Method for preparing natural lake based on phycocyanin, product and application thereof

Abstract

The application provides a method for preparing natural color lake based on phycocyanin, a product and application thereof, and relates to the technical field of color lake. The method comprises the following steps: step 1, mixing an aqueous solution of phycocyanin with an aqueous solution of stearoyl lactylate; step 2, after the aqueous solution of phycocyanin and the aqueous solution of stearoyl lactylate are uniformly mixed, a salt solution containing divalent metal ions is added, so that the negatively charged groups of phycocyanin and the stearoyl lactylate ions are combined by the bridging action of the divalent metal ions to form a color lake, and the color lake is precipitated from the aqueous solution; and step 3, collecting and processing the color lake to obtain a color lake product. The application can prepare natural blue color lake using phycocyanin, and can expand the application of water-soluble phycocyanin in oil-based food.

Description

Technical Field

[0001] This application relates to the field of lake technology, and in particular to a method, product and application for preparing natural lakes based on phycocyanin. Background Technology

[0002] Blue in food often gives consumers a fresh and refreshing impression. Compared to the orange-yellow color brought by natural pigments such as curcumin and beta-carotene, the green color brought by chlorophyll, and the red color brought by various anthocyanins, the scarcity of blue pigments in nature makes phycocyanin one of the limited choices for natural blue sources in the food industry.

[0003] The food additives standard GB2760-2024 allows the addition of synthetic indigo and brilliant blue and their lakes to food, as well as naturally derived gardenia blue and phycocyanin. Among these, phycocyanin has attracted the most attention because its upstream algae cultivation industry has achieved large-scale production globally, and its various physiological functions as a biological macromolecule, such as antioxidant, anti-inflammatory, anti-tumor, and immunomodulatory effects, are widely recognized. Extensive research data supports its anti-lung cancer activity, anti-melanoma activity, anti-colon cancer activity, and anti-liver cancer activity.

[0004] Currently, common lakes can be water-insoluble pigments obtained by precipitating or adsorbing water-soluble pigments onto an insoluble matrix (such as aluminum hydroxide). Generally, the formulation of lakes enhances the stability of the original pigments to light, heat, and salt ions. However, the binding of the pigment to the matrix may cause changes in the polarity of its chromophores and shifts in binding sites, resulting in changes in the pigment's chroma and hue. In the food additive standard GB2760-2024, pigments and their aluminum lakes are permitted as colorants added to various foods, such as aluminum hydroxide lakes like lemon yellow aluminum lake, erythrosine aluminum lake, and brilliant blue aluminum lake, which are prepared using aluminum hydroxide as the adsorption matrix.

[0005] Currently, scientific research has gradually revealed the mechanisms by which aluminum harms the human body: aluminum ions are absorbed through the digestive system, competitively inhibiting the absorption of minerals and trace elements such as calcium, iron, and zinc, easily causing electrolyte imbalances in the body's internal environment, increasing the burden on organs such as the liver and kidneys, and forming stones in local organ tissues. Aluminum ions may also be one of the important causes of Alzheimer's disease. Furthermore, the widespread popularity of aluminum-free baking powder on the market demonstrates consumers' strong preference for "clean" labels, and they favor "all-natural" and "less processed" foods. Considering consumers' increasing emphasis on healthy living, phycocyanin, positioned in the key area of ​​life and health, will have broader development prospects. Developing aluminum-free phycocyanin lakes based on natural macromolecules and expanding the application of water-soluble phycocyanin in oil-based foods meets practical needs and has broad development prospects. Summary of the Invention

[0006] This application provides a method, product, and application for preparing natural lakes based on phycocyanin. Natural blue lakes can be prepared using phycocyanin without the need for the addition of aluminum ions, which can expand the application of water-soluble phycocyanin in oil-based foods.

[0007] Existing lake preparation technologies generally rely on a combination of "small molecule pigments + macromolecular matrix" to generate lakes. In terms of the size of the monomer structures, the pigment monomers are smaller, while the matrix monomers are larger. This allows a large number of small molecule pigments to adhere to the periphery of the macromolecular matrix, thus enabling the preparation of lakes.

[0008] In this embodiment, a lake is prepared using large-molecule, water-soluble phycocyanin as the pigment source. From the perspective of monomeric structure size, phycocyanin particles are relatively large, making it difficult to produce phycocyanin lakes using the existing combination of "small-molecule pigment + large-molecule matrix".

[0009] Based on the macromolecular pigment characteristics of phycocyanin, this application uses a novel combination of "macromolecular pigment + small molecule emulsifier" to prepare lakes. Specifically, water-soluble phycocyanin is used as the source of macromolecular pigment, stearoyl lactate (such as calcium stearoyl lactate) is used as the small molecule emulsifier, and divalent metal salt (such as calcium chloride) is used as the crosslinking agent to produce phycocyanin lakes.

[0010] Compared to stearoyl lactate, phycocyanin has a larger particle size. Therefore, based on the combination of "large molecular weight pigment + small molecular weight emulsifier," and combined with the bridging effect of divalent metal ions, a large number of hydrophobic and oleophilic stearoyl lactate ions can be attached to the surface of the phycocyanin molecules. This results in the formation of a phycocyanin-calcium-stearoyl lactate complex (taking calcium ions as an example), which precipitates from the aqueous solution, thus yielding a lake. Both stearoyl lactate and the divalent metal salt are processing aids used in the preparation of lakes.

[0011] Furthermore, through experimental verification, the lake obtained in the embodiments of this application is an oil-dispersible lake, which can be stably dispersed in the oil phase, and can therefore be used for coloring oil-based products.

[0012] As can be seen, the "small molecule emulsifier" in this application embodiment is not used as the "macromolecule matrix" in the existing combination. That is, this application embodiment does not use the existing combination to prepare the lake, but prepares the phycocyanin lake by attaching a large amount of emulsifier to the surface of the macromolecule phycocyanin.

[0013] Based on the above, in a first aspect, embodiments of this application provide a method for preparing natural lakes based on phycocyanin. See also... Figure 1 The method may include the following steps:

[0014] Step 1: Mix the aqueous solution of phycocyanin with the aqueous solution of stearoyl lactate.

[0015] The aqueous solution of stearoyl lactate is used to provide stearoyl lactate ions. Thus, the aqueous solution of stearoyl lactate can be an aqueous solution of a single stearoyl lactate or a mixed aqueous solution of multiple stearoyl lactates.

[0016] To avoid the introduction of heavy metal ions, and taking into account factors such as the source of raw materials, stearoyl lactate can be, for example, calcium stearoyl lactate, sodium stearoyl lactate, etc.

[0017] By pre-mixing the aqueous solution of phycocyanin with the aqueous solution of stearoyl lactate, the mixed solution contains not only a large number of negatively charged phycocyanin groups but also a large number of stearoyl lactate ions. This supports the subsequent addition of a large number of divalent metal ions, which, based on the bridging effect of the divalent metal ions, can generate a large amount of phycocyanin-calcium-stearoyl lactate complex (taking calcium ions as an example) that precipitates from the aqueous solution, thus avoiding the generation of too many non-lake products (such as phycocyanin-calcium complexes formed when divalent metal ions only bind to phycocyanin groups).

[0018] In one embodiment, in step 1, 40-50 parts of phycocyanin and 50-60 parts of stearoyl lactate are used, by weight.

[0019] In one embodiment, in step 1, the mass ratio of phycocyanin to stearoyl lactate is 1:1 to 1:1.5. For example, 50 parts of phycocyanin and 50 parts of stearoyl lactate, or 40 parts of phycocyanin and 60 parts of stearoyl lactate.

[0020] Step 2: After the aqueous solution of phycocyanin and the aqueous solution of stearoyl lactate are mixed evenly (for example, this can be achieved by stirring at a constant speed for a certain period of time), a salt solution containing divalent metal ions is added. This allows the negatively charged groups of phycocyanin to combine with the stearoyl lactate ions through the bridging effect of the divalent metal ions to form a lake, which then precipitates from the aqueous solution.

[0021] Feasibly, after the aqueous solution of phycocyanin and the aqueous solution of stearoyl lactate are mixed evenly, a divalent metal salt solution can be added while stirring at a constant speed, and stirring can be continued to mix the three food additives evenly to promote the formation of the lake.

[0022] By adding a salt solution containing divalent metal ions to provide a large amount (or sufficient amount) of divalent metal ions, the phycocyanin groups can be stably bound to stearoyl lactate ions based on the bridging effect of the divalent metal ions. This allows a large number of hydrophobic stearoyl lactate ions to be attached to the surface of the phycocyanin molecules, causing them to precipitate from the aqueous solution.

[0023] To avoid introducing heavy metal ions, the divalent metal ion can, for example, be calcium ions, or other feasible ions such as magnesium ions. Taking calcium ions as an example, the salt solution containing divalent metal ions can be a calcium chloride solution, or other feasible solutions such as calcium sulfate solution.

[0024] In one embodiment, the raw material composition for preparing the lake includes the following food additives: 40-50 parts of phycocyanin, 50-60 parts of stearoyl lactate, and an appropriate amount of divalent metal salt (e.g., 50 parts, or other feasible parts), by mass. The amount of divalent metal salt used is sufficient to provide enough divalent metal ions to support lake formation.

[0025] Taking the preparation of lakes using sodium stearoyl lactylate and calcium chloride as an example, when sodium stearoyl lactylate solution is mixed with phycocyanin solution, during the dissociation and binding process of stearoyl lactylate groups (i.e., stearoyl lactylate ions) and sodium ions, some stearoyl lactylate ions can bind to the positively charged groups of phycocyanin, changing the solubility characteristics of phycocyanin. When calcium chloride solution is added, the large amount of newly added calcium ions causes the negatively charged groups of phycocyanin to bind stably with the stearoyl lactylate ions, resulting in a large number of hydrophobic stearoyl lactylate ions attached to the surface of the phycocyanin molecules (i.e., forming a lake), thus causing it to precipitate from the aqueous solution.

[0026] Taking the preparation of lakes using calcium stearoyl lactate and calcium chloride as an example, when the calcium stearoyl lactate solution is mixed with the phycocyanin solution, during the dissociation and binding process of the stearoyl lactate groups and calcium ions, on the one hand, the calcium ions (i.e., calcium ions derived from calcium stearoyl lactate) bind to the negatively charged groups of phycocyanin and the stearoyl lactate ions respectively based on the bridging effect of divalent metal ions (i.e., a small amount of lake has already been generated, and the formation of a light blue flocculent substance can be observed before the addition of calcium chloride). On the other hand, some stearoyl lactate ions can bind to the positively charged groups of phycocyanin, changing the solubility characteristics of phycocyanin. When the calcium chloride solution is added, a large number of newly added calcium ions (i.e., calcium ions derived from calcium chloride) cause the negatively charged groups of phycocyanin to bind stably with the stearoyl lactate ions, resulting in a large number of hydrophobic stearoyl lactate ions attached to the surface of the phycocyanin molecules (i.e., a large amount of lake is generated), thereby causing it to precipitate from the aqueous solution.

[0027] For example, the precipitation of lakes in some test examples of this application are as follows: Figure 2 As shown. See also Figure 2 The supernatant after centrifugation in each test case was light blue to pale blue, with high phycocyanin transfer rate and low loss, which met the expectations for industrial mass production and could be used to prepare high-quality phycocyanin lakes.

[0028] Step 3: Collect and process the lake to obtain the lake product.

[0029] In one embodiment, step 3 includes: collecting the precipitated lake in the aqueous solution by means of static sedimentation and / or centrifugation (e.g., static sedimentation only, or centrifugation only, or static sedimentation followed by centrifugation); washing the collected lake and drying the washed lake (e.g., hot air drying, vacuum drying, or freeze drying); and crushing and sieving the dried lake to obtain the lake product.

[0030] By crushing and sieving to maintain the uniformity of the lake particles, the particle size can be controlled between 50 mesh and 120 mesh (e.g., 80 mesh to 100 mesh), so that the lake can be stably dispersed in the oil phase later.

[0031] In a feasible way of collecting the lake by static sedimentation, the static sedimentation time can be set reasonably as needed. For example, in one embodiment, it can be set to 120min-360min.

[0032] In a feasible way of collecting the lake by centrifugation, the centrifugation conditions can be set reasonably as needed. For example, in one embodiment, it can be set to 8000rpm-12000rpm and 10min-20min.

[0033] In the implementation of collecting the lake by first allowing it to settle and then centrifuging, the relevant experimental conditions can be set reasonably as needed. For example, in one embodiment, it can be set to allow it to settle for 60 min-120 min and then centrifuge at 8000 rpm-12000 rpm for 5 min-10 min.

[0034] Feasibly, the washing conditions can be reasonably set as needed to achieve the purpose of thorough washing. For example, in one embodiment, it can be set to wash 3 to 5 times with pure water at a mass ratio of 1:50 to 1:100.

[0035] Feasibly, the conditions for hot air drying can be set reasonably as needed. For example, in one embodiment, the drying temperature can be set to 60℃-110℃ and the drying time to 6h-12h.

[0036] Feasibly, the conditions for vacuum drying can be set reasonably as needed. For example, in one embodiment, the drying temperature can be set to 25℃-50℃, the vacuum degree to about -0.1MPa (e.g. -0.08MPa), and the drying time to 6h-12h.

[0037] Feasibly, the freeze-drying conditions can be set reasonably as needed. For example, in one embodiment, the pre-cooling temperature can be set to -80°C to -60°C, the air pressure to ≤10Pa, and the drying time to 1-2 days.

[0038] For example, the appearance of the lake products obtained after drying and pulverizing in some test examples of this application is as follows: Figure 3 As shown. See also Figure 3 Although the lake products obtained from different test cases showed some differences in color due to the difference in the mass ratio of phycocyanin to emulsifier, the lake products from each test case basically showed a pure blue color.

[0039] The lake product obtained in this application embodiment is an oil-dispersible natural blue lake, which has the characteristics of pure blue color, good oil dispersibility, high stability, and low coloring dosage.

[0040] For example, the stability index (TSI) of some lake products in this application after dispersion in the oil phase is as follows: Figure 4 As shown. See also Figure 4 The lake products obtained from each test example showed a stability index below 2.0 under several days of dark standing conditions. Since a lower stability index indicates better stabilization, this means that no significant phase separation was observed in the lake products obtained from each test example, indicating that all lake products exhibited good physical stability.

[0041] For example, the condition of a lake product in one test example of this application after dispersion in the oil phase is as follows: Figure 5 As shown. See also Figure 5 Based on a coloring dosage of 1 g / L, after the lake product has been dispersed in the oil phase for several days, its blue color remains clear and pleasant to the naked eye, maintaining its original bright blue color. The dispersion system is uniform, stable, and transparent, with no aggregation or sedimentation.

[0042] To ensure the reaction effect of the raw materials, phycocyanin and divalent metal salts can be dissolved separately in room temperature purified water, and stearoyl lactate can be dissolved in hot water to prepare the solution. In one embodiment, the above preparation method further includes the following steps: adding purified water at a temperature of 40℃-80℃ (e.g., 75℃-80℃) to a mixer, adding stearoyl lactate to the mixer while stirring, continuing to stir and dissolve after the addition is completed, and then cooling to 25℃-60℃ (e.g., 50℃-60℃) to prepare an aqueous solution of stearoyl lactate; adding room temperature purified water to another mixer, adding phycocyanin to the mixer while stirring, continuing to stir and dissolve after the addition is completed to prepare an aqueous solution of phycocyanin; adding room temperature purified water to yet another mixer, adding the salt containing divalent metal ions to the mixer while stirring, continuing to stir and dissolve after the addition is completed to prepare a salt solution containing divalent metal ions.

[0043] For example, the cooling step described above can be cooling from 75°C to 50°C, or cooling from 80°C to 60°C, etc.

[0044] It is feasible to set the stirring speed as needed, for example, in one embodiment, it can be set to 200rpm-300rpm.

[0045] It is feasible to set the stirring and dissolution time of stearoyl lactate as needed after the feeding is completed, for example, in one embodiment it can be set to 30min-120min.

[0046] Feasibly, after the feeding is completed, the stirring and dissolving time of phycocyanin can be reasonably set as needed, for example, in one embodiment it can be set to 60min-360min.

[0047] Feasibly, after the feeding is completed, the stirring and dissolution time of the divalent metal salt can be reasonably set as needed, for example, in one embodiment it can be set to 30min-60min.

[0048] Based on the above, in one feasible implementation, the process of preparing a lake based on phycocyanin may include the following:

[0049] 1) Solution preparation process for various food additives used to prepare lakes

[0050] Add purified water at a temperature of 40℃-80℃ to the pre-cleaned mixer, then add the qualified calcium stearoyl lactylate and / or sodium stearoyl lactylate into the mixer while adding the materials and stirring at a speed of 200rpm-300rpm. After adding the materials, continue stirring to dissolve for 30min-120min.

[0051] Add room temperature purified water to another mixer after pre-cleaning, then add the qualified phycocyanin powder to this mixer while adding the material and stirring at a speed of 200 rpm-300 rpm. After adding the material, continue stirring to dissolve for 60 min-360 min.

[0052] After pre-cleaning, inject room temperature purified water into another mixer, and then add the qualified calcium chloride powder into this mixer while adding the material and stirring. The stirring speed is 200rpm-300rpm. After the material is added, continue stirring and dissolving for 30min-60min.

[0053] 2) The sequential mixing process of various food additive solutions

[0054] After the above food additives are completely dissolved, cool the calcium stearoyl lactylate and / or sodium stearoyl lactylate solution to 25℃-60℃, and mix it with the phycocyanin dispersion at the set mass ratio, stirring at 200rpm-300rpm. After the two are thoroughly mixed, add the calcium chloride solution at the set mass ratio while stirring at a constant speed of 200rpm-300rpm, and continue stirring for 30min-90min.

[0055] In this process, after the addition of calcium chloride solution, various food additives interact to produce a lake (i.e., the above-mentioned phycocyanin-calcium-stearoyl lactic acid complex), which precipitates from the aqueous solution.

[0056] 3) Sedimentation and collection process

[0057] After sufficient reaction, the blue lake can be collected by allowing it to settle for 120-360 minutes, or by centrifugation at 8000-12000 rpm for 10-20 minutes. The collected blue lake is then washed 3-5 times with purified water at a mass ratio of 1:50-1:100 to ensure thorough washing.

[0058] 4) Precipitation and drying process

[0059] The thoroughly washed blue lake is then subjected to hot air drying, vacuum drying, or freeze drying. Hot air drying is performed at 60℃-110℃ for 6-12 hours, with moisture content controlled at ≤8%. Vacuum drying is performed at 25℃-50℃ with a vacuum level of approximately -0.1MPa for 6-12 hours, with moisture content controlled at ≤5%. Freeze drying pre-cooling temperature is -80℃ to -60℃, with the freeze-drying chamber pressure ≤10Pa during the main drying stage, for 1-2 days.

[0060] 5) Sedimentation and pulverization process

[0061] The dried blue lake is pulverized and sieved. The particle size after sieving can be controlled between 50 mesh and 120 mesh, thereby obtaining a phycocyanin lake with pure blue color, good oil dispersibility, and high stability.

[0062] Secondly, embodiments of this application provide a lake product prepared using any of the methods described in the first aspect. Specifically, embodiments of this application provide an oil-dispersible natural blue lake based on phycocyanin, which has the characteristics of stable dispersion in the oil phase, pure blue color after oil dispersion, transparent and stable system, and low coloring dosage, thus possessing potential for industrial application.

[0063] Thirdly, embodiments of this application provide the application of the lake product described in the second aspect in oil-based food products. Exemplarily, the oil-based food product may be cream or chocolate, etc.

[0064] Based on the above, the embodiments of this application have at least the following characteristics:

[0065] (1) Unlike existing methods that use a combination of "small molecule pigments + macromolecular matrix" to generate lakes, the embodiments of this application use...

[0066] The new combination of "large molecular pigment + small molecular emulsifier" specifically uses water-soluble phycocyanin as the source of large molecular pigment, stearoyl lactate (such as calcium stearoyl lactate) as the small molecular emulsifier, and divalent metal salt (such as calcium chloride) as the cross-linking agent to produce phycocyanin lake.

[0067] (2) The lake prepared in the embodiments of this application is an oil-dispersible blue lake, which can be stably dispersed in the oil phase. It has the characteristics of pure blue color after oil dispersion, no aggregation and precipitation, transparent system and good color stability, and low coloring amount. When added to oil-based foods as a coloring agent, it can provide a refreshing and pleasant blue hue to the food.

[0068] (3) The lake prepared in the embodiments of this application is a blue lake, which can meet the urgent need of the food industry for blue colorants.

[0069] (4) The process flow of this application embodiment is simple and easy to operate, with a high phycocyanin precipitation rate (which can reach more than 85% according to the test) and high production efficiency, making it suitable for large-scale industrial production.

[0070] (5) In this application, phycocyanin is used as a pigment source to prepare natural lake. Based on the inherent physiological function of phycocyanin, the resulting lake can bring corresponding health benefits when used for food coloring.

[0071] (6) The lake preparation process of this application embodiment is simple, economical, safe and pollution-free. It does not involve the addition of aluminum ions, heavy metal ions, etc., and does not require the addition of organic reagents. It can increase consumers' acceptance of lakes, meet the market demand for "clean labels", and is more in line with market demand and has broad prospects.

[0072] (7) Based on the rapid development of the global marine economy and algae economy, and considering the inherent physiological functions of phycocyanin, the preparation process of preparing lakes using phycocyanin is suitable for large-scale industrial production. Attached Figure Description

[0073] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below.

[0074] Figure 1 A schematic flowchart illustrating a method for preparing natural lakes based on phycocyanin, provided as an embodiment of this application;

[0075] Figure 2 The figures show the experimental results of phycocyanin lake precipitation in Examples 1-3;

[0076] Figure 3 The images show the appearance of the dried and pulverized phycocyanin lake products from Examples 1-3.

[0077] Figure 4 The figures show the experimental results for evaluating the physical stability of the phycocyanin lakes in Examples 1-3 after dispersion in the oil phase.

[0078] Figure 5 This is a diagram illustrating the stability of the phycocyanin lake in Example 2 after it has been dispersed in the oil phase and left to stand for 5 days. Detailed Implementation

[0079] To better understand the technical solution of this application, the embodiments of this application will be described in detail below with reference to the accompanying drawings.

[0080] It should be understood that the described embodiments are merely some, not all, of the embodiments in this application. All other embodiments obtained by those skilled in the art based on the embodiments in this application without inventive effort are within the scope of protection of this application.

[0081] The terminology used in the embodiments of this application is for the purpose of describing particular embodiments only and is not intended to be limiting of this application. The singular forms “a,” “the,” and “the” used in the embodiments of this application and the appended claims are also intended to include the plural forms unless the context clearly indicates otherwise.

[0082] It should be understood that the term "at least one" as used in this document refers to one or more, and "more than one" refers to two or more. The term "and / or" as used in this document is merely a description of the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A alone, A and B simultaneously, and B alone. A and B can be singular or plural. Additionally, the character " / " in this document generally indicates that the preceding and following related objects are in an "or" relationship. "At least one of the following" and similar expressions refer to any combination of these items, including any combination of singular or plural items. For example, at least one of a, b, and c can represent: a, b, c, ab, ac, bc, or abc, where a, b, and c can be single or multiple.

[0083] Example 1

[0084] Raw material ratio: 50 parts phycocyanin, 50 parts sodium stearoyl lactylate, and 50 parts calcium chloride.

[0085] The preparation method is as follows:

[0086] 1. Preparation of food additive solutions

[0087] Add purified water at 75°C to the pre-cleaned mixer, then add qualified sodium stearoyl lactylate at a mass ratio of 1:150 (sodium stearoyl lactylate: purified water) to the mixer while adding the material and stirring at a speed of 300 rpm. Continue stirring after adding the material and dissolve for 60 minutes.

[0088] Add room temperature purified water to another mixer after pre-cleaning at a certain mass ratio. Then add qualified phycocyanin powder to this mixer at a mass ratio of 1:50 (phycocyanin: purified water). Stir while adding the material, and the stirring speed is 200 rpm. Continue stirring after adding the material, and the dissolution time is 360 minutes.

[0089] After pre-cleaning, add room temperature purified water to another mixer at a certain mass ratio. Then, add qualified calcium chloride powder to this mixer at a mass ratio of 1:20 (calcium chloride: purified water), stirring while adding the material. The stirring speed is 200 rpm. After adding the material, continue stirring. The dissolution time is 60 minutes.

[0090] 2. Mixing of food additive solutions

[0091] After the above food additives are completely dissolved, cool the sodium stearoyl lactylate solution to 50°C and mix it with the phycocyanin dispersion at a stirring speed of 300 rpm. After the two are thoroughly mixed, add the calcium chloride solution while stirring at a constant speed of 300 rpm.

[0092] 3. Sedimentation Collection

[0093] After mixing the three food additive solutions, continue stirring for 60 minutes, then collect the precipitate. Centrifuge at 10,000 rpm for 15 minutes to obtain a blue lake (see [link to precipitate]). Figure 2 The blue lake was washed three times with purified water at a ratio of 1:100 by mass.

[0094] 4. Drying

[0095] The thoroughly washed blue lake was freeze-dried at a pre-cooling temperature of -80℃, with the main drying stage at a freeze-drying chamber pressure of ≤10Pa and a time of ≥2 days.

[0096] 5. Crushing

[0097] The dried sample was pulverized and sieved. The particle size was controlled at 80 mesh after sieving to obtain a pure blue, highly oil-dispersible, and stable natural phycocyanin lake. The lake was then sealed and packaged with a desiccant to prevent the product from absorbing moisture and clumping.

[0098] The product in this example is a dark blue powder (see...). Figure 3It has no unpleasant odor. When added to the MCT oil phase at a concentration of 1 g / L and stirred for 60 min, the resulting blue MCT oil phase system was homogeneous, stable, and transparent, showing no significant changes after standing for 5 days (see...). Figure 4 ).

[0099] Example 2

[0100] Raw material ratio: 40 parts phycocyanin, 60 parts sodium stearoyl lactylate, and 50 parts calcium chloride.

[0101] The preparation method is as follows:

[0102] 1. Preparation of food additive solutions

[0103] Add purified water at 75°C to the pre-cleaned mixer, then add qualified sodium stearoyl lactylate at a mass ratio of 1:150 (sodium stearoyl lactylate: purified water) to the mixer while adding the material and stirring at a speed of 300 rpm. Continue stirring after adding the material and dissolve for 60 minutes.

[0104] Add room temperature purified water to another mixer after pre-cleaning at a certain mass ratio. Then add qualified phycocyanin powder to this mixer at a mass ratio of 1:50 (phycocyanin: purified water). Stir while adding the material, and the stirring speed is 200 rpm. Continue stirring after adding the material, and the dissolution time is 360 minutes.

[0105] After pre-cleaning, add room temperature purified water to another mixer at a certain mass ratio. Then, add qualified calcium chloride powder to this mixer at a mass ratio of 1:20 (calcium chloride: purified water), stirring while adding the material. The stirring speed is 200 rpm. After adding the material, continue stirring. The dissolution time is 60 minutes.

[0106] 2. Mixing of food additive solutions

[0107] After the above food additives are completely dissolved, cool the sodium stearoyl lactylate solution to 50°C and mix it with the phycocyanin dispersion at a stirring speed of 300 rpm. After the two are thoroughly mixed, add the calcium chloride solution while stirring at a constant speed of 300 rpm.

[0108] 3. Sedimentation Collection

[0109] After mixing the three food additive solutions, continue stirring for 60 minutes, then collect the precipitate. Centrifuge at 10,000 rpm for 15 minutes to obtain a blue lake (see [link to precipitate]). Figure 2 The blue lake was washed three times with purified water at a ratio of 1:100 by mass.

[0110] 4. Drying

[0111] The thoroughly washed blue lake was subjected to vacuum drying at a temperature of 40℃ and a vacuum degree of -0.08MPa for 12 hours to reduce the moisture content of the phycocyanin lake to less than 5%.

[0112] 5. Crushing

[0113] The dried sample was pulverized and sieved. The particle size was controlled at 80 mesh after sieving to obtain a pure blue, highly oil-dispersible, and stable natural phycocyanin lake. The lake was then sealed and packaged with a desiccant to prevent the product from absorbing moisture and clumping.

[0114] The product in this example is a blue powder (see...). Figure 3 It has no unpleasant odor. It was added to the MCT oil phase at a concentration of 1 g / L and stirred for 60 min. The resulting blue MCT oil phase system was homogeneous and stable, transparent blue, and showed no significant changes after standing for 5 days (see...). Figure 4 , Figure 5 ).

[0115] Example 3

[0116] Food additive ratio: 50 parts phycocyanin, 50 parts calcium stearoyl lactylate, and 50 parts calcium chloride.

[0117] The preparation method is as follows:

[0118] 1. Preparation of food additive solutions

[0119] Add purified water at 80℃ to the pre-cleaned mixer, then add qualified calcium stearoyl lactylate at a mass ratio of 1:150 (calcium stearoyl lactylate: purified water) to the mixer while adding the material and stirring at a speed of 300 rpm. Continue stirring after adding the material and dissolve for 120 minutes.

[0120] Add room temperature purified water to another mixer after pre-cleaning at a certain mass ratio. Then add qualified phycocyanin powder to this mixer at a mass ratio of 1:50 (phycocyanin: purified water). Stir while adding the material, and the stirring speed is 200 rpm. Continue stirring after adding the material, and the dissolution time is 360 minutes.

[0121] After pre-cleaning, add room temperature purified water to another mixer at a certain mass ratio. Then, add qualified calcium chloride powder to this mixer at a mass ratio of 1:20 (calcium chloride: purified water), stirring while adding the material. The stirring speed is 200 rpm. After adding the material, continue stirring. The dissolution time is 60 minutes.

[0122] 2. Mixing of food additive solutions

[0123] After the above food additives are completely dissolved, cool the calcium stearoyl lactylate solution to 60°C and mix it with the phycocyanin dispersion at a stirring speed of 300 rpm. After the two are thoroughly mixed, add the calcium chloride solution while stirring at a constant speed of 300 rpm.

[0124] 3. Sedimentation Collection

[0125] After mixing the three food additive solutions, continue stirring for 60 minutes, then collect the precipitate. Centrifuge at 10,000 rpm for 15 minutes to obtain a blue lake (see [link to precipitate]). Figure 2 The blue lake was washed three times with purified water at a ratio of 1:100 by mass.

[0126] 4. Drying

[0127] The thoroughly washed blue lake was then subjected to hot air drying at 80℃ for 12 hours, with the moisture content controlled to be ≤5%.

[0128] 5. Crushing

[0129] The dried sample was pulverized and sieved. The particle size was controlled at 80 mesh after sieving to obtain a pure blue, highly oil-dispersible, and stable natural phycocyanin lake. The lake was then sealed and packaged with a desiccant to prevent the product from absorbing moisture and clumping.

[0130] The product in this example is a dark blue powder (see...). Figure 3 It has no unpleasant odor. It was added to the MCT oil phase at a concentration of 1 g / L and stirred for 60 min. The resulting blue MCT oil phase system was homogeneous and stable, transparent blue, and showed no significant changes after standing for 5 days (see...). Figure 4 ).

[0131] Effect evaluation

[0132] 1) Phycocyanin precipitation rate

[0133] 1.1) Calculation of phycocyanin precipitation rate

[0134] The completely dissolved phycocyanin (i.e., the phycocyanin dispersion before lake preparation) and the supernatant collected by centrifugation of the lake in Examples 1-3 were subjected to UV-Vis full-wavelength scanning (if the absorbance value is too high, it must be diluted) to obtain the absorbance values ​​of the samples at 620 nm and 650 nm.

[0135] The concentration of phycocyanin (PC) before and after lake preparation was calculated using formula (1), and the concentration of PC before and after lake preparation was obtained (in mg / mL). Then, the precipitation rate of PC was calculated using formula (2).

[0136] C PC =0.162×A 620nm-0.098×A 650nm (1)

[0137]

[0138] Among them, “C” PC "Represents the concentration of phycocyanin, "A" 620nm "and "A 650nm "C" represents the absorbance values ​​of the phycocyanin solution / lake supernatant at 620 nm and 650 nm, respectively. PC "Original" and "C" PC "Lake supernatant" represents the concentration of phycocyanin in the phycocyanin dispersion before lake preparation and the supernatant after lake preparation, respectively. PC "original" and "V" PC "Supernatant" represents the volume of the phycocyanin dispersion before lake preparation and the supernatant after lake preparation, respectively; "Dilution factor" refers to the dilution factor of the solution during the determination of phycocyanin concentration; "R" PC "Represents the phycocyanin precipitation rate."

[0139] The calculation results of the lake precipitation rate in Examples 1-3 are shown in Table 1.

[0140] Table 1

[0141] sample Sedimentation rate (%) Example 1 <![CDATA[88.83±0.17 b ]]> Example 2 <![CDATA[89.46±0.53 b <!-- 9 -->]]> Example 3 <![CDATA[86.98±0.28 a ]]>

[0142] In Table 1, different superscript letters indicate significant differences in phycocyanin precipitation rates.

[0143] 1.2) Evaluation of phycocyanin precipitation rate

[0144] The precipitation of lakes in Examples 1-3 is as follows: Figure 2 As shown. Calculations show that the lake precipitation rate of Examples 1-3 is as high as 86%-90%, and the supernatant after centrifugation is light blue to pale blue. The phycocyanin transfer rate is high and the loss is low, which meets the expectations for industrial mass production.

[0145] See Figure 2 Examples 1-3 can all prepare high-quality phycocyanin lakes.

[0146] 2) Physical stability

[0147] 2.1) Physical stability test

[0148] The phycocyanin lakes from Examples 1-3 were added to the MCT oil phase at a concentration of 1 g / L and stirred continuously for 12 h to ensure thorough dispersion. The samples were then poured into Turbiscan bottles and placed in the Turbiscan test chamber. The instrument scanning interval was set to 100 min, and the samples were continuously tested for 5 days under completely static conditions. By continuously collecting changes in transmitted and backscattered light from the samples, the stability index of the samples over time was obtained through comprehensive calculation of the overall stability kinetics, thereby evaluating the stability of the samples under static conditions.

[0149] 2.2) Physical stability evaluation

[0150] See Figure 4 The phycocyanin lakes prepared in Examples 1-3 all exhibited good physical stability, with a stability index below 2.0 after 5 days of dark standing. Since a lower stability index indicates better stabilization, no significant phase separation was observed in the phycocyanin lakes prepared in the three examples.

[0151] also, Figure 5 The image shows a sample vial removed from the Turbiscan test chamber after the lake obtained in Example 2 was dispersed in the oil phase, allowed to stand for 5 days, and then scanned. See also Figure 5 When observed with the naked eye, its blue color remains clear and pleasant, and the dispersion system is uniform and stable.

[0152] See Figure 4 and Figure 5 Examples 1-3 can all prepare natural phycocyanin lakes that are pure blue, have good oil dispersibility, and high stability.

[0153] 3) Color of phycocyanin lake

[0154] 3.1) Colorimetric measurement of phycocyanin lake

[0155] The phycocyanin lakes obtained after drying and pulverizing in Examples 1-3 were measured using a handheld colorimeter (NH310, 3hn). The color values ​​in three parallel experiments were measured, and their standard deviation and significance of difference were analyzed.

[0156] The colorimetric measurement results of the lakes in Examples 1-3 are shown in Table 2.

[0157] Table 2

[0158] sample Example 1 Example 2 Example 3 L value <![CDATA[12.33±0.58 a ]]> <![CDATA[34.33±1.16 c ]]> <![CDATA[21.33±2.08 b ]]> value of a <![CDATA[14.67±2.52 c ]]> <![CDATA[-4.33±1.16 a ]]> <![CDATA[5.67±2.08 b ]]> b value <![CDATA[-41.67±1.53 a ]]> <![CDATA[-37.67±0.58 b ]]> <![CDATA[-39.33±1.16 b ]]>

[0159] In Table 2, within the same row, different superscript letters indicate significant differences in the product's chromaticity values. The L value represents brightness, the a value represents the red-green valence, and the b value represents the yellow-blue valence. A positive b value indicates that the object has a yellow tint, and a negative b value indicates that the object has a blue tint. The larger the absolute value, the higher the color saturation.

[0160] 3.2) Colorimetric evaluation of phycocyanin lake

[0161] The appearance of the lake products obtained in Examples 1-3 is as follows: Figure 3 As shown. Among them, the lake color of Example 1 is dark blue, the lake color of Example 2 is blue, and the lake color of Example 3 is dark blue.

[0162] See Figure 3 According to Table 2, compared with Examples 1 and 3, the proportion of phycocyanin in Example 2 is lower, while the proportion of sodium stearoyl lactylate is higher. In the colorimetric measurement results of the lake obtained in Example 2, the b value representing the blue color is relatively low, and the blue color of the lake product is slightly lighter.

[0163] Although the lake products obtained in Examples 1-3 showed some differences in color intensity due to variations in the mass ratio of phycocyanin to emulsifier, all lake products in Examples 1-3 essentially exhibited a pure blue color. Furthermore, see Table 1 and... Figure 4 Although the blue color of the lake product obtained in Example 2 is slightly lighter, the phycocyanin precipitation rate of Example 2 is relatively higher, and the physical stability of this phycocyanin lake after dispersion in the oil phase is relatively better.

[0164] Those skilled in the art will clearly understand that, for the sake of convenience and brevity, the same or similar parts between the various embodiments in this application can be referred to mutually.

[0165] The above description is merely a specific embodiment of this application and is not intended to limit this application. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the scope of protection of this application.

Claims

1. A process for the preparation of natural color lake based on phycocyanin characterized in that, The method comprises the following steps: Step 1, mixing the aqueous solution of phycocyanin with the aqueous solution of stearic acid lactylate; Step 2, after the aqueous solution of phycocyanin is mixed with the aqueous solution of stearic acid lactylate, a salt solution containing divalent metal ions is added, so that the negatively charged groups of phycocyanin and the stearic acid lactylate ions are combined by the bridging action of divalent metal ions to form a lake, and the lake is precipitated from the aqueous solution; Step 3, collecting and processing the lake to obtain a lake product; The salt solution containing divalent metal ions includes a calcium ion salt solution. In step 1, the phycocyanin is 40-50 parts, and the stearic acid lactylate is 50-60 parts by mass.

2. The method of claim 1, wherein, The stearic acid lactylate includes calcium stearic acid lactylate and / or sodium stearic acid lactylate.

3. The method of claim 1, wherein, The calcium ion salt solution includes a calcium chloride solution.

4. The method according to any one of claims 1 to 3, characterized in that, The method further comprises the following steps: A beater is added with pure water at a temperature of 40-80℃, the stearic acid lactylate is put into the beater while stirring, and after the feeding is completed, the stirring is continued for dissolution, and then the temperature is lowered to 25-60℃, so as to prepare the aqueous solution of stearic acid lactylate; Another beater is added with normal temperature pure water, the phycocyanin is put into the beater while stirring, and after the feeding is completed, the stirring is continued for dissolution, so as to prepare the aqueous solution of phycocyanin; Another beater is added with normal temperature pure water, the salt containing divalent metal ions is put into the beater while stirring, and after the feeding is completed, the stirring is continued for dissolution, so as to prepare the salt solution containing divalent metal ions.

5. The method according to any one of claims 1 to 3, characterized in that, Step 3 comprises: The lake precipitated in the aqueous solution is collected by means of static settling and / or centrifugation; The collected lake is washed, and the washed lake is dried; The dried lake is crushed and sieved to obtain a lake product.

6. A lake product characterized in that, The lake product is prepared by any one of the methods of claims 1-5.

7. The lake product of claim 6 is used in oil-based food.

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