CN115462493B - Composite nano particle based on plant polypeptide encapsulation taste masking and beverage obtained by composite nano particle - Google Patents
Composite nano particle based on plant polypeptide encapsulation taste masking and beverage obtained by composite nano particle Download PDFInfo
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- 229920001184 polypeptide Polymers 0.000 title claims abstract description 76
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 76
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 76
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 53
- 239000002131 composite material Substances 0.000 title claims abstract description 50
- 238000005538 encapsulation Methods 0.000 title claims abstract description 17
- 235000013361 beverage Nutrition 0.000 title claims abstract description 13
- 235000019640 taste Nutrition 0.000 title abstract description 19
- 230000000873 masking effect Effects 0.000 title abstract description 13
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- 239000013543 active substance Substances 0.000 claims abstract description 29
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 28
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- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
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- 229920002472 Starch Polymers 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
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- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
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- 230000000595 bitter masking effect Effects 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
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- SJWWTRQNNRNTPU-ABBNZJFMSA-N fucoxanthin Chemical compound C[C@@]1(O)C[C@@H](OC(=O)C)CC(C)(C)C1=C=C\C(C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)C(=O)C[C@]1(C(C[C@H](O)C2)(C)C)[C@]2(C)O1 SJWWTRQNNRNTPU-ABBNZJFMSA-N 0.000 description 1
- AQLRNQCFQNNMJA-UHFFFAOYSA-N fucoxanthin Natural products CC(=O)OC1CC(C)(C)C(=C=CC(=CC=CC(=CC=CC=C(/C)C=CC=C(/C)C(=O)CC23OC2(C)CC(O)CC3(C)C)C)CO)C(C)(O)C1 AQLRNQCFQNNMJA-UHFFFAOYSA-N 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 210000003026 hypopharynx Anatomy 0.000 description 1
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- 230000007246 mechanism Effects 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
- A23L2/66—Proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, 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
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/84—Flavour masking or reducing agents
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention provides a composite nanoparticle based on plant polypeptide encapsulation taste masking and a beverage obtained by the composite nanoparticle, and belongs to the technical field of food chemistry. The plant polypeptide encapsulation taste masking-based composite nano-particle provided by the invention is obtained by encapsulating hydrophilic active substance seaweed ferment powder by adopting plant polypeptides with different molecular weights. The encapsulation taste masking mode provided by the invention is original, and the encapsulation mode not only can mask unpleasant smell of active substances, but also provides good protection for chemical stability of the active substances in the storage process.
Description
Technical Field
The invention belongs to the technical field of food chemistry, and particularly relates to a composite nanoparticle based on plant polypeptide encapsulation taste masking and a beverage obtained by the composite nanoparticle.
Background
The development of bitter fishy smell can be understood from the point of view of the eater, for example, plants evolve unpleasant-smelling compounds to prevent consumption. The bitter and fishy smell compound products are various in variety and structure. At the same time, their receptor systems are very complex and can respond to such a wide range of potential stimuli at very low levels. Humans have about 25 intact bitter fishy receptors, 21 of which have been identified. With the development of modern medicine and food technology, these substances with bitter and fishy tastes prove to have strong biological activity and are of great benefit to human health.
Challenges faced by the pharmaceutical and food industries: the need for bitter masking formulations for pharmaceuticals is particularly acute, and european regulations require the development of pediatric development programs, where controlling bitter and unpleasant tastes is an important issue. Common taste masking schemes include chemical, central control, encapsulation, and the like.
The packaging method is characterized in that a stable raw material is used as a wall material, active substances with unstable chemical properties are packaged, and then the active substances are effectively delivered to the intestinal tract for release, so that the packaging method is a current research hot spot in the fields of modern medical foods and the like. Common wall materials are, for example, casein (Koo, mok et al 2016), starch (Oliyaei, moossavi-Nasab et al 2020), chitosan (Malgarim Cordenonsi, faccendini et al 2019), whey protein (Zhu, sun et al 2017), zein (Li, xu et al 2018), and the like. The packaging principle is as follows: the wall material generally has both hydrophilic groups and hydrophobic groups, and the hydrophilic groups and the water molecules are combined to form a circular cavity at one end of the hydrophobic groups. When it is combined with a hydrophobic active substance, the hydrophobic group of the active substance enters the inside of the cavity, forms a stable complex with the hydrophobic group of the wall material, such as a molecular bond, a chemical bond, and a hydrogen bond, and then dissolves in water together with the wall material, as shown in fig. 1. Therefore, most of the wall materials only encapsulate hydrophobic active substances, but not hydrophilic active substances, and the research is still blank at present.
The seaweed ferment powder is extracted from brown algae, red algae, green algae and the like, belongs to natural food, and has the main component of seaweed oligosaccharide and obvious functions in the aspects of resisting tumor, resisting inflammation, scavenging free radical activity, resisting obesity, resisting oxidation, resisting fatigue and the like (the seaweed ferment powder is prepared and has the functions of CN 201510159810.6). Therefore, the seaweed ferment powder has great potential value and application prospect in the food industry. However, as with many marine products, seaweed ferment powder has unpleasant fishy smell, which is particularly evident after long-term illumination, and largely limits its application in the beverage field. The trehalose is a hydrophilic active substance, and its solubility in water is 5%, so that it cannot be effectively encapsulated using conventional encapsulation methods.
Zein not only possesses more polar functional groups, but also has self-assembly properties, i.e., it can be induced to automatically form nanoparticles by changing the polarity of the solvent, for encapsulating the active substance (Wang and Padua 2010). Zein itself is not stable in performance, has an isoelectric point of 6.2, i.e., when the pH is in the range of 5-6, a large amount of precipitate is automatically generated and is irreversible, so that it cannot be used as a wall material alone to encapsulate an active substance. Often researchers use them with other proteins or polysaccharides as composite wall materials. Zhang Xin et al, 2022, used a single zein for the first time to encapsulate the hydrophobic active substance fucoxanthin, and was very effective (Zhang, fan et al 2022, journal of Industrial and Engineering Chemistry). In the experiment, zein is firstly hydrolyzed into polypeptides with different molecular weights, and then the amphipathy of the polypeptides is utilized to complete the encapsulation of the hydrophobic active substances. However, there is no report on the encapsulation of hydrophilic active materials. Therefore, it would be a considerable topic to develop an effective taste masking of hydrophilic active substances while at the same time guaranteeing their chemical stability.
Disclosure of Invention
The invention provides a plant polypeptide-based encapsulated taste-masking composite nanoparticle and a beverage obtained by the same, wherein plant polypeptides with different molecular weights are adopted in the composite nanoparticle to encapsulate hydrophilic active substance seaweed ferment powder to achieve the effect of masking the taste, so that unpleasant smell of active substances can be masked, and good protection is provided for chemical stability of the active substances in the storage process.
In order to achieve the aim, the invention provides a plant polypeptide encapsulation taste-masking based composite nanoparticle, which is obtained by encapsulating hydrophilic active substance seaweed ferment powder by adopting plant polypeptides with different molecular weights.
Preferably, the plant polypeptides with different molecular weights are prepared by the following method:
dissolving zein in deionized water to a concentration of 1% -3%, adjusting the pH value to 9.0-9.5, and controlling the pH value to be 9.0-9.5 all the time by using an automatic potentiometric titrator;
adding protease, and controlling the enzymolysis temperature to be 50 ℃;
the incomplete hydrolysis method is adopted, after hydrolysis is carried out for 1 to 1.5 hours, 1M hydrochloric acid is immediately used for adjusting the pH value of the enzymolysis liquid to 7.0 to 8.0, and the enzymolysis liquid is heated for 5 to 10 minutes at 95 ℃ to inactivate enzymes;
centrifuging the prepared polypeptide at 10000r/min for 20min to remove a small amount of insoluble substances, dialyzing the supernatant with a dialysis bag of 100Da for 24h to desalt, and freeze-drying the polypeptide liquid to obtain plant polypeptide.
Preferably, the protease is added in a mass ratio of 2:100 to the substrate, wherein the protease is of type 2.4L, P4860 (Sigma-Aldrich).
Preferably, the molecular weight of the plant polypeptide comprises a small molecular weight fragment selected from the range of molecular weights 200-599 and a large molecular weight fragment selected from the range of 600-1400.
Preferably, the diameter of the nano particles of the composite nano particles is less than or equal to 200nm, the dispersion coefficient PDI is less than or equal to 0.4, and the absolute value of the potential is more than or equal to 30mv.
Preferably, the composite nanoparticle exhibits the following functional groups in the infrared spectrum:
O-H stretching: 3303.+ -. 60cm -1 The method comprises the steps of carrying out a first treatment on the surface of the C-H stretching: 2924+ -50 cm -1 The method comprises the steps of carrying out a first treatment on the surface of the And characteristic peak amide bands of zein: 1651+ -30 cm -1 The method comprises the steps of carrying out a first treatment on the surface of the But is provided withCharacteristic peaks of seaweed ferment powder are not included: C-H bending vibration 1392+ -50 cm -1 And C-O stretching vibration 1024+ -50 cm -1 。
Preferably, the composite nanoparticle is prepared by the following method:
dissolving seaweed ferment powder in water at 1-3mg/mL, magnetically stirring for 0.5-2h, adding plant polypeptide in proportion after the seaweed ferment powder is fully dissolved, continuously stirring for 1-1.5h, centrifuging to remove insoluble substances, and filtering by using a 420 μm water-based membrane to obtain the composite particle nanometer.
Preferably, the mass ratio of the plant polypeptide to the seaweed ferment powder added into the composite nano particles is 1:5 < 1:2 or less.
The invention also provides a beverage, which comprises the composite nano particles based on plant polypeptide encapsulation taste masking according to any one of the technical schemes.
Preferably, when the mass ratio of the plant polypeptide to the seaweed ferment powder added into the composite nano particles is 1:5 < 1:2, the obtained beverage has no fishy smell and smooth and no astringent feel.
Compared with the prior art, the invention has the advantages and positive effects that:
the invention provides a composite nanoparticle based on encapsulation and taste masking of plant polypeptides and a beverage obtained by the composite nanoparticle, wherein plant polypeptides with different molecular weights are adopted in the composite nanoparticle to encapsulate hydrophilic active substance seaweed ferment powder to achieve the effect of taste masking.
Drawings
FIG. 1 is a schematic illustration of a single wall encapsulated hydrophobic active;
FIG. 2 is a mass spectrum of the plant polypeptide provided by the invention after 1-1.5 hours hydrolysis;
fig. 3 is a schematic diagram of 15 weeks monitoring results of particle size of composite nanoparticles prepared according to the A, B protocol provided by the present invention;
fig. 4 is a schematic diagram of 15 weeks monitoring results of PDI of the composite nanoparticle prepared according to the A, B scheme provided by the present invention;
FIG. 5 is a schematic diagram showing the 15-week monitoring result of the potential of the composite nanoparticle prepared according to the A, B scheme provided by the invention;
FIG. 6 is a Fourier transform infrared spectrum provided by the invention, wherein seaweed ferment powder, plant polypeptide, A scheme composite nano-particles and B scheme composite nano-particles are respectively arranged from bottom to top;
fig. 7 is a schematic diagram of a mechanism of the composite nanoparticle prepared according to the A, B scheme provided by the invention.
Detailed Description
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Experimental materials
Seaweed ferment powder is prepared according to the method in CN201510159810.6, and corn protein Zein (Sigma Aldrich).
Example 1 preparation of plant Polypeptides
Dissolving zein in deionized water to a concentration of 1% -3%, adjusting the pH value to 9.0-9.5, and controlling the pH value to be 9.0-9.5 all the time by using an automatic potentiometric titrator;
protease is added (the ratio of enzyme to substrate is 2:100), and the enzymolysis temperature is controlled to be 50 ℃;
adopting an incomplete hydrolysis method, hydrolyzing for 1-1.5 hours, immediately regulating the pH of the enzymolysis liquid to 7.0-8.0 by using 1M hydrochloric acid, and heating at 95 ℃ for 5-10min to inactivate enzymes;
centrifuging the prepared polypeptide at 10000r/min for 20min to remove a small amount of insoluble substances, and desalting the supernatant by dialysis for 24h with a dialysis bag of 100 Da;
and finally, freeze-drying the polypeptide liquid, and drying and preserving at 4 ℃ for later packaging experiments.
The hydrolyzed plant polypeptides were validated for different molecular weight fragments using a mass spectrometer (Q-exact, thermo Fisher Scientific). As shown in FIG. 2, after 1-1.5 hours of hydrolysis, it can be confirmed from the mass spectrum of zein plant polypeptide that the prepared plant polypeptide has various polypeptide fragments with different molecular weights, wherein the most content of the polypeptide fragments has a molecular weight of 639 area, and the next 534 area is the fragment comprising a small molecular weight fragment with a molecular weight of 200-599 and a large molecular weight fragment with a molecular weight of 600-1400, which meets the experimental requirements.
EXAMPLE 2 preparation of encapsulation compound
Magnetic stirring is used in this case to prepare the encapsulated composite particles. Specifically, two preparation schemes are adopted:
scheme a: dissolving seaweed enzyme powder in water at a concentration of 1-3mg/mL, stirring for 0.5-2h by using a magnetic stirrer, and fully dissolving the seaweed enzyme powder according to the ratio of plant polypeptide to seaweed enzyme powder: the plant polypeptide is added into the mixture according to the ratio of less than or equal to 1:5, the ratio of less than or equal to 1:5 and less than or equal to 1:2, the ratio of less than or equal to 1:1 and less than or equal to 2:1, the ratio of less than or equal to 2:1 and less than or equal to 5:1, and after continuously stirring for 1-1.5h, insoluble matters are removed by centrifugation for 20min at 10000g, and the composite nano particles A are obtained by filtering with a 420 mu m water-based film.
Scheme B: the plant polypeptide and seaweed ferment powder prepared by the method are prepared according to the following proportion: the proportion is less than or equal to 1:5, the proportion is less than or equal to 1:2, the proportion is less than or equal to 1:1, the proportion is less than or equal to 2:1, and after the proportion is less than or equal to 5:1, the mixture is dissolved in water, the mixture is stirred for 0.5 to 2 hours by using a magnetic stirrer, after the mixture is fully dissolved, seaweed ferment powder is added at 1 to 3mg/mL, the mixture is continuously stirred for 1 to 1.5 hours, insoluble matters are removed by centrifugation for 20 minutes at 10000g, and the mixture is filtered by using a 420 mu m water-based film, so that the composite nanoparticle B is obtained.
Example 3 characterization of composite nanoparticle Performance
3.1 particle diameter and zeta potential
The stability of colloidal particles can be characterized by the particle size of the particles and the zeta potential of the particles. In general, the smaller the particle size of the particles, the higher the zeta potential (positive or negative), indicating that the better the physical stability of the particles. The experiment adopts a Nano-ZS Nano-particle size potentiometer to measure the particle size distribution and zeta potential of the embedded composite Nano-particles. The test temperature was 25 ℃. The experimental results are shown in fig. 3-5.
As shown in fig. 3, stable chemical bonds are formed between the active material and the wall material, so that the composite nano particles form colloid after being dissolved in water, and the particle size can intuitively reflect whether the system is stable or not. From the particle size monitoring results in FIG. 3, it can be seen that the system area was stable from week 10, and each index remained unchanged. Based on 15 weeks of monitoring, in regimen a: when the ratio of the plant polypeptide to the seaweed ferment powder is 1:5 < 1:2 and 1:2 < 1:1, the nanometer grain diameter is stable and always kept below 200nm, and no aggregation phenomenon is found. In scheme B: when the ratio of the plant polypeptide to the seaweed ferment powder is 1:5 < 1:2, the ratio of the plant polypeptide to the seaweed ferment powder is 1:1 < 2:1, the nanometer grain diameter is stable and always kept below 200nm, and no aggregation phenomenon is found. Wherein when the ratio of the plant polypeptide to the seaweed ferment powder is less than or equal to 1:5, the A scheme and the B scheme are both beyond 200nm in the monitoring in the first week, so that no monitoring value exists in the following 15 weeks.
As shown in fig. 4, the PDI of the colloid is referred to as the system dispersion coefficient. Smaller PDI values represent more uniform particle sizes in the system. Typically when PDI is above 0.4, it represents large particle precipitation in the system and is one of the manifestations of colloid instability. As can be seen from the monitoring results of PDI in FIG. 4 for 15 weeks, in the scheme A, the PDI of the system is less than 0.4 only when the ratio of the plant polypeptide to the seaweed ferment powder is 1:5 < 1:2, and no aggregation phenomenon is found. In the scheme B, the PDI of the system is less than 0.4 only when the ratio of the plant polypeptide to the seaweed ferment powder is 2:1 < ratio less than or equal to 5:1. Wherein, when the ratio of the plant polypeptide to the seaweed ferment powder is less than or equal to 1:5, the monitoring of the first week is beyond 0.4 in both the A scheme and the B scheme, so that no monitoring value exists in the following 15 weeks.
As shown in FIG. 5, ZP, referred to as the potential of the system, is another key factor in assessing the stability of the nanodispersion. ZP is determined by the electrophoretic mobility of particles in a medium and can predict whether a system will have long-term stability characteristics. Negative ZP values indicate that the system is negatively charged, while positive ZP values indicate that the system is positively charged. Regardless of whether the ZP value of the dispersion is positive or negative, there is a greater repulsive force between the particles and less easy aggregation when the absolute value is higher. Whereas in the case of lower absolute ZP, aggregation or flocculation is liable to occur, possibly resulting in a decrease in stability. To achieve electrostatic stability, the dispersion should have an absolute value of ZP of greater than 30mV. That is, when the system is negatively charged, the measured ZP should be less than-30 mv, and when the system is positively charged, the measured value needs to be more than 30mv to be considered as a stable system. As shown in fig. 5, in scheme a: after 15 weeks, only when the absolute value of the system potential is above 30 and the ratio of the plant polypeptide to the seaweed ferment powder is 1:5 < 1:2 and 1:2 < 1:1, the scheme B is as follows: when the ratio of the plant polypeptide to the seaweed ferment powder is 1:5 < 1:2,1:1 < 2:1 and 2:1 < 5:1, the absolute value of the system potential is above 30 after 15 weeks.
As comprehensive evaluation, considering that the most stable state of the system is achieved by the three indexes at the same time, namely, the diameter of the nano particles is less than or equal to 200nm, the PDI (dispersion coefficient) is less than or equal to 0.4, and the absolute value of the potential is more than or equal to 30mv. Therefore, the scheme A is selected when the ratio of the plant polypeptide to the seaweed ferment powder is 1:5 < 1:2, which is the most stable colloid system.
3.2 Fourier transform Infrared Spectrometry (FT-IR)
FT-IR (Nicolet iS10, thermo Fisher, USA) was used to observe the functional groups of composite nanoparticles, plant polypeptides and seaweed ferment powder produced by the AB protocol, which iS an effective means to determine whether chemical bonds are formed between bioactive substances and wall materials and to determine whether the active substances enter the interior of the cavities formed by the wall materials. The operation steps are as follows: the sample was mixed with potassium bromide, ground in an agate mortar, and pressed into a sheet for measurement. FT-IR of pure potassium bromide was set as background for all samples and wavenumber range was set to 4000 to 400cm -1 . The experimental results are shown in FIG. 6.
As is clear from FIG. 6, the characteristic peak of the plant polypeptide is a broad and strong O-H stretch (3303 cm) -1 ) Shows the hydrophilicity of ZH; C-H stretching (2924 cm) -1 ) Representing its hydrophobicity; and a characteristic peak amide band of zein (1651 cm) -1 ). Likewise, O-H stretching (3315 cm) -1 ) With a small red shift; C-H stretching (2895 cm) -1 ) Characteristic peak amide bands (1630 cm) associated with small blue shift and zein -1 ). Characteristic peak C-H bending vibration 1392cm of seaweed ferment powder -1 And C-O stretching vibration 1024cm -1 None was detected in the A-regime, whereas the C-H bending vibrations 1354cm were clearly seen in the composite nanoparticles prepared in the B-regime -1 And C-O stretching vibration 1015cm -1 Both peaks belong to characteristic peaks of seaweed ferment powder. This can be an indication that the active substances of the nanocomposite particles prepared by the scheme A all enter the interior of the wall material, so that the characteristic peaks of the seaweed ferment powder are not detected when the nanocomposite particles are scanned by the Fourier transform infrared spectrum. The nano composite particles prepared by the scheme B do not enter the cavity of the wall material, but are attached to the periphery of the wall material, so that two characteristic peaks of seaweed ferment powder and plant polypeptide can be clearly seen in a Fourier transform infrared spectrogram of the scheme B. The experimental results are consistent with those of particle size, PDI and potential.
As the seaweed ferment powder belongs to hydrophilic active substances and has hydrophilic groups, the seaweed ferment powder can be combined with water molecules after being put into water. Therefore, if the conventional packaging mode is adopted, the packaging cannot be carried out. The zein can form plant polypeptide fragments with different molecular weights after hydrolysis for 1-1.5 hours. After the seaweed ferment powder is dissolved in water by adopting the scheme A, water molecules and the seaweed ferment powder can form weak intermolecular forces so as to be dissolved in water. After the plant polypeptide is fully dissolved, the plant polypeptide with nonuniform molecular weight is added to encapsulate the plant polypeptide, the hydrophilic group of the plant polypeptide with smaller molecular weight is combined with the hydrophilic active substance in the inner layer, the outside of the exposed hydrophobic group is combined with the hydrophobic group of the plant polypeptide with larger molecular weight, and finally the composite nano particle with stable performance is formed as shown in A in figure 7. When the scheme B is adopted, the plant polypeptides with different molecular weights are firstly dissolved in water, nano particles can be formed by self-assembly in the water, hydrophobic groups are encapsulated in the particles, and hydrophilic groups are exposed outside the particles. At this time, the seaweed ferment powder is added into water, the hydrophilic group of the seaweed ferment powder can be directly combined with the hydrophilic group of water molecules or plant polypeptides, a part of the seaweed ferment powder is directly dissolved in the water, and a part of the seaweed ferment powder is connected to the outer end of the plant polypeptides and cannot enter the cavity as shown in B in FIG. 7. Therefore, seaweed ferment powder can still be detected when Fourier transform infrared spectrum is scanned. It follows that the order of addition of the raw materials during the encapsulation process described above plays a critical role in the desired properties of the composite nanoparticle obtained.
According to the experimental results, the scheme A is determined to prepare the nano composite particles, the property of the nano composite particles is most stable when the ratio of the plant polypeptide to the seaweed ferment powder is 1:5 < 1:2, and the active substances are identified to be encapsulated in the plant polypeptide wall material through Fourier transform infrared spectroscopy.
Example 4 evaluation experiment
To further verify the validity of the above parameters, a qualification experiment was performed.
Panel of evaluations: 10 persons fixed, the diet is light, and the food is evaluated between 9:00 and 10:00 am;
sample classification: the proportion of the polypeptide and seaweed ferment powder prepared by the AB scheme is as follows: the ratio is less than or equal to 1:5, the ratio is more than or equal to 1:5 and less than or equal to 1:2, the ratio is more than or equal to 1:2 and less than or equal to 1:1, the ratio is more than or equal to 1:1 and less than or equal to 2:1, and the ratio is more than or equal to 2:1 and less than or equal to 5:1; pure water; untreated samples of the same concentration seaweed ferment powder solution. All samples were thermostated at 15 ℃.
And (3) evaluation flow: after being fully shaken, the liquid is drunk into a sample in a large mouth, flows in an oral cavity for more than 10 seconds, and feels the fishy smell and taste of the liquid in the flowing process of the oral cavity and the hypopharynx; carrying out evaluation experiments on samples according to random sequences, rinsing with clean water between samples, and staying for 1min between samples;
the fishy smell scoring principle is less than or equal to 1, and no fishy smell exists; score 1 < 2: less fishy smell; score 2 < 4: medium fishy smell; 4: obvious fishy smell.
The taste scoring principle is less than or equal to 1, smooth and non-astringent; score 1 < 2: has slight astringent feel; score 2 < 4: a moderate feeling of astringency; 4: obvious astringent feel.
Comprehensive score: the fishy smell score was combined with the mouthfeel score to obtain a composite score, and the scores of ten persons were averaged to obtain a final composite score, which was recorded in table 1. The lower the composite score value, the better the system evaluation index.
Table 1 sensory evaluation method analysis
From the experimental results of table 1, it can be seen that the fishy smell and astringent score of water are the lowest, which ensures the effectiveness of the experiment. The untreated seaweed ferment powder solution has prominent fishy smell and prominent astringent taste, so that the seaweed ferment powder solution is necessary to mask the smell as a drink. Comprehensively evaluating the fishy smell scores under both protocols A, B, we found that both protocols B can taste a slight fishy to fishy smell prominence, consistent with the results of the fourier transform infrared spectrum. The fishy smell caused by the active substances still exists because the active substances in the scheme B do not enter the cavity of the plant polypeptide wall material. In the scheme A, the fishy smell is moderately presented except that the ratio of the plant polypeptide to the seaweed ferment powder is less than or equal to 1:5, and the active substances are completely sealed in the cavity of the wall material along with the increase of the adding amount of the plant polypeptide, so that the fishy smell brought by the seaweed ferment powder can be covered. The experimental results of the evaluation in terms of astringency were consistent with the particle size, PDI and potential results. When there are large particles in the liquid, the astringency is enhanced. Therefore, when all indexes are integrated, the A scheme is used, and the proportion of the plant polypeptide to the seaweed ferment powder is ensured to be 1:5 < 1:2, the packaging effect is best, and the taste masking effect of the beverage can be achieved.
Through the verification of the evaluation experiment, the three comprehensive indexes of the particle size PDI and the potential can be further confirmed to accurately screen out the sample with stable performance and excellent taste, and a solid foundation is laid for mass production.
Claims (6)
1. The plant polypeptide-based encapsulated taste-masking composite nanoparticle is characterized in that plant polypeptides with different molecular weights are adopted to encapsulate hydrophilic active substance seaweed ferment powder;
wherein, the plant polypeptides with different molecular weights are prepared by the following method:
dissolving zein in deionized water to a concentration of 1% -3%, adjusting the pH value to 9.0-9.5, and controlling the pH value to be 9.0-9.5 all the time by using an automatic potentiometric titrator;
adding protease, controlling the enzymolysis temperature to be 50 ℃, and controlling the mass ratio of the added protease to zein to be 2:100;
the incomplete hydrolysis method is adopted, after hydrolysis is carried out for 1 to 1.5 hours, 1M hydrochloric acid is immediately used for adjusting the pH value of the enzymolysis liquid to 7.0 to 8.0, and the enzymolysis liquid is heated for 5 to 10 minutes at 95 ℃ to inactivate enzymes;
centrifuging the prepared polypeptide at 10000r/min for 20min to remove a small amount of insoluble substances, dialyzing the supernatant with a dialysis bag of 100Da for desalting at 24h, and lyophilizing the polypeptide liquid to obtain plant polypeptide;
the composite nano-particles are prepared by the following method:
dissolving seaweed ferment powder in water at a concentration of 1-3mg/mL, magnetically stirring for 0.5-2h, adding plant polypeptide according to a proportion after the seaweed ferment powder is fully dissolved, continuously stirring for 1-1.5h, centrifuging to remove insoluble substances, and filtering by using a 420 mu m water-based membrane to obtain composite particle nano-particles;
the mass ratio of the plant polypeptide to the seaweed ferment powder added into the composite nano particles is 1:5 < 1:2.
2. The composite nanoparticle of claim 1, wherein the molecular weight of the plant polypeptide comprises a small molecular weight fraction selected from the range of molecular weights 200-599 and a large molecular weight fraction selected from the range of 600-1400.
3. The composite nanoparticle according to claim 2, wherein the composite nanoparticle has a nanoparticle diameter of 200nm or less, a dispersion coefficient PDI of 0.4 or less, and an absolute value of potential of 30mv or more.
4. A composite nanoparticle according to any one of claims 1 to 3, wherein the composite nanoparticle exhibits the following functional groups in the infrared spectrum:
O-H stretching: 3303.+ -. 60cm -1 The method comprises the steps of carrying out a first treatment on the surface of the C-H stretching: 2924 + -50 cm -1 The method comprises the steps of carrying out a first treatment on the surface of the And characteristic peak amide bands of zein: 1651+ -30 cm -1 The method comprises the steps of carrying out a first treatment on the surface of the But does not include characteristic peaks of seaweed ferment powder: C-H bending vibration 1392+ -50 cm -1 And C-O stretching vibration 1024+ -50 cm -1 。
5. Beverage comprising the plant polypeptide encapsulation taste-masked composite nanoparticle according to any one of claims 1 to 4.
6. The beverage according to claim 5, wherein when the mass ratio of the plant polypeptide and the seaweed ferment powder added in the composite nano particles is 1:5 < 1:2, the obtained beverage has no fishy smell and is smooth and has no astringent feel.
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