Cocoa Liquor Research Articles

Tocopherols in cocoa butter obtained from cocoa bean roasted in different forms and under various process parameters

Obtaining good quality chocolate strongly depends on raw material, i.e. cocoa beans. The processing of cocoa beans consists of some important steps, including fermentation, drying fermented beans and roasting. Traditionally roasting is performed on whole beans but currently, roasting crushed cocoa beans or cocoa liquor becomes more and more popular. Many biologically active compounds may be found in the cocoa beans, including tocopherols. This work investigates the influence of the constant or variable roasting process parameters (temperature, velocity and relative humidity of roasting air) on the tocopherol concentration in cocoa butter (CB) extracted from cocoa beans originating from Togo and roasted in two different forms, namely as whole and crushed beans. Whole cocoa beans were roasted to a 2% moisture content and crushed cocoa beans were firstly partially dried which further enables easier dehusking, then ground, dehusked and roasted until their humidity decreased to around 2%. Roasting resulted in lowering the content of individual tocopherols in analyzed material. The degree of degradation of tocopherols in CBs was different, depending on the form of roasted beans from which these CBs were extracted. Higher concentrations were determined in CBs extracted from beans roasted in the form of crushed samples comparing to CBs obtained from beans roasted as a whole. The study investigates different roasting parameters of crushed beans, none of which drastically lowered the concentration of tocopherols in extracted CBs. Their concentration in CBs extracted from whole beans was, on the other hand, influenced by roasting air parameters. In case of whole beans roasted under constant parameters, application of 150°C proved to be more favorable than roasting at 135°C, as well as application of “dry” air and 1m/s roasting air velocity. Discussing the variable roasting process parameters, in case of applied roasting temperature it is more favorable to change it from 150°C to 135°C, than the other way round. Changing the relative humidity of roasting air from 5 to 0.3%, lower degradation of tocopherols in CB occurs when the process is conducted at 150°C. It may be further concluded that a direct dependence between the velocity of roasting air varied during the process and the concentration of tocopherols in extracted CB may not be indicated. In conclusion, it is stated that the temperature of the air applied during the roasting process has the decisive influence on the tocopherol content in CBs extracted from cocoa beans subjected to the process.

Total synthesis of (−)-bis-8,8′-catechinylmethane isolated from cocoa liquor

An efficient synthesis of bis-8,8′-catechinylmethane, a dimeric flavanol linked through a methylene bridge, is described. Our strategy involved a regioselective coupling via a trifluoroacetic anhydride condensation reaction followed by ketone reduction to the methylene employing new conditions (lithium aluminum hydride and hexafluorophosphoric acid).

Semiquantitative Determination of Mesophilic, Aerobic Microorganisms in Cocoa Products Using the Soleris® NF-TVC Method

The Soleris Non-fermenting Total Viable Count method was previously validated for a wide variety of food products, including cocoa powder. A matrix extension study was conducted to validate the method for use with cocoa butter and cocoa liquor. Test samples included naturally contaminated cocoa liquor and cocoa butter inoculated with natural microbial flora derived from cocoa liquor. A probability of detection statistical model was used to compare Soleris results at multiple test thresholds (dilutions) with aerobic plate counts determined using the AOAC Official Method 966.23 dilution plating method. Results of the two methods were not statistically different at any dilution level in any of the three trials conducted. The Soleris method offers the advantage of results within 24 h, compared to the 48 h required by standard dilution plating methods.

Effects of polar cosolvents on cocoa butter extraction using supercritical carbon dioxide

Cocoa butter was successfully extracted from cocoa liquor by supercritical carbon dioxide (SC–CO2) at 35 MPa, 60 °C and 2 mL/min with 5%, 15% and 25% cosolvents. The extraction yield of triglycerides (TG) and fatty acid (FA) compositions were significantly influenced by the concentration of polar cosolvents. The SC–CO2 extraction efficiency was increased with cosolvent significantly. Ethanol was found to be the best cosolvent for cocoa butter extraction using SC–CO2 followed by isopropanol and acetone. The triglycerides of 1,3-dipalmitoyl-2-oleoylglycerol (POP), 1-palmitoyl-2-oleoyl-3-stearoyl-glycerol (POS) and 1,3-distearoyl-2-oleoyl-glycerol (SOS) were contained in the extracted cocoa butter with POS being the major component. Where palmitic, stearic and oleic were the main fatty acids in the cocoa butter samples, with stearic being the highest component. The lower molecular weight (MW) of TGs and FAs showed the higher selectivity compared to the high MW of TGs and FAs. Thus, they were fractionated during the first stage of SC–CO2 process. The cocoa butter was successfully extracted from cocoa liquor by SC–CO2 at 35 MPa, 60 °C and 2 mL/min using different concentrations of polar cosolvents (ethanol, isopropanol and acetone). The extraction yield was significantly (p < 0.05) influenced by the concentration of polar cosolvents. Similarly, polar cosolvent concentration had significant (p < 0.05) effects on the TG and FA compositions. Ethanol was found to be the most efficient polar cosolvent for cocoa butter extraction compared to isopropanol and acetone. POS (42.2–45.9%) being the major triglycerdies component, followed by SOS (27.6–31.4%) and POP (20.3–22.7). Palmitic, stearic and oleic acids were the main fatty acids in the extracted cocoa butter, with stearic being the highest (34.9-37.8%), followed by oleic (30.3-31.8%) and palmitic (28.3-30.0%) acids, respectively. The choice of modifiers becomes a great challenge and ethanol was shown to be the best polar cosolvent, and it enhanced the solubility during the cocoa butter extraction by SC–CO2. This method can be feasibly implemented in the cocoa industry for the production of high quality cocoa butter.

Flavanol and Procyanidin Content (by Degree of Polymerization 1–10) of Chocolate, Cocoa Liquors, Cocoa Powders, and Cocoa Extracts: First Action 2012.24

An international collaborative study was conducted on an HPLC method with fluorescent detection for the determination of flavanols and procyanidins in chocolate and cocoa-containing materials. The sum of the oligomeric fractions with degree of polymerization 1-10 was the determined content value. Sample materials included dark and milk chocolates, cocoa powder, cocoa liquors, and cocoa extracts. The content ranged from approximately 2 to 500 mg/g (defatted basis). Thirteen laboratories--representing commercial, industrial, and academic institutions in six countries--participated in this interlaboratory study. Fourteen samples were sent as blind duplicates to the collaborators. Results for 12 laboratories yielded repeatability RSD (RSDr) values below 10% for all materials analyzed, ranging from 4.17 to 9.61%. Reproducibility RSD (RSDR) values ranged from 5.03 to 12.9% for samples containing 8.07 to 484.7 mg/g material analyzed. In one sample containing a low content of flavanols and procyanidins (approximately 2 mg/g), the RSDR was 17.68%.

Analysis of cocoa products for ochratoxin A and aflatoxins

Eighty-five samples of cocoa products sampled in Canada were analysed for ochratoxin A (OTA) and aflatoxins in 2011–2012. Inclusion of the aflatoxins in this survey required additional method development. Chocolate was extracted with methanol–water plus NaCl, while for cocoa two successive extractions with methanol and methanol–water were made. Extracts were cleaned on an AflaOchra immunoaffinity column (IAC). Determination was by reversed phase high performance liquid chromatography (HPLC). Detection of the aflatoxins was with a post-column photochemical reactor and of OTA by fluorescence detection. Mean limits of quantification (LOQ) of chocolate and cocoa powders were 0.16 ng/g (OTA) and 0.07 ng/g (aflatoxin B1), respectively. Survey results showed that the incidences of OTA above the LOQ in natural cocoa were 15/15 (mean 1.17 ng/g), 20/21 for alkalized cocoa (mean 1.06 ng/g), 9/9 for baking chocolate (mean 0.49 ng/g), 20/20 for dark chocolate (mean 0.39 ng/g), 7/10 for milk chocolate (mean 0.19 ng/g), 5/5 for cocoa liquor (mean 0.43 ng/g), and 0/5 for cocoa butter. These results confirm our previous work with OTA. In the same samples, incidences of aflatoxin B1 above the LOQ were 14/15 for natural cocoa (mean 0.86 ng/g), 20/21 for alkalized cocoa (mean 0.37 ng/g), 7/9 for baking chocolate (mean 0.22 ng/g), 16/20 for dark chocolate (mean 0.19 ng/g), 7/10 for milk chocolate (mean 0.09 ng/g), 4/5 for cocoa liquor (mean 0.43 ng/g), and 0/5 for cocoa butter. Both aflatoxins and OTA were confirmed by HPLC-MS/MS when OTA or aflatoxin levels found were above 2 ng/g in cocoa.

Chemical composition of cupuassu (Theobroma grandiflorum) and cocoa (Theobroma cacao) liquors and their effects on streptozotocin-induced diabetic rats

Cupuassu (Theobroma grandiflorum Willd. Ex Spreng) Shum. is a Brazilian Amazon rainforest fruit, phylogenetically close to cocoa (Theobroma cacao L.), with excellent flavor and high agro-economic potential, whose seeds can also be used to prepare a product similar to chocolate. Here, we compared the chemical composition and in vitro antioxidant capacity of cupuassu and cocoa liquors. In addition, we evaluated the effect of the daily intake of cupuassu and cocoa liquors (3.6 and 7.2g/kg body weight) on oxidative stress and lipid profile in streptozotocin (STZ)-induced diabetic rats. Although cocoa liquor presented higher polyphenol, caffeine contents and in vitro antioxidant capacity, saturated fatty acid contents were also higher than in cupuassu liquor. Daily administration of both cocoa and cupuassu liquors resulted in ameliorated body weight gain and liver weight/body weight ratio in STZ-diabetic rats. Plasma triacylglycerol levels were strongly reduced by both doses of liquors, while HDL-c was raised in 27% when the highest doses of cupuassu and cocoa were administrated. Plasma antioxidant capacity was improved, and lipid peroxidation in plasma and tissues decreased in a dose-dependent manner. In spite of the lower polyphenol content of cupuassu, these results indicate advantages of cupuassu compared to cocoa in relation to potential health benefits.

Evaluation of Corrosion Behaviour of Grey Cast Iron and Low Alloy Steel in Cocoa Liquor and Well Water

Corrosion behaviour of cast iron and low alloy steel in cocoa liquor and well water was investigated. The average weight losses of the specimens were measured using digital weighing balance. The results showed that the weight losses of both cast iron and low alloy steel in both media increases with time. Corrosion rate of cast iron in cocoa liquor increases rapidly with time for up to 336 hours (1000 μm/yr), but in well water the rapid rate of corrosion only lasted up to 187 hours (1160 μm/yr) thereafter it continuously dropping until 264 hours (667 μm/yr) after which it remains constant. Low alloy steel corroded faster in cocoa liquor up to 264 hours (200 μm/yr), whereas the initial rapid corrosion rate only lasted up to 168 hours (180 μm/yr) in well water environment. The results revealed that low alloy steel exhibited better corrosion resistance in both media, with cocoa liquor been more aggressive. Thus, low alloy steel will be a better material for piping and pumping system in cocoa processing industries.

Effects of Moisture and pH on Supercritical Fluid Extraction of Cocoa Butter

The effects of moisture content and alkali treatment on cocoa butter extraction using supercritical fluid were studied. Ground cocoa nibs were examined at moisture contents of 1.95 %, 3.91 %, 5.87 %, 9.79 % and 17.64 % and the pH of the cocoa liquor was maintained at 5.0–5.9, 6.8–7.2 and 7.5–7.9. Cocoa butter was successfully extracted, using supercritical carbon dioxide (SC-CO2) for the moisture content study and SC-CO2 with 25 % ethanol as a cosolvent for the pH-level alkali treatment study, at 35 MPa, 60 °C and 2 ml/min. The results showed that increases in moisture content and pH level significantly (p < 0.05) increase the yield efficiency. A moisture content of 9.79 % and pH-alkali treatment at 7.5–7.9 produced the highest yield (60.36 % and 73.70 % at 20 and 18 h extraction time, respectively). Triglycerides (TG) and fatty acids (FAs) were similar to those found in cocoa butter obtained using the Soxhlet method. Saturated and short-chain TG and FA constituents were more soluble than unsaturated and long-chain constituents.

Thermal, structural and rheological characteristics of dark chocolate with different compositions

The effect of different cocoa composition of dark chocolate samples and their ingredients on their thermal, structural and rheological characteristics was investigated. Thermal behavior was evaluated by means of differential scanning calorimetry (DSC), while the crystal morphology was observed by polarized light optical microscopy. The rheological measurements were carried out using both continuous and oscillatory experiments. The formation of more stable polymorphic structures was time and temperature dependent; and it was not affected by either cocoa composition or particle sizes. The kinetics of crystallization was determined by a step crystallization method and modeled by the Avrami equation, it was accelerated by solid particles concentration, lower particle size and lower crystallization temperatures. Negative spherulites with featherlike microstructures defined the time dependent crystal growth and were consistent with Avrami indexes of 3–4. Under continuous shear, cocoa butter was well described by the Bingham rheological model, while Casson and Carreau equations modeled the flow of cocoa liquor and chocolate samples. However the Carreau model was preferred for presenting better fittings and for predicting apparent viscosities at low and at high shear rates. From both, continuous and oscillatory experiments, it was found that composition of chocolate samples in terms of fat and nonfat cocoa solids, and sugar content, affected their rheological behavior. The solid liquid transition of chocolate samples and cocoa liquor was obtained at a yield stress of around 1Pa from both continuous and oscillatory shear experiments.

Characterization of cocoa liquors by GC–MS and LC–MS/MS: focus on alkylpyrazines and flavanols

Flavor is one of the most important characteristics of chocolate products and is due to a complex volatile fraction, depending both on the cocoa bean genotype and the several processes occurring during chocolate production (fermentation, drying, roasting and conching). Alkylpyrazines are among the most studied volatiles, being one of the main classes of odorant compounds in cocoa products. In this work, a mass spectrometric approach was used for the comparison of cocoa liquors from different countries. A headspace solid-phase microextraction gas chromatography-mass spectrometry method was developed for the qualitative study of the volatile fraction; the standard addition method was then used for the quantitative determination of five pyrazines (2-methylpyrazine, 2,3-dimethylpyrazine, 2,5-dimethylpyrazine, 2,3,5-trimethylpyrazine and tetramethylpyrazine). Satisfactory figures of merit were obtained: Limits of quantitation were in the range 0.1-2.7 ng/g; repeatability and reproducibility varied between 3% and 7% and between 8% and 14%, respectively. The total content of the pyrazines was remarkably different in the considered samples, ranging from 99 to 708 ng/g. Tetramethylpyrazine showed the highest concentration in all samples, with a maximum value of 585 ng/g. A preliminary study was also performed on the nonvolatile fraction using LC-MS/MS, identifying some flavanols such as catechin, epicatechin and procyanidins.

Determination of Flavanol and Procyanidin (by Degree of Polymerization 1–10) Content of Chocolate, Cocoa Liquors, Powder(s), and Cocoa Flavanol Extracts by Normal Phase High-Performance Liquid Chromatography: Collaborative Study

An international collaborative study was conducted on an HPLC method with fluorescent detection (FLD) for the determination of flavanols and procyanidins in materials containing chocolate and cocoa. The sum of the oligomeric fractions with degree of polymerization 1-10 was the determined content value. Sample materials included dark and milk chocolates, cocoa powder, cocoa liquors, and cocoa extracts. The content ranged from approximately 2 to 500 mg/g (defatted basis). Thirteen laboratories representing commercial, industrial, and academic institutions in six countries participated in the study. Fourteen samples were sent as blind duplicates to the collaborators. Results from 12 laboratories yielded repeatability relative standard deviation (RSDr) values that were below 10% for all materials analyzed, ranging from 4.17 to 9.61%. The reproducibility relative standard deviation (RSD(R)) values ranged from 5.03 to 12.9% for samples containing 8.07 to 484.7 mg/g. In one sample containing a low content of flavanols and procyanidins (approximately 2 mg/g), the RSD(R) was 17.68%. Based on these results, the method is recommended for Official First Action for the determination of flavanols and procyanidins in chocolate, cocoa liquors, powder(s), and cocoa extracts.

Exacerbation of facial acne vulgaris after consuming pure chocolate

To the Editor: The effect of diet, specifically chocolate, on acne exacerbation is controversial. 1 Wolf R. Matz H. Orion E. Acne and diet. Clin Dermatol. 2004; 22: 387-393 Abstract Full Text Full Text PDF PubMed Scopus (41) Google Scholar Two studies suggested that chocolate does not cause an exacerbation of acne. Fulton et al 2 Fulton Jr., J.E. Plewig G. Kligman A.M. Effect of chocolate on acne vulgaris. J Am Acad Dermatol. 1969; 210: 2071-2074 Google Scholar had subjects consume a chocolate bar containing 10 times the amount of cocoa as a normal chocolate candy bar, or a placebo bar without chocolate liquor or cocoa butter. No significant difference in lesion count, sebum production, or sebum composition between groups existed. 2 Fulton Jr., J.E. Plewig G. Kligman A.M. Effect of chocolate on acne vulgaris. J Am Acad Dermatol. 1969; 210: 2071-2074 Google Scholar Similarly, Anderson 3 Anderson P.C. Foods as the cause of acne. Am Fam Physician. 1971; 3: 102-103 PubMed Google Scholar had subjects consume six servings of 39-gram chocolate bars for 7 days. A limited relationship between chocolate and acne was determined; only one-third of subjects developed new lesions. 3 Anderson P.C. Foods as the cause of acne. Am Fam Physician. 1971; 3: 102-103 PubMed Google Scholar

Phenolic compounds, flavonoids and antioxidant activity in different cocoa samples from organic and conventional cultivation

Purpose – Due to the importance attributed to the phenolic compounds present in cocoa samples for their beneficial effects on health, the purpose of this paper was to analyze four samples of organically and conventionally cultivated cocoa from the south area of Bahia for their composition of phenolics, flavonoids and their antioxidant activity.Design/methodology/approach – Non‐fermented beans, fermented beans, roasted nibs and cocoa liquor were analyzed using spectrophotometry.Findings – In general, the samples that contained a higher level of phenolics and flavonoids were the roasted nibs and the non‐fermented samples in both cultivation systems. The fermented beans and the liquor contained a lower level.Practical implications – The relationship between the concentration of total phenols and the capacity to “mop up” free radicals from the cocoa extracts appears to be highly significant. The extracts with a higher concentration of phenols also show higher antioxidant activity (non‐fermented beans extracts...

Optimisation of chocolate formulation using dehydrated peanut–cowpea milk to replace dairy milk

The rheological properties of chocolate, based upon its acceptability by consumers, are determined largely by the ingredients and their proportions used in the formulations. Milk chocolates are very popular because milk provides flavour and smooth texture to the product. This study aimed to determine the optimal ingredient formulation for vegetable milk chocolate using peanut-cowpea milk as a substitute for dairy milk. The study followed a four-component constrained mixture design, with cocoa liquor, vegetable milk, cocoa butter and sugar as the components. Lecithin and vanillin were added at a constant amount to all formulations. Critical attributes of the chocolates were evaluated using descriptive sensory tests and instrumental techniques. Regression models were fitted to the data, and the optimum ingredient formulation for acceptable vegetable milk chocolate was determined. The vegetable milk had significant (P = 0.05) influence on flavour, mouth feel, hardness and after taste of chocolates. The optimum ingredient formulation for acceptable vegetable milk chocolates was determined to be cocoa liquor (18.00%), sugar (30.75%), peanut-cowpea milk (28.93%), and cocoa butter (22.32%). The results demonstrate that it is feasible to use vegetable source milk for chocolate. The findings also provide clues for scale-up criteria for large-scale production of vegetable milk chocolate.

Bioactive and Sensory Properties of Herbal Spirit Enriched with Cocoa (Theobroma cacao L.) Polyphenolics

In the Southeast European and Mediterranean regions, consumption of various alcoholic beverages in the form of plain and aromatized spirits and liqueurs is an important part of traditional local gastronomy. Especially herbal spirits and liquors are widely spread and consumed. The objective of this study was to evaluate the potential of using cocoa raw materials (cocoa beans, cocoa nibs or cocoa liquor) in order to enhance the bioactive profile of traditional herbal liqueur, and to produce an appealing cocoa-flavoured alcoholic beverage. Therefore, the polyphenolic profile, antioxidative and sensory properties of herbal liquors containing the macerated cocoa raw materials were monitored during 6 months, in order to evaluate the kinetics of cocoa polyphenols release and optimise the maceration procedure for obtaining the best cocoa-enriched herbal spirit. Compared to plain herbal liquor without the additions, the obtained herbal liqueurs exhibited significantly higher content of polyphenolic antioxidants. Among the used cocoa raw materials for the maceration, cocoa nibs were evaluated as the best source of polyphenolic compounds (herbal liqueur containing cocoa nibs, 3,081.82 mg GAE/L), followed by cocoa liquor (1,897.73 mg GAE/L) and cocoa beans (1,358.64 mg GAE/L). Prolonged maceration time (up to 6 months) resulted with an increase in the content of polyphenolic compounds and the antioxidant capacity of herbal liqueurs. Herbal liqueurs containing the cocoa raw materials of bigger particle size (cocoa beans and cocoa nibs) exhibited better sensory properties (overall acceptability and chocolate-like flavour) than herbal liqueur containing cocoa liquor.

Nutritional Aspects of Second Generation Soy Foods

Samples of 15 second generation soy-based products (n = 3), commercially available, were analyzed for their protein and isoflavone contents and in vitro antioxidant activity, by means of the Folin-Ciocalteu reducing ability, DPPH radical scavenging capacity, and oxygen radical absorbance capacity. Isoflavone identification and quantification were performed by high-performance liquid chromatography. Products containing soy and/or soy-based ingredients represent important sources of protein in addition to the low fat amounts. However, a large variation in isoflavone content and in vitro antioxidant capacity was observed. The isoflavone content varied from 2.4 to 18.1 mg/100 g (FW), and soy kibe and soy sausage presented the highest amounts. Chocolate had the highest antioxidant capacity, but this fact was probably associated with the addition of cocoa liquor, a well-known source of polyphenolics. This study showed that the soy-based foods do not present a significant content of isoflavones when compared with the grain, and their in vitro antioxidant capacity is not related with these compounds but rather to the presence of other phenolics and synthetic antioxidants, such as sodium erythorbate. However, they may represent alternative sources and provide soy protein, isoflavones, and vegetable fat for those who are not ready to eat traditional soy foods.

Regular Dark Chocolate Consumption’s Reduction of Oxidative Stress and Increase of Free-Fatty-Acid Mobilization in Response to Prolonged Cycling

This study investigated the effects of regular consumption of dark chocolate (DC), rich in cocoa polyphenols, on plasma metabolites, hormones, and markers of oxidative stress after prolonged exhaustive exercise. Twenty active men cycled at 60% maximal oxygen uptake (VO2max) for 1.5 hr, with the intensity increased to 90% VO2max for a 30-s period every 10 min, followed by a ride to exhaustion at 90% VO2max. In the 2 wk before exercise participants consumed 40 g of DC or an isocarbohydrate-fat control cocoa liquor-free chocolate (CON) twice daily and once 2 hr before exercise in a randomized, counterbalanced, crossover design. Venous blood samples were taken immediately before exercise, postexercise (fixed duration), postexhaustion, and after 1 hr of recovery. F2-isoprostanes were significantly lower (post hoc tests: p < .001) at exhaustion and after 1 hr of recovery with DC. Oxidized low-density lipoproteins were significantly lower with DC (p < .001) both before and after exercise and at exhaustion. DC was also associated with ~21% greater rises in free fatty acids during exercise (main effect: p < .05). Changes in circulating glucose, insulin, glucagon, cortisol, and interleukin (IL)-6, IL-10, and IL-1ra were unaffected by treatment. Time to exhaustion at 90% VO2max was not significantly different between trials (398 ± 204 and 374 ± 194 s for DC and CON, respectively). These results suggest that regular DC intake is associated with reduced oxidative-stress markers and increased mobilization of free fatty acids after exercise but has no observed effect on exercise performance.

High cocoa polyphenol rich chocolate may reduce the burden of the symptoms in chronic fatigue syndrome

BackgroundChocolate is rich in flavonoids that have been shown to be of benefit in disparate conditions including cardiovascular disease and cancer. The effect of polyphenol rich chocolate in subjects with chronic fatigue syndrome (CFS) has not been studied previously.MethodsWe conducted a double blinded, randomised, clinical pilot crossover study comparing high cocoa liquor/polyphenol rich chocolate (HCL/PR) in comparison to simulated iso-calorific chocolate (cocoa liquor free/low polyphenols(CLF/LP)) on fatigue and residual function in subjects with chronic fatigue syndrome. Subjects with CFS having severe fatigue of at least 10 out of 11 on the Chalder Fatigue Scale were enrolled. Subjects had either 8 weeks of intervention in the form of HCL/PR or CLF/LP, with a 2 week wash out period followed by 8 weeks of intervention with the other chocolate.ResultsTen subjects were enrolled in the study. The Chalder Fatigue Scale score improved significantly after 8 weeks of the HCL/PR chocolate arm [median (range) Exact Sig. (2-tailed)] [33 (25 - 38) vs. 21.5 (6 - 35) 0.01], but that deteriorated significantly when subjects were given simulated iso-calorific chocolate (CLF/CP) [ 28.5 (17 - 20) vs. 34.5 (13-26) 0.03]. The residual function, as assessed by the London Handicap scale, also improved significantly after the HCL/PR arm [0.49 (0.33 - 0.62) vs. 0.64 (0.44 - 0.83) 0.01] and deteriorated after iso-calorific chocolate [00.44 (0.43 - 0.68) vs. 0.36 (0.33 - 0.62)0.03]. Likewise the Hospital Anxiety and Depression score also improved after the HCL/PR arm, but deteriorated after CLF/CP. Mean weight remained unchanged throughout the trial.ConclusionThis study suggests that HCL/PR chocolate may improve symptoms in subjects with chronic fatigue syndrome.

Stilbenic Profile of Cocoa Liquors from Different Origins Determined by RP-HPLC-APCI(+)-MS/MS. Detection of a New Resveratrol Hexoside

trans-Resveratrol and trans-piceid were recently discovered in chocolate. In the present work, both were quantified by RP-HPLC-APCI(+)-MS/MS in 22 cocoa liquors from 11 different countries. A very large range of concentrations was observed for trans-piceid. The most concentrated sample (Arriba 06) reached 0.4 and 2.6 mg/kg of trans-resveratrol and trans-piceid, respectively, but in other cultivars stilbene levels were five times lower. Neither cis-resveratrol nor cis-piceid was found in cocoa liquors. An unknown compound eluting 0.5 min before trans-piceid and present at concentrations up to 0.8 mg/kg of trans-piceid equivalents in cocoa liquors was tentatively identified by HRMS as a trans-piceid-like hexoside.