A jelly containing a carotenoid ester

Abstract

This invention provides a jelly containing carotenoid esters, prepared from raw materials comprising the following mass percentages: 0.01-0.02% carotenoid esters, 9% sweetener, 1% thickener, 9-15% fruit juice, and the balance being water; the carotenoid esters are added in one or more of the following forms: chromopeptide extract, carotenoid ester crystals, and carotenoid ester microcapsule powder. The jelly containing carotenoid esters proposed in this invention optimizes the proportions of the added components and the preparation process; the textural parameters of the prepared jelly sample are essentially not significantly different from those of commercially available jellies.

Description

Technical Field

[0001] This invention belongs to the field of food processing technology, and specifically relates to a jelly containing carotenoid esters. Background Technology

[0002] Naturally derived carotenoid esters are a class of natural derivatives formed by the esterification of oxygenated carotenoids such as lutein, zeaxanthin, and β-cryptoxanthin with fatty acids. They are widely distributed in the fruit tissues of plants such as wolfberry, physalis, citrus, sea buckthorn, and yellow peach, as well as in the petal tissues of plants such as marigold. These components possess both coloring properties and nutritional functions, exhibiting antioxidant activity and provitamin A activity. They also demonstrate certain biological effects in areas such as vision protection, nerve development, brain health regulation, and anti-tumor activity (Duan Hao et al., *Food Industry Technology*, 2023). Therefore, they have significant application value in food, health food, and related products.

[0003] Because carotenoid esters are highly hydrophobic, they are difficult to disperse stably in aqueous systems. Current food processing and formulation technologies typically employ microencapsulation, emulsification, or oil dissolution to prepare them as microcapsule powders, crystals, or oil formulations before adding them to food. For example, Gao Jiangtao et al. applied lutein ester CWS microcapsule powder to beverages and yogurt systems (Food Industry, 2021); patent CN106085586A discloses a fortified edible vegetable oil rich in natural carotenoids and its preparation method, which increases the carotenoid content in the edible oil through ultrasonic, microwave, or a combination of ultrasonic and microwave extraction. All of these technical routes rely on exogenous wall materials, emulsifiers, or oil carriers to construct artificially embedded or emulsified structures, enabling carotenoid esters to exist in liquid or semi-solid food formulations.

[0004] Chromoplasts are natural organelles in plant cells specifically responsible for the synthesis, accumulation, and storage of carotenoids. Their internal complex structures, composed of lipids and proteins, can stably support fat-soluble pigments such as carotenoid esters. Chromoplast particles can form stable dispersions in aqueous systems, exhibiting excellent water dispersibility. Essentially, they are naturally occurring carotenoid delivery and stabilization carriers that can be directly introduced into aqueous food systems without the need for exogenous wall materials or artificial encapsulation.

[0005] However, current technologies lack a technical solution for directly using chromopeptides rich in carotenoid esters as structurally and functionally integrated raw materials in gel-type foods such as jellies, and for constructing stable and uniform functional gel systems by utilizing their natural water dispersibility and organelle microstructure. How to maintain the gel structure and sensory quality of jellies while ensuring the uniform distribution of chromopeptide particles and preserving their natural structural characteristics, thereby achieving the stable existence and structured delivery of carotenoid esters without artificial encapsulation, still requires further research and development.

[0006] Therefore, developing a chromopeptide jelly with plant chromopeptides as the core functional structural unit, which can achieve stable dispersion of carotenoid esters in aqueous gel foods without artificial encapsulation, and its preparation method, has important technological innovation and application value. Summary of the Invention

[0007] In view of the shortcomings of the existing technology, the purpose of this invention is to propose a jelly containing carotenoid esters, which has superior performance in terms of texture, taste and bioavailability.

[0008] The technical solution for achieving the above-mentioned objective of this invention is as follows: A jelly containing carotenoid esters is prepared from the following raw materials in the indicated weight percentages: 0.01-0.02% carotenoid esters, 8-10% sweetener, 1% thickener, 9-15% fruit juice, and the remainder being water; wherein the thickener is one or more of carrageenan, konjac gum, sodium alginate, and agar. The carotenoid esters are added in one or more of the following forms: chromoplast extracts, carotenoid ester crystals, and carotenoid ester microcapsule powders.

[0009] The chromopeptide extract is made from one of the following plant tissues rich in carotenoid esters: wolfberry, pumpkin, papaya, marigold, sweet pepper, apricot, yellow-fleshed kiwi, loquat, yellow peach, persimmon, sea buckthorn, citrus fruits, Physalis fruits, and Rosa fruits. The extraction method includes the following steps: 1) Crushing and centrifugation: Add extraction solvent at a ratio of 1:10 (g / mL) to extract, then crush and filter, collect the filtrate and centrifuge, discarding the supernatant; 2) Chromoplastics: The precipitate obtained by centrifugation is resuspended in water to obtain chromoplastics extract. The crude extract obtained in this step is used for jelly making to ensure extraction efficiency.

[0010] In step 2), the extraction solvent is selected from one of the following: a sucrose solution with a concentration of 0.2-0.4 mol / L, a PBS buffer solution with a concentration of 0.01-0.02 mol / L, or a sorbitol solution with a concentration of 0.3-0.4 mol / L. Preferably, the extraction solvent is a sucrose solution with a concentration of 0.33 mol / L.

[0011] The extraction solvent is added to the raw material, and then the mixture is crushed for 10-15 seconds using a juicer at 1300 W. After filtering through two layers of filter cloth, the filtrate is collected and centrifuged at 3000-5000 g for 10-20 minutes at 2-7°C. The supernatant is then discarded.

[0012] Goji berries ( Lycium barbarumLycium barbarum (L.) is a traditional medicinal and edible plant. Its fruit is rich in carotenoids, primarily stored in tubular chromoplasts, thus possessing potential for the development and utilization of plant-derived chromoplasts. The raw material for chromoplast extraction can be fresh or dried Lycium barbarum. Dried Lycium barbarum has a lower cost, and its extraction rate is not significantly different from that of fresh Lycium barbarum, both being approximately 50%. Therefore, this invention preferably uses dried Lycium barbarum after rehydration for extraction.

[0013] A preferred embodiment of the present invention is that the raw material for extraction is dried wolfberry. The dried wolfberry is first rehydrated at a temperature of 2-7°C for 3-5 hours, then an extraction solvent is added, and the mixture is crushed using a juicer.

[0014] Step 2) The ratio of water to dried goji berries used for resuspension can be 5~15mL:10g. For example, if the raw material of dried goji berries is 10g, it can be resuspended in 10mL of water.

[0015] The thickener is a mixture of konjac gum and carrageenan in a mass ratio of 2-4:2; and / or The sweetener is selected from one or more of maltitol, sorbitol, xylitol, lactitol, mannitol, and erythritol.

[0016] More preferably, the thickener is a mixture of konjac gum and carrageenan in a mass ratio of 3:2.

[0017] Another preferred embodiment of the present invention is that the juice is selected from one of the following juices: concentrated apple juice, concentrated orange juice, concentrated grape juice, concentrated pineapple juice, concentrated mango juice, concentrated pomegranate juice, concentrated jujube juice, concentrated peach juice, concentrated pear juice, concentrated apricot juice, etc.; the sugar content of the juice is 60-72% by mass.

[0018] The jelly is prepared through the following steps: S1 Dry Powder Mixing: The thickener and sweetener are pre-dry mixed evenly, and then added to the water while stirring; S2 swelling and boiling: Stop stirring, let it stand at room temperature for 25~40 min to swell, then place it in a 75~85℃ water bath and stir gently for 4~6 min to obtain the glue solution; S3. Mixing and blending: First, mix the carotenoid esters with water, then add them together with the fruit juice into the gel, and let the gel cool naturally.

[0019] S2 is better prepared by allowing it to swell at room temperature for 30 minutes, and then placing it in an 80°C water bath and stirring gently for 5 minutes. This avoids introducing too many air bubbles and makes the solution clear and transparent. However, if the gel is boiled for too long, it will not gel properly.

[0020] If excessive foam is generated, trace impurities and foam can be removed by filtration. During water bath operation, the adhesive solution should be kept below the water surface in the bath to prevent failure to gel properly.

[0021] Further, in step S3, the carotenoid ester is added in the form of a chromopeptide extract. The chromopeptide extract is mixed with water at a volume ratio of 1:2~8 and sheared at 7000~10000 rpm for 2~4 min. Previous experiments have shown that uncuttered chromopeptides cannot be evenly added to the jelly.

[0022] More preferably, in step S3, when the adhesive solution cools to 58~62°C, the chromophore extract and fruit juice are added while stirring.

[0023] The beneficial effects of this invention are as follows: The jelly containing carotenoid esters proposed in this invention has optimized the addition ratio of ingredients and the preparation process. The textural parameters of the prepared jelly sample are basically not significantly different from those of commercially available jelly.

[0024] The present invention proposes a jelly containing carotenoid esters. The jelly prepared with different forms of carotenoid esters was compared. The jelly prepared with chromopeptide extract was preferred. The jelly prepared with chromopeptide extract had the best antioxidant activity, with a scavenging rate of 26.40% at a concentration of 1.0 mg / mL.

[0025] The bioavailability of jelly prepared from chromopeptide extracts is 13.3%, and since chromopeptides are naturally extracted, the process is simple and the raw material cost is lower. Attached Figure Description

[0026] Figure 1 Photographs of samples of jellies made with different forms of added carotenoid esters; Figure 2 A comparison of the ABTS antioxidant activity of four types of jelly; Figure 3 This is a radar image of an electronic nose. Figure 4 This is a PCA diagram of the electronic nose.

[0027] Figure 5 The in vitro release curves of carotenoid ester microcapsule powder and chromopeptide are shown.

[0028] Figure 6 The hemolysis rate of microcapsules containing different concentrations of chromopeptides and carotenoid esters under different concentrations of blood cells was measured.

[0029] Figure 7 Comparison of single-factor experimental results on the amount of carotenoid esters added to jelly. Detailed Implementation

[0030] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention.

[0031] Unless otherwise specified, all technical means used in this instruction manual are known in the field, and all raw materials used are commercially available.

[0032] Antioxidant activity assay: ABTS reacts with potassium persulfate, becoming oxidized to generate stable blue-green cationic free radicals ABTS⁺. ABTS⁺ exhibits maximum absorbance at a specific wavelength of 734 nm. When an antioxidant is added to the ABTS solution, the antioxidant reacts with ABTS⁺, reducing it and causing it to fade, resulting in a decrease in absorbance. The degree of absorbance decrease is positively correlated with the activity of the antioxidant in the sample (i.e., its ability to scavenge free radicals).

[0033] The fruit juice used in this experiment was Andeli brand apple concentrate, with a sugar content of 70.2%.

[0034] Example 1: Extraction of Chromosomes This embodiment uses dried wolfberry as the extraction raw material to extract chromophores. The extraction method includes the following steps: 1) Pretreatment: Rehydrate dried goji berries at 4℃ for 3.5 h.

[0035] 2) Crushing and centrifugation: Add 0.33 mol / L sucrose solution as extraction solvent at a material-to-liquid ratio of 1:10 (g / mL), and crush using a juicer at 1300 W for 10 s. After filtering through two layers of filter cloth, collect the filtrate. Centrifuge at 3000 g for 15 min at 4℃ to separate the filtrate, and discard the supernatant.

[0036] 3) Chromoplasts: The precipitate was resuspended in a small amount of water to obtain the chromoplast extract. The ratio of water to dried wolfberries used for resuscitation was 10 mL: 10 g.

[0037] The obtained chromoplast extract contained 16.21 ± 0.32 g / 100 g of carotenoid esters on a dry basis.

[0038] Example 2 Preparation of Jelly This embodiment uses thickeners, fruit juice, water, etc. to prepare jelly, in order to obtain an optimized process for jelly preparation. The preparation process is as follows: S1. Dry Powder Mixing: Pre-mix carrageenan, konjac gum, and maltitol evenly. While stirring, slowly and in batches add the mixed dry powder to cold water to ensure that the powder is fully dispersed and free of lumps.

[0039] S2, swelling and gel cooking: Stop stirring and let it swell at room temperature for 30 minutes. Then place it in an 80°C water bath and stir gently for 5 minutes (avoid introducing too many air bubbles, as excessive cooking time will prevent normal gelation). The solution should be clear and transparent.

[0040] If excessive foam is generated, trace impurities and foam can be removed by filtration. During water bath operation, the adhesive solution should be kept below the water surface in the bath to prevent failure to gel properly.

[0041] S3. Mixing and blending: When the glue solution cools to 60℃, add apple concentrate while stirring.

[0042] In this embodiment, the sum of carrageenan and konjac gum is 1% of the total mass of raw materials, and the ratio of the two is set to 1~6:1~7; maltitol is 9%; and the amount of apple concentrate added is 9~15% of the total mass of raw materials. The jelly obtained in different proportions was evaluated for taste (the number of people conducting the sensory evaluation was 10), and the average scores are listed in the table below.

[0043] Table 1: Jelly Solution Optimization plan Konjac gum: Carrageenan (1%) Apple concentrate addition amount (%) Sensory evaluation score 1 1:1 9 72.1 2 1:1 12 75.3 3 1:1 15 70.2 4 6:4 9 70.1 5 6:4 12 87.6 6 6:4 15 68.2 7 3:7 9 62.1 8 3:7 12 66.1 9 3:7 15 68.2 Based on comprehensive sensory evaluation, the jelly with a konjac gum to carrageenan ratio of 6:4 and 12% apple concentrate received the highest sensory evaluation. This jelly was clear and transparent, with good elasticity and a moderate balance of acidity and sweetness. A jelly prepared according to the same konjac gum to carrageenan ratio of 6:4 and 12% apple concentrate was labeled as a blank jelly.

[0044] Example 3: Single-factor experiment on the amount of carotenoid esters added This embodiment provides a jelly containing carotenoid esters, and its preparation steps are as follows: S1. Dry Powder Mixing: Pre-mix carrageenan, konjac gum (3:2, 1%), and maltitol (9%) until homogeneous. While stirring, slowly and in batches add the mixed dry powder to cold water to ensure that the powder is fully dispersed and free of lumps.

[0045] S2, swelling and boiling: Stop stirring, let it swell at room temperature for 30 minutes, then place it in an 80℃ water bath and stir gently for 5 minutes until the solution is clear and transparent.

[0046] S3, Mixing and blending: Carotenoid ester formulation: The chromopeptide extract was mixed with a small amount of water and sheared at 8000 rpm for 3 min to obtain a liquid phase containing carotenoid esters.

[0047] During the glue boiling process in step S2 (i.e., at a temperature of 80°C), a liquid phase containing carotenoid esters is added. The mixture is then poured into a mold and allowed to cool naturally.

[0048] See Figure 5When the amount of carotenoids added to the colored jelly was 10 mg / 40 g, obvious flocculent aggregation and poor dispersion occurred. When the amount of carotenoids added to the goji berry puree was 8 mg / 40 g, the jelly texture was significantly affected, and it failed to set.

[0049] Through single-factor optimization experiments, it was determined that 8 mg of carotenoid esters should be added to every 40 g of jelly. This amount of carotenoid esters was set based on the recommended daily intake and sensory evaluation, taking into account both the recommended daily intake and taste. 4 mg per 40 g of jelly is equivalent to 0.01% (by mass) of carotenoid esters.

[0050] Example 4 This embodiment provides a jelly containing carotenoid esters, and its preparation steps are as follows: S1. Dry Powder Mixing: Pre-mix carrageenan, konjac gum (3:2, 1%), and maltitol (9%) until homogeneous. While stirring, slowly and in batches add the mixed dry powder to cold water to ensure that the powder is fully dispersed and free of lumps.

[0051] S2, swelling and gel cooking: Stop stirring and let it swell at room temperature for 30 minutes. Then place it in an 80°C water bath and stir gently for 5 minutes (avoid introducing too many air bubbles, as excessive cooking time will prevent normal gelation). The solution should be clear and transparent.

[0052] If excessive foam is generated, trace impurities and foam can be removed by filtration. During water bath operation, the adhesive solution should be kept below the water surface in the bath to prevent failure to gel properly.

[0053] S3, Mixing and blending: Preparation of carotenoid ester components: The chromopeptide extract was mixed with a small amount of water (the ratio of chromopeptide extract to water was 1:5 v / v), and sheared at 8000 rpm for 3 min (unsheared chromopeptide extract could not be evenly added to the jelly) to obtain a liquid phase containing carotenoid esters.

[0054] During the glue boiling process in step S2 (i.e., at a temperature of 80°C), a liquid phase containing carotenoid esters is added. The mixture is then poured into a mold and allowed to cool naturally. Example 5

[0055] This embodiment provides a jelly containing carotenoid esters. The preparation steps are basically the same as in Example 4, except that: after the gel is cooled to 70°C, the liquid phase containing carotenoid esters is added. Example 6

[0056] This embodiment provides a jelly containing carotenoid esters, and its preparation steps are basically the same as those in Example 4, except that: after the gel is cooled to 60°C, the liquid phase containing carotenoid esters is added. The resulting product was labeled as chromopeptide jelly.

[0057] The jelly yielded 120 g. 5 g of jelly was taken, and water was added to bring the total to 50 g. After mixing, 5 g was used to extract carotenoid esters, and the loss was calculated. The results are shown in Table 2. Comparing the jellies prepared in Examples 4-6, the jelly prepared according to the process in Example 6, which cooled to 60°C before adding the chromophore, had the lowest loss rate. At lower temperatures, the jelly solution had already partially solidified, making it impossible to add more chromophore. 60°C will be used as the operating parameter in subsequent tests. Example 7

[0058] This embodiment provides a jelly containing carotenoid esters. The preparation steps are basically the same as in Example 6, except that: in step S3, the carotenoid esters are added in the form of carotenoid ester crystals (Chenguang Biotech). The carotenoid ester crystals are made into a solution according to the ratio of 10mg of carotenoid ester crystals to 10mL of water, so that each 40g of the resulting jelly contains 4mg of carotenoid esters.

[0059] The resulting product was labeled as carotenoid ester crystal jelly. Example 8

[0060] This embodiment provides a jelly containing carotenoid esters. The preparation steps are basically the same as in Example 6, except that: in step S3, the carotenoid esters are added in the form of carotenoid ester microcapsule powder (Chenguang Biotech) at a ratio of 10mg of carotenoid ester microcapsule powder to 10mL of water to make a solution, so that each 40g of the resulting jelly contains 4mg of carotenoid esters.

[0061] The resulting product was labeled as carotenoid ester microcapsule powder jelly.

[0062] The jelly weighed 120 g. 5 g of jelly was taken, and water was added to make up to 50 g. After mixing well, 5 g was taken to extract carotenoids, and the loss was calculated. The results are shown in Table 2.

[0063] Table 2. Loss of carotenoids during processing

[0064] Different letters represent significant differences. p <0.05 The jelly samples obtained in each example were tested using a texture analyzer, and the results are shown in Table 3 (the chromopy jelly in the table below was prepared by the method in Example 6).

[0065] Table 3 Texture parameters of different jellies Blank Jelly Chromoplastics Carotenoid ester crystal jelly Carotenoid ester microcapsule powder jelly hardness 544.65±81.421a 437.21±32.36a 528.98±92.72a 465.17±24.68a viscosity -47.13±14.29a -39.37±1.43a -47.10±12.69a -37.32±8.73a elasticity 0.68±0.09b 0.68±0.03b 0.68±0.02b 0.69±0.05b cohesion 0.44±0.05b 0.48±0.07b 0.41±0.03b 0.48±0.07b Adhesion 237.04±43.08a 148.32±30.17b 164.42±31.44b 166.20±12.81b chewing 163.92±50.13a 134.46±36.34a 148.24±25.34a 127.73±20.50a responsive 0.09±0.01b 0.12±0.03b 0.14±0.04b 0.15±0.04b There were no significant differences in the textural parameters of the four types of jelly samples.

[0066] See photos of jelly samples Figure 1 The chromopy jelly in the table below was prepared using the method of Example 6.

[0067] Table 4 Color difference parameters of different jellies

[0068] Table 4 L* Represents brightness (0-100, the higher the value, the brighter). a* Represents the red-green tint (positive values ​​indicate redness, negative values ​​indicate greenness). b* Represents the degree of yellow-blue (positive values ​​indicate yellowishness, negative values ​​indicate bluishness). c* Represents chroma (saturation; the higher the value, the more vibrant the color). h° Represents the hue angle (0 ° -360° (representing different colors). As can be seen from the results in Table 4, carotenoid ester crystals cannot dissolve well in the jelly solution, and the values ​​are close to those of the blank group, with a large error.

[0069] Antioxidant activity assay: Results are shown in Figure 2 The antioxidant activity of ABTS in the four types of jelly was dose-dependent, with the chromoplastic jelly showing the best antioxidant activity, achieving a clearance rate of 26.40% at a concentration of 1.0 mg / mL.

[0070] Electronic nose detection: Samples from Examples 6-8 were cut into small pieces and placed approximately 1 / 3 of the volume into headspace vials. The vials were sealed with the headspace cap and allowed to stand for 60 minutes. The electronic nose preparation time was 10 s, sample injection time was 60 s, cleaning time was 120 s, and detection time was 120 s. The gas flow rate was 1 L / min. Each sample was tested in triplicate. After the tests, the obtained sample data were processed and analyzed. Results are shown below. Figure 3 , Figure 4 (The chromosome jelly in the figure was prepared using the method of Example 6). According to the radar chart and PCA chart analysis, the aroma characteristics of the chromosome jelly are significantly different from those of other jellies (the elliptical regions are completely separated), while the aroma characteristics of the other three jellies are highly similar.

[0071] The table below shows the types and properties of representative materials for electronic nose sensors.

[0072] Table 5. Representative Material Types and Performance Descriptions of Sensors

[0073] Electronic nose results analysis: The main aroma characteristics of the chromopeptide jelly are methyl and sulfide compounds, while the other three jelly types are aromatic. Goji berries themselves contain abundant methyl and sulfide aroma components, such as 3-methylbutyraldehyde and methylthiopropanol. These two aroma components are more easily released during processing, which is the main reason for the significant difference between the chromopeptide jelly and the other three types. The core components of the other three jelly types are basic esters and simple alcohols, with the main aroma provided by apple concentrate. The addition of crystal powder and microcapsule powder has a negligible impact on the aroma and is insufficient to create a unique aroma profile. In vitro release experiment:

[0074] 1. Sample preparation: Take 1 mL of sample and place it in a dialysis tube with a molecular weight cutoff of 10 kDa.

[0075] 2. Release Experiment (1) Stomach Immerse the sealed dialysis bag into an Erlenmeyer flask containing 120 mL of SGF. Place the flask in a constant temperature water bath shaker at 37°C and 100 rpm, and take samples at time points of 15, 30, and 60 min. Each time, take 1 mL of solution from the external medium and immediately add 1 mL of fresh, preheated SGF to maintain a constant medium volume.

[0076] (2) Intestines Two hours later, remove the dialysis bag from the SGF and blot dry the surface liquid with filter paper. Quickly transfer the dialysis bag to an Erlenmeyer flask containing an equal volume of fresh SIF. Continue incubation at 37°C and 100 rpm. Take samples at time points of 15 / 30 / 60 / 90 / 120 / 240 / 360 / 480 min. Each time, take 1 mL of solution from the external medium and immediately add 1 mL of fresh, preheated SIF to maintain a constant medium volume.

[0077] 3. Sample testing The extracted sample solution was centrifuged at 3000 g for 5 minutes, and the supernatant was collected. The carotenoid content was determined using a liquid chromatography method.

[0078] Calculation method: The concentration of each sample is measured to determine the cumulative release. M n = C n × V total + Σ (C i × V sample ) C n : The drug concentration measured in the supernatant after centrifugation of a sample taken from the external medium at the nth time point.

[0079] V totalTotal volume of the release medium (120 mL for both the gastric and intestinal stages).

[0080] V sample : The volume of each sample taken (5 mL).

[0081] Σ (C i × V sample ): Sum the amount of drug in the samples taken at the first n-1 time points.

[0082] like Figure 5 As shown, considering that both carotenoid ester microcapsule powder and chromoplasts are encapsulation-type delivery systems, while carotenoid ester crystal powder is in a free state and lacks clear structural encapsulation protection, this study further compared and analyzed the release dynamics of the two encapsulation systems during in vitro digestion to evaluate the regulatory effect of different delivery structures on the release behavior of carotenoid esters. The analysis results showed that both systems exhibited low release levels and slow release rates in the gastric phase (0–60 min), indicating that both microcapsule powder and chromoplast structures can, to some extent, resist the damage of the gastric environment, thereby playing a protective role for carotenoids. After entering the intestinal phase, the release amount of both groups gradually increased and tended to stabilize in the later stages of digestion, with a final release rate of approximately 54%, and no significant difference was observed. p > 0.05). However, from the perspective of release kinetics, the chromopeptide system exhibits a smoother release curve throughout the intestinal phase, and its release rate is significantly lower than that of the microcapsule powder system, showing typical sustained-release behavior.

[0083] This difference indicates that although the two delivery systems have similar final release amounts, chromoplasts, through their natural lipid-protein complex structure, provide a more stable spatial constraint on carotenoids, thereby delaying their release in the digestive system. Compared to the looser or more easily destroyed artificial encapsulation structures in microcapsules, the substructures within chromoplasts (such as lipid droplets or membrane structures) may gradually deconstruct during digestion, achieving sustained release.

[0084] This "delayed release-supply" kinetic characteristic helps prolong the time window for carotenoids to exist in the intestine, which is more conducive to their formation with mixed micelles and their contact and absorption with intestinal epithelial cells, thus providing more favorable conditions for improving their bioavailability and exerting their functional activity. In vitro digestion experiment:

[0085] 1. Preparation of digestive fluid: Thaw the digestive fluid on a constant temperature water bath shaker at 95 rpm and 37°C, and adjust the pH of SSF, SGF and SIF to 7, 3 and 7 respectively with 1M HCl and 1M NaOH.

[0086] 2. Preparation of α-amylase solution and pepsin solution: All enzyme solutions must be prepared with the corresponding digestive fluids, and oral and gastric solutions should be prepared at the same time.

[0087] α-Amylase solution: SSF simulated saliva + "thermally stable α-amylase" 10 mL ~ 161 mg (~ 161 mg indicates the amount of amylase added) Pepsin solution: SGF mimics gastric juice + "Pepsin" 20 mL ~ 3516 mg 3. Oral digestion: Add 3 mL of simulated saliva (SSF), 1 mL of α-amylase solution (mix well before use), 25 μL of CaCl2 (0.3 M) solution and 975 μL of ultrapure water to 5 g of sample, and shake the mixture in a water bath for 2 min (95 rpm, 37℃).

[0088] 4. Gastric phase digestion: Add 6 mL of simulated gastric juice (SGF) to the mixture after oral phase digestion, and adjust the pH to 4 with 1 M HCl. Add 2 mL of pepsin solution (mix well before use), 10 μL of CaCl2 (0.15 M) solution, and adjust the pH to 3 with 1 M HCl. Add 1.99 mL of ultrapure water (minus the amount of hydrochloric acid added). Cover the mixture with nitrogen and purge air for 1–2 min. Shake in a water bath for 2 h at 37 °C and 95 rpm.

[0089] 5. Preparation of pancreatic enzymes and bile: Dissolve intestinal digestive fluid (SIF) at 37℃ and 95 rpm, and adjust the pH to 7.

[0090] Pancreatic enzyme: 60 mL (SIF) ~3.7398 g (prepared in a beaker with magnetic stirring); Bile: 30 mL (SIF) ~1.7001 g (prepared in a 50 mL centrifuge tube with thorough vortex dispersion) 6. Intestinal Digestion: Adjust the pH of the digested chyme from the mouth and stomach to 6 using 1M NaOH solution. Then add 10 mL of porcine pancreatic enzyme, 5 mL of bile, and 40 µL of 0.3 M CaCl2 solution. Adjust the pH to 7 again using 1M NaOH solution. Make up to 40 mL with distilled water (3.96 mL added). Cover the mixture with nitrogen and purge air for 1-2 minutes. Incubate in a water bath with shaking for 2 hours at 37°C and 95 rpm. After intestinal digestion, make up to 50 mL with distilled water. Shake thoroughly to obtain the final digestive solution.

[0091] 7. Determination of carotenoids: Take 3 × 2.5 mL of the digestion solution after volume adjustment and shaking, and measure the recovery and hydrolysis. Centrifuge the remaining digestion solution at 4℃, 10000 g, and 60 min. After centrifugation, take 5 mL of the supernatant and measure the release rate. Pass the remaining solution through a membrane (0.22 µm cellulose acetate membrane) to obtain 5 mL of micelle phase and measure the accessibility.

[0092] Table 6 Results of in vitro digestion of jelly

[0093] Under the same carotenoid ester addition level (4 mg / 40 g), the chromopeptide jelly exhibited significantly superior properties compared to the carotenoid ester crystalline powder jelly during in vitro digestion. Specifically, the recovery rate of the chromopeptide jelly (68.21±3.92%) was significantly higher than that of the crystalline powder jelly (35.64±3.22%), indicating better stability in the gastrointestinal digestive environment. Simultaneously, its release rate and bioavailability reached 27.12±1.21% and 13.34±1.54%, respectively, both significantly higher than those of the crystalline powder jelly (19.23±2.65% and 5.11±0.89%), demonstrating that the chromopeptide structure not only helps reduce the degradation loss of carotenoid esters but also promotes their effective release from the jelly matrix and further entry into the absorbable phase.

[0094] Furthermore, we selected commercially available carotenoid ester microencapsulated powder jelly as a control for comparison. The results showed that the recovery rate, release rate, and bioavailability of a certain brand of blueberry carotenoid ester jelly (2.5 mg / 40 g) were 42.92±2.45%, 21.45±2.61%, and 6.10±1.12%, respectively, which were generally lower than the chromopeptide jelly system constructed in this study. Although this commercially available product used microencapsulation technology to improve the stability and dispersibility of carotenoid esters, its bioavailability was still significantly lower than that of the chromopeptide jelly, indicating that the artificially embedded structure has limited effect on promoting the release and micellization of carotenoid esters during digestion. In contrast, chromopeptides, as a natural lipid-protein complex structure, not only provide effective digestive protection but also achieve a more favorable dynamic structural response for release and transport during digestion, thus exhibiting superior in vitro digestion behavior and potential absorption and utilization advantages overall. These results further support the advantages of natural chromoplast structures in carotenoid delivery, showing greater potential in terms of structural adaptability and functional synergy compared to traditional microencapsulation systems. Biocompatibility assessment:

[0095] This experiment compared the ability of chromoplasts and microcapsule powders to lyse red blood cells.

[0096] Blood samples (purchased) were centrifuged at 3000 rpm for 5 min to separate red blood cells (RBCs). The obtained RBCs were washed approximately 6 times with PBS buffer at pH 7.4 until the supernatant was clear. Subsequently, the RBCs were resuspended in PBS to prepare a 4% (v / v) RBC suspension.

[0097] Chromosomal bodies (CHR, 100 µL) dispersed in PBS were added to 900 µL of erythrocyte suspension to achieve final concentrations of 25, 50, 100, 200, and 400 µg / mL. PBS was used as a negative control, and deionized water (ddH2O) was used as a positive control.

[0098] The above mixture was incubated at 37 °C for 3 h, followed by centrifugation at 8000 rpm for 10 min, and the supernatant was collected. The absorbance of the released hemoglobin in the supernatant was measured at 545 nm using a microplate reader.

[0099] The hemolysis rate is calculated using the following formula: A sample =A (RBC+CHR) -A (CHR blank)

[0100] in, : A suspension of red blood cells containing chromoplasms; : Only different concentrations of chromoplasts, no blood cells; : Absorbance of sample group.

[0101] In in vitro digestion experiments, the carotenoid ester crystal powder system has shown poor stability and bioaccessibility, making it difficult to use as an effective delivery form for subsequent biological evaluation. Therefore, this study further selected two systems with structural embedding characteristics—chromosomes and carotenoid ester microcapsule powders—and compared their biocompatibility. The results showed that ( Figure 6When the concentration exceeded 100 μg / mL, both systems exhibited some degree of hemolysis, which increased with increasing concentration. Within the tested concentration range (maximum 400 μg / mL), the maximum hemolysis rate of the chromopeptide system was 0.42%, while that of the carotenoid ester microcapsule powder was 0.63%. Overall, both systems showed low hemolysis levels within this concentration range, indicating good blood compatibility. Further comparison revealed that the hemolysis rate of the chromopeptide system was significantly lower than that of the microcapsule powder, indicating superior biocompatibility under the same conditions. Loading capacity and system compatibility of chromopeptide jelly:

[0102] Based on the advantages of chromopeptides in terms of digestibility, bioavailability, and biocompatibility, their application potential is further evaluated from the perspective of food system construction. (See also...) Figure 7 Different levels of carotenoid addition significantly affected the jelly-forming ability of the system. With increasing addition, the chromopeptide system consistently produced structurally intact and uniformly shaped jellies over a wide range, indicating good system compatibility and high loading capacity. In contrast, the wolfberry pulp system was more sensitive to the amount of carotenoid added; when the carotenoid addition exceeded 8 mg / 40 g, the jelly structure was significantly disrupted, making it difficult to form a stable gel.

[0103] The above results indicate that, compared to wolfberry puree, chromopeptides, as structured delivery units, can maintain the structural integrity of the jelly system at higher addition levels, thus significantly broadening the application scope of carotenoids in gel foods.

[0104] Although the present invention has been described above through embodiments, those skilled in the art should understand that any improvements and modifications made to the present invention without departing from its spirit and essence should fall within the protection scope of the present invention.

Claims

1. A jelly containing carotenoid esters, characterized in that, It is prepared from raw materials comprising the following mass percentages: 0.01-0.02% carotenoid esters, 8-10% sweetener, 1% thickener, 9-15% fruit juice, and the balance being water; wherein the thickener is one or more of carrageenan, konjac gum, sodium alginate, and agar; The carotenoid esters are added in one or more of the following forms: chromoplast extracts, carotenoid ester crystals, and carotenoid ester microcapsule powders.

2. The jelly containing carotenoid esters according to claim 1, characterized in that, The raw materials for the chromopeptide extract are selected from one or more of the following: wolfberry, pumpkin, papaya, marigold, sweet pepper, apricot, yellow-fleshed kiwi, loquat, yellow peach, persimmon, sea buckthorn, citrus fruits, fruits of plants in the genus Physalis, and fruits of plants in the genus Rosa. The extraction method includes the following steps: 1) Crushing and centrifugation: Add extraction solvent to the raw material at a ratio of 1:10 (g / mL) for extraction, then crush and filter, collect the filtrate and centrifuge, discarding the supernatant; 2) Chromoplasts: The precipitate obtained by centrifugation was resuspended in water to obtain chromoplast extract.

3. The jelly containing carotenoid esters according to claim 2, characterized in that, In step 2), the extraction solvent is selected from one of the following: sucrose solution with a concentration of 0.2~0.4 mol / L, PBS buffer with a concentration of 0.01~0.02 mol / L, or sorbitol solution with a concentration of 0.3~0.4 mol / L.

4. The jelly containing carotenoid esters according to claim 2, characterized in that, Add extraction solvent to the raw material, then use a juicer to crush it at 1300 W for 10-15 s. After filtering through two layers of filter cloth, collect the filtrate and centrifuge it at 3000-5000 g for 10-20 min at 2-7℃. Discard the supernatant.

5. The jelly containing carotenoid esters according to claim 4, characterized in that, The raw material for extraction is dried goji berries. The dried goji berries are first rehydrated at 2-7℃ for 3-5 hours, then the extraction solvent is added, and the mixture is crushed using a juicer.

6. The jelly containing carotenoid esters according to claim 1, characterized in that, The thickener is a mixture of konjac gum and carrageenan in a mass ratio of 2-4:2; and / or The sweetener is selected from one or more of maltitol, sorbitol, xylitol, lactitol, mannitol, and erythritol.

7. The jelly containing carotenoid esters according to claim 1, characterized in that, The juice is selected from one of the following: concentrated apple juice, concentrated orange juice, concentrated grape juice, concentrated pineapple juice, concentrated mango juice, concentrated pomegranate juice, concentrated jujube juice, concentrated peach juice, concentrated pear juice, and concentrated apricot juice; the sugar content of the juice is 60-72% by mass.

8. The jelly containing carotenoid esters according to any one of claims 1 to 7, characterized in that, The jelly is prepared by the following steps: S1 Dry Powder Mixing: The thickener and sweetener are pre-dry mixed evenly, and then added to the water while stirring; S2 swelling and boiling: Stop stirring, let it stand at room temperature for 25~40 minutes to swell, then place it in a 75~85℃ water bath and stir gently for 4~6 minutes to obtain the glue solution; S3. Mixing and blending: First, mix the carotenoid esters with water, then add them together with the fruit juice into the gel and let the gel cool naturally.

9. The jelly containing carotenoid esters according to claim 8, characterized in that, In step S3, the carotenoid ester is added in the form of a chromopeptide extract. The chromopeptide extract is mixed with water at a volume ratio of 1:2 to 8 and sheared at 7000 to 10000 rpm for 2 to 4 minutes.

10. The jelly containing carotenoid esters according to claim 8, characterized in that, In step S3, when the gel solution cools to 58~62℃, add the chromopeptide extract and fruit juice while stirring.

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