Issue 2 (202)/2025

ALSE and ACS Style
Alhassan, M.; Kumah, P.; Tandoh, P.K.; Idun, I.A. Effect of pod size and fermentation method on temperature changes and the chemical quality of cocoa (Theobroma cacao) beans. Journal of Applied Life Sciences and Environment 2025, 58 (2), 233-244. https://doi.org/10.46909/alse-582174

AMA Style
Alhassan M, Kumah P, Tandoh PK, Idun IA. Effect of pod size and fermentation method on temperature changes and the chemical quality of cocoa (Theobroma cacao) beans. Journal of Applied Life Sciences and Environment. 2025; 58 (2): 233-244.
https://doi.org/10.46909/alse-582174

Chicago/Turabian Style
Alhassan, Mohammed, Patrick Kumah, Paul Kweku Tandoh, and Irene Akua Idun. 2025. “Effect of pod size and fermentation method on temperature changes and the chemical quality of cocoa (Theobroma cacao) beans.” Journal of Applied Life Sciences and Environment 58, no. 2: 233-244.
https://doi.org/10.46909/alse-582174

Effect of pod size and fermentation method on temperature changes and the chemical quality of cocoa (Theobroma cacao) beans

Mohammed ALHASSAN¹, Patrick KUMAH², Paul Kweku TANDOH^2* and Irene Akua IDUN²

¹Quality Control Division of Ghana Cocoa Board, Kumasi, Ghana; email: mbadahala2017@yahoo.com

²Department of Horticulture, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana; email: patrickumah@gmail.com; reneidun@gmail.com

*Correspondence: pktandoh.canr@knust.edu.gh 

Received: Jan. 21, 2025. Revised: Apr. 06, 2025. Accepted: Apr. 08, 2025. Published online: Jun. 26, 2025

ABSTRACT. Cocoa is a highly valuable economic crop that generates income and foreign exchange for Ghana. However, the rejection of cocoa beans on the international market due mainly to inappropriate fermentation methods and postharvest practices is a major problem confronting this sector. The objective of this study was to determine the effect of the pod size and fermentation method on temperature changes and chemical attributes of the beans. The experiment was arranged in a 3 × 3 factorial with a completely randomised design and three replicates. Pod size (small, medium and large) and fermentation method (heap, tray and basket fermentation) were each evaluated at three levels. There were changes in temperature using all three fermentation methods, such that temperature increased from an average of 37.8 to 41.7°C in the first 24 h. The heap fermentation method had the highest temperature (41.7°C) after 4 days. The pod size and fermentation method did not influence the fat content, total titratable acidity, or free fatty acids in the sampled beans. Small pods fermented using the heap fermentation method had the highest pH. The study concluded that the heap fermentation method enhances temperature changes without compromising the chemical quality of cocoa beans, making it the preferred technique for high-quality cocoa production.

Keywords: bean quality; fermentation methods; pod sizes.

 

INTRODUCTION

Cocoa (Theobroma cacao), belonging to the Sterculiaceae family (Nair, 2010), provides food, income, employment, industrial raw material and resources for poverty reduction (Peprah, 2015). Peprah (2019) reported that cocoa provides raw material for the multibillion-dollar global chocolate industry, in addition to providing a livelihood for millions of smallholder farmers.

The cocoa market has experienced record-breaking growth in global supply over the last decade (TRIDGE, 2024). Sasu (2025) reported that in 2023, cocoa in Ghana was forecasted to contribute GHS 3.15 billion, or around USD 262.8 million, to the country’s gross domestic product (GDP).

Additionally, this agricultural product was projected to contribute GHS 3.75 billion (approximately USD 312.9 million) to the country’s GDP by 2027, marking the highest contribution during the observed period. In the 2021/2022 crop season, Ghana produced an estimated 689,000 metric tonnes of cocoa beans.

There has been a growing interest in researching cocoa and its byproducts as superfoods, not only for their distinct sensory properties but also for their nutritional benefits to human health. Cocoa is rich in phenolic compounds, mainly methylxanthines, such as theobromine and caffeine, and polyphenols, such as flavan-3-ol monomers, with anthocyanins and proanthocyanidins being the most prevalent (Carrillo et al., 2014; Dugo et al., 2018).

In recent years, Ghanaian cocoa beans have been frequently rejected on the international market because of poor bean quality resulting from improper fermentation methods coupled with other post-harvest practices (Lass and Wood, 2020). Additionally, the International Cocoa Organization (ICCO, 2021) highlighted how improper fermentation and drying methods contribute to quality deterioration, leading to the rejection of Ghanaian cocoa beans in global markets.

These developments have adverse social and economic implications. The multi-buying system, which allows for competition among licenced buying companies in the purchase of cocoa beans, has been identified as one of the major causes of poor fermentation, as it compels farmers to present stocks of cocoa for sale at any stage of fermentation, thus affecting quality. The fermentation method plays a vital role in achieving good-quality cocoa beans with the best chocolate flavour (De Vuyst and Weckx, 2016). Cocoa beans that undergo inadequate or improper fermentation exhibit several quality defects that affect their marketability and usability in chocolate production. Poorly fermented beans tend to be excessively bitter and have a high polyphenol content and undesirable acidic flavours (Jinap et al., 1995).

Furthermore, defective beans often contain high levels of undesirable compounds, such as acrid tannins, which contribute to off-flavours (Aprotosoaie et al., 2016). A study by Oliveira et al. (2006) demonstrated that improperly fermented beans had lower concentrations of desirable volatile compounds, such as aldehydes and pyrazines, which are responsible for the characteristic chocolate aroma.

Furthermore, Arulmari and Visvanathan (2024) opined that turning at 12 h intervals in the heap method resulted in a less acidic nature (titratable acidity) and desirable quality attributes, which was considered optimum.

This highlights the importance of proper fermentation control in achieving premium-quality cocoa beans. During fermentation, enzymatic activity from microorganisms (yeasts, lactic acid bacteria and acetic acid bacteria) contributes to lipid hydrolysis.

Afoakwa and Paterson (2011) mentioned that larger pods tend to produce higher mucilage content, creating anaerobic environments that extend microbial activity and the enzymatic breakdown of lipids, which facilitates the formation of free fatty acids (FFAs).

However, in Ghana, the overreliance on the traditional heap method for fermentation with its attendant problems necessitate extensive research into alternative methods, such as basket, tray and box fermentation.

Although several studies have determined the impact of different fermentation methods on cocoa quality and found that factors such as the stage of pod ripening, duration of pod storage, duration of fermentation, turning and environmental conditions, little is known about the interaction between the pod size and fermentation method on the chemical quality of cocoa beans.

Alhassan et al. (2024) reported that pod size and fermentation method (tray, basket and heap) did not influence the physical properties (slaty, germinated and mouldy) of cocoa beans.

Therefore, the aim of this study was to determine the influence of pod size and fermentation method on the temperature changes and chemical attributes of cocoa beans.

 

MATERIALS AND METHODS

Sources of cocoa pods and beans

Cocoa pods of the hybrid variety ‘Akokorabedi’ were obtained from a 4-acre farm located in Bronikrom in the Ahafo Ano South District of the Ashanti region (Latitude: 6.98791° N, Longitude: -1.95266° W with an of elevation of approximately 387 meters (1,270 feet) above sea level). Fermented and dried cocoa beans were obtained from these pods and sent to the Quality Control district office at Mankranso and the Quality Control Company laboratories at Tema for physical and chemical analyses, respectively.

Sample preparation of pods and fresh beans

Beans obtained from a total of 50 uniformly ripened cocoa pods, which were divided into small, medium and large pods, were used for the study. Pods were considered small when they weighed between 100 and 450 g, medium when they weighed between 451 and 800 g, and large when they weighed more than 800 g (choosing 50 for 3 sizes means that the sample size was unequal between groups). Each sampled pod was broken and the number of beans in each counted and recorded. Fresh beans from each pod were also weighed and recorded.

Experimental design and procedure

The experiment was arranged in a 3 × 3 factorial with a completely randomised design. The experiment comprised of two factors: pod size at three levels (small, medium and large) and fermentation method at three levels (heap, tray and basket fermentation). Each of the resulting 9 treatment combinations was replicated 3 times. Hybrid cocoa pods of uniform ripeness and not more than 16 years old (Class A and B trees) were harvested, assembled and segregated based on their sizes. Pods were broken separately and according to their sizes using blunt machetes to facilitate easy removal of beans from pods. From each pod size category, 2 kg of extracted cocoa beans were subjected to fermentation using one of three methods: basket, tray, heap fermentation (Figure 1).

Fermentation methods

Basket fermentation

Three sets of plastic baskets of dimensions L = 55 cm × B = 37 cm × H = 24 cm with a maximum capacity of 30 kg were used for fermentation. Each basket was filled with fresh cocoa beans of the large, medium and small pods and left uncovered for 24 h prior to the start of fermentation. They were labelled as depicted in Figure 2. The three main sides, bottom and top surfaces, were lined with banana leaves to prevent the beans dehydration and as a heat conservation mechanism. Fermentation spanned a period of 6 days (144 h). The baskets were opened and the beans turned every 48 h.

Tray fermentation

Wooden trays of dimensions L = 90 cm × B = 60 cm × H = 13 cm were used for fermentation (Figure 3). They were arranged in three sets, one on top of the other, so that each stand had three trays, each constituting a replication. The trays were labelled TR₁, TR₂ and TR₃ and filled trays were covered with plantain leaves. Tray fermentation also lasted for 6 days (144 h).

Heap fermentation

The traditional heap fermentation method was also carried out in each pod size category. A flat and dry spot was located, and cocoa beans were heaped and covered with banana leaves. This method also lasted for 6 days (144 h).

Monitoring of temperature in trays, heaps and baskets

During fermentation, the temperature of the fermenting mass of beans was measured every morning (6 am) and evening (5 pm) in all treatments at a minimum depth of 15 cm, using a thermometer obtained from the Department of Horticulture, KNUST (Food Network 984732). Temperature readings were collected for all samples during heap, basket and tray fermentation.

Drying of beans

The sun drying technique was used to dry the fermented beans on raised platforms, which were made of raffia palm fronds and raised 1 m high using bamboo as stands.

The platform was divided into nine sections based on the three pod size categories and three fermentation methods. Sun drying was performed at 30–40°C with a relative humidity of 50–70%.

Sampling of beans for chemical analysis

Using a moisture meter (KPM Aqua Boy, London, UK), the moisture loss of dried cocoa beans from each pod size and fermentation method was monitored for 6 days until a moisture content of 7% was attained.

The beans were mixed thoroughly to ensure uniformity. Cocoa beans from each fermentation method and pod size were dried separately and sampled for cut tests and laboratory analysis.

 

 

 

 

Data collection

Chemical quality attribute determination

All laboratory work was performed at the laboratories of the Quality Control Division of the Ghana Cocoa Board located in Tema, Ghana. Samples were analysed based on AOAC (2012) procedures for chemical properties, such as fat content, FFA content, pH and titratable acidity.

Determination of the fat content

All samples were dried in an oven at 105°C to facilitate peeling of the external layer of the beans, followed by grinding. A 5.0-g sample of powdered cocoa beans was collected and transferred into an extraction thimble, and the fat was removed using the Soxhlet method of cocoa powder fat extraction standard procedure.

Petroleum ether was used as a solvent, and after being dissipated through evaporation, the extracted fat in the Erlenmeyer flask was dried for 2 h in an oven at 105°C. Using a desiccator, the fat in the flask was cooled for 30 min and reweighed with the residue. The fat content was expressed as a percentage of the weight of 5.0 g using the following formula (AOAC, 2012) (Equation 1):

% Fat = (M3 − M1) / M2 × 100

(1)

where: M₁ = mass of the Erlenmeyer flask (grams); M₂ = mass of the cocoa powder sample (grams); M₃ = mass of the flask with the fat residue (grams)

Determination of free fatty acids (FFA)

To determine the FFA content in each sample, 25 mL of 95% ethanol and 25 mL diethyl ether were added to 2 g of already extracted fat. A uniform mixture was obtained by swirling it gently and titrating it using a phenolphthalein indicator with 0.1 M NaOH. The average titre values were noted. The formula below (AOAC, 2012) was used to calculate the FFA content as oleic acid and expressed as a percentage (Equation 2):

where Vol. Base = volume of the base; Vol. Blank = blank titre value; M. Base = base molarity; Equivalent = equivalence of oleic acid = 282 × 100

pH determination

Cocoa nibs from each treatment sample were finely milled, and 10 g of each sample was placed into a kilner jar. Clean distilled water was boiled to 100°C, and 100 mL was poured into the kilner jars.

The solution was vigorously stirred for 5 min for all samples. Using Whatman filter paper, the stirred solution was filtered into another kilner jar, and the filtrate was left to cool to a tem-perature of 20°C. A pH meter with three-point calibration was then used to deter-mine the pH in each sample (AOAC, 2012).

Titratable acidity (TTA) determination

A 10-g sample of ground nibs was added to 100 mL of distilled water and swirled occasionally for 30 min. The mixture was filtered, and a 20 mL of the filtrate was titrated with 0.1 M sodium hydroxide (NaOH) solution using phenolphthalein as an indicator. Titration was stopped when a faint pink colour appeared, which faded after a few seconds. The following formula was used to calculate the titratable acidity (%TA) (Equation 3):

where Equivalent as oleic acid = 282; Wt. of sample = 2.00 g; MBase = Molarity of the base.

Data analysis

Data were subjected to analysis of variance (ANOVA) using Statistix 10 statistical software.

The differences between means were separated using Tukey’s honestly significant difference (HSD) at a 1% (0.01) probability level.

 

RESULTS

Temperature changes during fermentation

Figure 4 shows the temperature changes using the three fermentation methods. The temperature increased from 37.8 to 41.7°C in the first 24 h. The highest temperature (44.17°C) was observed after 4 days using the heap fermentation method, and the lowest tem-peratures were recorded using the basket (41.17°C) and tray methods (41.43°C).

Chemical attributes of cocoa beans after analysis

Fat content of cocoa beans

No significant effects (p ≥ 0.01) of pod size, fermentation method or their interaction were observed on the fat content (Table 1).

Free fatty acids of cocoa beans

No significant effects (p ≥ 0.01) of pod size, fermentation method or their interaction were observed on the FFA content (Table 2).

pH of cocoa beans

There was significant interaction (p ≤ 0.01) between pod size and fermentation method for the pH of cocoa beans (Table 3). The highest pH (5.51) was produced by beans obtained from small pod sizes and fermented using the heap fermentation method. This was similar to beans obtained from large pods fermented using the heap method and medium and large pods fermented using the tray method. The lowest pH (5.10) was produced by large pods fermented using the basket fermentation method.

 

 

In terms of the fermentation method, the highest pH (5.47) was produced using the heap method, and the lowest (5.18) was obtained using the basket fermentation method. There was no significant effect of pod size.

Total titratable acidity (TTA) of cocoa beans

No significant effects (p ≥ 0.01) of pod size, fermentation method or their interaction were observed on TTA (Table 4).

 

Table 1
Effects of the pod size and fermentation method on the fat content (%) of cocoa beans

Fermentation Method (FM)
Pod size (PS)	Heap		Tray	Basket	Means
Small	55.00	52.07		57.00		54.69
Medium	54.00		54.13	51.07	53.07
Large	52.60		53.93	56.53	54.36
Mean	53.87		53.38	54.87
CV (%) = 5.85; HSD (0.01): FM = 3.036, PS = 3.036, FM × PS = 7.251

CV: Coefficient of variation; HSD: Honestly significant differences

 

Table 2
Effects of the pod size and fermentation method on the free fatty acid content (%) of cocoa beans

Fermentation Method (FM)
Pod size (PS)	Heap	Tray	Basket	Means
Small	0.75ab	0.69ab	0.79ab	0.75a
Medium	0.60b	0.73ab	0.92a	0.75a
Large	0.78ab	0.79ab	0.69ab	0.76a
Mean	0.71a	0.74a	0.80a
CV (%) = 5.05; HSD (0.01): FM = 0.134, PS = 0.134, FM × PS = 0.319

Means followed by different lowercase letters are significantly different from each other (p < 0.01).
CV: Coefficient of variation; HSD: Honestly significant differences

 

Table 3
Effects of the pod size and fermentation method on the pH of cocoa beans

Fermentation Method (FM)
Pod size (PS)	Heap	Tray	Basket	Means
Small	5.51a	5.28b	5.24b	5.35a
Medium	5.40a	5.42a	5.20bc	5.34a
Large	5.48a	5.45a	5.10c	5.34a
Mean	5.47a	5.38b	5.18c
CV (%) = 5.85; HSD (0.01): FM = 0.048, PS = 0.048, FM × PS = 0.115

Means followed by different lowercase letters are significantly different from each other
(p < 0.01). CV-Coefficient of variation; HSD-Honestly significant differences

 

Table 4
Effects of pod size and fermentation method on the total titratable acidity (TTA) of cocoa beans

Fermentation Method (FM)
Pod size (PS)	Heap	Tray	Basket	Means
Small	4.98	5.52	5.18	5.23
Medium	5.26	4.97	4.99	5.03
Large	5.74	5.71	4.87	5.48
Mean	5.33	5.40	5.02
CV (%) = 5.85; HSD (0.05): FM = 0.811, PS = 0.811, FM × PS = 1.937

CV: Coefficient of variation; HSD: Honestly significant differences

 

DISCUSSION

The results indicate that there was a general rise in the temperature of the beans from one to four days of fermentation. This could be attributed to the conversion of sugars in the pulp surrounding the beans to ethanol by yeast. Bacteria start to oxidise the ethanol to acetic acid and eventually to carbon dioxide and water, producing more heat, thereby increasing the temperature. Ordoñez-Araque et al. (2020) reported that the breakage and eventual drainage of the mucilage commenced after 24 h of fermentation. Under anaerobic conditions, lactic acid bacteria transform alcohol into lactic acid. However, this is halted when acetic acid begins to oxidise the alcohol into acetic acid under aerobic conditions. The temperature of the fermenting mass of beans increases from about 40 to 45°C during the first 2 days of fermentation (Bobiles et al., 2022). The remaining days are characterised by bacterial activity, which proceeds under an oxygenated environment. Pulp drainage and tempera-ture can be constant at this stage (Díaz-Muñoz et al., 2021). These chemical reactions are responsible for the develop-ment of chocolate flavour and colour. The duration of fermentation varies based on the type of bean; Forastero beans require approximately 5 days, while Criollo beans only need 2–3 days (Calvo et al., 2021).

Although there were no significant effects of pod size, fermentation method or their interaction on the fat content, the values obtained for all treatment samples were fairly satisfactory for the environmental conditions prevalent in the study area (Torres-Moreno et al., 2015). Both the highest and lowest values of 57.00 and 51.07% were obtained from small and medium pods, respectively, that were fermented using basket fermentation. According to Thompson et al. (2007), the FFA content can increase by over 30% if fermentation is prolonged beyond optimal limits, especially under high-humidity conditions. In the current study, no significant effects of pod size, fermentation method or their interaction were observed on the FFA content. To understand the degree of oxidation of the fat component of cocoa beans, the amount of FFAs available is a major determinant, as it shows the degree of mouldiness caused by microbial activity (Houphouet et al., 2023). The FFA content in a given sample of cocoa beans can be used to determine its degree of rancidity. Despite the absence of significant differences among samples, the observed values fell within the acceptable limit of less than 1.75%. Houphouet et al. (2023) reported that FFA levels exceeding the minimum standard of 1.75% are unacceptable and added that recommended postharvest practices should be followed to improve the quality of fermented and dried cocoa beans with an FFA content below the international standard of 1.75%. An FFA content above 2% in cocoa beans can lead to quality rejection by premium buyers, particularly in Europe and North America (ICCO, 2021).

The results indicate that the highest pH was produced by beans obtained from small pod sizes and fermented using the heap fermentation method. This was similar to large-sized pods fermented using the heap method and medium and large pods fermented using the tray method. The lowest pH was produced by large pod sizes fermented using the basket method. This observation could be due to the increased aeration of the mass from turning and the loss of mucilage, which enables the proliferation of acetic acid bacteria (Barel, 1998; Camu et al., 2007). Barel (1998) noted that lactic acid negatively affects the quality of raw cocoa beans because it remains in the beans due to its low volatility. In the present study, the fermentation method increased the chances of obtaining beans with the desired chocolate flavour and reduced the level of acidity as was also reported by Afoakwa and Paterson (2011). Biehl et al. (1990) revealed that the duration of fermentation changed the acidity of the beans. Furthermore, the turning of beans and the fermentation technique have a significant influence on the acidity and physical quality of raw cocoa beans (Guehi et al., 2010). This suggests that the heap fermentation method leads to higher acid accumulation. Tsegay (2020) stated that the organic acid composition and fermentation process are input factors directly and jointly affecting TTA. In the current study, TTA was not affected by the interaction between the pod size and fermentation method. This could be due to the equal levels of lactic acid produced among the fermentation processes. Unlike pH, which measures the concentration of hydrogen ions in a solution, TTA measures the total acidity as an approximate value by giving the sum of free protons and undissociated acids in a solution. It is another quantitative measure akin to pH and has been proposed as a more accurate representation of perceived acidity because it makes it possible to taste both free hydrogen ions and those that are bound to organic acids (Rogers et al., 1999). Studying the relationship between the pod size and fermentation method on the physical attributes of cocoa beans, Alhassan et al. (2024) reported that the highest bean purity was produced by medium-sized pods fermented using tray fermentation, while medium pod sizes produced the least germinated beans under all three fermentation methods.

 

CONCLUSIONS

This study demonstrated that the heap fermentation method consistently resulted in higher temperature changes during fermentation, with no detrimental impact on the key chemical quality attributes of cocoa beans, such as fat content, FFAs, pH and TTA. Among the fermentation methods analysed, the heap method resulted in the highest pH level, especially when applied to small pods, but maintained the chemical quality within acceptable industry standards. For cocoa farmers, the heap fermentation method is recommended, as it supports the development of optimal chocolate fla-vour without compromising the chemical integrity of the beans, making it a viable practice for producing high-quality cocoa beans suitable for international markets.

 

Author contributions: Conceptualization: MA and PK; Methodology: MA, PK, PKT and IAI; Analysis: MA, PKT; Investigation: MA and PKT; Resources: MA, PK, PKT and IAI; Data curation: MA, PK, PKT and IAI; Writing, review, supervision: MA, PK, PKT and IAI. All authors declare that they have read and approved the publication of the manuscript in this present form.

Funding: There was no external funding for this study.

Conflicts of interest: There are no conflicts of interest regarding this article.

 

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