Determination of aflatoxins in by-products of industrial processing of cocoa beans

This study has examined the occurrence of aflatoxins in 168 samples of different fractions obtained during the processing of cocoa in manufacturing plants (shell, nibs, mass, butter, cake and powder) using an optimised methodology for cocoa by-products. The method validation was based on selectivity, linearity, limit of detection and recovery. The method was shown to be adequate for use in quantifying the contamination of cocoa by aflatoxins B(1), B(2), G(1) and G(2). Furthermore, the method was easier to use than other methods available in the literature. For aflatoxin extraction from cocoa samples, a methanol-water solution was used, and then immunoaffinity columns were employed for clean-up before the determination by high-performance liquid chromatography. A survey demonstrated a widespread occurrence of aflatoxins in cocoa by-products, although in general the levels of aflatoxins present in the fractions from industrial processing of cocoa were low. A maximum aflatoxin contamination of 13.3 ng g(-1) was found in a nib sample. The lowest contamination levels were found in cocoa butter. Continued monitoring of aflatoxins in cocoa by-products is nevertheless necessary because these toxins have a high toxicity to humans and cocoa is widely consumed by children through cocoa-containing products, like candies.     

Changes of phenolic compounds and antioxidant capacity in cocoa beans processing

A number of studies have shown that the consumption of cocoa and chocolate products has positive health effects on humans. The object of this research was to monitor changes of total and individual phenolics and antioxidant capacity during the cocoa bean manufacturing. The loss of cocoa phenolics and their antioxidant activity vary with the degree of technological process. The process of roasting and cocoa nib alkalisation had the major influence on phenolic compounds as well as on antioxidant capacity. The roasting treatment resulted in 14% loss of the total phenolics content, while alkalisation resulted in 64% loss of total phenolics content. Procyanidins B1 and B2 as well as (?)‐epigallocatechin were the unstable components, while caffeic acid derivate showed the greatest stability in all technological process. Furthermore, PCA showed that phenolic contents, antioxidant capacity and non‐fat cocoa solids parts of the samples were classified in groups according technological conditions.     

Evaluation of the effect of processing on cocoa polyphenols: antiradical activity,anthocyanins and procyanidins profiling from raw beans to chocolate

The content and composition of anthocyanins and procyanidins in fermented cocoa beans (from different geographic origins: Ecuador, Cameroon, Ivory Coast, Ghana and Nigeria), roasted nibs, cocoa mass and chocolate were determined, beside the determination of the total antiradical capacity. Concerning geographic origin, cocoa beans and processed products from Ecuador showed the highest levels of anthocyanins, followed by Nigeria and Cameroon. Generally, as cocoa beans were further processed, the levels of anthocyanins and flavan‐3‐ols decreased. The largest observed losses of phenolics occurred during roasting. A progressive decreasing trend in polyphenol concentration was observed in the other processed samples as well. Despite the original content of polyphenols in raw cocoa beans, technological processes imply a significant impact on cocoa quality, confirming the need of specific optimisation to obtain high value chocolate.     

Impact of industrial treatments on ochratoxin A content in artificially contaminated cocoa beans

Ochratoxin A (OTA) is a mycotoxin mainly produced by mould species of the genera Aspergillus and Penicillium, which grow on a variety of agricultural products. OTA-contaminated foodstuffs pose a major health hazard to consumers, including human and animal. In Côte d’Ivoire, numerous studies are being carried out to find the best way of preventing OTA contamination of cocoa raw material. The objectives of this investigation were to assess the impact of industrial treatment on OTA content in cocoa-derived products. Samples of cocoa pods were prepared under specific conditions promoting fungal proliferation on cocoa beans before processing. The beans underwent the usual industrial treatments – roasting, shelling, crushing, pressing and additive addition – and samples were taken at each stage. OTA was extracted with a methanol/3% sodium hydrogen carbonate solution and purified using an immunoaffinity column prior to HPLC analysis with fluorescence detection. OTA was detected in artificially contaminated cocoa beans at levels ranging from 3.4 to 44.7 µg kg^?1 with a mean value of 22.9 ± 3.6 µg kg^?1. OTA was mainly concentrated in the shell (93%). Roasting, shelling and additive addition significantly decreased levels of OTA by 24–40, 76 and 52%, respectively, with an overall reduction of ～91%. These results indicate that industrial processing of cocoa has a real impact on the reduction of OTA in final cocoa products.     

Dark chocolate acceptability: influence of cocoa origin and processing conditions

BACKGROUND: Chocolate properties can vary depending on cocoa origin, composition and manufacturing procedure, which affect consumer acceptability. The aim of this work was to study the effect of two cocoa origins (Ghana and Ecuador) and two processing conditions (roasting time and conching time) on dark chocolate acceptability. RESULTS: Overall acceptability and acceptability for different attributes (colour, flavour, odour and texture) were evaluated by 95 consumers. Differences in acceptability among dark chocolates were mainly related to differences in flavour acceptability. The use of a long roasting time lowered chocolate acceptability in Ghanaian samples while it had no effect on acceptability of Ecuadorian chocolates. This response was observed for most consumers (two subgroups with different frequency consumption of dark chocolate). However, for a third group of consumers identified as distinguishers, the most acceptable dark chocolate samples were those produced with specific combinations of roasting time and conching time for each of the cocoa geographical origin considered. CONCLUSION: To produce dark chocolates from a single origin it is important to know the target market preferences and to select the appropriate roasting and conching conditions. Copyright © 2011 Society of Chemical Industry     

Effect of post-harvest treatments on the occurrence of ochratoxin A in raw cocoa beans

Cocoa beans are the principal raw material for chocolate manufacture. Moulds have an important place in the change in the quality of cocoa beans due to their role in the production of free fatty acids and mycotoxins, namely ochratoxin A (OTA). This study investigated the impact of the key post-harvest treatments, namely the fermentation and drying methods on OTA contamination of raw cocoa beans. Analytical methods for OTA detection were based on solid–liquid extraction, clean-up using an immunoaffinity column, and identification by reversed-phase HPLC with fluorescence detection. Of a total of 104 randomly selected cocoa samples analysed, 32% had OTA contents above 2 µg kg^–1. Cocoa sourced from pods in a bad state of health had a maximum OTA content of 39.2 µg kg^–1, while that obtained from healthy pods recorded 11.2 µg kg^–1. The production of OTA in cocoa beans increased according to the pod-opening delay and reached 39.2 µg kg^–1 after an opening delay of 7 days after harvest, while 6.1 and 11.2 µg kg^–1 were observed when pods were opened after 0 and 4 days. OTA production also seemed to depend considerably to the cocoa fermentation materials. When using plastic boxes for bean fermentation, the OTA production was enhanced and reached an average OTA content of about 4.9 µg kg^–1, while the raw cocoa treated in banana leaves and wooden boxes recorded 1.6 and 2.2 µg kg^–1 on average respectively. In parallel, the OTA production was not really influenced by either the mixing or the duration of the fermentation or the drying materials.     

Some quality characteristics of solar-dried cocoa beans in St Lucia

A field comparison of solar drying and open-air sun-drying of cocoa (Theobroma cocoa L) beans was carried out in St Lucia. Four methods of drying (indirect solar drier, direct solar drier, open air/perforated steel surface and open air/non-perforated wooden surface) were examined at three loading rates: 13·7, 26·9 and 40·4 kg m⁻². Beans from the open air had a higher incidence of external mould and poorer external appearance, though differences were minor. Beans dried at the lower rate of 13 kg m⁻² showed the best colour, but the highest titratable acidity. Conversely, beans dried at the higher loading rate of 40·4 kg m⁻² showed significantly lower titratable acidity, but poorer colour. Differences in cut-test score, colour, pH and titratable acidity between the open air and closed driers were small or not significant. While not significant, the indirect drier did show the highest cut-test score and the direct drier the poorest. Beans from the indirect drier were darker and more purple, while those from the direct drier were lighter coloured and less purple. The beans from the direct drier, dried to 6% moisture (WB) were, though not significantly so, more brittle and higher in titratable acidity than those from either the open air or indirect drier. Overall the beans from the indirect drier showed the highest quality and those from the direct drier the poorest. Whether the modest improvement of the indirect drier over the open air driers is sufficient to warrant investment in such a drier is unclear. © 1998 SCI.     

Fermentation of cocoa beans: influence of microbial activities and polyphenol concentrations on the flavour of chocolate

BACKGROUND: Spontaneous cocoa bean fermentation is characterised by a succession of microbial activities. Cocoa flavour precursors are developed during fermentation and drying of cocoa beans. Polyphenols and alkaloids contribute to astringency and bitterness of cocoa and chocolate. RESULTS: Population dynamics, metabolite target analyses, and chocolate production were performed for seven independent spontaneous cocoa bean heap fermentations in Ghana. Although the same micro‐organisms were involved in these heaps, carried out at different farms or in different seasons, heap temperatures and microbial metabolite concentrations were different. This could be due to heterogeneity and size of the heaps, but was mainly ascribed to microbial variability. Indeed, differences in microbial activity could be linked with the flavour of chocolates made from the corresponding dried, fermented cocoa beans. Whereas the polyphenol and alkaloid contents of cocoa beans were crop‐ and heap‐dependent, epicatechin and theobromine levels decreased during fermentation due to diffusion out of the bean cotyledons and polyphenol oxidation and condensation. Residual levels were responsible for the degree of bitterness of the final chocolates. CONCLUSION: Differences in microbial activities between different heap fermentations can result in dried fermented cocoa beans and chocolates with different flavour characteristics. Hence, fermentation control may direct the flavour of chocolate. Copyright © 2008 Society of Chemical Industry     

Effect of farm and industrial processing on the amino acid profile of cocoa beans

An investigation into the amino acid profiles of unfermented and fermented cocoa nibs, as well as process-line cocoa nibs (P-LCN) and processed cocoa cake samples (PCCS) was carried out. In the unfermented cocoa nibs, Glu (128 mg/g crude protein i.e. 128 mg/gcp) was the most abundant amino acid whilst the most concentrated essential amino acid in the same sample was Leu (72.2 mg/g crude protein); in the fermented cocoa nibs, a similar trend was observed, with respective values of Glu (153 mg/gcp) and Leu (62.4 mg/gcp). Lys (181 mg/gcp) was most abundant amino acid in P-LCN and Ile (63.3 mg/gcp) was the second most abundant; also, in PCCS, Lys (52.7 mg/gcp) was the most abundant amino acid but Asp (43.7 mg/gcp) was the second most abundant. The total amino acid content was (mg/gcp), 641 (unfermented nibs), 708 (fermented nibs), 635 (P-LCN) and 368 (PCCS), with corresponding essential amino acids of 300, 287, 478 and 185 (all with His), respectively. Based on whole hen’s egg, the limiting amino acids for the samples were: Ser (unfermented nibs), Met (fermented nibs), Ala (P-LCN) and Val (PCCS), whereas under provisional amino acid scoring pattern, they were: Met + Cys (unfermented nibs), Met + Cys (fermented nibs), Thr (P-LCN) and Val (PCCS). Prolonged and high heat treatments appeared to have reduced the essential amino acids of the PCCS as compared to the P-LCN. Significant differences existed between contents of essential amino acids and non-essential amino acids at p < 0.05 in unfermented cocoa nibs, fermented cocoa nibs and P-LCN.     

Aflatoxins and ochratoxin A: occurrence and contamination levels in cocoa beans from Brazil

In the present study, the occurrence of aflatoxins (AFs) and ochratoxin A (OTA) was evaluated in 123 samples of cocoa beans produced in five Brazilian states. The presence of these mycotoxins was determined by high-performance liquid chromatography with fluorescence detection (HPLC-FLD) after immunoaffinity column clean-up. The mean level of total AFs in cocoa beans samples was 5.7 μg.kg^?1. Four (3.3%) samples exceeded the maximum limit of 10 μg.kg^?1 established by the Brazilian legislation for total AFs. The mean level of OTA contamination was 1.2 μg.kg^?1, and none of the samples exceeded the maximum limit established by the Brazilian legislation. The co-occurrence of AFs and OTA was observed in 4.9% of the samples. The results of the present study demonstrated that, in relation to the levels of AFs and OTA established by the Brazilian legislation, most samples of cocoa beans analyzed are safe for consumption. This is the first report on the occurrence and levels of AFs and OTA in cocoa beans from the five main Brazilian states producing cocoa. The data in this study provide important information for farmers, traders, industry, consumers and law enforcement agencies.     

The effect of high-pressure jet processing on cocoa stability in chocolate milk

Fat-free chocolate milk formulations containing skim milk, cocoa powder, and sugar were thermally treated and then processed using high-pressure jet (HPJ) technology from 125 to 500 MPa. The rheological properties and stability of HPJ-treated chocolate milks were compared with controls (no HPJ processing) prepared both with and without added κ-carrageenan. As expected, carrageenan-free chocolate milk exhibited immediate phase separation of the cocoa powder, whereas formulations containing κ-carrageenan were stable for 14 d. An increased stability was observed with increasing HPJ processing pressure, with a maximum observed when chocolate milk was processed at 500 MPa. The apparent viscosity at 50 s⁻¹ of HPJ-processed samples increased from ~3 mPa·s to ~9 mPa·s with increasing pressure, and shear-thinning behavior (n < 0.9) was observed for samples processed at HPJ pressures ≥250 MPa. We suggest that HPJ-induced structural changes in casein micelles and new casein-cocoa interactions increased cocoa stability in the chocolate milk. Because casein seemed to be the major component enhancing cocoa stability in HPJ-treated samples, a second study was conducted to determine the effect of additional micellar casein (1, 2, or 4%) and HPJ processing (0–500 MPa) on the stability of fat-free chocolate milk. Formulations with 4% micellar casein processed at 375 and 500 MPa showed no phase separation over a 14-d storage period at 4°C. The addition of micellar casein together with HPJ processing at 500 MPa resulted in a higher apparent viscosity (~17 mPa·s at 50s⁻¹) and more pronounced shear-thinning behavior (n ≤ 0.81) compared with that without added micellar casein. The use of HPJ technology to improve the dispersion stability of cocoa provides the industry with a processing alternative to produce clean-label, yet stable, chocolate milk.     

Occurrence of ochratoxin A in cocoa beans: Effect of shelling

The aim of this study was to investigate the influence of the shelling process on the presence of ochratoxin A (OTA) in cocoa samples. Twenty-two cocoa samples were analysed for the determination of OTA before (cocoa bean) and after undergoing manual shelling process (cocoa nib). In order to determine OTA contamination in cocoa samples, a validated high-performance liquid chromatography (HPLC) method with fluorescence detection was used for the quantitative analysis of ochratoxin A (OTA). In both types of samples, OTA was extracted with methanol-3% sodium hydrogen carbonate solution and then purified using immunoaffinity columns prior to HPLC analysis. Due to the fact that different recovery values were obtained for OTA from both types of samples, a revalidation of the method in the case of cocoa nibs was needed. Revalidation was based on the following criteria: Selectivity, limits of detection and quantification (0.03 and 0.1 µg kg^-1, respectively), precision (within-day and between-day variability) and recovery 84.2% (RSD = 7.1%), and uncertainty (30%). Fourteen of the twenty-two cocoa bean samples (64%) suffered a loss of OTA of more than 95% due to shelling, six samples suffered a loss of OTA in the range 65-95%, and only one sample presented a reduction of less than 50%. The principal conclusion derived from this study is that OTA contamination in cocoa beans is concentrated in the shell; therefore, improvements of the industrial shelling process could prevent OTA occurrence in cocoa final products.     

生产工艺对可可奶稳定性的影响

研究了可可粉预处理及均质条件对可可奶稳定性的影响。结果表明，可可粉先在90℃保温30min后过胶体磨可明显提高可可奶的稳定性和口感。可可奶的较佳均质工艺为：65℃，25MPa下均质两次。     

Application of kokum (Garcinia indica) fat as cocoa butter improver in chocolate

Although cocoa butter (CB) is an ideal fat for use in chocolate, it softens with heat and is not suitable for use in warm climates. CB extenders or improvers, preferably from stearic acid‐rich fats, are good candidates to increase the heat‐resistance property of CB and chocolate. In the present investigation, one such fat, kokum, is used as an improver to increase the hardness of chocolate. Kokum fat is added in various proportions replacing CB in dark and milk chocolate formulations and its effects on rheology, hardness and triglyceride composition were studied. The results revealed that up to 5% kokum fat addition by weight of the product did not significantly affect the plastic viscosity or yield stress of milk or dark chocolate. Hardness of both dark and milk chocolate increased with increase in addition of kokum fat. The solids fat content at and above 30 °C increased with increase in level of kokum fat with CB, especially at and above 15%. These physical properties are due to increase in 2‐oleodistearin triglycerides with addition of kokum fat with CB. The results revealed that kokum fat could be used up to 5% by wt of the product to increase the heat‐resistance property of chocolate so that it can be used in warm climates. Copyright © 2004 Society of Chemical Industry     

Comparison of enzyme activities involved in flavour precursor formation in unfermented beans of different cocoa genotypes

The activities of endoprotease, aminopeptidase, carboxypeptidase and invertase (cotyledon and pulp) were studied in unfermented beans of 10 genotype samples with different flavour characteristics (high and low cocoa flavour). Analysis of variance showed that significant differences in enzyme activities exist between certain genotypes. Aminopeptidase and endoprotease activities in beans of the PA7 genotype were higher than in all others. Principal component analysis (PCA) showed that the PA7 genotype (high cocoa flavour) was very different from the UIT1 genotype (low cocoa flavour). Although significant differences exist, no simple and general relationship is established between the flavour potential of a genotype and the level of key enzyme activities in unfermented beans. Carboxypeptidase is of key importance for peptide and free amino acid formation, but differences in enzyme activity could not be correlated to flavour potential of the genotype. It is suggested that the level of enzyme activities present in unfermented beans is not a limiting factor for optimal formation of flavour precursors during the fermentation process. © 2000 Society of Chemical Industry     

Metal contents of some cocoa beans produced in Ondo State,Nigeria

The metal contents in 11 cocoa samples collected from different parts of Ondo State, Nigeria, were determined by the use of atomic absorption spectrophotometry. The moisture and ash contents of the samples were in the range 2.0–3.2% and 2.0–3.8%, respectively, the means being 2.5% and 2.9%, respectively. The mean contents of calcium, magnesium, potassium, sodium, manganese, iron, copper and zinc were 25.5, 5170, 2480, 21, 17, 15.8, 8.2 and 62 μg g^?1, respectively of cocoa bean as purchased. Magnesium was present at much higher levels than the other metals. In some cases the difference between the mean metal content of samples was significant.     

The effects of the organic acids in cocoa on the flavour of chocolate

Samples (54) of dried fermented cocoa beans from different world regions were analysed for levels of organic acids, pH and titratable acidity. The effects of the organic acids on the flavour characteristics of cocoa were examined by sensory evaluation of chocolate made from samples of cocoa beans. Concentrations (g kg^?1) of acids ranged from 1.3 to 11.8 for acetic, 1-6 to 9-9 for citric, 0.6 to 11.1 for lactic and 2.1 to 6.5 for oxalic. pH values ranged from 4.6 to 5.8, while titratable acidity ranged from 0.08 to 0.31 equivalents of sodium hydroxide per kg sample. Cocoas from South East Asia and the South Pacific tended to be more acidic than West African beans in terms of both chemical and sensory characteristics. Lactic and acetic acids were found to be in greater concentrations in cocoas from the former regions and were considered to be largely responsible for higher acid flavour scores. In contrast, citric and oxalic acids were generally lower in these beans. Flavour assessments of cocoas with and without added organic acids indicated that oxalic acid played an important role in chocolate flavour. These results suggest that a reduction in the levels of acetic and lactic acids only, may not be sufficient to produce a desirable flavour balance.     

复配乳化剂对月桂酸型代可可脂巧克力物化性质影响研究

通过制备添加由磷脂、单甘油脂、Span 60两两复配(1∶3/1∶1/3∶1,w/w)月桂酸型代可可脂(CBS)模型巧克力,研究添加复配乳化剂对CBS巧克力物化性质影响。结果表明,复配乳化剂种类和添加量均对CBS巧克力物性产生显著影响;其中,磷脂/Span 60复配添加使巧克力表面WI值增加趋势延缓,具有较好延缓起霜作用。     

制备类可可脂原料用油的研究进展

近年来,巧克力市场逐年壮大,可可脂的需求也随之不断增加。而可可脂产量有限,加之成本较高,可可脂替代品成为学者和商家研究的热点。其中,类可可脂以其特有的优势,成为可可脂替代品中独具特色的一种。对制备类可可脂的原料用油进行了归纳总结,分别从类可可脂的制备方法、原料油脂组成及其相关研究方面对原料油脂进行了概述,从而加深对类可可脂的了解。