Aflatoxin production by Aspergillus flavus in Brazil nuts

Experiments were conducted to evaluate the effects of relative humidity (r.h.; 75%, 80%, 85%, 97%) and temperature (10, 13, 15, 25, 30 °C) on aflatoxin production in previously dried (3.5% moisture content; m.c.) Brazil nuts. Initially Aspergillus spp. were isolated from the surfaces of whole in-shell (WIS) Brazil nuts imported from Peru using A. flavus and A. parasiticus agar (AFPA). Isolates were subsequently screened for aflatoxin production using yeast extract sucrose medium. Total aflatoxin (B₁+B₂+G₁+G₂+M₁) was analyzed using an immunoassay technique while the presence of aflatoxin was confirmed using thin-layer chromatography. The surface of shelled half-nuts (simulating damaged or trimmed nuts), shelled whole (SW) nuts, and WIS nuts following a chlorine wash and water rinse, served as sites for inoculation (10 μl; 10⁵/ml) using an aflatoxigenic isolate. Maximum concentrations of total aflatoxin and B₁ were detected in nuts stored at 97% r.h. and at temperatures of 25–30 °C. Shelled half-nuts contained the highest total (6817 ng/g) and B₁ (4483 ng/g) aflatoxin. WIS nuts contained the least total and B₁ toxin with maximum concentrations of 93 and 49 ng/g, respectively. Aflatoxin was not detected (detection limit of 1.75 ng/g) in nuts maintained at either 10 °C (97% r.h.) or at 30 °C (75% r.h.) for up to 60 d. Maximal moisture contents (%) and water activity values (a_W) for nuts stored at these conditions were 4.50 and 0.78, and 9.14 and 0.92, respectively. Results of this study indicate that the limiting moisture content and a_W values required to control aflatoxin production () in SW and WIS stored at 30 °C for up to 60 d are 4.5, 0.68, 5.0, and 0.75, respectively. Overall, increasing the relative humidity and temperature during storage resulted in an increase in aflatoxin and these were shown to be the most significant variables influencing toxin production in Brazil nuts.     

Enthalpy-Entropy compensation in food vapor adsorption

Enthalpy-entropy compensation was analysed for sorption isotherms of potatoes, macadamia nuts, apricots, figs, currants, prunes and raisins. Plots of (ΔH_dif)_T vs (ΔS_dif)_T for potatoes and macadamia nuts presented two isokinetic temperatures: T_B1 = 272.0 ± 57.7 K (− 1 °C) for potatoes, T_B1 = 265.0 ± 18.8 K (− 8 °C) for macadamia nuts and T_B2 = 382.5 ± 7.3 K (109.5 °C) for both products. The first isokinetic temperature (T_B1) appeared only at the upper portion of the temperature range tested (50, 60 and 70 °C for potatoes and 50 and 60 °C for macadamia nuts). The two isokinetic temperatures observed for potatoes and macadamia nuts suggested that during the initial stages at low a_w T_B1 is controlled by changes in the entropy of water, whereas the second isokinetic temperature (T_B2) is considered to be enthalpy-controlled. Dried fruits presented only one isokinetic curve T_B = 315.7 ± 3.5 K (42.7 °C), for raisins, currants and figs (75.2–82.3% d.b. sugars) and T_B = 317.7 ± 4.6 K (44.7 °C) for prunes and apricots (51.5–54.5% d.b. sugars), indicating an enthalpy-controlled adsorption process for the whole range of moisture contents covered.     

Theoretical analysis of efficient microwave processing of oil–water emulsions attached with various ceramic plates

A detailed theoretical analysis has been carried out to study efficient microwave processing of 1D emulsion samples placed on ceramic plates (alumina, SiC). The effective dielectric property of an emulsion is a strong function of the continuous medium of the emulsion (o/w or w/o). A preliminary study has been carried out via average power within an emulsion vs emulsion thickness diagram for various cases. The maxima in average power, also termed as ‘resonances’, are observed for specific emulsion thicknesses and the two consecutive resonances are termed as R₁ and R₂ modes. The heating scenarios have been analyzed at the dominant resonance mode. Based on spatial distributions of power and temperature for various cases, alumina support at the left side may be recommended as the optimal heating strategy due to greater heating rates with controlled thermal runaway for both o/w and w/o emulsion samples whereas SiC support may be favored for o/w emulsion samples due to lesser thermal runaway. A comparison of the heating characteristics has been illustrated for 50% o/w and 50% w/o emulsion samples to analyze the role of continuous medium of an emulsion on heating effects. The distribution of microwave incidences also plays an important role to minimize thermal runaway for specific o/w and w/o emulsions.     

NMR relaxometry and differential scanning calorimetry during meat cooking

By combining simultaneous nuclear magnetic resonance (NMR) T₂ relaxometry and differential scanning calorimetry (DSC) on pork samples heated to nine temperature levels between 25 and 75 °C, the present study investigates the relationship between thermal denaturation of meat proteins and heat-induced changes in water characteristics. Principal component analysis (PCA) on the distributed ¹H NMR T₂ relaxation data revealed that the major changes in water characteristics during heating occur between 40 and 50 °C. This is probably initiated by denaturation of myosin heads, which however, could not be detected in the DSC thermograms obtained directly on the meat. In contrast, the DSC thermograms revealed endothermic transitions at 54, 65 and 77 °C, probably reflecting the denaturation of myosin (rods and light chain), sarcoplasmic proteins together with collagen and actin, respectively. Simultaneous modelling of DSC and NMR data by partial least squares regression (PLSR) revealed a correlation between denaturation of myosin rods and light chains at 53–58 °C and heat-induced changes in myofibrillar water (T₂ relaxation time 10–60 ms) as well as between actin denaturation at 80–82 °C and expulsion of water from the meat. Accordingly, the present study demonstrates a direct relationship between thermal denaturation of specific proteins/protein structures and heat-induced changes in water mobility during heating of pork.     

Water desorption thermodynamic properties of pineapple

The water desorption isotherms of pineapple were determined at 5, 15, 25, 35 and 45 °C by using a gravimetric technique within the water activity range of 0.1–0.9. The effect of the temperature showed a cross-over of the desorption isotherms at water activity of 0.75. The experimental data was fitted to the GAB model taking into account the effect of the temperature. The obtained simulation could be considered satisfactory (mean relative error <5.0% and percentage of explained variance >99.8%). The net isosteric heat of sorption () and the differential entropy (S_d) were estimated as functions of the moisture content, the enthalpy–entropy compensation theory was applied to sorption isotherms and plots of Q^st vs. S_d provided the isokinetic temperature, suggesting an enthalpy controlled sorption process.     

Dielectric properties of Indian Basmati rice flour slurry

Dielectric properties of Indian Basmati rice samples (dry ground flour and wet aqueous slurry) were measured over the frequency range 500–2500 MHz. Temperature dependence of dielectric properties of rice flour slurries (20, 30 and 50 g flour per 100 g water yielding approximately 1:5, 1:3 and 1:2 flour to water ratio) were studied between 30 and 80 °C. Results indicated that dielectric constants (ε′) generally did not vary with frequency while the loss factor (ε″) showed an increasing trend. A sharp change in dielectric parameters were noted above 70 °C attributable to rice starch gelatinization, an observation confirmed later by differential scanning calorimetry. An increase in flour slurry concentration systematically reduced ε′ during the entire frequency range while variations in ε″ values were mixed. Addition of 1% salt markedly increased ε″ of slurries whereas butter resulted in significant reduction in ε′ values. Rice flour slurry containing both salt and butter exhibited intermediate values of dielectric properties. Both ε′ and ε″ data of rice slurries generally fitted a 2nd order polynomial relationship with temperature. These results suggest that dielectric measurements could be used to study frequency, concentration, ingredient and temperature dependent changes in starch–water systems during heating.     

Thermal inactivation of the frozen thawed traditional meat starter culture, Pediococcus pentosaceus, in a meat model system

The equation, y_(t) = y₍₀₎e^kt, was fitted (R = 0.9281, 0.9220 and 0.9117, respectively) to thermal inactivation data (55, 60 and 65 °C) of the traditional meat starter culture Pediococcus pentosaceus (10⁷ cfu/ml) in a meat model system. The population reduction constant (‘k’) increased (about 2.5- and 3-fold) with an increase in the treatment temperature (from 55 to 60 °C and from 60 to 65 °C, respectively). The Q₁₀ (55–65 °C) for ‘k’ was 7.63. Thermal treatments of 19.1, 9.0 and 3.1 min (55, 60 and 65 °C, respectively) reduced the population of P. pentosaceus by 2.0 logs. The value of ‘k’ and the duration of the thermal treatment played an important role in the extent of the inactivation of the culture. The “zero inactivation” temperature (T₀) for P. pentosaceus was 49.9 °C. About 5 logs of the culture would be destroyed at 63 and 68 °C within about 15.5 and 6.5 min, respectively.     

Impact of sorbitol on the thermostability and heat-induced gelation of bovine serum albumin

The influence of sorbitol (0–40 wt.%) on the thermal denaturation and gelation of bovine serum albumin (BSA, pH 7.0) in aqueous solution has been studied. The effect of sorbitol on heat denaturation of 0.5 wt.% BSA solutions was measured using ultrasensitive differential scanning calorimetry. The unfolding process was irreversible and was characterized by the thermal denaturation temperature (T_m). As the sorbitol concentration increased from 0 to 40 wt.%, T_m increased from 73.0 to 80.9 °C. The rise in T_m was attributed to the increased thermal stability of the globular state of BSA relative to its native state. The dynamic shear rheology of 4 wt.% BSA solutions containing 200 mM NaCl was monitored as they were heated from 30 to 90 °C at 1.5 °C min⁻¹, held at 90 °C for 120 min, and then cooled back to 30 °C at −1.5 °C min⁻¹. Sorbitol increased the protein gelation temperature (ΔT_gel +10 °C for 40 wt.% sorbitol), decreased the isothermal gelation rate at 90 °C, but increased the final shear modulus of the gels cooled to 30 °C. The impact of sorbitol on gel characteristics was attributed to its ability to increase protein thermal stability, increase the attractive force between proteins and decrease the protein–protein collision frequency.     

Influence of various preservatives on the quality of minced beef under modified atmosphere at chilled storage

The effect of preservatives on microbial quality, pH, drip-loss, roasting-loss, colour, and sensorial properties of modified atmosphere packaged (70% O₂ and 30% CO₂) minced beef (M. semimembranosus) stored at (2 ± 0.5 °C) for 12 days was investigated. Beef cubes (approx. 20 × 20 × 20 mm size) were immersed in solutions of 2% and 5% lactic acid, 2% lactic acid combined with 0.5% sodium ascorbate, 20% potassium lactate and 20% potassium sorbate before mincing. Addition of lactic acid was associated with pH drop, which increased drip-loss and roasting-loss. Application of all additives inhibited aerobic micro-organisms (10³–10⁴ CFU g⁻¹ on day 12) compared to reference sample (9 × 10⁵ CFU g⁻¹ on day 12). Lactic acid discoloured samples, while sodium ascorbate seemed to improve colour stability. Despite good visual colour characteristics, potassium sorbate treated samples were organoleptically unacceptable with massive off-flavour.     

Effect of season on color of Hanwoo (Korean native cattle) beef

A total of 1278 head of Hanwoo (Korean native cattle) slaughtered over four seasons were used to evaluate the effect of season on color characteristics of beef longissimus dorsi (LD) muscle. CIE L^*, a^*, b^*, C^* values and hue angle were significantly lower (P<0.05) in cattle slaughtered in the winter season. Meat color was darker in the winter than in the spring and autumn seasons. The L^* values among three average daily temperature (T_a) categories were different (P<0.05) in order of: [5 °CT_a<25 °C] > [T_a25 °C] > [T_a<5 °C], indicating that the meat color of cattle slaughtered at T_a<5 °C was darker. The a^*, b^*, C^* values and hue angle were significantly lower (P<0.05) in cattle slaughtered at T_a<5 °C. Season at slaughter is of great importance for meat color. Namely, meat color of Hanwoo beef was influenced by environmental temperature. Overall, cattle slaughtered in the winter season of T_a<5 °C produced beef with more undesirable meat color properties.     

Influence of weak organic acids and salts on the denaturation characteristics of intramuscular connective tissue. A differential scanning calorimetry study

Intramuscular connective tissue obtained from Longissimus dorsi muscle of a 4-year-old beef carcass was treated with NaCl solutions of 2, 4, and 6% (w/v), and CaCl₂ solutions of 50, 100, and 150 mM, and citric and lactic acid solutions of 0.5, 1.0, and 1.5% for three different marinating periods (24, 46, and 72 h). Changes in denaturation characteristics were investigated using differential scanning calorimetry and it was found that the denaturation onset temperature (T_o) and the denaturation peak temperature (T_p) increased as the NaCl concentration increased but decreased as the CaCl₂ concentration increased, irrespective of marinating time. Lactic and citric acid decreased T_o to about 39°C, from over 60°C breaking the structure of fibrils.     

A novel use of modified atmospheres: Storage insect population control

The research described here aimed to establish the feasibility of using modified atmospheres (MA) to protect commodities throughout their storage life by using oxygen (O₂) levels that disrupt the life cycles of the target beetle species. Rather than achieving complete mortality of all stages, the aim was to identify more easily obtainable MAs that would kill the most susceptible stage and prevent population growth. Simulated burner gas and nitrogen (N₂) atmospheres with O₂ contents between 3% and 6%, were tested, along with a N₂-based MA with elevated carbon dioxide (CO₂) (10–20%).

Laboratory tests were carried out on five species of stored-product beetles, Cryptolestes ferrugineus, Oryzaephilus surinamensis, Sitophilus granarius, S. oryzae and Tribolium castaneum. After exposure to the MAs for 28 d an assessment was made of the mortality of adults, the number of adults from progeny produced under the MAs and, for the simulated burner gas, the number of adults from progeny produced in a 28-d period after exposure to the MA. The tests were carried out at 20 and 25 °C with 75% and 85% r.h. at each temperature.

The O₂ content preventing population growth varied with species and temperature. For simulated burner gas or N₂ it was about 4% for O. surinamensis, S. granarius and S. oryzae, and about 3% for C. ferrugineus and T. castaneum at 25 °C. At 20 °C it was about 3% for all species tested. When CO₂ was increased to 10% or 20%, reducing O₂ to 5% was sufficient to eliminate emergence of S. granarius at 20°C, but a few individuals emerged at 25 °C. For C. ferrugineus there was a 95% reduction with 5% O₂ plus 20% CO₂ at 20 °C, but not at 25 °C.     

Design, fabrication, and testing of a returnable, insulated, nitrogen-refrigerated shipping container for distribution of fresh red meat under controlled CO2 atmosphere

In today's market, fresh red meat is cut and packaged at both the wholesale and retail level. Greater economies could result if the wholesaler prepared all consumer cuts centrally, but the short storage life of meat limits distribution. Use of CO₂-controlled atmosphere, master packaging, and strict temperature control (−1.5±0.5°C) can enhance storage life and, therefore, distribution ease. An insulated shipping and storage container was designed and tested for its suitability to distribute master-packaged meat. Shelves in the container supported 36 master trays (508 × 381 × 60 mm), with the source of refrigeration being injected liquid nitrogen (N₂). Electric fans dispersed the N₂ gas throughout the container. To reduce costs, 36 saline water bags (10% w/v NaCl) were used to thermally simulate the meat. Temperatures of 20 bags were recorded during storage experiments. The container was tested at outside temperatures of 15, 0 and −15°C with 4 internal fans and at 30°C with 2, 4 and 6 fans. In all instances, bags cooled from 10°C to an equilibrium temperature of −1.5°C within 5.5 h. Minimum equilibrium temperatures during any 8 h trial were −2.6, −2.0 and −2.0°C for 2, 4 and 6 fans, respectively. Correspondingly, maximum temperatures were −0.2, −0.7 and −0.3°C. Initial chilling of the product required, on average, 19 kg of N₂, while equilibrium was maintained at a N₂ consumption rate of 5.5, 4.0, 2.6 and 0.93 kg/h at outside temperatures of 30, 15 0 and −15°C, respectively, with 4 fans. The N₂ use for 2 and 6 fans was 5 and 6.3 kg/h, respectively, at an outside temperature of 30°C. During simulated power failure or when the N₂-tank ‘ran dry', temperatures in the container rose 0.9 and 2.0°C/h, respectively. When the door to the container was opened long enough to remove three trays, temperature was restored within 5 min. Convective heat transfer coefficients between saline water bags and circulating N₂ were in the range of 80–100, 115–135, and 140–155 W/(m²·K) for 2, 4 and 6 fans, respectively. Heat transfer to meat will be limited by conduction in master packaged meat if similar convection coefficients prevail.     

Modified stainless steel surfaces targeted to reduce fouling – Evaluation of fouling by milk components

Several stainless steel based surfaces with different properties were evaluated according to their fouling behaviour for different dairy products under different conditions. Surface properties were obtained by the following modification techniques: , and TiC ion implantation; diamond-like carbon (DLC) sputtering; DLC, DLC–Si–O and SiO_x, plasma enhanced chemical vapor Deposition (PECVD); autocatalytic Ni–P–PTFE and silica coating. Aqueous solutions that simulate milk (SMUF – simulated milk ultrafiltrate for the mineral components, β-lactoglobulin for the protein components and FMF – fouling model fluid for complex milk systems) were used to study the fouling behaviour during pasteurisation. Bacteriological deposition studies were also performed with two heat resistant strains of Bacillus. The experiments were carried out at laboratory scale for the evaluation of calcium phosphate and protein deposition, and at pilot scale for adhesion of bacteria and deposits from complex milk systems.

In all cases, the fouling behaviour was affected by the surface material, although in different ways for the deposition or the cleaning phases. For the non-microbiological deposits (calcium phosphate, whey protein and FMF milk-based product), the Ni–P–PTFE surface was the most promising one, since it generally promoted less deposit build up and, in all cases, was the easiest to clean. On the other hand, for bacterial adhesion, the most suitable surface was the ion implanted (TiC) surface, which also showed less spores after the cleaning process.     

Inactivation of polyphenol oxidases in cloudy apple juice exposed to supercritical carbon dioxide

The inactivation of polyphenol oxidase (PPO) in cloudy apple juice exposed to supercritical carbon dioxide (SCCO₂) treatment was investigated. Higher pressure, higher temperature, and longer treatment time caused more inactivation of PPO. The maximum reduction of PPO activity reached more than 60% at 30 MPa and 55 °C for 60 min. The experimental data followed first-order reaction kinetics; the kinetic rate constant k and the decimal reduction time D were closely related to the pressure and temperature of SCCO₂ treatment. Higher pressures or higher temperatures resulted in lower D values (higher k), the D value of PPO was minimized to 145 min treated by the combination of 30 MPa and 55 °C. Activation energy of 18.00 kJ /mol, was significantly reduced by SCCO₂ treatment at 30 MPa, as compared to activation energy of 72.0 kJ/mol for identical treatment at atmospheric pressure. Pressure and temperature sensitivity of kinetic parameters were studied. Z_P at 55 °C was 66.7 MPa and Z_T at 30 MPa was 108 °C.     

Thermal-death kinetics of fifth-instar Amyelois transitella (Walker) (Lepidoptera: Pyralidae)

Information on kinetics for thermal mortality of navel orangeworm, Amyelois transitella (Walker) (Lepidoptera: Pyralidae), is needed for developing post-harvest phytosanitation thermal treatments of walnuts. Thermal-death kinetics for fifth-instar navel orangeworms were determined at temperatures between 46°C and 54°C at a heating rate of 18°C min⁻¹ using a heating block system. Thermal-death curves for fifth-instar navel orangeworms followed a 0.5th-order of kinetic reaction. The time required to achieve 100% mortality (N₀=600) decreased with increasing temperature in a logarithmic manner. Complete kill of 600 insects required a minimum exposure time of 140, 50, 15, 6, and 1 min at 46°C, 48°C, 50°C, 52°C, and 54°C, respectively. The reaction rate (k) was affected by treatment temperatures following an Arrhenius relationship. The activation energy for thermal kill of fifth-instar navel orangeworms was estimated to be between 510 and 520 kJ mol⁻¹.     

Softness in dry-cured porcine biceps femoris muscles in relation to meat quality characteristics and processing conditions

The aim of the study was to quantify the effect of meat quality characteristics and some processing conditions on the softness of dry-cured biceps femoris (BF) muscles. The BF muscles were dissected from forty hams and classified according to their pH_BF into three groups: LpH (pH < 5.66), MpH (5.66  pH  6.00) and HpH (pH > 6.00). BF muscles within each pH_BF group were distributed into three different Salting levels (1%, 2% or 4% of added NaCl). Muscles were salted, vacuum-packed and stored at 3 °C for 30 days. The post-salting BF muscles were classified into two intramuscular fat (IMF) levels: Low (IMF < 4%) and High (IMF  4%). Thereafter, the muscles were divided into two pieces and dried at two of the three different Drying levels (1.5, 2 and 2.5 g H₂O/g desalted dry matter). Then, each piece was divided into two samples that were packed in N₂ and stored at 5 °C or 30 °C for 1 month. Stress Relaxation was used to evaluate texture. Dry-cured BF muscles with initial pH > 6.0, with IMF > 4% or with added NaCl levels less than 2% were more prone to show soft texture. Softness in dry-cured muscles can be reduced by applying an ageing temperature of 30 °C for 30 days, despite increasing proteolysis. The softness reduction by ageing at 30 °C compared with 5 °C is expected to be higher when applied to drier samples, which show a smaller increase in proteolysis.     

The effects of residual oxygen concentration and temperature on the degradation of the colour of beef packaged under oxygen-depleted atmospheres

Samples of beef longissimus dorsi (LD), approximately 5 × 5 × 1 cm, were packaged in pairs under 10 litre volumes of N₂ or CO₂ containing O₂ at concentrations between 100 and 1000 ppm. The packaged samples were stored at temperatures of 5, 1, 0 or −1·5°C, for times between 4 and 48 h. Samples of beef psoas major (PM) were packaged under N₂ or CO₂ containing O₂ at between 100 and 600 ppm, and stored at −1·5°C for 24 or 48 h. After storage, each sample was assessed for colour deterioration and discoloration, and for the fraction of metmyoglobin in the surface pigment.

The results obtained with N₂ and CO₂ atmospheres were similar. The colours of all LD samples had deteriorated after 4 h storage at 5 or 1°C, although the degree of deterioration increased with increasing O₂ concentration. All LD samples stored for 12 h at 5 or 1°C were extensively discoloured, with metmyoglobin fractions generally exceeding 60%, but those stored at −1·5°C for 48 h or less, under O₂ concentrations ≤ 400 ppm had undergraded colours. The colours of some LD samples stored at −1·5°C under about 600 ppm of O₂ were also undergraded, but the colours of samples stored under 800 or 1000 ppm had deteriorated by 24 h. The colours of LD samples stored at 0°C under > 200 ppm had deteriorated after 24 h storage, and the colours of samples stored under 100 ppm O₂ had deteriorated after 48 h storage. All PM samples were wholly discoloured after storage at −1·5°C. Evidently, the colour of beef muscle of high colour stability is resistant to degradation by atmospheres containing < 600 ppm of O₂ when the meat is stored at sub-zero temperatures, but not when the storage temperature is at or above 0°C. Beef muscle of low colour stability, such as the PM, will discolour at all low concentrations of O₂ irrespective of the storage temperature.     

Effect of carbon monoxide in modified atmosphere packaging, storage time and endpoint cooking temperature on the internal color of enhanced pork

The objective of this research was to evaluate the effects of gas atmosphere, refrigerated storage time, and endpoint temperature on internal cooked color of injection-enhanced pork chops. Enhanced chops were packaged in 0.36% CO/20.34% CO₂ (CO-MAP), 80% O₂/20% CO₂ (HO-MAP), or PVC-overwrapped (PVC-OW; controls), stored at 4 °C for 0, 12, 19 or 26 days, displayed for 2 days then cooked to six endpoint temperatures (54, 60, 63, 71, 77, and 82 °C). L^*, a^*, and b^* values, hue angle and chroma were determined on the internal cut surface of cooked chops. Chops packaged in CO-MAP had the highest a^* values; a^* value began increasing on day 14. The lowest hue angles occurred in chops cooked to lower endpoint temperatures. Chops in CO-MAP had lower hue angles and higher chroma than those in HO-MAP and PVC-OW. Above 71 °C, hue angle and chroma increased. Overall, CO-MAP packaged chops stored for longer time periods then cooked to lower endpoint temperatures appeared reddest. HO-MAP packaged chops were less red, did not change over time, and appeared more well done at lower endpoint temperatures than those in other gas atmospheres. CO-MAP packaged chops retained redness even after cooking at 82 °C.     

Enzymatic synthesis of cocoa butter analog through interesterification of lard and tristearin in supercritical carbon dioxide by lipase

Substrate oil composition, reaction time, acyl donor, temperature, and pressure affected the triacylglycerol (TG) content of cocoa butter analog during the interesterification reaction catalyzed by lipase in a supercritical carbon dioxide (SC-CO₂) system. Among oil sources used to interact with tristearin, the content of 1(3)-palmitoyl-3(1)-stearoyl-2-monoolein (POS) (P, palmitate; O, oleate; S, stearate) and 1-palmitoyl-2, 3-dioleoylglycerol (POO) in analog was most similar to the corresponding TG content of cocoa butter when analog was prepared with lard. The optimized interesterification reaction using lard and tristearin (at a mole ratio of 1.4) as substrates to produce cocoa butter analog in a SC-CO₂ system was at 17 MPa, 50 °C, pH 9, for 3 h with an immobilized lipase, Lipozyme IM-20, from Mucor miehei. The lyophilized enzyme facilitated the production of cocoa butter analog in anhydrous substrates (a_w 0.33). The yield and melting point of the purified cocoa butter analog by a silica column was 63% and 34.5 °C, respectively, when the analog was produced under optimal conditions.