Cocoa Butter Equivalents Research Articles

Depiction of 2-oleic acid-1,3-saturated fatty acid triacylglycerols intended for cocoa butter equivalents from Rhodosporidium toruloides

Cocoa butter equivalent (CBE) serves as a viable substitute for cocoa butter (CB) in the confectionary industry. In this study, four types of CBE were produced via fermentation of Rhodosporidium toruloides and systematically characterized. Fatty acid analysis revealed similarities in both total and sn-2 fatty acid composition compared to cocoa butter. Specifically, palmitic acid, stearic acid and oleic acid constituted 87 % to 91 % of the total fatty acid, with palmitic acid predominating at the sn-2 position (80 %). Analysis of triacylglycerol species indicated that the combined proportion of POP, POSt, and StOSt was lower (50 %) than that found in cocoa butter. Solid fat content analysis showed that CBE were soft than cocoa butter. X-ray analysis confirmed that both CBE and CB predominantly exhibited the desired β crystal form. Additionally, spherical-shaped crystals were observed in CBE during polarized light microscopy analysis.

Development of a novel cocoa butter equivalent from mahua oil - palm stearin blend through two-stage solvent fractionation

Cocoa butter (CB) plays a pivotal role in confectionery, facing escalating demand and associated challenges, necessitating alternatives. A novel cocoa butter equivalent (CBE) was developed from mahua oil (Madhuca longifolia) through two-stage solvent fractionation. Neutralized mahua oil first underwent acetone fractionation at 4°C for 4 h. The resulting stearin was blended with 5% palm stearin before undergoing a second acetone fractionation at 4 °C for 3 h. Fractionation increased the symmetrical triacylglycerols by 35% in mahua CBE, comprising 19% POP, 35% POS, and 22% SOS (76%), meeting regulatory requirements. The melting range of CBE fell within the CB range where peak melting temperature shifted from 2 to 18 °C, ensuring close resemblance. CBE blended with Malaysian CB across varying proportions (10–90%) demonstrated up to 40% full compatibility with CB regarding solid fat content, crystalline structure (β2 crystals), and thermal profile. Mahua CBE offers a commercially viable and globally utilized substitute for exotic fat-based CBEs, notably beneficial in confectionery. Thus, mahua oil's transition from household to commercial applications enhances its overall value proposition, delivering versatility and economic benefits across sectors.

Development of symmetric monounsaturated triacylglycerols by Rhodosporidium toruloides cell factory for cocoa butter equivalent

AbstractIn the present study, various factors including carbon source, nitrogen source, Mg2+, and pH value were investigated to determine the optimal conditions for producing cocoa butter equivalent using R. toruloides. The maximum of biomass, lipid content, and lipid production were achieved at levels of 7.8 g/L, 61.0%, and 4.4 g/L, respectively. The oil from R. toruloides mainly consisted of 16:0, 18:0, and 18:1 with 18:1 dominating at the sn‐2 position, which was further confirmed by 13C NMR analysis. UPLC‐MS analysis showed that the sum of sn‐POP, sn‐POS, and sn‐SOS accounted for up to 27.11% in the triacylglycerol species of the oil produced, which allows those triacylglycerols to be further separated and isolated from other triacylglycerols and used for cocoa butter equivalent. Notably, the production of cocoa butter equivalent reached 1.13 g/L, demonstrating the scientific feasibility of producing CBE through fermentation. These findings indicate a promising future for the fermentation‐based production of cocoa butter equivalent.

A synchrotron X-ray scattering study of the crystallization behavior of mixtures of confectionary triacylglycerides: Effect of chemical composition and shear on polymorphism and kinetics

Cocoa butter equivalents (CBE) are mixtures of triglycerides from multiple sources (e.g., sunflower oil, mango kernel and sal), which resemble cocoa butter (CB) in both physical and chemical properties. Despite being widely used to replace CB in chocolate products, the crystallization behavior of many CBEs is still poorly understood. The aim of this work was to develop a fundamental understanding, at the molecular level, of the crystallization behavior of selected CBEs, and compare it with that of CB. Chromatography was used to determine the composition of CBEs, in terms of fatty acids and triacylglycerides (TAGs), while their thermodynamic behavior and crystallization kinetics were studied using polarized microscopy, differential calorimetry and three different synchrotron X-ray scattering setups. CBEs of different origin and chemical composition (e.g., different ratios of the main CB TAGs, namely POP, SOS and POS) crystallized in different polymorphs and with different kinetics of nucleation, growth and polymorphic transformation. SOS rich CBEs presented showed more polymorphs than CB and POP rich samples; whereas, CBEs with high concentration of POP showed slow kinetic of polymorphic transformation towards the stable β(3L) form.Additionally, it was observed that the presence of small amounts (<1% w/w) of specific TAGs, such as OOO, PPP or SSS, could significantly affect the crystallization behavior of CBEs and CBs in terms of kinetics of polymorphic transformation and number of phases detected (multiple high melting β(2L) polymorphs were identified in all samples studied). Finally, it was found that, regardless of the CBE composition, the presence of shear could promote the formation of stable β polymorphs over metastable β’ and γ forms, and reduced the size of the crystal agglomerates formed due to increased secondary nucleation.

Crystallization and polymorphic behaviors of cocoa butter alternatives: A review

AbstractAlthough cocoa butter (CB) has remarkable physical properties, its high price, growing difficulties, and increased consumption have been the main incentive to explore alternatives to replace or improve it. The potential of fats systems obtained from tropical butters, mixtures of them with vegetable oils, or marine fats as cocoa butter equivalent (CBE), extender (CBEx), substitute (CBS), replacer (CBR), or improver (CBI) have been deeply investigated and their physical chemical properties have been compared to those of CB. The TAGs composition of fats systems is a key factor that determines fats crystallization and polymorphic behaviors, and the suitability for an application. Fats with high concentrations of the TAGs StOSt, StOA, or StOB and low concentrations of POP compared to CB show some incompatibility with CB as analyzed by iso‐solid diagrams and a more complex polymorphic behavior. The presence of low melting TAGs such as POO, StOO, and AOO also leads to significant differences in physical behavior compared to CB. In those fats systems, co‐crystallization and polymorphic transitions of co‐existing solid solutions were reported. A few of the studied fat systems may behave as CBE. However, most of them have potential as CBS, CBR, CBEx or CBI in confectionery products. Studies reported the relevance of fractionation and interesterification processes to modify TAGs composition and the need of finding the right processing conditions and additives to extend fats applications.

Physical Blending of Fractionated Bambangan Kernel Fat Stearin and Palm Oil Mid-Fraction to Formulate Cocoa Butter Equivalent.

In this study, the physicochemical properties, composition, thermal properties, and crystal microstructure of fractionated bambangan kernel fat stearin and palm oil mid-fraction blends were investigated with respect to a potential cocoa butter equivalent. The blends were prepared in five ratios, and all of the blends exhibited similar physicochemical properties to cocoa butter. Although all of the blends had similar physicochemical properties, the blend containing 70% bambangan kernel fat stearin and 30% palm oil mid-fraction showed remarkable similarity to cocoa butter. The blend had similar fatty acid and triacylglycerol content to cocoa butter with 18.74% palmitic acids, 38.26% stearic acids, 34.05% oleic acids, 15.20% 1,3-dipalmitoyl-2-oleoyl glycerol, and 29.74% 1,3-disteroyl-2-oleoyl glycerol with improved thermostability (high solid fat content at 30 °C but reaching 0% at 40 °C). It also exhibited spherulite crystals with a needle-like crystal structure of 50 µm. This mixture showed good compatibility with cocoa butter at all mixing ratios; hence, it is suggested as a potential cocoa butter equivalent.

Effect of tempering and cocoa butter equivalents on crystallization kinetics, polymorphism, melting, and physical properties of dark chocolates

The high price and scarcity of cocoa butter (CB) motivate the search for new cocoa butter equivalents (CBE) for manufacturing dark chocolates. Using CBEs in combination with tempering regimens is an option to produce fine chocolates. Thus, CBEs' thermal properties and crystallization kinetics were studied, and the effect of CBE, and tempering regimes on the crystallization kinetics, polymorphism, melting, and physical properties of dark chocolates were determined. The CBE consisted of CB mixtures with coconut oil (CB/CNO) and sacha inchi oil (CB/SIO) in concentrations of 55/45 and 65/35, respectively. CBEs combined with two tempering regimes (high and low) were used to produce Ch-CNO and Ch-SIO dark chocolates, whose thermal data were obtained by Differential Scanning Calorimetry. The adjustment of thermal data to the Avrami equation permitted us to determine that both factors accelerated the crystallization process up to 7.25 ± 0.18 min in Ch-CNO and 7.08 ± 0.15 min in Ch-SIO and produced spherical and plate-like geometries crystallized nuclei. Also, they generated 90.3 ± 1.18 of form β2 in Ch-CNO and 92.56 ± 1.49 in Ch-SIO. Moreover, the chocolates' texture, rheology, color, and melting properties were within acceptable limits. Therefore, using coconut oil and sacha inchi oil as an alternative to traditional CBEs in dark chocolates is feasible.

Novel application of 1H NMR spectroscopy coupled with chemometrics for the authentication of dark chocolate

The applicability of 1H NMR spectroscopy coupled with chemometric in the quality control of dark chocolate was investigated for the first time to detect cocoa-butter equivalents (CBEs) above the allowed limit by European regulation. Blends of chocolate-fats with CBEs in the range 0–50 % were prepared and analyzed by 1H NMR spectroscopy. Datasets composed of peaks’ areas or spectral variables (fingerprinting) in glycerol region were tested for the creation of multivariate statistical models. Partial least-squares discriminant analysis (PLS-DA) and regression (PLS-R) methods were used to correctly identify the type of CBE and quantify its concentration respectively. The performances of the models created on the two datasets were evaluated in terms of chemometric indicators and compared. The robustness of models was investigated through the analysis of test sets and random permutation tests. Fingerprinting models revealed fruitful results in classifying and quantifying CBEs in blends demonstrating the applicability of NMR in chocolate quality control.

Supercritical fluid chromatography coupled to high-resolution tandem mass spectrometry: an innovative one-run method for the comprehensive assessment of chocolate quality and authenticity.

To assess chocolate quality and authenticity comprehensively, a combination of various analytical procedures is involved, thereby making the process time-consuming and costly. Thus, we investigated the potential of ultra-high performance supercritical fluid chromatography coupled to quadrupole-time of flight mass spectrometry (UHPSFC-QTOF-MS) as an alternative to "classic" methods. By combining hexane and aqueous extracts from sequential extraction, a single 8-min analytical run enabled us (i) to determine cocoa butter equivalents (CBEs) and milk fat content based on the detection of selected triacylglycerols, (ii) to calculate dry non-fat cocoa solids based on determined theobromine and caffeine content, and (iii) to profile contained sugars. To obtain the most comprehensive information about sample composition, the MS method comprised a full MS scan for non-target screening and several time-scheduled targeted MS/MS functions ("parallel reaction monitoring") optimized according to the possible concentration ranges of the analytes. For 40 different chocolate samples, our results and those obtained by using standard methods (LC-UV for non-fat cocoa solids, and GC-FID for CBEs) were in good agreement. Compared to the conventional approach for chocolate quality and authenticity control, the presented SFC-MS method is a fast, cost-effective, and efficient alternative, and only samples suspicious for the presence of CBE should be referred to the standard GC-FID method for exact CBE quantification. In the study, also some challenges offered by SFC-MS have been addressed.

Cocoa butter equivalent from Kpangnan butter and Pequi oil

AbstractNovel cocoa butter equivalents were designed using dry fractionated Pequi oil and solvent fractionated Kpangnan butter. Static crystallization of binary mixtures into the triclinic form (β2) was achieved after 12 days of static crystallization at room temperature for all mixtures and after 4 days only for the 80:20 w/w and 90:10 w/w fractionated Kpangnan: Pequi oil mixtures. However, after 60 days of storage at 22°C, all binary blends (except 100% fractionated pequi oil and 100% fractionated Kpangnan butter) were crystallized in the most stable triclinic crystal form (β1). Here we also reported a higher melting behavior for the fractionated Pequi oil: fractionated Kpangnan (80:20 w/w and 90:10 w/w), after 4 days of static crystallization at room 22°C, which, based on the x‐ray results, could be addressed to completion of crystal polymorphic transition to the triclinic β2 form during 4 days of storage. Our results suggest that the 70:30 w/w fractionated Pequi oil: Kpangnan mixture after 60 days of storage at 22°C showed a melting point of 34°C, a stable triclinic β2 form, and a triglyceride composition of 28% POP, 4.6% POS, and 33% SOS displayed solid‐state characteristics, melting point, and crystal structure, of a novel cocoa butter equivalent.

Structural and Vibrational Investigations of Mixtures of Cocoa Butter (CB), Cocoa Butter Equivalent (CBE) and Anhydrous Milk Fat (AMF) to Understand Fat Bloom Process

Some studies found that the proportions of cocoa butter (CB), cocoa butter equivalent (CBE) and milk fatty acid (AMF) tend to influence the blooming delay when mixing them. The goal of our research is to determine the effects of the proportion of CB, CBE and AMF on the structural organization of the final mixtures. X-ray, DSC, MIR and Raman spectroscopy were used to analyze the structural features and the vibrational modes of four mixtures: CB + 0.5AMF, CB + AMF, CB + 0.5AMF + CBE and CB + AMF + CBE. At room temperature, the triglycerides are ingredients of CB, and CBE and AMF do not fully exhibit the known crystalline forms V or VI, unlike a recent CB sample. Part of these triglycerides is in the form IV instead. The presence of the latter seems to be a key parameter that favors the deceleration of the transformation to the form VI, which is responsible for the development of fat bloom.

Crystallization and Polymorphism of Cocoa Butter Equivalents from Blends of Palm Mid Fraction and Hard Stearins Produced by Enzymatic Acidolysis of High Oleic Sunflower Oil

AbstractCocoa butter equivalents (CBEs) are formulated by mixing palm mid fraction (PMF) with silica‐treated hard stearin fractions produced via enzymatic acidolysis of high oleic sunflower oil with various mixtures of stearic and palmitic acids. The CBE blends always contain ≈30% 1,3‐distearoyl‐2‐oleoyl‐sn‐glycerol, but vary in 1,3‐dipalmitoyl‐2‐oleoyl‐sn‐glycerol (POP; 40–45%) and rac‐1‐palmitoyl‐2‐oleoyl‐3‐stearoyl glycerol (POSt; 10‐16%) content. Overall symmetric disaturated monounsaturated (Sat‐O‐Sat) levels range from 81–86%. The effects of varying POP/POSt content on the non‐isothermal and isothermal crystallization (at 20 °C) of the CBEs are then studied using differential scanning calorimetry (DSC), stop and return DSC, and X‐ray diffraction (XRD), and compared with CB. Four polymorphs (I/sub‐α, II/α, IV/β′, and V/β) are identified which appear at different stages. The addition of PMF (raising POP levels) slows crystallization rates compared to the original stearins, but higher levels of POP also produce a small amount of V/β in under 4 h at 20 °C via apparent transformation from II/α. This does not occur with (high POSt) CB samples. However, after storage at 20 °C for 7 days, CB demonstrates greater transformation to V/β (from IV/β′) as confirmed via XRD. Solid fat contents of CBE blends are higher than CB due to higher Sat‐O‐Sat levels.Practical applications: The paper demonstrates a new route for formulating CBEs. The relative proportions of StOSt, POSt and POP in the formulation can be controlled by changing the proportions of palmitic and stearic acid used in the enzymatic acidolysis and the blend ratio with the PMF. This can then be used to fine‐tune the crystallization behavior.

Physicochemical and oxidative stability of indigenous traditional tengkawang butter as potential cocoa butter equivalent (CBE)

ABSTRACT This study identified the quality, physicochemical properties, and oxidation stability of tengkawang butter (TB) from indigenous plants in various areas of Kalimantan, Indonesia. The fatty acid profile was dominated by palmitic acid, stearic acid, and oleic acid. Acidity ranged from 6.88 to 15.94%, while the peroxide number was 0.41 and 8.27 meq O2/kg. TB had a melting point of 36–37°C. The solid fat content (SFC) varied from 0.03 to 3%. The Oxidation Stability Index (OSI22) was more than 10,000 h. The composition and properties show the potential of TB to become a cocoa butter equivalent (CBE).

An innovative approach for real-time authentication of cocoa butter using a combination of rapid evaporative ionization mass spectrometry and chemometrics

Cocoa butter (CB) is one of the most valuable raw materials in the chocolate industry, and its authenticity is essential to guarantee the safety and quality of chocolate products. For the first time, a real-time and high-throughput method using rapid evaporative ionization mass spectrometry (REIMS) was developed to determine the authenticity of CB. Various types of CB and cocoa butter equivalents (CBEs), as well as CB adulterated by the addition of CBEs, were measured, and different chemometric approaches were implemented for different adulteration scenarios. Hierarchical cluster analysis (HCA) was performed to obtain an overview of the dataset, exploring similarities and differences between CB and CBEs. An orthogonal partial least squares-discriminant analysis (OPLS-DA) model was developed to differentiate between authentic and adulterated CB. A classification accuracy of 100% was achieved, and CBEs could be detected at a level of 10%. A one-class support vector machine (OCSVM) model capable of detecting unsuspected adulterants was successfully built. This model had 95% specificity and 100% sensitivity. Single-origin CB was successfully differentiated from blended CB. This study demonstrated that REIMS could be used as a rapid and sensitive approach for authentication and adulteration analysis of CB. The methodology could enhance the ability of chocolate manufacturers in ensuring the authenticity of their products, helping to avoid complex and ever-changing fraudulent activities.

Production of cocoa butter equivalent from blending of illipé butter and palm mid-fraction

Cocoa butter equivalent (CBE) was prepared from illipé butter (IB) and palm mid-fraction (PMF) blends in different wt ratios. The solid fat content (SFC), compatibility, crystallization/melting thermograms and crystal microstructure of the blends were studied. Softening occurred with IB/PMF blends due to eutectic formation and resulted in blend 75/25 (IB/PMF) exhibiting SFC profile and crystallization/melting behavior comparable to CB and hence it was selected to be used as a CBE. The CBE was fully compatible with CB and displayed similar polymorphic structure (β2) and isothermal crystallization curve to CB. When it was used to partially replace CB in chocolate (5 %wt of the finished product), the hardness, SFC and the bloom formation behavior of the chocolate were not significantly different from the chocolate made with CB. Overall, the 75/25 blend met all criteria to be considered as CBE, hence, it can be used as an alternative to commercially available CBE.

Altering the fatty acid profile of Yarrowia lipolytica to mimic cocoa butter by genetic engineering of desaturases

BackgroundDemand for Cocoa butter is steadily increasing, but the supply of cocoa beans is naturally limited and under threat from global warming. One route to meeting the future demand for cocoa butter equivalent (CBE) could be to utilize microbial cell factories such as the oleaginous yeast Yarrowia lipolytica.ResultsThe main goal was to achieve triacyl-glycerol (TAG) storage lipids in Y. lipolytica mimicking cocoa butter. This was accomplished by replacing the native Δ9 fatty acid desaturase (Ole1p) with homologs from other species and changing the expression of both Ole1p and the Δ12 fatty acid desaturase (Fad2p). We thereby abolished the palmitoleic acid and reduced the linoleic acid content in TAG, while the oleic acid content was reduced to approximately 40 percent of the total fatty acids. The proportion of fatty acids in TAG changed dramatically over time during growth, and the fatty acid composition of TAG, free fatty acids and phospholipids was found to be very different.ConclusionsWe show that the fatty acid profile in the TAG of Y. lipolytica can be altered to mimic cocoa butter. We also demonstrate that a wide range of fatty acid profiles can be achieved while maintaining good growth and high lipid accumulation, which, together with the ability of Y. lipolytica to utilize a wide variety of carbon sources, opens up the path toward sustainable production of CBE and other food oils.

The oleaginous yeast Pichia manshurica isolated from Lansium domesticum fruit in Thailand and its fatty acid composition of single cell oil

Abstract. Planonth S, Chantarasiri A. 2022. The oleaginous yeast Pichia manshurica isolated from Lansium domesticum fruit in Thailand and its fatty acid composition of single cell oil. Biodiversitas 23: 801-809. Oleaginous microbes can accumulate intracellular lipids or single cell oils (SCOs) in quantities higher than 20% of their biomass. They can be a sustainable alternative to fossil fuels, biofuels, and oleochemicals. Studies concerning efficient oleaginous yeasts isolated from the natural environments remain scarce. Therefore, this study isolated and screened for efficient oleaginous yeasts from the surfaces of longkong fruit (Lansium domesticum) samples in Thailand. Their intracellular SCOs were extracted using an ultrasonic-assisted extraction (UAE) method and quantitatively analyzed. The SCO-accumulating yeasts (produce the amount of SCOs &lt;20 % of their biomass) genetically identified were Candida jaroonii, Meyerozyma caribbica, Kodamaea ohmeri, and Pichia sp., while the oleaginous yeasts (produce the amount of SCOs &gt;20% of their biomass) identified were Pichia manshurica and Hanseniaspora opuntiae. Several isolated yeasts were designated as rare oleaginous microbes. The P. manshurica strain Y2 was considered as the most effective oleaginous yeast with a SCO content of 43.03% (w/w). The fatty acids in the accumulated SCO of this strain were analyzed by gas chromatography (GC) that consisted of palmitic, stearic, oleic, linoleic, and palmitoleic acids. These fatty acids could be further applied in the production of third-generation biodiesel, cocoa butter equivalents, and related high-value oleochemicals.

Structural and Vibrational Investigations of Mixtures of Cocoa Butter (CB), Cocoa Butter Equivalent (CBE) and Anhydrous Milk Fat (AMF) to Understand Bloom Chocolate Process

Structural and Vibrational Investigations of Mixtures of Cocoa Butter (CB), Cocoa Butter Equivalent (CBE) and Anhydrous Milk Fat (AMF) to Understand Bloom Chocolate Process

The Comparison of the Effectiveness of Cocoa Butter (CB) and Cocoa Butter Alternatives (CBA) in Chocolate Manufacture

This study aimed at (a) comparing the effectiveness of the use of Cocoa Butter (CB) and Cocoa Butter Alternatives (CBA) on food products and (b) measuring the effectiveness of the sub-groups of the Cocoa Butter Alternatives. In this study, the researchers used Structural Equation Modelling (SEM) with the Partial Least Square (PLS) approach. The analysis showed that (a) CB was more effective in the utilization than CBA and (b) after comparing sub-groups of the CBA examined in this study, Cocoa Butter Substitute (CBS) was more effective than Cocoa Butter Replacer (CBR) and Cocoa Butter Equivalent (CBE). This study is a practical recommendation for the use of CBA so that it can be widely applied in the chocolate industry.

Evaluation of the Miscibility of Novel Cocoa Butter Equivalents by Raman Mapping and Multivariate Curve Resolution-Alternating Least Squares.

Cocoa butter (CB) is an ingredient traditionally used in the manufacturing of chocolates, but its availability is decreasing due to its scarcity and high cost. For this reason, other vegetable oils, known as cocoa butter equivalents (CBE), are used to replace CB partially or wholly. In the present work, two Peruvian vegetable oils, coconut oil (CNO) and sacha inchi oil (SIO), are proposed as novel CBEs. Confocal Raman microscopy (CRM) was used for the chemical differentiation and polymorphism of these oils with CB based on their Raman spectra. To analyze their miscibility, two types of blends were prepared: CB with CNO, and CB with SIO. Both were prepared at 5 different concentrations (5%, 15%, 25%, 35%, and 45%). Raman mapping was used to obtain the chemical maps of the blends and analyze their miscibility through distribution maps, histograms and relative standard deviation (RSD). These values were obtained with multivariate curve resolution–alternating least squares. The results show that both vegetable oils are miscible with CB at high concentrations: 45% for CNO and 35% for SIO. At low concentrations, their miscibility decreases. This shows that it is possible to consider these vegetable oils as novel CBEs in the manufacturing of chocolates.