Lauric Fat Cocoa Butter Replacer from Krabok (Irvingia Malayana) Seed Fat and Coconut Oil

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1 Journal of Oleo Science Copyright 2015 by Japan Oil Chemists Society doi : /jos.ess14244 Lauric Fat Cocoa Butter Replacer from Krabok (Irvingia Malayana) Seed Fat and Coconut Oil Sopark Sonwai *, Pimwalan Ornla-ied and Tanapa Aneknun Department of Food Technology, Faculty of Engineering and Industrial Technology, Silpakorn University (6 Rajmakkanai Road), Nakhonpathom, 73000, Thailand Abstract: Lauric fat cocoa butter replacer (LCBR) was produced from a blend of krabok seed fat (KSF) and coconut oil (CO). Four fat blends with different ratios of KSF/CO (20/80, 40/60, 60/40 and 80/20 (%wt)), CO, KSF and a commercial LCBR (C-LCBR) were characterized using various techniques. It was found that blend 60/40 exhibited SFC curve and crystallization/melting behavior most similar to that of C-LCBR. The blend met the requirements to be considered as LCBR and has potential as an alternative to commercial LCBR that are being used nowadays and hence it was recommended as LCBR (called R-LCBR). The polymorphic behavior of both C-LCBR and R-LCBR was investigated and both fats displayed mainly short spacing pattern associated with b polymorph, a required polymorph for LCBR. The compatibility between R-LCBR and CB was investigated by mixing the R-LCBR with CB in different proportions and softening due to the eutectic effect was observed in the mixed fats. This limits the proportion of CB and the R-LCBR in compound coatings to no more than 5% of CB in the total fat phase. Key words: krabok seed fat, coconut oil, lauric fat cocoa butter replacer 1 INTRODUCTION Cocoa butter CB is predominantly composed of three monounsaturated triglycerides - POP, POS and SOS where P palmitic acid, O oleic acid, and S stearic acid. CB is mainly solid at temperatures below 25 but is almost entirely liquid at the body temperature The fat is an important major constituent of chocolate and other confectionery products. CB has a complex polymorphism which is manifested in six polymorphs called forms I to VI 2. However, only form V gives the optimal melting behavior for CB in chocolate 1. It also confers chocolate products proper snap ability to break apart easily, good demoulding properties contraction and a good quality finish in terms of color and gloss. Shortage of supply and poor CB quality of individual harvests have prompted the chocolate and confectionery industries to look for alternative fats to partially or fully replace CB. Lauric fat cocoa butter replacers LCBR are modified lauric fats which are used extensively in the chocolate and confectionery industry 3. LCBR are made primarily from fats that are high in lauric acid such as palm kernel oil and coconut oil. LCBR melt in the same temperature range as CB, and so have a similar texture and mouthfeel, but which crystallize in a very different way 1. However, one factor identified with the LCBR is their very low tolerance for CB and hence can be used only when very little CB is present or if almost all CB is replaced. In addition, unlike CB, they crystallize directly into one crystal form, β polymorph, which is their stable polymorph and therefore no pre-crystallization is required 4. LCBR generally contain of lauric acid in combination with lesser amount of relatively low molecular weight fatty acids. Modification techniques, including hydrogenation, interesterification and fractionation, are used to enhance the solid fat content and melting properties of palm kernel oil and blends of palm kernel, coconut and palm oils 5, resulting in LCBR which are inexpensive and have eating qualities virtually identical to cocoa butter. Krabok Irvingia Malayana is a large-grown and woody wild almond tree widely distributed in tropical and subtropical areas. The most interesting part of the krabok nut is its fat, which is the most abundant component of kernels. It has been reported that krabok seed fat KSF is rich in lauric acid 6, 7, indicating its potential uses as crude oil feed stock for confectionery fats. The fat exhibited solid fat content SFC 80 at 30 and below and a slip melting point that was much higher than that of CB 6. However, despite all these promising properties, KSF has so far been * Correspondence to: Sopark Sonwai, Department of Food Technology, Faculty of Engineering and Industrial Technology, Silpakorn University (6 Rajmakkanai Road), Nakhonpathom, 73000, Thailand ssonwai@gmail.com, ssonwai@su.ac.th Accepted December 2, 2014 (received for review October 30, 2014) Journal of Oleo Science ISSN print / ISSN online

2 S. Sonwai, P. Ornla-ied and T. Aneknun underutilized. The objective of the present work was to investigate the possible utilization of KSF as LCBR. KSF was mixed with coconut oil CO, one of the most important crude oil feed stocks for LCBR, in different proportions and the physicochemical properties and crystallization and melting characteristic of the blends was studied and compared with a commercial LCBR. The blend that exhibited the characteristics most similar to those of the commercial LCBR would be selected as LCBR. 2 EXPERIMENTAL PROCEDURES 2.1 Materials Mature krabok seeds were collected from the northeastern region of Thailand. Refined, bleached and deodorized CO was purchased from Katevanich Indurtry Co., Ltd. Bangkok, Thailand. The krabok seeds were cut open and the seed kernels were removed. The almond-like seed kernels were crushed by a hand mill and dried in a vacuum oven for 4 h at 60. Then, the dried kernels were finely ground and KSF was extracted using the Soxhlet extraction method at 60 for 6 h with n-hexane as a solvent and the crude fat was purified 6. A commercial LCBR C-LCBR was purchased from Sino-Pacific Trading Thailand Co., Ltd. Bangkok, Thailand. This C-LCBR has been used extensively by many confectionery companies in Thailand to produce compound coatings. The standard fatty acid methyl esters for fatty acid analysis using gas chromatography GC were purchased from AccuStandard, Inc. USA. Acetone and acetonitrile were of GC grade from Burdick and Jackson Muskegon, MI, USA. All other chemicals and solvents used were obtained commercially and were of the highest purity available. 2.2 Fat blending The purified KSF was blended with CO into four proportions of KSK/CO: 20/80, 40/60, 60/40 and 80/20 wt. Many different properties of the fat blends, CO, KSF and C-LCBR were characterized using various techniques and methods as described below. 2.3 Characterization of fatty acid composition CO, KSF and commercial LCBR were converted into fatty acid methyl esters FAMEs using AOAC official method The FAMEs analysis was performed in a Shimadzu GC with flame ionization detector Model GC The system had an VertiBond TM wax capillary column 50 m long, 0.25 mm internal diameter and 0.20 mm film thickness. Identifications of the compounds were carried out by external standardization method. Helium was used as a carrier gas with a flow rate of 1 ml/min and with a controlled initial pressure of 93.2 kpa at 120. N 2 and air were makeup gases. The injection temperature was 210, and the oven temperature program was holding at 120 for 3 min before increasing at a rate of 10 /min to 220, holding at this temperature for 30 min, increasing at a rate of 5 /min to 240, followed by holding at 240 for 30 min. The split ratio was 100:1, the injection volume was 1 μl, and the detector temperature was Characterization of physicochemical properties Iodine value Iv and slip melting point SMP of CO, KSF, KSF/CO blends and C-LCBR were analyzed following American Oil Chemists Society AOCS Methods Cd 1d-92 and Cc 3-25, respectively Characterization of solid fat content Changes in SFC as a function of temperature between 15 and 40 and the melting behavior of CO, KSF, KSF/ CO blends and C-LCBR were determined by pulse-nuclear magnetic resonance spectrometer p-nmr Minispecmq20, BRUKER, Karlsruhe, Germany following a method for measuring solid fat content of hardened fats and fats for bakery/confectionery use developed by the Joint Committee for the Analysis of Fats, Oils, Fatty Products, Related Products and Raw Materials GA FETT as described by Fiebig and Luttke 10. Briefly, the fat samples were heated to 80 for at least 10 min, cooled to 0 and held for 60 min, then heated to measuring temperature and held for min prior to measurement. 2.6 Characterization of crystallization and melting profiles The crystallization and melting profiles of all fat samples were determined with a Perkin-Elmer differential scanning calorimeter DSC model DSC 8000, PerkinElmer Co., Norwalk, CT following the procedure Cj 1-94 recommended by AOCS 9. The heat flow of the instrument was calibrated with indium mp as a reference standard. A fat sample of 3-5 mg was placed in an aluminum pan 20 μl capacity and hermetically sealed. An empty pan served as reference. The samples were heated from room temperature to 80 at 30 /min and maintained for 10 min in order to destroy any crystal memory. Then, the samples were cooled to 60 at the rate of 5 /min and held at this temperature for 30 min, followed by heating at the rate of 5 /min to 80. The crystallization and melting profiles were generated during the cooling and heating, respectively. The crystallization onset T CO and melting completion temperatures T MC were obtained from the peaks located at the highest temperature of each fat sample. T CO and T MC were considered to be the temperatures at which the crystallization began and the melting ended, respectively. 2.7 Crystal morphology The observation of the crystal network microstructure of all fats crystallized under static conditions at 25 was per- 358

3 Lauric Fat Cocoa Butter Replacer from Krabok (Irvingia Malayana) Seed Fat and Coconut Oil formed by a polarized light microscopy Olympus BX51, Olympus Optical Co., Ltd., Tokyo, Japan equipped with a digital camera Olympus C-7070, Olympus Optical Co., Ltd., Tokyo, Japan. All fat samples were melted at 80 for 10 min to completely eliminate the memory effect. 20 μl of each molten sample was placed on a glass slide, which was heated to 80 prior use. A preheated glass cover slip was carefully placed over the sample to produce a film of uniform thickness. Then, the samples were incubated at for 48 h in a temperature-controlled cabinet. A 4 lens was employed to image the gray scale photographs of the fat crystals. 2.8 Selection of LCBR from the fat blends All the results obtained from the studies in sections were used to decide which of the fat blends would have the highest potential to be used as a LCBR fat, e.g. the blend that exhibited the SFC curve and crystallization and melting temperatures/behavior closest to those of the C-LCBR. 2.9 Polymorphic characterization The polymorphic form of the fat crystals in the C-LCBR and the selected or recommended LCBR called R-LCBR was determined with an x-ray diffractometer XRD Rigaku TTRAX III, Rigaku Corporation, Tokyo, Japan. Scans were made in wide-angle x-ray scattering from 18 2θ to 26 2θ with a scan speed and a step width of 4.5 2θ/min and θ, respectively. The molten fat samples were put inside XRD capillary sample holders using a pipette. The samples were heated by dipping the capillaries in a water bath set at 80 for 10 min before being stored at 0 for 90 min. After that they were stabilized at 26 for 40 h before XRD measurement Compatibility study The compatibility between the R-LCBR and CB was tested by mixing it with CB in different ratios between 0/100 and 100/0 wt for R-LCBR/CB and the SFC of each ratio was measured at using the p-nmr following the procedure given by Smith RESULTS AND DISCUSSION 3.1 Fatty acid composition analysis Table 1 exhibits the fatty acid compositions of CO and KSF compared with C-LCBR. All three samples consisted mainly of lauric acid and 90 of their fatty acid compositions was saturated fatty acids. CO consisted mainly of lauric 45.2 g/100g, myristic 17.9 g/100g, palmitic 9.4 g/100g, caprylic 8.2 g/100g and oleic 7.5 g/100g acids. The main fatty acids in KSF were lauric 47.3 g/100g and myristic 42.0 g/100g acids whereas the major fatty acids Table 1 Fatty acid compositions of coconut oil (CO), krabok seed fat (KSF) and commercial lauric cocoa butter replacer (C-LCBR). Fatty acids Concentration (g/100g) CO KSF C-LCBR Caproic acid (C 6 ) Caprylic acid (C 8 ) Capric acid (C 10 ) Lauric acid (C 12 ) Tridecanoic acid (C 13 ) Myristic acid (C 14 ) Tetradecenenoic acid (C14:1) Pentadecanoic acid (C 15 ) Palmitic acid (C 16 ) Palmitoleic acid (C 16:1 ) Margaric acid (C 17 ) Stearic acid (C 18 ) Oleic acid (C 18:1 ) Elaidic acid (C18:1, trans) Linoleic acid (C 18:2 ) Linolenic acid (C 18:3 ) < Arachidic acid (C 20 ) Eicosenoic acid (C 20:1 ) Arachidonic acid (C20:4) Behenic acid (C 22 ) Lignoceric acid (C 24 ) ΣSFA ΣUSFA in C-LCBR were lauric 43.5 g/100g, stearic 21.2 g/100g, myristic 13.1 g/100g and palmitic 8.2 g/100g acids. Among the three samples, CO contained highest amount of caprylic, capric, palmitic and oleic acids, KSF exhibited highest content of lauric and myristic acids and C-LCBR consisted of the highest content of stearic acid. KSF and C-LCBR exhibited highest total amount of saturated fatty acids 96.9 and 96.2 g/100g, respectively whilst CO contained highest total content of unsaturated fatty acids 9.54 g/100 g. All three samples contained relatively low contents of arachidic C 20, behenic C 22 and lignoceric C 24 acids. These are long-chain saturated fatty acids with high melting temperatures. The fatty acid composition of CO reported here was in agreement with the data published earlier 13. Finally, only C-LCBR and CO contained transfatty acid elaidic acid, which was present for 3.12 g/100 g and 0.04 g/100 g in C-LCBR and CO, respectively. The high content of trans-fatty acid in C-LCBR could be the result of hydrogenation process that is commonly used for the production of plastic fats and CB-like fats

4 S. Sonwai, P. Ornla-ied and T. Aneknun 3.2 Physicochemical properties Table 2 shows the Iv and the SMP of CO, KSF, different KSF/CO blends and C-LCBR. The Iv provides information on the degree of unsaturation and hardness of fats. The higher the Iv, the more unsaturated fatty acid bonds present in a fat. CO exhibited the highest Iv g I 2 /g fat and KSF and C-LCBR has the lowest Iv 4.44 and 3.96 g I 2 /g fat, respectively. This indicates that CO had higher level of unsaturation or higher total content of unsaturated fatty acids than KSF and C-LCBR Table 1. As the amount of KSF in the fat blends increased, Iv decreased, implying that less unsaturated fatty acid bonds were present with increasing amount of KSF. Iv of all fat blends was higher than that of the C- LCBR. SMP of a fat is the temperature at which a column of fat in an open capillary tube becomes sufficiently fluid to slip up the tube when it is subjected to a controlled heating. At 39.5, SMP of both KSF and C- LCBR were the highest and SMP of CO was the lowest at The less content of unsaturated fatty acids in both KSF and C- LCBR when compared with CO Table 1 could have contributed to their high SMP. As the quantity of KSF in the fat blends increased, SMP also increased. Again, this could be related to the fact that the content of unsaturated fatty acids in the blends decreased as the KSF content increased. 3.3 Characterization of solid fat content SFC versus temperature plots generally provide not only the information on how hard or soft fats are at different temperatures but also the melting behavior of the fats. Figure 1 shows SFC of CO, KSF, different KSF/CO blends and C-LCBR measured at temperatures between The figure showed that the SFC of all samples decreased as the temperature increased. Among all fats, SFC of KSF was the highest between while CO showed the lowest SFC between The highest content of saturated fatty acids, most of which were medium-chain fatty Table 2 Physicochemical properties of coconut oil (CO), krabok seed fat (KSF), different KSF/CO blends and commercial lauric cocoa butter replacer (C-LCBR). Samples Iodine value (g I 2 /g fat) Slip melting point ( ) 0/100 (CO) 10.15± ± / ± ± / ± ± / ± ± / ± ± /0 (KSF) 4.44± ±0.20 C-LCBR 3.96± ±0.71 Fig. 1 Solid fat content measured at of coconut oil (CO), krabok seed fat (KSF), different KSF/CO blends and commercial lauric cocoa butter replacer (C-LCBR). acids C 12 and C 14, and lowest content of unsaturated fatty acids contained in KSF Table 1 could be the key contribution to its high SFC throughout the temperature range studied. At any given temperature, as the content of KSF in the fat blends increased, the SFC also increased almost proportionally. The SFC of blends 20/80 and 40/60 was lower than C-LCBR at all temperatures. On the other hand, the SFC of blend 80/20 was significantly higher than C-LCBR at all temperatures except at 15 and 40. It can be seen from the figure that blend 60/40 exhibited SFC curve that was most similar to C-LCBR. The blend exhibited slightly lower SFC than C-LCBR at 20 and below with a fast drop in SFC between 25 and 35, indicating a sharp melting characteristic. This would allow it to melt quickly with a pronounced cooling effect when consumed 14. At 35 the SFC of blend 60/40 dropped to a much lower value than C-LCBR and the fat melted completely as the temperature approached 40, indicating that it would give no waxy mouthfeel. In order to eliminate waxy mouthfeel, a fat should have SFC 3.5 at temperatures above On the other hand, the SFC at 40 of C-LCBR was still as high as 8 and this would make the commercial fat waxy when consumed. 3.4 Analysis of crystallization and melting profiles The DSC crystallization thermograms for CO, KSF, blends of KSF/CO and C-LCBR are given in Fig. 2a and the temperature of phase transition points for the samples were summarized in Table 3. The thermograms of CO showed two distinct peaks; a sharp peak at and a broader peak at , suggesting that heterogeneous types of triacylglycerols in CO were solidified at dif- 360

5 Lauric Fat Cocoa Butter Replacer from Krabok (Irvingia Malayana) Seed Fat and Coconut Oil Fig. 2 DSC crystallization thermograms (a) and melting thermograms (b) of coconut oil (CO), krabok seed fat (KSF), different KSF/CO blends and commercial lauric cocoa butter replacer (C-LCBR). T CO and T MC indicate crystallization onset temperature and melting completion temperature, respectively. 1-4 indicates temperatures of phase transition as shown in Table 3. ferent temperatures. The crystallization thermogram of KSF showed two overlapping exothermic peaks at and whilst the crystallization thermogram of C-LCBR exhibited two broad and non-distinct exotherms at and When 20 of KSF was blended with 80 CO 20/80 blend, the original crystallization peak of CO at 1.99 disappeared and the sharp crystallization peak at 5.25 was replaced by much broader and overlapping peaks at and As the amount of KSF in the fat blend increased to 40 and 60, the crystallization thermogram exhibited a single exotherm which was centered at a higher temperature, and for blends 40/60 and 60/40, respectively. For blend 80/20, the thermogram showed a major crystallization peak centered at with a small overlapping peak on the lower temperature side at The crystallization onset temperature, T CO, was highest for C-LCBR 29.66, possibly due to its high stearic acid content Table 1. At 24.58, T CO of KSF was significantly lower than that of C-LCBR, and at 8.18, T CO of CO was the lowest. T CO of the fat blends increased when the content of KSF in the blend increased. Similarly, the crystallization enthalpy ΔH increased with the increasing amount of KSF in the blends and KSF exhibited the highest crystallization enthalpy. The DSC thermograms for melting of the fat samples are given in Fig. 2b. CO had two overlapping endothermic peaks at and and KSF showed one sharp but asymmetric melting peak at The melting of C-LCBR displayed extra broad melting range with four overlapping peaks between and , implying that the fat consisted of heterogeneous type of triacylglycerols. When 20 KSF was mixed with 80 CO, the melting thermogram exhibited one main peak at , which was overlapping with two smaller peaks at and When the KSF content in the blends increased from 20 to 40, 60 and 80, the two small peaks disappeared gradually and eventually the thermogram showed only one melting peak, the position of which moved gradually towards the highertemperature side. This suggested that the blends contained more homogeneous type of triacylglycerols with higher degree of saturation and/or longer fatty acid chains as more KSF in the blends increased. Similar to T CO, the melting completion temperatures T MC was lowest for CO and highest for C-LCBR It was likely that the high content of stearic acid and trans-fatty acid in the C-LCBR Table 1 had contributed to its high T MC. For the KSF/CO blends, T MC increased as the KSF content in the blends increased. Finally, the melting enthalpy was highest for KSF and the enthalpy of the fat blends increased with an increase in their KSF content. 3.5 Morphology study The polarized light microscopy micrographs of the fat samples obtained after static crystallization at 25 for 48 h are given in Fig. 3. The solid phase appears white or gray while the liquid phase appears black. The microstructure of CO crystals was densly-packed spherulites consisting of fine needle-like crystals radiating and branching outward from central nuclei Fig. 3a. The crystal size of the fat ranged from 400 to 1000 μm. The microstructure of KSF crystals was small but densely-packed spherulites that strongly aggregated to one another Fig. 3f and the morphology of C-LCBR crystals was continuous granular data 361

6 S. Sonwai, P. Ornla-ied and T. Aneknun Table 3 Temperature of phase transition points (T) and crystallization and melting enthalpies (ΔH) for coconut oil (CO), krabok seed fat (KSF), different KSF/CO blends and commercial lauric cocoa butter replacer (C-LCBR). T CO and T MC represent crystallization onset and melting completion temperatures, respectively. 1-4 indicates temperatures of phase transition based on Fig. 2. Samples ΔH (J/g) Transition point temperature ( ) T CO T MC Crystallization 0/100 (CO) / / / / /0 (KSF) C-LCBR Melting 0/100 (CO) / / / / /0 (KSF) C-LCBR not shown. When 20 KSF was mixed with 80 CO, the microstructure of the blend was still spherulites but the crystals were smaller, less densely-packed and consisting of larger needle-like crystals. As the content of KSF increased further, the the crystal morphology remained relatively the same but the crystal size decreased substantially and the crystal number increased significantly. Some extent of crystal aggregation was also observed in all the blends. 3.6 Selection of LCBR from the fat blends All binary fat blends exhibited SMP that were lower than the C-LCBR Table 2. However, the SMP of blends 60/40 and 80/20 were closer to C- LCBR than blends 20/80 and 40/60. From the SFC versus temperature plots, blend 60/40 displayed the SFC curve that was most similar to the C-LCBR Fig. 1 and in good agreement with SFC curves of LCBR reported in the literature 5, 16. The blend had high SFC at 15-25, giving it a hard consistency within this temperature range, and melted rapidly between 25 and 35 and finally became almost completely liquid at the body temperature 37. In addition, the blend exhibited the DSC melting peak that situated well within the melting range of C-LCBR Fig. 2, giving it a similar texture and mouthfeel to that of C-LCBR. As a result of this, blend 60/40 was recommended as LCBR in this work. 3.7 Polymorphic characterization The polymorphic structure of the C-LCBR and the recommended LCBR called R-LCBR after being stored at 0 for 90 min and then stabilized at 26 for 40 h was given in Fig. 4. The C-LCBR crystallized mainly into β structure with two diffraction peaks at 3.85 Å 1 and 4.24 Å Additionally, there was a small diffraction peak at 4.15 Å 2 which could be related to either α structure 18 or pseudo-β structure 19. Similarly, the R-LCBR exhibited mainly β structure with two main diffraction peak at 3.84 Å 1 and 4.24 Å The sample displayed two additional diffraction peaks at 4.04 Å 2 and 4.41 Å 4. The 4.04 Å peak was close to peak range of β Å reported by Ghotra et al. 20. The peak at 4.41 Å matched one of the diffraction peaks of pseudo-β structure reported by D Souza et al. 19 and O Brien 21. It can be seen here that, under the same crystallization conditions, the polymorphic behavior of R-LCBR was the same as C-LCBR. Both fat exhibited mainly β structure after the crystallization at 26 for 40 h. β is a required polymorph for LCBR so that the fat does not need to be tempered, which is useful in many applications

7 Lauric Fat Cocoa Butter Replacer from Krabok (Irvingia Malayana) Seed Fat and Coconut Oil Fig. 3 P olarized light microscope images showing crystal morphology of coconut oil (a), krabok seed fat (f) and different KSF/CO blends: 20/80 (b), 40/60 (c), 60/40 (d), 80/20 (e), obtained during static isothermal crystallization at 25 for 48 h. 3.8 Compatibility between the recommended LCBR and CB The compatibility between the R-LCBR and CB was studied and the results are given in Fig. 5, which exhibits the SFC at 15, 20, 25 and 30 of the mixtures of the R-LCBR and CB. The figure shows that at all temperatures, except 15, the SFC of the fat mixtures did not lie on the straight line that connected the SFC of the individual fats. The results indicate that the two fats were not fully compatible and because of this their mixtures could result in a softening or eutectic effect. For example, the addition of 0 to 40 of R-LCBR to CB at 20 resulted in a decrease in SFC from 57.4 to This softening was even more obvious at 25 and 30. Nonetheless, this is a typical 363

8 S. Sonwai, P. Ornla-ied and T. Aneknun and CO with a weight ratio of 60 to 40 exhibited the SFC profile and crystallization/melting behavior most similar to that of C-LCBE. The blend met all the requirements to be considered as LCBR and hence it was recommended as LCBR. The polymorphic behavior of R-LCBR was studied and compared with C-LCBR and both fats crystallized into β structure, a required crystal structure for LCBR. The compatibility between R-LCBR and CB was investigated by mixing R-LCBR with CB in different proportions and softening due to the eutectic effect was observed in the mixed fats. This limits the proportion of CB and the R-LCBR in compound coatings to no more than 5 of CB as a portion of the total fat phase. Fig. 4 Polymorphic structure of commercial lauric cocoa butter replacer (C-LCBR) and the recommended lauric cocoa butter replacer (R-LCBR) obtained after static isothermal crystallization at 26 for 40 h. ACKNOWLEDGEMENT The authors would like to thank Auttapol Jarearnkaew, Pongsakorn Sankot and Pawitchaya Podchong for their technical assistance. Fig. 5 Solid fat content of mixtures of cocoa butter (CB) and the recommended lauric cocoa butter replacer (R-LCBR) measured at SFC characteristic for LCBR when mixed with CB as the triacylglycerol composition of LCBR is normally different from that of CB 12. In general, it is recommended that the proportion between CB and LCBR in compound coatings should not exceed 5 of CB in the total fat phase and that LCBR is normally used with cocoa powder, where little CB is present, for the production of compound coatings 1. 4 CONCLUSION From the results obtained in this study, a blend of KSF REFERENCES 1 Beckett, S. T. The Science of Chocolate. 2 nd ed. The Royal Society of Chemistry, Cambridge Wille, R. L.; Lutton, E. S. Polymorphism of Cocoa Butter. J. Am. Oil Chem. Soc. 43, Smith, K. W.; Cain, F. W.; Talbot, G. Nature and composition of fat bloom from palm kernel stearin and hydrogenated palm kernel stearin compound chocolates. J. Agric. Food Chem. 52, Stewart, I. M.; Timms, R. E. Fats for chocolate and sugar confectionery. in Fats in Food Technology Rajah, K. K. ed. Sheffield Academy Press, Sheffield, pp Pease, J. J. Confectionery fats from palm oil and lauric oil. J. Am. Oil Chem. Soc. 62, Sonwai, S.; Ponprachanuvut, P. Characterization of physicochemical and thermal properties and crystallization behavior of Krabok Irvingia Malayana and rambutan seed fats. J. Oleo Sci. 61, Bandelier, J.; Chunhieng, T.; Olle, M.; Montet, D. Original study of the biochemical and oil composition of the Cambodia nut Irvingia malayana. J. Agri. Food Chem. 50, AOAC. Official Methods of Analysis of AOAC International. AOAC International, Arlington, VA AOCS. Official Methods and Recommended Practices of the American Oil Chemists Society. 6th ed. American Oil Chemists Society. Champaign, USA Fiebig, H. J.; Luttke J. Solid fat content in fats and oils determination by pulsed nuclear magnetic resonance 364

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