Cocoa Butter Equivalents Research Articles

Alternative method for the quantification by gas chromatography triacylglycerol class analysis of cocoa butter equivalent added to chocolate bars.

Directive 2000/36/EC allows chocolate makers to add up to 5% of only six specific cocoa butter equivalents (CBEs) to cocoa butter (CB). A quantification method based on triacylglycerol (TAG) class analysis by gas chromatography with an unpolar column was set up for routine control purposes of chocolate bars. Mixtures of CBEs/CB were elaborated according to a Placket-Burman experiment design and analyzed by gas chromatography. A matrix was built with the normalized values of TAG classes (C50, C52, C54, and C56) of pure CBs of various origins, homemade CB/CBE mixtures (1 CB type), and mixtures containing CBE with CBs of various origins. A multivariate calibration equation was computed from this matrix using a partial least-squares regression technique. CBE addition can be detected at a minimum level of 2%, and the mathematical model allows its quantification with an uncertainty of 2% with respect to the cocoa butter fats. The model has also been applied for deconvolution and quantification of each CBE of a CBE mixture in chocolate bars.

Triacylglycerol analysis for the quantification of cocoa butter equivalents (CBE) in chocolate: feasibility study and validation.

A new European legislation (2000/36/CE) has allowed the use of vegetable fats other than cocoa butter (CB) in chocolate up to a maximum value of 5% in the product. The vegetable fats used in chocolate are designated as cocoa butter replacements and are called cocoa butter equivalents (CBE). The feasibility of CBE quantification in chocolate using triacylglycerol (TAG) profiles was conducted by analyzing 55 samples of CBs and 31 samples of CBEs using a liquid chromatograph equipped with an evaporative light scattering detector (HPLC-ELSD). Statistical evaluation of the data obtained has been performed, and a simulation study has been carried out to assess the viability to use this method for quantifying the amount of CBE in real mixtures and in chocolates. The TAGs POP, POS, PLS, and the ratios POP/PLS, POS/PLP (P, palmityl; O, oleyl; S, stearyl; L, linoleyl) are particularly significant to discriminate between CB and CBE. Analysis of 50 mixtures between 5 different CBEs and 10 different CBs at 2 different concentration levels is presented. The data are visualized and interpreted. A mathematical model has been developed to assess the amount of CBE in real mixtures. This predictive model has been successfully applied and validated on dark chocolates including authorized CBE. The results are affected by +/-2.1% absolute average error. In particular, estimations between 10 and 20% of CBE show a very good match. On the other hand, values equal to or smaller than 5% show a larger prediction error (detection limit of the method). For the main purpose of this method (i.e., quantification of CBE at 5% max in chocolate, which represents about 15% of the total fat) this model shows very good results. For milk chocolate, the mathematical model can also be used if TAG are integrated from partition number (PN) 46 to 54. Consequently, the model proposed provides sufficient information to verify the real application of the European legislation.

Supercritical Reduction of Lauric Acid in Palm Kernel Oil (PKO) to Produce Cocoa Butter Equivalent (CBE) Fat

The extraction of dehulled ground palm kernel using supercritical carbon dioxide (SC-CO 2 ) as a solvent at a temperature of 40°C and pressures range from 34.5 to 48.3 MPa was studied. At 48.3 MPa and 40°C the solubility of oil was about 21.1 g/100 g carbon dioxide, however maintaining constant temperature of 40°C the solubility of oil increased with pressure. The fatty acid composition of oil extracted varied with fraction time. The first fraction had 52.2 percent lauric acid content. The lauric acid content decreased to 34.3 percent in the third fraction with an increase to more than 40 percent fatty acid with 16 carbons or more. The breakdown was 0.1 percent palmitic acid (C16), 8.9 percent stearic acid (C18:0), 20 percent oleic acid (C18:1) and 4 percent linoleic acid (C18:2). Blending of palm oil (PO), palm kernel oil (PKO), stearic acid (C18:0) and oleic acid (C18:1) at different ratios were carried out to obtain blends as cocoa butter replacers (CBR) fat. The lauric and myristic acids were reduced to 9.03 and 4.44 percent in the CBE blend, although there are no lauric and a trace of myristic acid in CB. The percentages of other fatty acids in CBE were found to correlate well to the CB except C18:0 which was only 16.15 percent. The slip melting point (SMP) increased with the increase in C18:0. The value of SMP was found to be 36.1°C for the CBE blend whereas for the CB it is 35°C. The iodine value for the CBE blend was found 38.3 where as for CB it is 36. The solid fat content (SFC) of CBE at 20, 35 and 37.5°C was found to be 59.3, 8.5 and 0 percent, respectively. Whereas, the SFC values of CB are 72, 11 and 0 percent, respectively. Although the percent of the C18:0 in the CBE was lower than the commercial CB but it did not affect to the SMP and SFC.

Accumulation of a cocoa-butter-like lipid by Yarrowia lipolytica cultivated on agro-industrial residues.

Yarrowia lipolytica was cultivated on mixtures of saturated free fatty acids (an industrial derivative of animal fat called stearin), technical glycerol (the main by-product of bio-diesel production facilities), and glucose. The utilization of technical glycerol and stearin as co-substrates resulted in higher lipid synthesis and increased citric acid production than the combination of glucose and stearin. The lipids produced contained significant amounts of stearic acid (50-70%, wt/wt) and lower ones of palmitic (15-20%, wt/wt), oleic (7-20%, wt/wt), and linoleic (2-7%, wt/wt) acid. Single-cell oil having a composition similar to cocoa-butter up to 3.4 g/L was produced, whereas in some cases relatively increased citric acid quantities (up to 14 g/L) were excreted into the growth medium. The microorganism presented a high specificity for lauric, myristic, and palmitic acid, while a discrimination for the stearic acid was observed. As a conclusion, microbial metabolism could be directed by using mixtures of inexpensive saturated fats, glycerol, and glucose as co-substrates, in order to accumulate lipids with predetermined composition, e.g., cocoa-butter equivalents.

Analytical platforms to assess the authenticity of cocoa butter

AbstractThis paper reviews strategies to detect and analyse cocoa butter equivalents added to genuine cocoa butter or to chocolate products. The legal background of the issue is that current European legislation allows the addition of vegetable fats other than cocoa butter to chocolate up to a level of 5% of the product weight, provided that the addition is correctly indicated on the label. However, the Directive fails to specify a method of analysis to enforce compliance.The principal themes highlight compositional data (triglyceride and fatty acid composition, components of the unsaponifiable matter) of cocoa butter from different origins and suitable raw materials used for the formulation of cocoa butter equivalents, and explains how analytical techniques (gas‐liquid chromatography, high‐performance liquid chromatography, infrared spectroscopy, pyrolysis mass spectrometry) make use of subtle differences between the two commodities to detect commingling.

Characterisation of some underutilised vegetable oils and their evaluation as starting materials for lipase-catalysed production of cocoa butter equivalents

Oils were extracted from the inner skin of Strychnos madagascariensis fruit shell, Trichelia emetica seed and Ximenia caffra seed. Gas chromatography analysis of methyl esters of the glycerides of these oils was used to determine the fatty acid profiles of the oils. S. madagascariensis and X. caffra oils were found to contain high oleic acid levels, 68 and 59% respectively, while T. emetica oil was found to contain 53% palmitic acid. Pancreatic lipase digestion was used to perform structural analyses on the triglycerides of oils. Oleic acid was found to be concentrated at the 2-position of the triglycerides of the X. caffra and T. emetica oils. The three oils were evaluated for use as starting materials in lipase-catalysed production of cocoa butter equivalents using transesterification reactions. A 1,3-specific lipase from Rhizomucor miehei was used to incorporate stearic acid and palmitic acid into the 1- and 3-positions of the triglycerides. The products of the transesterifications in the case of X. caffra and S. madagascariensis compare well with those produced using lipase-catalysed transesterifications of other oils.

Rapid MS method for analysis of cocoa butter TAG

AbstractAmmonia negative ion CI‐MS was applied to analyze the M.W. distribution and regioisomeric structure of TAG in cocoa butter and in cocoa butter equivalents. The M.W. distribution results obtained for a reference cocoa butter were consistent with corresponding results obtained in an intercomparison study by chromatographic methods. Minor but statistically significant differences were observed when proportions of the three major M.W. species (52∶1, 54∶1, and 50∶1; acyl carbon number/number of double bonds) in a mixture of nine cocoa butters and in mixtures containing 10 or 20% (w/w) of specific cocoa butter equivalents were compared. Tandem MS was used to determine the regioisomeric structure of the three major TAG M.W. species in cocoa butter and in cocoa butter equivalents. The regioisomeric structure in cocoa butter and in all the equivalents analyzed were nearly identical, oleic acid being located primarily in the sn‐2 position. These results cannot be exploited in detecting added foreign fats in this case. However, the present study shows that useful TAG composition data, which may be used to detect foreign fats in cocoa butter by applying chemometric data evaluation, can be obtained by MS in a significantly shorter time compared to chromatographic methods.

Fourier transform infrared spectroscopy and chemometrics as a tool for the rapid detection of other vegetable fats mixed in cocoa butter

AbstractThe Fourier transform infrared (FTIR) technique in combination with multivariate data evaluation was used to analyze a wide variety of cocoa butters (CB), cocoa butter equivalents (CBE), and mixtures thereof. The sample set consisted of 14 CB (10 pure from various geographical origins and 4 commercial mixtures), 18 CBE (12 mixtures and 6 pure CBE from kokum, illipé, and palm midfraction), and 154 mixtures of CB with CBE at various concentrations (ranging from 5 to 20%). A total of 192 samples were analyzed in triplicate. All CB and CBE were shown to have very characteristic FTIR spectra that gave highly reproducible fingerprints. The main vibrational modes were also elucidated. FTIR can easily be employed to distinguish between pure CB and pure CBE. With prior knowledge of which cocoa butter is present in mixtures, FTIR can be applied to distinguish between CB mixed with CBE at the 10 and 20% levels (corresponding to about 2 and 5% of CBE in chocolate). However, the study revealed that a single “global” statistical model (multilayer perceptron, radial basis functions, or partial least square regression) was not able to predict the precise level of addition. The FTIR approach detailed here shows great potential as a rapid screening method for distinguishing between pure vegetable fats and, we believe, could be extended to investigate mixtures of CB and CBE by the establishment of a database.

Development of a Rapid Method for the Detection of Cocoa Butter Equivalents in Mixtures with Cocoa Butter

A simple and rapid gas chromatographic (GC) method was developed for the detection of cocoa butter equivalents (CBEs) in cocoa buffer (CB). It is based on the use of a 5 m nonpolar capillary column for the separation of the main triglycerides of CB according to their acyl/carbon numbers. The GC procedure was optimized to avoid thermal degradation of the triglycerides. By computing the ratio C54/C50 and (C54/C50) x C52 and by 2-dimensional plotting of these values, authentic CB samples were clearly distinguished from samples containing various CBEs. The detection of little as 1% CBE in CB (corresponding to about 0.3% CBE in chocolate) in a model system was shown to be possible. Under real conditions, for a wide range of CBs, about 2.5% CBEs in CB were detected. With this method, quantitation was possible at a concentration of 5% CBEs in CB mixtures, which corresponds to around 1% in chocolate; this value is far below the maximum level of 5% CBEs allowed to be added to chocolate.

Characterization of cocoa butter and cocoa butter equivalents by bulk and molecular carbon isotope analyses: implications for vegetable fat quantification in chocolate.

The fatty acids from cocoa butters of different origins, varieties, and suppliers and a number of cocoa butter equivalents (Illexao 30-61, Illexao 30-71, Illexao 30-96, Choclin, Coberine, Chocosine-Illipé, Chocosine-Shea, Shokao, Akomax, Akonord, and Ertina) were investigated by bulk stable carbon isotope analysis and compound specific isotope analysis. The interpretation is based on principal component analysis combining the fatty acid concentrations and the bulk and molecular isotopic data. The scatterplot of the two first principal components allowed detection of the addition of vegetable fats to cocoa butters. Enrichment in heavy carbon isotope ((13)C) of the bulk cocoa butter and of the individual fatty acids is related to mixing with other vegetable fats and possibly to thermally or oxidatively induced degradation during processing (e.g., drying and roasting of the cocoa beans or deodorization of the pressed fat) or storage. The feasibility of the analytical approach for authenticity assessment is discussed.

Composition of Genuine Cocoa Butter and Cocoa Butter Equivalents

The compositional variation of the fatty acid, triacylglycerol, tocopherol, tocotrienol and sterene composition of genuine cocoa butter (CB) due to different geographical growing regions and breeding lines as well as other vegetable fats used in confectionery manufacturing (cocoa butter equivalents, CBE) have been investigated. Genuine CB showed quite narrow ranges of compositional variation compared to CBEs. CBEs are prepared by blending different tropical fats to suit various technological requirements of confectionery manufacturers. Consequently, no consistent compositional patterns were observed within the latter group. The fatty acid and the triacylglycerol data presented could be utilized for the detection as well as the quantification of CBEs in plain chocolate, by using appropriate calibration models. However, the minor constituents (tocopherols, -trienols, and sterene data) are of limited use for quantitative purposes, but may be additional indicators for the presence of other vegetable fats in chocolate.

Cocoa butter equivalent through enzymic interesterification of palm oil midfraction

The production of a cocoa butter equivalent (CBE) through enzymic interesterification of palm oil midfraction (POMF) with stearic acid in a solvent free system using Novo lipase Lipozyme™ as a catalyst was analyzed. A two level factorial design was used to study the effect of the initial ratio of stearic acid–POMF, initial humidity of the enzyme preparation and the enzyme–substrate ratio on the yield, mass productivity and specific productivity. Studies were carried out both in batch and in a continuous packed bed reactor. The highest specific productivity obtained in shake flask was 0.0393 g/Batch Interesterification Unit (BIU) h at a stearic acid–POMF ratio of 1.6 and enzyme–substrate ratio of 23 BIU/g. In the continuous packed bed reactor the highest mass productivity observed was 1.54 g/g·h, using an enzymic load of 73 BIU/g. Unreacted fatty acids were separated from the intereresterified products by short path distillation at 0.2 mbar and 140°C obtaining a product practically without free fatty acids. Thermograms of the products obtained by scanning differential calorimetry were similar to cocoa butter (CB), but exhibited several distinct peaks, due presumably to the presence of diglycerides and trisaturated triglycerides.

An improved method for the measurement of added vegetable fats in chocolate

A method for identifying refined vegetable fats added to chocolate (cocoa butter equivalents, CBEs) was combined with established quantitative methods for determining the level of vegetable fat added to cocoa butter with the aim of providing improved precision. The identification of fats was based on the analysis of sterol and triterpene alcohol degradation products formed during the processing of the fat. The procedure was able to successfully discriminate between 95% of pairs of fats from a set (33) of CBE-type vegetable fats. Subsequent analysis of 80 mixtures of four CBEs with chocolate successfully identified, on cross-validation, 94% of the samples. Combining the qualitative procedure with established quantitative methodology, based on the analysis of triacylglycerols, improved the method precision from ± 2.1% to ± 0.3% (5% addition of CBE at 95% confidence). Identifying the fat analytically permits the use of quantitative methods for determining the level of added fat in chocolate that have improved precision in comparison with the measurement of an unidentified fat. This may obviate the need to use factory inspection as a means to identify the ingredients of a product and monitor compliance with proposed legislation.

Quantification of cocoa butter equivalents in mixtures with cocoa butter by chromatographic methods and multivariate data evaluation

Quantification of cocoa butter equivalents in mixtures with cocoa butter by chromatographic methods and multivariate data evaluation

Perfil sensorial de ovos de Páscoa

Ovo de Páscoa é um produto de chocolate na forma de ovo comercializado no Brasil durante a Páscoa. No presente estudo, Análise Descritiva Quantitativa foi aplicada para levantar atributos sensoriais que melhor definem modificações na aparência, aroma, sabor e textura, quando sucedâneos da manteiga de cacau (SUMC) são adicionados ao ovo de Páscoa. Amostras com e sem a adição de SUMC foram avaliadas por uma equipe selecionada de provadores e quatorze atributos sensorial foram definidos. Após um período de treinamento, os provadores avaliaram as amostras através de delineamento de blocos completos balanceadas usando escala não estruturada de 9cm. Os resultados foram analisados através da análise do componente principal, ANOVA e teste de Tukey (p<0,05). As amostras apresentaram diferença significativa (p<0,05) em todos os atributos analisados. Ovo de Páscoa sem SUMC apresentou maior intensidade dos atributos: cor marrom, aroma e sabor característico, aroma e sabor de massa de cacau, aroma de manteiga de cacau, dureza e fraturabilidade.

Production of specific-structured triacylglycerols by lipase-catalyzed reactions: a review

Regiospecificity is one of the major advantages of using lipase technology for the modification of oils and fats to produce high-value added products, such as cocoa butter equivalents, human milk fat substitutes, and other specific-structured lipids. Due to the high cost of biocatalysts, the mainstream applications of lipases for normal oils and fats are still limited. Therefore, positional specificity of lipases has the priority and will be the target property to be exploited for commercial and industrial developments, because no chemical method has such a specificity and is promising or possible for this task. In this paper, encouraging products resulting from this regiospecificity are reviewed together with the critical evaluation of their reaction schemes, side reactions and by-products, sources of substrate oils and acyl donors, and production processes.

Identification of cocoa butter equivalents added to cocoa butter by fatty acid composition of the triacylglycerol sub-fractions separated by Ag + -HPLC - II -

In a previous study the fatty acid compositions of triacylglycerol sub-fractions, separated by Ag+-HPLC, were studied in samples of cocoa butter, cocoa butter equivalents (CBE) and some of their mixtures. In this paper similar research is described which was carried out on three other CBE (Illexao 30–61, Illexao 30–63, Illexao 30–71). The results show the particular significance of the fully saturated sub-fraction, in which notable percentage changes in C16 : 0 and C18 : 0 contents were also detectable in the two mixtures with the lowest CBE content (5% or 15%, w/w) relative to genuine cocoa butter. Multiple regression analysis was used to verify the percentages changes of fatty acid contents in the fully saturated sub-fraction as a function of the percentage of CBE added to genuine cocoa butter.

Detection and quantification of cocoa butter equivalents in chocolate model systems: analysis of triglyceride profiles by high resolution GC

The detection of vegetable fats added to cocoa butters in chocolate formulations was investigated in model mixtures. Cocoa butters varying in origins, crop and treatment were analysed alone and combined at levels of 5, 10, 15 or 20% to a variety of cocoa butter equivalents. Triglyceride profiles were obtained by high resolution GC. The areas of the major triglycerides present were plotted one vs the other. The plots of percentages of specific triglycerides allowed detection of vegetable fats added to cocoa butters in these model systems, and quantification of the addition could be done down to a 5% level on a cocoa butter basis. The detection was most difficult in the case of the addition of illipe fat and the quantification could not be done below 10%. The results suggest that, in chocolate, which contains 20–25% cocoa butter, this detection system works down to 1–2%, and thus well below the proposed 5% regulatory level. ©

Identification of cocoa butter equivalents added to cocoa butter

Separation of the six sub-fractions of triacylglycerols (TAG) from cocoa butter, three cocoa butter equivalents (CBEs: calvetta, choclin and coberine) and their mixtures was carried out by silver ion high-performance liquid chromatography. The percentage fatty acid composition of all these sub-fractions was then measured by high-resolution gas chromatographic analysis of the methyl esters of the constitutive fatty acids. In particular, the percentage quantities of all acids in the fully saturated sub-fraction of pure fats and some of their mixtures were evaluated as a function of the percentage CBEs added to cocoa butter in order to devise multiple regression models. The aim was to obtain statistical tools for the identification and/or quantification of added CBEs. The results obtained show the effectiveness of TAG sub-fraction percentage fatty acid composition data for the purposes of research, as well as the potential use of stereospecific analysis of the same sub-fractions. In fact, the stereospecific analysis of TAG fractions from cocoa butter and CBEs already carried out has enabled observation of the different distributions of various fatty acids in the glycerol backbone of TAG from cocoa butter and the CBEs under study, and this will certainly result in the development of more powerful and discriminating statistical models.

Physical properties of Malaysian cocoa butter as affected by addition of milkfat and cocoa butter equivalent

Commercial samples of Malaysian cocoa butter (MCB), anhydrous milkfat (AMF), high melting fraction milkfat42 (HMF42) and cocoa butter equivalent (CBE) were blended in binary and ternary blends. All the fats were then evaluated for their solid fat content, thermal analyses and polymorphic stability. MCB possessed the highest solid followed by CBE, HMF42 and AMF. Data on thermal analyses showed that eutectic interaction was more noticeable when AMF and HMF42 were present in the MCB and CBE system. Moreover, X-ray diffraction patterns also showed that AMF and HMF42 exhibit the presence of β polymorph while MCB and CBE exhibit β polymorph.