CN110547464A - Antioxidant emulsion capable of improving bioavailability of curcumin and preparation method and application thereof - Google Patents

Antioxidant emulsion capable of improving bioavailability of curcumin and preparation method and application thereof Download PDF

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CN110547464A
CN110547464A CN201910899625.9A CN201910899625A CN110547464A CN 110547464 A CN110547464 A CN 110547464A CN 201910899625 A CN201910899625 A CN 201910899625A CN 110547464 A CN110547464 A CN 110547464A
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curcumin
emulsion
parts
bioavailability
oil
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王鹏
方艾虎
王越溪
徐幸莲
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Nanjing Agricultural University
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Nanjing Agricultural University
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105Plant extracts, their artificial duplicates or their derivatives
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/115Fatty acids or derivatives thereof; Fats or oils
    • A23L33/12Fatty acids or derivatives thereof
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17Amino acids, peptides or proteins
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs

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  • Food Science & Technology (AREA)
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  • Oil, Petroleum & Natural Gas (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

the invention discloses an antioxidant emulsion for improving the bioavailability of curcumin, and a preparation method and application thereof. The invention utilizes the ultrasonic dissolving-assisting method to dissolve curcumin in vegetable oil and construct high-concentration emulsion with non-meat protein, thereby achieving the effects of improving the bioavailability of curcumin and optimizing the fatty acid composition in emulsified intestines; the method effectively improves the condition that the emulsified sausage is easy to oxidize in the storage process, simultaneously meets the requirement of the current consumers for seeking fatty acid balanced emulsified sausage products, provides a method for improving the product formula for enterprises, has simple required equipment, simple and convenient operation and strong operability, and is suitable for wide popularization and application.

Description

Antioxidant emulsion capable of improving bioavailability of curcumin and preparation method and application thereof
Technical Field
The invention relates to an antioxidant emulsion capable of improving the bioavailability of curcumin, and a preparation method and application thereof, and belongs to the technical field of food processing.
Background
Curcumin is derived from plant turmeric and is a hydrophobic phenolic compound, and researches show that an enol structure and phenolic hydroxyl of curcumin play an important role in oxidation resistance. But because the curcumin is insoluble in water, insoluble in acidic conditions and unstable in neutral and alkaline conditions, the human body absorption rate is greatly reduced, thereby limiting the application of the curcumin in the industries of food and the like. Researches show that the stability and bioavailability of curcumin can be greatly improved by using an embedding system to prepare compounds such as gel, emulsion, particles, nanoparticles and the like. The biodegradability, biocompatibility, safety, sustainability, economic characteristics and the like of the protein provide favorable conditions for preparing the bioactive compound. Therefore, the protein is used as a carrier to carry curcumin, and the solubility, stability and bioavailability of curcumin can be effectively improved. At present, the selection of materials aiming at the embedding and conveying of curcumin in China mainly focuses on milk protein and plant protein, such as whey protein and soybean protein, and the egg white separated protein is not well applied as a by-product in the production of egg yolk processing products. In addition, with the increasing pursuit of healthy diet, the research of low-fat and non-fat foods has received more and more attention in recent years, and fat substitutes have gradually become important additives in the production process of low-fat and non-fat foods. The high-concentration emulsion prepared by using protein, water and vegetable oil as raw materials has excellent stability, solubility and water-retaining property, and is widely used for processing frankfurters and fermented sausages at foreign countries to improve the fatty acid composition. However, just because the product is rich in unsaturated fatty acid, the oxidation of the oil is more likely to occur.
a traditional nano emulsion embedding and conveying system has certain limitation, and the oil phase concentration is low, so that the embedding amount is limited, and the oxidation resistance is weak. In addition, fat substitutes do not have antioxidant factors, resulting in poor product storage properties, and lower concentrations of the internal phase in the emulsion can also affect the mouthfeel and flavor of the product.
Disclosure of Invention
The technical problem to be solved by the invention is to overcome the defects of the prior art and provide the antioxidant emulsion for improving the bioavailability of curcumin and the preparation method and application thereof, thereby solving the technical problems of low bioavailability of curcumin, easy oxidation and instability of fat substitutes.
in order to solve the technical problems, the invention provides an antioxidant emulsion for improving the bioavailability of curcumin, which is characterized by comprising the following raw materials in parts by weight:
the above-mentioned proportion can form an emulsion with high viscosity, good stability and good oxidation resistance.
Preferably, the vegetable oil is soybean oil, olive oil or corn oil
the invention also provides a preparation method of the antioxidant emulsion for improving the bioavailability of curcumin, which is characterized by comprising the following steps:
Preparation of dispersed phase: dissolving powdery curcumin in vegetable oil, performing ice bath ultrasonic dissolution promotion by using an ultrasonic cell disruptor, centrifuging to remove undissolved curcumin to obtain an oil phase in which curcumin is dissolved, and keeping out of the sun for later use;
Preparation of continuous phase: mixing the egg white separated protein powder with deionized water to ensure that the egg white separated protein powder is fully hydrated and dissolved to obtain a continuous phase of an emulsion;
preparation of an emulsion: adding the dispersed phase into the continuous phase, uniformly mixing the two phases to obtain a high-concentration emulsion, quickly cooling the obtained emulsion to room temperature, and storing the emulsion at low temperature in the dark.
Preferably, the concentration of the oil phase dissolved with curcumin is 1100-1300 mg/L, and the curcumin has better oxidation resistance in the range.
Preferably, the mass percentage of the egg white separated protein powder and the deionized water is 4% -6%, the protein concentration is higher than that of a common emulsion, the oil-water interface of the emulsion can be better stabilized, and the emulsifying property of the emulsion is enhanced.
Preferably, the volume fraction of the dispersed phase in the total emulsion is 65-85%, and the proportion endows the emulsion with very high viscosity, so that the emulsion becomes a purer elastic system, and the problems of juice loss, poor mouthfeel and the like in later application in an emulsion sausage are avoided.
Preferably, the two phases are mixed homogeneously by means of a shaker and a high-concentration emulsion is produced by shearing with a dispersion homogenizer at a speed of 13500r/min for 2.5 min. Under such high shear forces, the vegetable oil is broken up into uniform small droplets, increasing the surface area in contact with the protein, and thus the surface tension can be reduced more significantly, resulting in a more stable emulsion system.
The invention also provides a low-fat emulsified sausage which is characterized by comprising the following raw materials in parts by weight: 52-55 parts of lean pork, 4.7-9.4 parts of pig backfat, 18-20 parts of ice water, 14.1-18.7 parts of fat substitute, 1.41-1.61 parts of salt, 0.1-0.5 part of composite phosphate, 0.1-0.5 part of monosodium glutamate, 0.2-0.5 part of white pepper powder and 0.0005-0.005 part of sodium nitrite; the fat substitute is the emulsion of claim 1 or 2.
preferably, the feed comprises the following raw materials in parts by weight: 54.9 parts of lean pork, 5.8 parts of pig backfat, 19.5 parts of ice water, 17.6 parts of fat substitute, 1.61 parts of salt, 0.24 part of composite phosphate, 0.2 part of monosodium glutamate, 0.2 part of white pepper powder and 0.001 part of sodium nitrite.
the invention also provides application of the antioxidant emulsion for improving the bioavailability of curcumin in preparation of low-fat emulsified sausage.
The invention also provides a preparation method of the antioxidant low-fat emulsified sausage for improving the bioavailability of curcumin, and the specific production process is as follows:
1. Preparing raw and auxiliary materials:
Firstly, 54.9 parts by weight of lean pork is cleaned, connective tissue is removed, the lean pork is cut into pieces, 1.41-1.61 parts by weight of salt and 0.1-0.5 part by weight of composite phosphate are added, the mixture is pickled at 4 ℃ for 8-12h and then passes through a 10mm sieve meat grinder, 4.7-9.4 parts by weight of pork backfat the same time is cut into pieces of 1cm 3, the cut pieces are rinsed with warm water and then placed at 4 ℃ for 8-12 h;
2. Preparing a fat substitute:
Dissolving 0.096 part of powdery curcumin in 7.904 parts of linseed oil, performing ice-bath ultrasonic dissolution promotion (350-. Weighing 0.6 part of egg white separated protein powder, quantitatively mixing with 1.4 parts of water by weight, uniformly dispersing the protein powder in deionized water by a uniformly mixing instrument, and standing for 1 hour to fully hydrate and dissolve the protein powder; this solution served as the continuous phase of the high concentration emulsion. Adding 8 parts of oleum Lini containing curcumin into the continuous phase, and mixing the two phases with a shaker; shearing with dispersion homogenizer at 13500r/min for 2.5min (every 30s for 10s) to obtain high concentration emulsion, and refrigerating at 4 deg.C in dark place.
3. Chopping and mixing the ingredients:
cutting and mixing in three steps, namely, firstly, cutting the salted and twisted lean meat, uniformly scattering 0.1-0.5 part by weight of monosodium glutamate, 0.2-0.5 part by weight of white pepper and 40% of ice water into the cut meat, and cutting the cut meat into paste for 2 min; secondly, adding 4.7-9.4 parts by weight of pig backfat, 14.1-18.7 parts by weight of the fat substitute and 30% of ice water mixture, starting high-speed chopping, and chopping the pig backfat into fine particles for 2 min; thirdly, adding the rest 30 percent of ice water to begin high-speed chopping to prepare meat paste, and chopping until the meat paste is uniform, fine, sticky and glossy for 2 min;
4. standing the meat paste at 4 deg.C for 10 min;
5. Performing clysis:
filling by using a collagen casing with the diameter of 24mm, and manually controlling the filling amount to be 50g per section.
6. Low-temperature cooking:
(1) and (3) drying: keeping the drying temperature at 60-70 ℃ for 10-20 min;
(2) Smoking: the fuming temperature is 160-200 ℃, the temperature in the box is 60-70 ℃, and the time is 15-35 min;
(3) And (3) cooking: the cooking temperature is 70-90 ℃, and the time is 25-45 min.
7. heat dissipation and cooling:
The product is cooled in a refrigerator at 4 ℃ overnight after being cooled to room temperature.
8. And (3) vacuum packaging:
Quantitatively packaging the product by a continuous packaging machine and carrying out vacuum packaging; and producing products with different specifications according to market demands. The net content is strictly controlled and no negative bias is allowed to occur. And (4) adjusting the pattern position of the packaging bag to complete the packaging pattern of the product. Note that the longitudinal sealing temperature and the transverse sealing temperature are adjusted according to the thickness type of the packaging film, so that the packaging bag is firmly sealed in a heat sealing way. Cooling to room temperature, labeling, warehousing at 4 deg.C, and storing in dark place.
The invention achieves the following beneficial effects:
(1) the protein and oil concentration of the emulsion is higher than that of the emulsion of the common fat substitute, so that the effects of stabilizing an oil-water interface and enhancing the emulsifying performance are better embodied; the curcumin is embedded in the vegetable oil, and the oxidation resistance of the curcumin can effectively protect unsaturated fatty acid in the vegetable oil from being oxidized.
(2) The dissolution of the curcumin is promoted by a physical means, namely, the ultrasonic cell disruptor is used for ice bath ultrasonic dissolution promotion, the solubility of the curcumin in the vegetable oil reaches a saturated state, the curcumin can be brought into an emulsification system to the maximum extent, the solubility of the curcumin is greatly enhanced, and the high-concentration emulsion can well protect the antioxidant activity of the curcumin, so that the utilization rate of the curcumin is improved.
(3) the emulsion in the low-fat emulsified intestines and the myofibrillar protein in the lean pork form emulsion-myofibrillar protein mixed gel, and the three-dimensional network structure in the mixed gel increases the crosslinking degree of the interface layer protein around the dispersed liquid drops, so that the digestive effect of digestive juice on oil drops can be better prevented, the release of curcumin in the oil drops is delayed, and the decomposition amount of the curcumin in a digestive system is reduced.
(4) The curcumin-egg white protein isolate high-concentration emulsion prepared by the invention is used as a fat substitute, so that the fat content of a product can be reduced, the fatty acid composition of the product can be optimized, the intake of a natural antioxidant (curcumin) in a human body can be increased, and the health of the human body can be enhanced.
(5) The low-fat emulsified sausage prepared by the invention has the characteristics of high protein concentration, good oil phase quality, high concentration and addition of curcumin. The high protein concentration can enhance the ability of stabilizing the oil-water interface and enhance the stability of the emulsion; the oil phase concentration is increased to form a gel system with better texture, so that the emulsion is more stable, and the emulsion sausage is finally endowed with better mouthfeel; curcumin is a physiological active substance which prevents the added vegetable oil from generating oxidation reaction and provides health for human body.
Drawings
FIG. 1 shows the effect of different curcumin concentrations on DPPH in a high concentration emulsion;
FIG. 2 is a graph of the effect of different curcumin concentrations on the digestion products of a high concentration emulsion;
Figure 3 is a graph of the effect of different curcumin concentrations on egg white isolate-myofibrillar protein emulsion mixed gel digest products.
Detailed Description
The invention is further described below with reference to the accompanying drawings. The following examples are only for illustrating the technical solutions of the present invention more clearly, and the protection scope of the present invention is not limited thereby.
Example 1:
The lean pork is hind leg lean pork, 1400g of pork backfat, 500g of ice water, 480g of fat substitute (wherein 28.8g of ovalbumin, 383.5392g of linseed oil, 0.4608g of curcumin and 67.2g of water), 42g of salt, 6g of composite phosphate, 4g of monosodium glutamate, 6g of white pepper powder and 0.004g of sodium nitrite are weighed after connective tissues and surface fat are removed.
preparing a curcumin-egg white isolated protein high-concentration emulsion:
Dissolving 28.8g of the egg white isolated protein powder in 67.2g of water, fully dissolving, adding 0.4608g of curcumin into 383.5392g of linseed oil, dissolving by an ultrasonic cell disruptor (390W, 60min), then mixing the oil phase and the water phase in a stirrer at 13500r/min for 2min to prepare an egg white isolated protein-curcumin high-concentration emulsion, and refrigerating at 4 ℃ (<24h) for later use;
the preparation method of the sausage is completed according to the following steps:
pretreating raw meat, cleaning lean meat, removing connective tissue, cutting into pieces, adding above salt and compound phosphate, pickling at 4 deg.C for 8-12 hr, sieving with 10mm sieve meat grinder, cutting pig back fat into pieces of 1cm 3, rinsing with warm water, and standing at 4 deg.C for 8-12 hr;
Chopping and mixing the ingredients: firstly, cutting salted and twisted lean meat, uniformly scattering monosodium glutamate, white pepper powder and 40% ice water into a low-speed cutting knife and a cutting plate, and cutting the lean meat into paste for 2 min; secondly, adding pig backfat, the egg white isolated protein-curcumin high-concentration emulsion and 30% ice water mixture, starting high-speed chopping, and chopping the pig backfat into fine particles for 2 min; thirdly, adding the rest 30 percent of ice water to begin high-speed chopping until the meat stuffing is uniform, fine, sticky and glossy for 2 min;
Performing clysis: a collagen casing with the diameter of 24mm is adopted, and the amount of the casing is manually controlled, wherein each section is 50 g.
low-temperature cooking: the first step is as follows: drying at 65 deg.C for 15 min; the second step is that: steaming at 80 deg.C for 40 min.
Heat dissipation and cooling: the product is cooled in a refrigerator at 4 ℃ overnight after being cooled to room temperature.
vacuum packaging with a continuous packaging machine; cooling to room temperature, labeling, warehousing at 4 deg.C, and storing in dark place.
example 2:
the lean pork is hind leg lean pork, 1400g of pig backfat 180g, 500g of ice water, 420g of fat substitute (wherein 25.2g of ovalbumin, 335.5968g of linseed oil, 0.4032g of curcumin and 58.8g of water), 42g of salt, 6g of composite phosphate, 4g of monosodium glutamate, 6g of white pepper powder and 0.004g of sodium nitrite are weighed after connective tissues and surface fat are removed.
Preparing a curcumin-egg white isolated protein high-concentration emulsion:
dissolving 25.2g of the egg white isolated protein powder in 58.8g of water, fully dissolving, adding 0..4032g of curcumin into 335.5968g of linseed oil, dissolving by using an ultrasonic cell disruptor (390W, 60min), then mixing an oil phase and a water phase in a stirrer at 13500r/min for 2min to prepare an egg white isolated protein-curcumin high-concentration emulsion, and refrigerating at 4 ℃ (<24h) for later use;
The sausage was prepared in the same manner as in example 1.
example 3:
The lean pork is hind leg lean pork, 1400g of pork backfat, 500g of ice water, 480g of fat substitute (wherein 28.8g of ovalbumin, 383.5392g of linseed oil, 0.4608g of curcumin and 67.2g of water), 42g of salt, 6g of composite phosphate, 4g of monosodium glutamate, 6g of white pepper powder and 0.004g of sodium nitrite are weighed after connective tissues and surface fat are removed.
Preparing a curcumin-egg white isolated protein high-concentration emulsion:
Dissolving 28.8g of the egg white isolated protein powder in 67.2g of water, fully dissolving, adding 0.4608g of curcumin into 383.5392g of linseed oil, dissolving by an ultrasonic cell disruptor (390W, 60min), then mixing the oil phase and the water phase in a stirrer at 13500r/min for 2min to prepare an egg white isolated protein-curcumin high-concentration emulsion, and refrigerating at 4 ℃ (<24h) for later use;
The sausage was prepared in the same manner as in example 1.
Comparative example 4:
the lean pork is hind leg lean pork, 1400g of the lean pork is removed from connective tissues and surface fat, 600g of pig backfat, 500g of ice water, 42g of salt, 6g of composite phosphate, 4g of monosodium glutamate, 6g of white pepper powder and 0.004g of sodium nitrite are weighed.
the sausage was prepared in the same manner as in example 1.
TABLE 1 texture Properties of different groups of emulsion sausages
Note: different lower case shoulder letters indicate significant differences (P < 0.05).
and (3) analyzing a test result: in examples 1, 2 and 3, 33.33%, 30% and 20% of the backfat fat was replaced with the high concentration emulsion, respectively. From the texture measurement results of the different groups of emulsion intestines, the elasticity and the recovery of the emulsion intestines are improved after the high-concentration emulsion replacement, and the hardness and the chewiness in the example 3 are not obviously different from those of the control group without fat replacement. Therefore, the texture of the emulsified sausage after being replaced by the high-concentration emulsion can reach the level before replacement, and the elasticity and the resilience are enhanced.
Example 5: determination of curcumin antioxidant Properties in high concentration emulsion of ovalbumin (DPPH free radical scavenging ability)
Weighing 2mg of DPPH powder, dissolving in 10mL of absolute ethanol, diluting to 50mL of constant volume, preparing a DPPH ethanol solution with the final concentration of 0.1mM, storing in a brown bottle in a dark place, and using as soon as possible. After 1g of the curcumin high-concentration emulsion sample in example 1 was sufficiently dissolved in 20mL of anhydrous ethanol, the solution was centrifuged (3000Xg, 20min), and the supernatant was collected and used. The DPPH solution was diluted 1.75 times with absolute ethanol. Measuring 0.5mL of supernatant sample solution into a 10mL test tube, adding 3.5mL of diluted DPPH ethanol solution, mixing by vortex, standing in the dark for 30min, and measuring the absorbance value of the sample at 517 nm. Absolute ethanol was used as a blank control and the DPPH radical clearance was calculated according to the following formula:
DPPH radical scavenging ratio (%) - (1-A/A0). times.100
(1) wherein, A is the light absorption value of the supernatant sample at 517 nm;
(2) A0-absorbance of blank at 517 nm.
Curcumin has strong DPPH scavenging ability, but the solubility of curcumin in water is very poor, so that curcumin in the emulsion is extracted by absolute ethyl alcohol in an experiment to exert the capability of scavenging free radicals, and the DPPH free radical scavenging ability of high-concentration emulsions with different curcumin addition amounts is convenient to compare.
As shown in figure 1, the DPPH free radical clearance of the curcumin-embedded ovalbumin high-concentration emulsion is improved along with the increase of the curcumin concentration, and the free radical clearance of 26.01 percent of 800mg/L is improved to 63.84 percent of 1200 mg/L. In order to fully exert the antioxidant activity of curcumin, it is necessary to allow it to be released and dissolved from the food matrix. Early experiments showed that without curcumin being processed and encapsulated in the oil phase, DPPH clearance was much lower than that of the curcumin-ovalbumin high concentration emulsion after processing. Research shows that the curcumin can be protected from being degraded by embedding the protein, and the antioxidant activity of the curcumin can be maintained.
Example 6 determination of bioavailability of curcumin-Encapsulated emulsion of high concentration of ovalbumin (F B)
Accurately weighing 10mg of a curcumin sample, dissolving the curcumin sample in 100mL of chloroform, then 1, taking 25mL of 0.01mg/mL curcumin stock solution, mixing the curcumin stock solution with 25mL of chloroform to obtain a 0.005mg/L curcumin solution, taking 0, 2, 4, 6, 8 and 10mL of the curcumin solution with 0.005mg/L, diluting the curcumin solution to 10mL with chloroform to prepare a standard curcumin solution with mass concentration, measuring the absorbance of the standard solution at a wavelength of 419nm, and drawing a standard curve of the mass concentration of the standard curcumin sample by using the obtained absorbance (y is 26.448x +0.027, R 2 is 0.9983), wherein y represents absorbance, and x represents the curcumin concentration.
0.23g of the curcumin high-concentration emulsion of example 1 was added to 30mL of phosphate buffer solution to give a final oil phase concentration of 0.5 wt%, pH 7 of the solution was adjusted, the mixture was stirred at 100r/min in a water bath at 37 ℃ for 30min, and 1.5mL of NaCl (0.329g of phosphate buffer solution at pH 7, 37 ℃), 1mL of CaCl 2 (0.08325g of buffer solution at pH 7, 37 ℃), 2mL of bile salt (187.5mg of buffer solution at pH 7, 37 ℃), 2mL of pancreatic lipase (60mg of phosphate buffer solution at pH 7, 37 ℃), final volume of 37.5mL, 150mmol/L of NaCl, 20mmol/L of CaCl 2 20, 5mg/mL of bile salt, 1.6mg/mL of pancreatic lipase, pH 7, pH stat titration was performed in the emulsion using a pH-stat titrator, and the end point time was adjusted to be 7, titration time was adjusted to be 2.05M.
after in vitro digestion, the digest was centrifuged (12500rpm, 6 ℃, 30 min). After centrifugation the digest formed an opaque precipitate phase at the bottom, separated into a middle clear micellar phase, sometimes separated into an oily or creamy phase at the top. 4mL of the middle clear micellar phase was collected using a syringe, vortexed well with 4mL of chloroform, and then centrifuged at room temperature (1750rpm, 10min), collecting the centrifuged bottom chloroform layer while the top fraction was vortexed with 4mL of chloroform and centrifuged at room temperature (1750rpm, 10 min). The bottom chloroform layer of the second portion mixture was added to the first previously-retained chloroform layer, vortexed, and analyzed with a M2e multifunctional microplate reader at 425 nm. The absorbance values were zeroed using a cuvette with pure chloroform as a reference and the absorbance values of curcumin in the chloroform layer were determined. Finally, the concentration of curcumin extracted from the sample was determined according to a standard curve of curcumin concentration in chloroform.
As can be seen from figure 2, when the curcumin concentration is less than 1100mg/L, the curcumin content in the mixed micelle phase after digestion of the ovalbumin isolated protein emulsion gradually increases along with the increase of the curcumin concentration in the oil phase, and the trend is very obvious. When the curcumin concentration of the oil phase in the emulsion exceeds 1100mg/L, the increase trend of the curcumin content in the mixed micelle phase becomes slow, and the curcumin utilization rate in the sample is far higher than that of the curcumin which is not embedded in the emulsion.
Example 7 determination of bioacceptability of an ovalbumin high concentration emulsion-myofibrillar protein Mixed gel to entrap curcumin (F B)
1. Preparation of an ovalbumin high-concentration emulsion-myofibrillar protein mixed gel:
(1) 300g of chicken breast was weighed, connective tissue was removed, cut into small pieces and placed in crushed ice.
(2) mincing with a meat mincer into meat paste, adding 1.2L of standard salt solution (20mmol/L K2HPO4/KH2PO4, 100mmol/LKCl, 2mmol/LMgCl2, 1mmol/LEGTA, pH 7.0).
(3) Homogenizing the meat paste solution for 30S by a homogenizer, filtering the obtained meat paste with a 20-mesh filter screen, centrifuging (1500Xg for 10min), taking the centrifuged precipitate, and repeating the same steps twice.
(4) Adding 1.2L KCl (0.1M) into the precipitate obtained in the final treatment in (3), homogenizing with a homogenizer for 30S, centrifuging (2000Xg, 10min), collecting the centrifuged precipitate, repeating the same steps twice to obtain 115mg/mL myofibrillar protein solution, and placing in a refrigerator at 4 deg.C for use.
(5) 30g of the curcumin-embedded ovalbumin high-concentration emulsion was mixed with 18mL of a myofibrillar protein solution, homogenized twice with a homogenizer (7000r/min, 30S,) and centrifuged (2000Xg, 10 min.) to remove air bubbles from the mixture.
(6) the emulsion mixture was transferred to a 100mL beaker and heated in a water bath (2 ℃/min, 80 ℃ for 10min) and the resulting gel sample was transferred to a 4 ℃ freezer for use.
2. Determination of bioacceptability of an ovalbumin high concentration emulsion-myofibrillar protein Mixed gel to entrap curcumin (F B):
A digest was prepared and the gel subjected to simulated digestion experiments using a slightly modified protocol from Minekus et al, gel sample 0.25g was added to 6mL of artificial saliva to give a final concentration of oil in solution of 0.5 wt%, the pH of the solution was adjusted to 7, the solution was stirred in a 37 ℃ water bath at 100r/min for 30min, 1.5mL of NaCl (0.329g in phosphate buffer at pH 7, 37 ℃), 1mL of CaCl2(0.08325g in buffer at pH 7, 37 ℃), 2mL of bile salt (187.5mg in buffer, pH 7, 37 ℃), 2mL of pancreatic lipase (60mg in phosphate buffer, pH 7, 37 ℃), a final volume of 37.5mL, NaCl 150mmol/L, CaCl 2 20mmol/L, 5mg/mL of bile salt, pancreatic lipase 1.6mg/mL, pH 7, and the pH was titrated using a-t titrator to achieve in vitro dissolution of oil in the pH material at pH 7, an endpoint of 2.05M, and a titration time of NaOH was 0.05M.
After in vitro digestion, the digest was centrifuged (12500rpm, 6 ℃, 30 min). After centrifugation the digest formed an opaque precipitate phase at the bottom, separated into a middle clear micellar phase, sometimes separated into an oily or creamy phase at the top. 4mL of the middle clear micellar phase was collected using a syringe, vortexed well with 4mL of chloroform, and then centrifuged at room temperature (1750rpm, 10min), collecting the centrifuged bottom chloroform layer while the top fraction was vortexed with 4mL of chloroform and centrifuged at room temperature (1750rpm, 10 min). The bottom chloroform layer of the second portion mixture was added to the first previously-retained chloroform layer, vortexed, and analyzed with a M2e multifunctional microplate reader at 425 nm. The absorbance values were zeroed using a cuvette containing pure chloroform as a reference and the absorbance values of curcumin in the chloroform layer were determined. Finally, the concentration of curcumin extracted from the sample was determined according to a standard curve of curcumin concentration in chloroform.
By comparing fig. 3 with fig. 2, it can be found that the utilization rate of curcumin can be improved, the release time in the gastrointestinal tract digestion process can be delayed, and the utilization rate can be increased by preparing the ovalbumin-myofibrillar protein mixed gel. As shown in the graphs in FIGS. 2 and 3, the curcumin contents in the neutralized mixed micelle and the emulsified mixed micelle in the protein gel with the addition of 800mg/L of curcumin are 0.214mg/g and 0.200mg/g respectively, which indicates that the gel structure can increase the bioavailability of curcumin in human body.
since the present invention is primarily applied to the manufacture of emulsified sausages, for conventional emulsions, lipid digestion depends on interfacial properties, oil phase surface area and the ratio of oil to lipase/bile salts. In this case, the preparation of the gel is very important. From the experimental results it can be seen that the gel structure is better able to regulate the digestion of fats in the digestive system. The preparation of the gel increases the formation of an internal three-dimensional network structure, increases the crosslinking degree of interface layer protein around dispersed liquid drops, and avoids premature digestion of grease. Meanwhile, the gel network structure can effectively prevent solute, bile salt or digestive enzyme from diffusing to the surface of oil drops embedded in the gel, thereby delaying the decomposition of the solute, bile salt or digestive enzyme and enabling the solute, bile salt or digestive enzyme to reach the small intestine as far as possible. The results, in contrast to emulsion digestion, show that the curcumin content in the digestion product mixed micelles is higher under the gel structure, which is proving that the gel structure has stronger protective effect on curcumin.
The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.

Claims (10)

1. An antioxidant emulsion for improving the bioavailability of curcumin is characterized by comprising the following raw materials in parts by weight:
0.5 to 0.7 portion of egg white protein isolate powder,
7.5 to 8.5 portions of vegetable oil,
0.09 to 0.098 portion of curcumin,
1.2-1.5 parts of water.
2. The antioxidant emulsion for improving bioavailability of curcumin of claim 1, wherein said vegetable oil is soybean oil, olive oil or corn oil.
3. A preparation method of an antioxidant emulsion for improving the bioavailability of curcumin is characterized by comprising the following steps:
Preparation of dispersed phase: dissolving powdery curcumin in vegetable oil, performing ice bath ultrasonic dissolution promotion by using an ultrasonic cell disruptor, centrifuging to remove undissolved curcumin to obtain an oil phase in which curcumin is dissolved, and keeping out of the sun for later use;
Preparation of continuous phase: mixing the egg white separated protein powder with deionized water to ensure that the egg white separated protein powder is fully hydrated and dissolved to obtain a continuous phase of an emulsion;
preparation of an emulsion: adding the dispersed phase into the continuous phase, uniformly mixing the two phases to obtain a high-concentration emulsion, quickly cooling the obtained emulsion to room temperature, and storing the emulsion at low temperature in the dark.
4. the method for preparing an antioxidant emulsion with enhanced bioavailability of curcumin as claimed in claim 3, wherein the concentration of the oil phase with curcumin dissolved therein is 1100-1300 mg/L.
5. The method for preparing an antioxidant emulsion capable of improving the bioavailability of curcumin as claimed in claim 3, wherein the mass percentage of the egg white isolated protein powder to the deionized water is 4% -6%.
6. the method for preparing an antioxidant emulsion with enhanced bioavailability of curcumin as claimed in claim 3, wherein the volume fraction of the dispersed phase in the total emulsion is 65% -85%.
7. The method for preparing an antioxidant emulsion with improved bioavailability of curcumin as claimed in claim 3, wherein the two phases are mixed uniformly by using a shaker, and the mixture is sheared by using a dispersion homogenizer at 13500r/min for 2.5min to obtain a high concentration emulsion.
8. A low-fat emulsified sausage is characterized by comprising the following raw materials in parts by weight: 52-55 parts of lean pork, 4.7-9.4 parts of pig backfat, 18-20 parts of ice water, 14.1-18.7 parts of fat substitute, 1.41-1.61 parts of salt, 0.1-0.5 part of composite phosphate, 0.1-0.5 part of monosodium glutamate, 0.2-0.5 part of white pepper powder and 0.0005-0.005 part of sodium nitrite; the fat substitute is the emulsion of claim 1 or 2.
9. The low-fat emulsion sausage according to claim 8, which comprises the following raw materials in parts by weight: 54.9 parts of lean pork, 5.8 parts of pig backfat, 19.5 parts of ice water, 17.6 parts of fat substitute, 1.61 parts of salt, 0.24 part of composite phosphate, 0.2 part of monosodium glutamate, 0.2 part of white pepper powder and 0.001 part of sodium nitrite.
10. use of the antioxidant emulsion of claim 1 or 2 for increasing curcumin bioavailability in the preparation of a low fat emulsified sausage.
CN201910899625.9A 2019-09-23 2019-09-23 Antioxidant emulsion capable of improving bioavailability of curcumin and preparation method and application thereof Pending CN110547464A (en)

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Application publication date: 20191210