The main aim of this study was the development of lactase microcapsules, which are normally hydrolyzed only in the small intestine, using an enteric coating microencapsulation technique. These could be used to supplement into milk, for the prevention of lactose intolerance. The microcapsules develop...
The main aim of this study was the development of lactase microcapsules, which are normally hydrolyzed only in the small intestine, using an enteric coating microencapsulation technique. These could be used to supplement into milk, for the prevention of lactose intolerance. The microcapsules developed contained a core material of lactase, a medium-chain triglyceride (MCT) as a primary coating material, and hydroxypropyl methylcellulose phthalate (HPMCP), shellac, and zein secondary enteric coating materials. In addition, polyglycerol polyricinoleate (PGPR) and polyoxyethylene sorbitan monolaurate (PSML) were used as primary and secondary emulsifiers, respectively.
First, the optimum conditions for lactase microencapsulation and physicochemical properties of the capsules were investigated. The amount of plasticizer required was investigated by dynamic vapor sorption (DVS). The most efficient conditions for enteric-coated microencapsulation of lactase, yield of microencapsulation, in vitro release rate of core material, and hydrolysis rate of capsules in milk during storage (stability) were investigated by response surface methodology (RSM). The optimum plasticizer to enteric coating material ratio was determined to be 0.5:1. The optimum concentration of coating material and the amount of core material required were found to be 4.85 g/100 mL and 1.85 mL, respectively. The average size of the microcapsules obtained under optimal conditions was revealed to be approximately 2~5 µm. All microcapsules were discovered to evenly distribute the encapsulated lactase.
Second, the effect of moisture on lactase microcapsules during storage was evaluated. This was determined by moisture absorption and desorption analysis using an automated DVS analyzer under conditions of 0~90% relative humidity. The storage period of the capsules was predicted by an accelerated shelf life test (ASLT) conducted at 30°C, 40°C, and 50°C for 90 days, with the storage period being calculated by the Arrhenius equation. The isotherm profiles of the lactase microcapsule powder coated with HPMCP and shellac were revealed to belong to types II and III, respectively. The percentage of absorbed moisture of lactase microcapsule powder coated with HPMCP and shellac were determined to be approximately 2.88% and 38.06%, respectively. The desorption behaviors were also observed to be distinct for each type of coating. The absorbed moisture was completely desorbed from the microcapsules when coated with HPMCP (0%), whereas a small percentage of the absorbed moisture remained in the microcapsules coated with shellac (3.78%). The activation energies of HPMCP- and shellac-coated, and uncapsulated lactase were calculated (using the Arrhenius equation) to be 12,964.0667, -12,134.4570, and -6,786.1699 Kcal/mol, while the predicted half-lives at 4°C and 25°C were calculated to be 27.95, 12.25, and 3.33 months, and 5.31, 2.59, and 1.52 months, respectively. The storage periods of lactase microcapsules coated with HPMCP and shellac were approximately 9 and 4-times (at 4°C), and 3.4 and 1.7-times (at 25°C) longer than that of uncapsulated lactase, respectively.
The third part of the study focused on determining the physicochemical properties of milk containing the lactase microcapsules. In order to determine the stability of the microcapsules in milk during storage, the residual lactose content and pH of the milk was determined. In addition, the changes in color and sensory characteristics of milk during storage were evaluated, in order to determine the properties of the microcapsule-supplemented milk. The pH values of the supplemented milk were similar to those of the control; however, the pH of the HPMCP-treated milk was slightly lower than that in the other samples. The release of lactase (core material) from the microcapsules to the milk led to hydrolysis of approximately 18.82% (HPMCP-coated) and 35.09% (shellac-coated) of lactose during the 12 d storage period. A sensory analysis revealed that the sweetness and off-taste of milk supplemented with microcapsules coated with HPMCP was similar to that of the control up to 8 d (p < 0.05). It was also determined that the addition of enteric-coated lactase did not affect the color of the supplemented milk.
Finally, the in vitro and in vivo release properties of the capsules were investigated in this study. In order to determine the in vitro release properties of the lactase microcapsules in simulated gastric and intestinal conditions, the microcapsules were hydrolyzed with simulated gastric fluid (SGF) and simulated intestinal fluid (SIF), respectively, at 37°C and 100 rpm for 2 h in a shaking water bath. Twenty subjects with lactose intolerance were selected for the clinical (in vivo) study. The blood glucose of the subjects was measured at 30 min intervals post-ingestion of 500 mL of the capsule-supplemented milk after overnight fasting. The capsules released only a small quantity of the core material in the simulated gastric conditions. On the other hand, the capsules coated with HPMCP showed an approximate 70~90% release of lactase, while the shellac-coated capsules released 80~90% of the core material in the simulated intestinal conditions. The blood glucose concentration showed an increase of approximately 30 mg/dL (from fasting levels) or more within 60 min after ingestion of the milk in the clinical test. The subjects rarely exhibited the symptoms caused by lactose intolerance.
Based on the results of this study, the use of enteric-coated lactase microcapsule-supplemented milk was determined to effectively prevent lactose intolerance. Therefore, this method appears to be a promising strategy for the prevention of lactose intolerance.
The main aim of this study was the development of lactase microcapsules, which are normally hydrolyzed only in the small intestine, using an enteric coating microencapsulation technique. These could be used to supplement into milk, for the prevention of lactose intolerance. The microcapsules developed contained a core material of lactase, a medium-chain triglyceride (MCT) as a primary coating material, and hydroxypropyl methylcellulose phthalate (HPMCP), shellac, and zein secondary enteric coating materials. In addition, polyglycerol polyricinoleate (PGPR) and polyoxyethylene sorbitan monolaurate (PSML) were used as primary and secondary emulsifiers, respectively.
First, the optimum conditions for lactase microencapsulation and physicochemical properties of the capsules were investigated. The amount of plasticizer required was investigated by dynamic vapor sorption (DVS). The most efficient conditions for enteric-coated microencapsulation of lactase, yield of microencapsulation, in vitro release rate of core material, and hydrolysis rate of capsules in milk during storage (stability) were investigated by response surface methodology (RSM). The optimum plasticizer to enteric coating material ratio was determined to be 0.5:1. The optimum concentration of coating material and the amount of core material required were found to be 4.85 g/100 mL and 1.85 mL, respectively. The average size of the microcapsules obtained under optimal conditions was revealed to be approximately 2~5 µm. All microcapsules were discovered to evenly distribute the encapsulated lactase.
Second, the effect of moisture on lactase microcapsules during storage was evaluated. This was determined by moisture absorption and desorption analysis using an automated DVS analyzer under conditions of 0~90% relative humidity. The storage period of the capsules was predicted by an accelerated shelf life test (ASLT) conducted at 30°C, 40°C, and 50°C for 90 days, with the storage period being calculated by the Arrhenius equation. The isotherm profiles of the lactase microcapsule powder coated with HPMCP and shellac were revealed to belong to types II and III, respectively. The percentage of absorbed moisture of lactase microcapsule powder coated with HPMCP and shellac were determined to be approximately 2.88% and 38.06%, respectively. The desorption behaviors were also observed to be distinct for each type of coating. The absorbed moisture was completely desorbed from the microcapsules when coated with HPMCP (0%), whereas a small percentage of the absorbed moisture remained in the microcapsules coated with shellac (3.78%). The activation energies of HPMCP- and shellac-coated, and uncapsulated lactase were calculated (using the Arrhenius equation) to be 12,964.0667, -12,134.4570, and -6,786.1699 Kcal/mol, while the predicted half-lives at 4°C and 25°C were calculated to be 27.95, 12.25, and 3.33 months, and 5.31, 2.59, and 1.52 months, respectively. The storage periods of lactase microcapsules coated with HPMCP and shellac were approximately 9 and 4-times (at 4°C), and 3.4 and 1.7-times (at 25°C) longer than that of uncapsulated lactase, respectively.
The third part of the study focused on determining the physicochemical properties of milk containing the lactase microcapsules. In order to determine the stability of the microcapsules in milk during storage, the residual lactose content and pH of the milk was determined. In addition, the changes in color and sensory characteristics of milk during storage were evaluated, in order to determine the properties of the microcapsule-supplemented milk. The pH values of the supplemented milk were similar to those of the control; however, the pH of the HPMCP-treated milk was slightly lower than that in the other samples. The release of lactase (core material) from the microcapsules to the milk led to hydrolysis of approximately 18.82% (HPMCP-coated) and 35.09% (shellac-coated) of lactose during the 12 d storage period. A sensory analysis revealed that the sweetness and off-taste of milk supplemented with microcapsules coated with HPMCP was similar to that of the control up to 8 d (p < 0.05). It was also determined that the addition of enteric-coated lactase did not affect the color of the supplemented milk.
Finally, the in vitro and in vivo release properties of the capsules were investigated in this study. In order to determine the in vitro release properties of the lactase microcapsules in simulated gastric and intestinal conditions, the microcapsules were hydrolyzed with simulated gastric fluid (SGF) and simulated intestinal fluid (SIF), respectively, at 37°C and 100 rpm for 2 h in a shaking water bath. Twenty subjects with lactose intolerance were selected for the clinical (in vivo) study. The blood glucose of the subjects was measured at 30 min intervals post-ingestion of 500 mL of the capsule-supplemented milk after overnight fasting. The capsules released only a small quantity of the core material in the simulated gastric conditions. On the other hand, the capsules coated with HPMCP showed an approximate 70~90% release of lactase, while the shellac-coated capsules released 80~90% of the core material in the simulated intestinal conditions. The blood glucose concentration showed an increase of approximately 30 mg/dL (from fasting levels) or more within 60 min after ingestion of the milk in the clinical test. The subjects rarely exhibited the symptoms caused by lactose intolerance.
Based on the results of this study, the use of enteric-coated lactase microcapsule-supplemented milk was determined to effectively prevent lactose intolerance. Therefore, this method appears to be a promising strategy for the prevention of lactose intolerance.
주제어
#β-D-galactosidase
#lactose intolerance
#microencapsulation
#enteric coating
#RSM
#storage prediction
#glucose blood test
#clinical study
#in vitro
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