenish and brownish. Quality and flavor evaluated as fine, good
to fine, good, fair to good FAQ fully, FAQ, poor to fair, poor and very poor. Acidity and body evaluated as
pointed/ full, medium, light medium, light and lacking. The overall quality of the coffee was then evaluated as
fine, good to fine, good, fair to good FAQ fully, FAQ, poor to fair, poor and very poor. The overall quality
given infers to the class of the coffee sample.

III. DataAnalysis and Presentation

Data was analyzed using Statistical Packages for Social Scientist (SPSS) software version 20.
Measures of variability of means among the samples was done using ANOVA. The means were subjected to
Duncan Multiple Range test. The significance level was established atp≤0.05. Sensory data was subjected to
analysis of variance and multivariate analysis. Mean seperation was done using Student-Newman-Keuls
(SNK5%) test by Costat. Discriminant Function analysis was done using XL-STAT 2017.

IV. Results And Discussion

4.1 Physicochemical Composition of Coffee Processed Using Different Fermentation Methods
The levels of crude protein, pH, and titratable acidity and biochemical compounds in the coffee
samples are presented in Table 1. In this study, the protein content of the green beans ranged between 10.62 ±
0.20 and 10.82± 0.15 for all the treatments. The results were within the range reported in literature. Green coffee
beans contain about 9 to 16 % of protein in dry weight basis [15]. The results showed no significant differences
between the treatments used in regard to the protein content (p ≤ 0.05). Therefore, the wet fermentation method
which allows the mucilage to degrade slowly did not cause any changes in the protein content of the beans and
did not vary with the level in the dry fermented samples. However, [16] reported that free amino acids could be
released in the bean following the degradation of proteins during the fermentation process in coffee.

Table 1: physicochemical composition of coffee processed using different fermentation methods (%)
Parameter Treatment
Dry fermentation in Dry fermentation in Dry fermentation in a Wet fermentation in a
cement tank a sack plastic bucket plastic bucket
Crude Protein 10.67 ± 0.81 a 10.82± 0.15a 10.79 ± 0.54a 10.62 ± 0.20a
Titratable acidity 1.79± 0.2 a 1.66 ± 0.17 a 1.96 ± 0.58 a 1.62 ± 0.12 a
pH 6.06 ± 0.01 a 6.06 ± 0.01 a 6.05 ± 0.06a 6.00 ± 0.20 b
Sucrose 7.76 ± 1.15 a 7.92 ± 1.27 a
7.76 ± 1.11 a 8.03± 1.51 a
Caffeine 1.12± 0.44 a 1.13 ± 0.13 a 1.19± 0.41 a 1.17 ± 0.56 a
Chlorogenic acids 5.48± 1.03 a 6.34 ± 1.20 a
5.94±1.24 a 6.09± 0.74 a
Trigonelline 0.82 ± 0.14 a 0.87 ± 0.6 a 0.91± 0.43 a 1.01± 0.24 a

Values are means (± SD) of triplicate determinations. Means with the same letter are not significantly different
at p ≤ 0.05
It could be expected that the wet fermentation method causes more degradation of coffee than the dry
fermentation due to the soaking by water and the long period of fermentation involved but this was not noted in
the study. Losses of components such as protein in green beans could be minimal since protein and peptides are
reported to complex with polyphenols and hence remain intact in the cell structure[16]. A study by [17]noted
that the total concentration of free amino acids and protein content does not change significantly with the
chemical reaction occurring during the harvest season and the postharvest processing steps, such as
fermentation, drying and storage. Protein, peptides, and free amino acids are vital for coffee flavor since they are
needed for the Maillard reaction [18]. They serve as flavor precursors as they play major roles during roasting of
coffee to produced aroma compounds such as furans, pyrroles, pyrazines [15].
The mean titratable acidity of the green coffee beans that were fermented in a cement tank and in a
sack was 1.79 ± 0.2and 1.66 ± 0.17 respectively. Titratable acidity of green beans for dry and wet fermented
samples in a plastic bucket was 1.96 ± 0.58and 1.62 ± 0.12, respectively (Table 1). Similar results were obtained
by[19]who reported titratable acidity of 1.32 to 1.60 mL NaOHg-1[20]noted that dry processed coffee has higher
titratable acidity as compared to wet processed coffee due to high levels of fermentation. However there were no
significant differences between the fermentation methods used in terms of titratable acidity. Regarding the pH
level, the wet fermentation method showed a significantly lower pH level. This could be attributed to the longer
fermentation time taken during wet fermentation and the view that formation of aliphatic acids is increased by
underwater fermentation[21]. Wet fermentation method takes longer because the water lowers the temperature
and reduce the microbial growth hence degradation of mucilage from the beans proceeds at a slower phase.
Hence acids generated by the microorganisms during fermentation accumulates in the beans. A report by
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 Effect Of Different Fermentation Methods On Physicochemical Composition And Sensory Quality Of

[22]indicate that microbial metabolites such as acetic acids produced during post-harvest processing accumulate
in the coffee beans. The presence of these acids could lower pH levels.
Sucrose is an important biochemical component in coffee beans since their high levels in coffee beans
correlate with better cup quality. Sucrose levels in the coffee samples as analyzed in this study, were found
within the range of 7.76% and 8.03% for all the treatments (Table 1). These findings concur with [10] who
reported that sucrose level of Arabica coffee ranges between 5- 8.5%. There were no significant differences
between the treatments used in relation to the sucrose level. Although sucrose is soluble in water, these results
indicate no loss of sucrose to the fermenting water as in the case of wet fermentation where coffee parchment is
immersed in water and possible leaching of sucrose could occur. Studies on the effect of coffee processing on
content of sugars showed that only reducing sugars such as glucose and fructose may be lost to the processing
water [23]. Therefore sucrose as a disaccharide is not significantly affected by coffee fermentation and remains
intact as a storage compound [24].The level of caffeine in all the coffee samples for all the treatments used
ranged from 1.12% to 1.19% (Table 1). The level of caffeine in the samples were therefore within the range
reported in literature. Arabica coffee contains about 1.2% of caffeine with a range of 0.6 to 1.9%[20]. From the
study, there were no significant differences in the levels of caffeine in the coffee samples processed by the
different fermentation methods. The levels of caffeine mainly vary between coffee species and genotypes [25].
There was no indication of loss of caffeine to the water used during fermentation as in the case of wet
fermentation. This could be because caffeine is less soluble or it’s strongly bound to the alkaloid and other
compounds in the coffee bean [26].Caffeine plays an important role in coffee quality as it contributes to the
perceived strength and to a small extent the bitterness of the brew[27]. Chlorogenic acids are among some of the
aroma precursors in green coffee that are considered to contribute to acidity and formation of aroma compounds
during roasting[28]. From the results, the chlorogenic acid content values of the samples varied from 5.48% to
6.34% (Table 1). The values of chlorogenic acid content obtained were in agreement with what is reported in the
literature. The levels of chlorogenic acids range from 4% to 8.4 %[27]. Similarly, in the study, there were no
significant differences (p ≤ 0.05) between the different treatments used in relation to the level of chlorogenic
acids content. Hence different fermentation methods does not affect the level of chlorogenic acids in green
coffee beans. The wet fermentation method did not show any loss of this compound to the processing water.

4.2.1Sensory Quality of Coffee Processed By Different Fermentation Methods

Sensory quality evaluation was done to assess the influence of various fermentation methods on the
quality of coffee. The method of Devonshire, [14]is commonly used for the assessment of the raw bean color,
roast quality and liquor quality of coffee which aid in categorizing coffee into various classes. The attributes are
given scores that range from 1 to 7 with the lowest score indicating the best level of the attribute. From the
results, the color of the green coffee beans that were fermented in a cement tank and in a sack showed a mean
score of 4.00 ± 0.87and 4.33 ± 0.50,respectively. The color of green beans that were obtained by dry and wet
fermentation in a plastic bucket showed a mean score of 4.33 ± 0.50and 2.67 ± 0.0respectively (Table 2). There
were significant differences (p ≤ 0.05) between the treatments in terms of color. The wet fermented samples in a
plastic bucket showed a significantly lower score than the other treatments in terms of color. This was described
as a good greyish green color compared to the greyish green with brown tinge color shown by the other coffee
samples obtained by the dry fermentation method. The dry fermented samples also showed more silver skin
discoloration as compared to wet fermented coffee (Table 3). Considering overall quality, the wet fermented
coffees samples was categorize as class 4+ (fully FAQ), followed by dry fermented coffee in a cement tank;
class 4- , (About FAQ). The samples that were dry fermented in a sack and a plastic buckets both scored class 5-
, (Poor to fair quality). From the study, it was noted that there were significant differences (p ≤ 0.05) among the
treatments in terms of color and overall quality (Table 2). Wet fermentation method have shown better overall
quality and produces better color and with less silver skin discoloration. This improved overall quality may be
attributed to the controlled longer fermentation time which allows increased complexity of acids and the
possible leaching of some soluble substances bitter compounds from the beans to the water [29]. Moreover, it
allows controlled fermentation to occur for longer which results in coffee with desirable attributes in terms of
raw bean color and overall quality[2].
The acidity and body was described as medium for all the coffee samples for the different treatments.
Hence no variations in the level of the attributes for all the treatments used (Table 3). This indicates that the
different fermentation methods did not affect the perceived acidity and body of the brew. Although the pH level
of the green beans was found to be significantly different between the treatments, the titratable acidity on the
other hand was not significantly different (p ≤ 0.05) between the treatments. Hence the level of acids in the brew
for all the cofffee samples in the brew after roasting could be similar.The acids present in green beans include
citric, malic, chlorogenic acids and quinic acids. The level of acids such as citric, malic and chlorogenic acid are
shown to decrease during roasting while quinic acids increase due to the breakdown of chlorogenic acids[30].

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 Effect Of Different Fermentation Methods On Physicochemical Composition And Sensory Quality Of

Table 2: Mean sensory variables of raw bean color and coffee brew of coffee subjected to different
fermentation treatments
Treatment Color of green bean Flavor Overall class
Dry fermentation in cement tank 4.00 ± 0.87 b 5.00± 0.00 a 4- b
Dry fermentation in plastic bucket 4.33± 0.50 b 5.33± 0.50 a 5+ b
Dry fermentation in sack 4.33± 0.50 b 5.22± 0.44 a 5+ b
Wet fermentation in plastic bucket 2.67± 0.0a 5.00± 0.00 a 4+ a
Values are means (± SD) of triplicate determinations. Means with the same letter are not significantly different
at p ≤ 0.05.

Table 3: Description of Raw and Overall quality of coffee processed using dry and wet fermentation
methods
Treatment Raw quality Body, Acidity Overall quality
Dry fermentation in cement Greyish green with brown tinge, Rather Medium About FAQ
tank(Control) coated
Dry fermentation in sack Greyish green with brown tinge, Rather Medium Poor to fair
coated
Dry fermentation in plastic bucket Greyish green with brown tinge, Rather Medium Poor to fair
coated
Wet fermentation in plastic bucket Greyish green Medium Fully FAQ
Key: FAQ- Fair Average Quality

4.2.2 Discriminant Function Analysis of Sensory Quality Variables of Coffees Samples In Different
Containers
The quality data was subjected to discriminant function analysis (DFA). Results of the discriminant
function analysis showed that the first discriminant factor explained 92.56 % total variation (Fig. 1). Using
sensory variables, the coffees were not distinctly separated since overlapping of points were observed for the
samples obtained from different containers used. (Fig. 1). This shows that the different containers used during
the fermentation process did not have effect on the color, flavor and overall quality of coffee. This indicate that
the different containers did not affect or created different conditions for the fermentation of parchment. Hence it
can be deduce that, the use of different containers during fermentation process does not affect the color of green
bean, flavor and over quality of coffee brew.

Figure 1: Discriminant factor analysis (DFA) based on quality data of dry fermented coffee in different
containers.
V. Conclusion
From the study, the different fermentation methods used did not affect the level of crude protein,
titratable acidity, sucrose, caffeine, and chlorogenic acids content of green coffee beans. However, pH was
significantly different between the treatments. The wet fermentation method of coffee processing showed better
quality than the other methods used. The use of different containers during fermentation showed non-significant
differences on quality parameters such as color, flavor and overall class. Hence using different containers for
fermentation under good practice does not affect the quality of coffee.
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 Effect Of Different Fermentation Methods On Physicochemical Composition And Sensory Quality Of

Acknowledgement
This work was sponsored by Dedan Kimathi University of Technology. We are also grateful to Coffee Research
Institute for the facilitation of the laboratory analysis.

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