A chitosan membrane loaded with kaempferia galanga essential oil, its preparation method and application

Abstract

This invention relates to the field of food additives and food safety control, specifically disclosing a chitosan membrane loaded with galangal essential oil, its preparation method, and its application. The method includes: (1) first contacting an ethanol-water solution of moringa alcohol-soluble protein with a pectin-water solution to obtain a protein-pectin dispersion; (2) second contacting galangal essential oil with the protein-pectin dispersion to obtain a galangal essential oil emulsion; (3) dissolving chitosan, corn starch, and citric acid in water I, followed by gelatinization, and then adding glycerol for a third contact to obtain a membrane solution; (4) fourth contacting the galangal essential oil emulsion with the membrane solution, casting it into a film, and then drying and equilibrating it to obtain a chitosan membrane loaded with galangal essential oil. The method provided by this invention can improve the loading and protective capacity of essential oils, significantly enhance the inhibitory effect on bound heterocyclic amines, and improve the edible safety of heat-processed plant-based meat products.

Description

Technical Field

[0001] This invention relates to the field of food additives and food safety control, specifically to a chitosan membrane loaded with kaempferol essential oil, its preparation method, and its application. Background Technology

[0002] Plant-based meat is a vegetarian food that has a flavor and texture similar to meat. It typically uses plant proteins (soy protein, peanut protein, wheat gluten, etc.) as the main raw material and is processed using extrusion technology to achieve a meat-like texture. Currently, many food products based on plant-based meat have been developed, such as vegetarian meatballs, vegetarian patties, vegetarian sausages, and vegetarian chicken. However, plant-based meat products, rich in protein, are prone to forming various harmful substances during high-temperature processing, including heterocyclic amines and advanced glycation end products (AGEs). These harmful substances mainly exist in bound forms and accumulate in food. Studies have shown that excessive intake of these harmful substances has very adverse effects on human health.

[0003] Current technologies commonly used to inhibit the formation of harmful substances during meat processing mainly involve adding exogenous additives with specific antioxidant capabilities to the minced meat. However, for plant-based meat products, which require maintaining a specific product shape, the product cannot be minced. Furthermore, considering that during thermal processing, the surface temperature is much higher than the core temperature, causing harmful substances to mainly form and accumulate on the surface of the raw material, traditional methods of adding inhibitors are not applicable.

[0004] In recent years, plant essential oils, such as galangal oil, have received widespread attention due to their rich content of various antioxidants and their safety and significant antioxidant activity. Meanwhile, edible films have gained increasing attention due to their simple manufacturing process, low cost, and non-toxicity, making them widely used in various food preservation fields. However, it is worth noting that plant essential oils are highly hydrophobic and volatile. If used directly on food or mixed directly with hydrophilic films, they cannot exert their full effect, as their active ingredients will be rapidly lost, significantly reducing the film's efficacy.

[0005] Therefore, it is of great significance to provide an edible film that can stably load kaempferol essential oil, is suitable for the thermal processing of plant-based meat products, and can reduce the content of harmful bound heterocyclic amines and advanced glycation end products without affecting the sensory quality of the product. Summary of the Invention

[0006] The purpose of this invention is to overcome the problems of existing technologies where traditional additive inhibitors are not suitable for the thermal processing of plant-based meat products, and cannot reduce the content of harmful bound heterocyclic amines and advanced glycation end products without affecting the sensory quality of the products.

[0007] To achieve the above objectives, a first aspect of the present invention provides a method for preparing a chitosan membrane loaded with kaempferia galanga essential oil, the method comprising:

[0008] (1) The ethanol-water solution of Moringa alcohol-soluble protein was first contacted with the pectin water solution to obtain a protein-pectin dispersion solution.

[0009] (2) Kaempferia galanga essential oil is brought into a second contact with the protein-pectin dispersion solution to obtain Kaempferia galanga essential oil emulsion;

[0010] (3) Chitosan, corn starch and citric acid are dissolved in water I and gelatinized. Then glycerol is added for the third contact to obtain the membrane solution.

[0011] (4) The galangal essential oil emulsion is brought into a fourth contact with the membrane liquid, and after casting into a film, it is dried and balanced to obtain a chitosan membrane loaded with galangal essential oil.

[0012] The moringa prolysin is a non-water-soluble protein extracted from moringa leaf residue, and the protein content of the moringa prolysin is 65-73 wt%.

[0013] The concentration of moringa prolysin in the ethanol-water solution is 3-5 wt%, and the concentration of the pectin water solution is 1.5-5 wt%.

[0014] The second aspect of the present invention provides a chitosan membrane loaded with kaempferia oil prepared by the method described in the first aspect above.

[0015] The third aspect of this invention provides the application of the chitosan membrane loaded with kaempferol essential oil as described in the second aspect above in the field of food safety control.

[0016] Compared with the prior art, the present invention has at least the following advantages:

[0017] (1) In the method of the present invention, pectin, as an anionic polysaccharide, can greatly improve the hydrophobicity of proteins and enhance the stability of emulsions. The unique three-dimensional network structure of the binding interface in the emulsion can significantly improve the loading and protective capacity of essential oils. At the same time, under the encapsulation of colloidal particles, plant essential oils can better exert the function of quenching free radicals.

[0018] (2) Compared with adding galangal essential oil directly to the membrane solution, the method provided by the present invention significantly reduces the amount of galangal essential oil used, while significantly enhancing the inhibition effect on bound heterocyclic amines and significantly reducing the adverse effects on sensory quality. Detailed Implementation

[0019] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0020] It should be noted that, in all aspects of the present invention, the same components or terms in each aspect are described only once in one aspect and not repeatedly, and those skilled in the art should not understand this as a limitation of the present invention.

[0021] As mentioned above, the first aspect of the present invention provides a method for preparing a chitosan membrane loaded with kaempferia galanga essential oil, the method comprising:

[0022] (1) The ethanol-water solution of Moringa alcohol-soluble protein was first contacted with the pectin water solution to obtain a protein-pectin dispersion solution.

[0023] (2) Kaempferia galanga essential oil is brought into a second contact with the protein-pectin dispersion solution to obtain Kaempferia galanga essential oil emulsion;

[0024] (3) Chitosan, corn starch and citric acid are dissolved in water I and gelatinized. Then glycerol is added for the third contact to obtain the membrane solution.

[0025] (4) The galangal essential oil emulsion is brought into a fourth contact with the membrane liquid, and after casting into a film, it is dried and balanced to obtain a chitosan membrane loaded with galangal essential oil.

[0026] The moringa prolysin is a non-water-soluble protein extracted from moringa leaf residue, and the protein content of the moringa prolysin is 65-73 wt%.

[0027] The concentration of moringa prolysin in the ethanol-water solution is 3-5 wt%, and the concentration of the pectin water solution is 1.5-5 wt%.

[0028] In a preferred embodiment, in step (1), the ethanol-water solution of moringa prolysin is a product prepared by a method comprising the following steps: mixing moringa prolysin with 70wt% ethanol-water solution at a rotation speed of 800-1200 rpm for 15-30 min to obtain the ethanol-water solution of moringa prolysin.

[0029] In a preferred embodiment, in step (1), the pectin aqueous solution is a product prepared by a method comprising the following steps: pectin and water II are mixed for a second time at 68-72°C and 800-1200 rpm for 1-2 hours, and the precipitate is removed to obtain the pectin aqueous solution. Studies have found that this preferred embodiment facilitates the formation of a pectin aqueous solution with good dispersibility and stability, thereby improving the moisturizing and mechanical properties of the chitosan membrane loaded with kaempferol essential oil.

[0030] Preferably, the separation is carried out by centrifugation, and at least the following conditions must be met: centrifugal force of 4800-5200g and time of 50-70min.

[0031] It should be noted that in this invention, both water I and water II are water.

[0032] Preferably, in step (1), the volume ratio of the ethanol-water solution of the moringa alcoholic protein to the pectin water solution is 2.5-4:1.

[0033] In a preferred embodiment, the method of the present invention further includes: in step (2), before performing the second contact, adjusting the pH of the protein-pectin dispersion solution to 5.0-6.5, and then applying it to the second contact. Studies have found that in this preferred embodiment, the interaction between pectin molecules can be weakened, promoting the formation of the protein-pectin network.

[0034] Preferably, in step (2), the volume ratio of the galangal essential oil to the protein-pectin dispersion solution is 1-5:100.

[0035] Preferably, in step (1), the first contact is carried out under stirring conditions, and at least the following conditions are met: the rotation speed is 800-1200 rpm and the time is 0.8-1.5 h.

[0036] In a preferred embodiment, in step (2), the second contact is performed in a high-speed homogenization manner, and at least the following conditions are met: the rotation speed is 15000-18000 rpm, the time for each high-speed homogenization is 30-45 s, the time interval between each two adjacent high-speed homogenizations is 30 s, and the high-speed homogenization is performed a total of 3-5 times.

[0037] In a preferred embodiment, in step (3), the dissolution is carried out under stirring conditions, and at least the following conditions are met: temperature is 58-62°C, rotation speed is 800-1200 rpm, and time is 30-45 min.

[0038] Preferably, in step (3), the gelatinization conditions are at least: a temperature of 80-90°C and a time of 1-2.5h.

[0039] In a preferred embodiment, in step (3), the third contact is carried out under stirring conditions, and at least the following conditions are met: temperature is 32-38°C, rotation speed is 800-1200 rpm, and time is 30-45 min.

[0040] In a preferred embodiment, in step (3), the weight ratio of chitosan, corn starch, citric acid, glycerol, and water I is 3-5:1-3:1-1.5:5-8:100.

[0041] Preferably, in step (4), the volume ratio of the galangal essential oil emulsion to the film liquid is 5-8:100.

[0042] Preferably, in step (4), the fourth contact is carried out under stirring conditions, and at least the following conditions are met: the rotation speed is 1300-1700 rpm and the time is 15-30 min.

[0043] Preferably, the method of the present invention further includes: in step (4), before casting the film, the product of the fourth contact is first homogenized and ultrasonically treated in sequence, and then applied to the casting film.

[0044] More preferably, the homogenization conditions shall at least satisfy the following: pressure of 10-15 MPa and time of 3-5 min.

[0045] More preferably, the conditions for the ultrasonic treatment are at least: the power is 380-420W, the time interval between two adjacent ultrasonic treatments is 5s, the duration of each ultrasonic treatment is 20-30s, and a total of 15-30 ultrasonic treatments are performed.

[0046] Preferably, before casting the film, the ultrasonically treated product is degassed using a vacuum degassing instrument before being applied to the casting film.

[0047] In a preferred embodiment, in step (4), the casting amount of the film is 0.2-0.35 mL / cm. 2 .

[0048] Preferably, in step (4), the drying temperature is 48-52°C and the time is 6-8 hours.

[0049] Preferably, in step (4), the equilibration is carried out in a constant temperature and humidity chamber for 10-12 hours, and the temperature is controlled at 20-25℃ and the humidity at 35-50wt%.

[0050] As previously stated, the second aspect of the present invention provides a chitosan membrane loaded with kaempferia oil prepared by the method described in the first aspect above.

[0051] As mentioned above, the third aspect of the present invention provides the application of the chitosan membrane loaded with kaempferol essential oil described in the second aspect above in the field of food safety control.

[0052] Preferably, the food is a plant-based meat product.

[0053] It should be noted that there are no specific limitations on the instruments and parameters for vacuum degassing in this invention, and conventional equipment and techniques in the field can be used.

[0054] The present invention will be described in detail below through examples. In the following examples, unless otherwise specified, the raw materials used are all commercially available products.

[0055] Unless otherwise specified, "room temperature" in this invention means 25±3℃.

[0056] Moringa prolysin:

[0057] Moringa prolysin I: a non-water-soluble protein extracted from moringa leaf residue, with a protein content of 70 wt%.

[0058] Moringa leaf protein: crude protein extracted from moringa leaf residue, with a protein content of 70 wt%, containing glutenin, prolysin and globulin, and the glutenin content of the protein is 80 wt%.

[0059] Pectin: weight average molecular weight 50-100 kDa, purchased from Guangdong Osman Biotechnology Co., Ltd., brand name 0135;

[0060] Kaempferia galanga essential oil: purchased from Hubei Xinyuhong Biomedical Technology Co., Ltd.;

[0061] Chitosan: Food-grade chitosan, purchased from Qingdao Bozhi Huili Biotechnology Co., Ltd., brand name 20200609;

[0062] Corn starch: Food-grade high amylose corn starch, purchased from Quanyinxiangyu (Beijing) Biotechnology Co., Ltd., with an amylose content of 72wt%.

[0063] Citric acid: Food-grade anhydrous citric acid, purchased from Weifang Yingxuan Industrial Co., Ltd.

[0064] Vacuum degassing instrument: Model ZKT-25, purchased from Tianjin Ruiside Technology Co., Ltd.;

[0065] The constant temperature and humidity chamber model is DHTHM-16, purchased from Duohe Experimental Equipment (Shanghai) Co., Ltd.

[0066] Homogenizer: Model ULTRA-TURRAX, purchased from IKA GmbH, Germany.

[0067] Example 1

[0068] This embodiment illustrates that the method for preparing chitosan membranes loaded with kaempferia galanga essential oil provided by the present invention is carried out according to the following steps:

[0069] (1) At a speed of 1000 rpm, 3.75 g of moringa prolysin (moringa prolysin I) was mixed with 70 wt% ethanol-water solution for 20 min to obtain 3 wt% moringa prolysin ethanol-water solution.

[0070] At 70°C and 1000 rpm, 1.0 g of pectin was mixed with water II for 1 h, and then centrifuged at 5000 g for 60 min to remove the precipitate, to obtain the pectin aqueous solution with a concentration of 2 wt%.

[0071] The ethanol-water solution of Moringa prolysin and the pectin water solution were first contacted at 1000 rpm for 60 min at a volume ratio of 2.5:1 to obtain a protein-pectin dispersion solution.

[0072] (2) At 15000 rpm, the galangal essential oil and the protein-pectin dispersion solution with pH adjusted to 5.0 were homogenized at a volume ratio of 1:100 using a high-speed homogenizer for the second contact. The time for each high-speed homogenization was 30 s, and the time interval between each two adjacent high-speed homogenizations was 30 s. The high-speed stirring was carried out 5 times in total to obtain the galangal essential oil emulsion.

[0073] (3) Chitosan, corn starch, and citric acid were dissolved in water I at 60℃ and 1000rpm for 45min, then gelatinized at 80℃ for 1.5h, cooled to 35℃, and then glycerol was added at 1000rpm for a third contact for 30min to obtain a membrane solution; wherein the weight ratio of chitosan, corn starch, citric acid, glycerol, and water I was 4:3:1.5:7:100.

[0074] (4) At 1500 rpm, the galangal essential oil emulsion and the membrane liquid were contacted for the fourth time at a volume ratio of 5:100 for 15 min, then homogenized at 10 MPa for 5 min, and then ultrasonically treated at 400 W power: a total of 30 ultrasonic treatments were performed, with a time interval of 5 s between each two adjacent ultrasonic treatments and a time of 20 s for each ultrasonic treatment.

[0075] Vacuum degassing was then performed using a vacuum degassing instrument, followed by degassing at 0.2 mL / cm³. 2 The film was cast by casting the film, then dried in an oven at 50°C for 6 hours. After cooling to room temperature, the film was peeled off and then equilibrated at 25°C and 40 wt% humidity for 10 hours to obtain a chitosan film loaded with kaempferol essential oil, named P1.

[0076] Example 2

[0077] This embodiment illustrates that the method for preparing chitosan membranes loaded with kaempferia galanga essential oil provided by the present invention is carried out according to the following steps:

[0078] (1) At a speed of 1000 rpm, 6.25 g of Moringa prolysin was mixed with 70 wt% ethanol-water solution for 20 min to obtain Moringa prolysin ethanol-water solution with a concentration of 5 wt%.

[0079] At 70°C and 1000 rpm, 1.5 g of pectin was mixed with water II for 1 h, and then centrifuged at 5000 g for 60 min to remove the precipitate, resulting in a pectin aqueous solution with a concentration of 3 wt%.

[0080] The ethanol-water solution of Moringa prolysin and the pectin water solution were first contacted at 1000 rpm for 60 min at a volume ratio of 2.5:1 to obtain a protein-pectin dispersion solution.

[0081] (2) At 17000 rpm, the galangal essential oil and the protein-pectin dispersion solution with pH adjusted to 5.0 were homogenized at a volume ratio of 2:100 using a high-speed homogenizer for the second contact. The time for each high-speed homogenization was 30 s, and the time interval between each two adjacent high-speed homogenizations was 30 s. The high-speed homogenization was performed a total of 3 times to obtain the galangal essential oil emulsion.

[0082] (3) Chitosan, corn starch, and citric acid were dissolved in water I at 60℃ and 1000rpm for 35min, then gelatinized at 90℃ for 1.5h, cooled to 35℃, and then glycerol was added at 1000rpm for a third contact for 30min to obtain a membrane solution; wherein the weight ratio of chitosan, corn starch, citric acid, glycerol, and water I was 3:3:1:6:100.

[0083] (4) At 1500 rpm, the galangal essential oil emulsion and the membrane solution were subjected to a fourth contact at a volume ratio of 7:100 for 15 min. Then, homogenization was performed at 10 MPa for 4 min, followed by 20 ultrasonic treatments at 400 W, with a 5-second interval between each ultrasonic treatment and a duration of 25 seconds per treatment. Vacuum degassing was then performed using a vacuum degassing device, followed by degassing at 0.25 mL / cm³. 2 The film was cast by casting the film, then dried in an oven at 50°C for 7 hours. After cooling to room temperature, the film was peeled off and then equilibrated at 20°C and 40 wt% humidity for 10 hours to obtain a chitosan film loaded with kaempferol essential oil, named P2.

[0084] Example 3

[0085] This embodiment illustrates that the method for preparing chitosan membranes loaded with kaempferia galanga essential oil provided by the present invention is carried out according to the following steps:

[0086] (1) At a speed of 1000 rpm, 5g of Moringa prolysin was mixed with 70wt% ethanol-water solution for 20min to obtain Moringa prolysin ethanol-water solution with a concentration of 4wt%.

[0087] At 70°C and 1000 rpm, 2 g of pectin was mixed with water II for 1 h, and then centrifuged at 5000 g for 60 min to remove the precipitate, resulting in a pectin aqueous solution with a concentration of 4 wt%.

[0088] The ethanol-water solution of Moringa prolysin and the pectin water solution were first contacted at 1000 rpm for 60 min at a volume ratio of 2.5:1 to obtain a protein-pectin dispersion solution.

[0089] (2) At 18000 rpm, the galangal essential oil and the protein-pectin dispersion solution with pH adjusted to 5.0 were homogenized at a volume ratio of 1:100 using a high-speed homogenizer for the second contact. The time for each high-speed homogenization was 30 s, and the time interval between each two adjacent high-speed homogenizations was 30 s. The high-speed stirring was carried out 5 times in total to obtain the galangal essential oil emulsion.

[0090] (3) Chitosan, corn starch, and citric acid are dissolved in water I at 60℃ and 1000rpm for 30min, then gelatinized at 80℃ for 1.5h, cooled to 32℃, and then glycerol is added at 1000rpm for a third contact for 30min to obtain a membrane solution; wherein the weight ratio of chitosan, corn starch, citric acid, glycerol, and water I is 5:3:1.5:8:100;

[0091] (4) At 1500 rpm, the galangal essential oil emulsion and the membrane solution were subjected to a fourth contact at a volume ratio of 8:100 for 15 min. Then, homogenization was performed at 15 MPa for 4 min, followed by 15 ultrasonic treatments at 400 W, with a 5-second interval between each ultrasonic treatment and a duration of 30 seconds per ultrasonic treatment. Vacuum degassing was then performed using a vacuum degassing device, followed by degassing at 0.35 mL / cm³. 2 The film was cast by casting the film, then dried in an oven at 50°C for 8 hours. After cooling to room temperature, the film was peeled off and then equilibrated at 25°C and 40 wt% humidity for 12 hours to obtain a chitosan film loaded with kaempferol essential oil, named P3.

[0092] Example 4

[0093] This embodiment is carried out using a method similar to that of Embodiment 1. The difference is that in step (2), the volume ratio of the galangal essential oil to the protein-pectin dispersion solution is 5:100.

[0094] Finally, a chitosan membrane loaded with galangal essential oil was obtained and named P4.

[0095] Example 5

[0096] This embodiment uses a method similar to that of Embodiment 1, except that in step (4), the volume ratio of the galangal essential oil emulsion to the film liquid is 1:100;

[0097] Finally, a chitosan membrane loaded with galangal essential oil was obtained and named P5.

[0098] Example 6

[0099] This embodiment uses a method similar to that of Embodiment 1, except that in step (4), the volume ratio of the galangal essential oil emulsion to the film liquid is 15:100.

[0100] Finally, a chitosan membrane loaded with galangal essential oil was obtained and named P6.

[0101] Example 7

[0102] This embodiment uses a method similar to that of Embodiment 1. The difference is that in step (4), the ultrasonic treatment is not performed. That is, the homogenized product is directly degassed under vacuum and then cast into a film.

[0103] Finally, a chitosan membrane loaded with galangal essential oil was obtained and named P7.

[0104] Example 8

[0105] This embodiment uses a method similar to that of Embodiment 1, except that in step (4), the casting amount is 0.58 mL / cm. 2 ;

[0106] Finally, a chitosan membrane loaded with galangal essential oil was obtained and named P8.

[0107] Comparative Example 1

[0108] This comparative example was carried out using a method similar to that of Example 1, except that in step (2), an equal volume of Torreya grandis essential oil was used instead of the Kaempferia galanga essential oil for the second contact.

[0109] Finally, a chitosan membrane was obtained and named DP1.

[0110] Comparative Example 2

[0111] This comparative example was carried out using a method similar to that of Example 1, except that in step (1), the protein used was Moringa leaf protein;

[0112] Finally, a chitosan membrane was obtained and named DP2.

[0113] Test Example 1

[0114] The inhibitory effects of the chitosan membrane loaded with kaempferol essential oil prepared in the above examples on the total heterocyclic amines in the plant-based meat after heat processing were tested, including:

[0115] S1. Cover the surface of the plant-based meat product with a chitosan membrane or chitosan membrane loaded with galangal essential oil, and bake it in an oven at 170℃ for 45 minutes to obtain the heat-processed plant-based meat.

[0116] S2. Add 4.0g of heat-processed plant-based meat to 5mL of deionized water, shake to mix, centrifuge and collect the supernatant. Add 5mL of 50wt% sodium hydroxide solution to the precipitate, then transfer the meat paste to an Erlenmeyer flask containing 14g of diatomaceous earth, add 50mL of ethyl acetate, mix well, and ultrasonically extract twice for 30min each time. After centrifugation, discard the supernatant and transfer the remaining precipitate to a pressure-resistant bottle. Then add 40mL of 6mol / L hydrochloric acid, blow with nitrogen for 2min, and hydrolyze at 110℃ for 24h to obtain a hydrolyzed mixture.

[0117] S3. Filter the hydrolysate mixture and obtain the clear liquid as the bound HAs extract. Take 10 mL of the bound HAs extract and perform solid-phase extraction purification on an Oasis MCX column. After the purified eluent is dried under nitrogen, it is redissolved in 400 μL of methanol and then the total heterocyclic amine content is analyzed by UHPLC-MS / MS (ultra-high performance liquid chromatography-tandem mass spectrometry).

[0118] The UHPLC-MS / MS detection conditions include:

[0119] The chromatographic column was a Waters Atlantics dC18 column (250 × 4.6 mm id, 3.0 μm), the column temperature was 35 °C, the mobile phase was methanol (A), 0.1% formic acid aqueous solution (B), the flow rate was 0.5 mL / min, and the gradient elution program was: 0–0.1 min, 5 vol% (A); 0.1–10 min, 5–100 vol% (A); 10–10.5 min, 100–5 vol% (A). Before the next injection, 10 vol% (A) was held for 5 min to equilibrate the column; the single injection volume was 1 μL.

[0120] The mass spectrometry analysis was performed using an electrospray ionization source (ESI source), positive ion mode, and multiple reaction monitoring (MRM) mode; source temperature: 130℃; capillary voltage: 3.5V; desolvation gas (nitrogen, 99.0% purity) flow rate: 800L / h; desolvation temperature: 350℃; conical gas (nitrogen, 99.0% purity) flow rate: 50L / h; collision gas (argon, 99.9% purity) flow rate: 0.13mL / min.

[0121] The total heterocyclic amine content (ng / g) obtained from S4 and S3 is denoted as C. t The total heterocyclic amine content in the blank control is recorded as C0. The inhibition rate (%) of the test subject against total heterocyclic amines is calculated as follows: = ((C0 - C0) / ( ... t () / C0)*100%, see Table 1 for specific results.

[0122] Table 1

[0123]

[0124] Table 1 (continued)

[0125]

[0126] Test Example 2

[0127] The inhibitory effects of the chitosan membrane loaded with kaempferol essential oil prepared in the above examples on the advanced glycation end products (total AGEs) in plant-based meat after heat processing were tested, including:

[0128] SS1. Cover the surface of the plant-based meat product with a chitosan membrane or chitosan membrane loaded with galangal essential oil, and bake it in an oven at 170℃ for 45 minutes to obtain the heat-processed plant-based meat.

[0129] SS2. Take approximately 5 mg of heat-processed plant-based meat containing protein equivalents, add 3 mL of n-hexane, shake vigorously, and then centrifuge at 10000 g for 15 min. Remove the supernatant and repeat this step three times to completely defatted the meat. Place the defatted powder sample in a fume hood to evaporate any residual n-hexane. Add 1.5 mL of borate buffer (0.2 mol / L, pH = 9.2), then dissolve it in 0.01 mol / L sodium hydroxide solution to obtain 1 mL of sodium borohydride solution. Add 1-octanol as an antifoaming agent and reduce the solution overnight at 4 °C. Add an appropriate amount of TCA solution (60%, w / v) to the reduced sample solution to make the final TCA concentration in the sample solution 20% (w / v). Let it stand in a refrigerator at 4 °C for 2 h, then centrifuge at 15000 g for 25 min. Discard the supernatant and retain the precipitate for later use.

[0130] SS3. Add 5 mL of hydrochloric acid solution (6 mol / L) to the precipitate, seal under vacuum, and then place in an oven at 110 °C for hydrolysis for 24 h. After hydrolysis, filter the hydrolysate to remove the precipitate and make up to 10 mL. Accurately pipette 600 μL of the hydrolysate, blow dry at 60 °C, add 150 μL of internal standard solution containing 0.0891 μg of d4-CML and 0.0825 μg of d4-CEL, and redissolve the sample in 2 mL of 5 mmol / L nonafluoropentanoic acid for later use.

[0131] SS4. The total AGEs content was then analyzed using UHPLC-MS / MS (ultra-high performance liquid chromatography-tandem mass spectrometry).

[0132] The H2P column was activated and equilibrated sequentially with 3 mL of methanol and 3 mL of 5 mmol / L nonafluorovalerate, then loaded with the sample. The column was then eluted sequentially with 2 mL of 5 mmol / L nonafluorovalerate and 2 mL of methanol-nonafluorovalerate solution I (methanol / 5 mM nonafluorovalerate = 5 / 95, v / v), and finally eluted with 6 mL of methanol-nonafluorovalerate solution II (methanol / 5 mM nonafluorovalerate = 50 / 50, v / v). The flow rate was controlled at approximately 2 mL / min during activation and elution, and less than 0.5 mL / min during loading and elution.

[0133] The HPLC-MS detection column used was an X-Bridge C18 column (3.5 μm, 100 mm × 2.1 mm); mobile phase A: 5 mmol / L NPFA, mobile phase B: acetonitrile; sample chamber temperature: 8℃; column temperature: 35℃; injection volume: 5 μL.

[0134] The mass spectrometry analysis was set to use electrospray ionization (ESI+) as the ionization source; detection mode: multiple reaction detection (MRM); capillary voltage: 3.55 kV; ion source temperature: 110℃; desolvation gas temperature: 350℃; conical gas (nitrogen, purity 99.9%, flow rate: 50 L / h); desolvation gas (nitrogen, purity 99.9%) flow rate: 500 L / h; collision gas (argon, purity 99.9%) flow rate: 0.15 mL / min.

[0135] The total AGEs content (ng / g) obtained from SS5 and SS43 is denoted as C. 2t The total AGEs content in the blank control is denoted as C. 20 Calculate the inhibition rate (%) of total AGEs in the test subjects = ((C 20 -C 2t ) / C 20 ()*100%, see Table 2 for specific results.

[0136] Table 2

[0137]

[0138] Table 2 (continued)

[0139]

[0140] Test Example 3

[0141] Chitosan membranes loaded with galangal essential oil were applied to the surface of plant-based meat products, and then baked in an oven at 170°C for 45 minutes to obtain heat-processed plant-based meat products. The sensory quality of the heat-processed plant-based meat products was then tested using an expert sensory evaluation method and with reference to the scoring criteria in Table 3. The results are shown in Table 4.

[0142] Table 3

[0143]

[0144] Table 4

[0145]

[0146]

[0147] As can be seen from the above results, the method and application of preparing chitosan membrane loaded with galangal essential oil provided by the present invention can effectively reduce the content of bound heterocyclic amines and AGEs in plant-based meat products, without affecting the sensory quality of plant-based meat products, and can be well applied to the processing and production of plant-based meat products.

[0148] The preferred embodiments of the present invention have been described in detail above; however, the present invention is not limited thereto. Within the scope of the inventive concept, various simple modifications can be made to the technical solutions of the present invention, including combinations of various technical features in any other suitable manner. These simple modifications and combinations should also be considered as the content disclosed in the present invention and are all within the protection scope of the present invention.

Claims

1. A method for preparing a chitosan membrane loaded with kaempferia galanga essential oil, characterized in that, The method includes: (1) The ethanol-water solution of Moringa alcohol-soluble protein was first contacted with the pectin water solution to obtain a protein-pectin dispersion solution; (2) The galangal essential oil is brought into a second contact with the protein-pectin dispersion solution to obtain a galangal essential oil emulsion; (3) Chitosan, corn starch and citric acid are dissolved in water I and gelatinized, and then glycerol is added for the third contact to obtain the membrane solution; (4) The galangal essential oil emulsion is brought into a fourth contact with the membrane liquid, and after casting into a film, it is dried and balanced to obtain a chitosan membrane loaded with galangal essential oil. The moringa prolysin is a non-water-soluble protein extracted from moringa leaf residue, and the protein content of the moringa prolysin is 65-73 wt%. The concentration of moringa prolysin in the ethanol-water solution is 3-5 wt%, and the concentration of the pectin water solution is 1.5-5 wt%. In step (2), the volume ratio of the galangal essential oil to the protein-pectin dispersion solution is 1-5:100; In step (4), the volume ratio of the galangal essential oil emulsion to the film liquid is 5-8:

100.

2. The method according to claim 1, wherein, The method further includes: in step (2), before the second contact is performed, the pH of the protein-pectin dispersion solution is adjusted to 5.0-6.5 and then applied to the second contact.

3. The method according to claim 1 or 2, wherein, In step (1), the ethanol-water solution of moringa prolysin is a product prepared by a method comprising the following steps: at a speed of 800-1200 rpm, moringa prolysin is first mixed with 70 wt% ethanol-water solution for 15-30 min to obtain the ethanol-water solution of moringa prolysin.

4. The method according to claim 1 or 2, wherein, In step (1), the pectin aqueous solution is a product prepared by a method including the following steps: pectin and water II are mixed for a second time at 68-72℃ and 800-1200 rpm for 1-2 hours, and the precipitate is separated and removed to obtain the pectin aqueous solution.

5. The method according to claim 1 or 2, wherein, In step (1), the volume ratio of the ethanol-water solution of moringa prolysin to the pectin water solution is 2.5-4:1; and / or In step (3), the weight ratio of chitosan, corn starch, citric acid, glycerol, and water I is 3-5:1-3:1-1.5:5-8:

100.

6. The method according to claim 1 or 2, wherein, In step (2), the second contact is performed in a high-speed homogenization manner, and at least the following conditions are met: the rotation speed is 15000-18000 rpm, the time for each high-speed homogenization is 30-45 s, the time interval between each two adjacent high-speed homogenizations is 30 s, and the high-speed homogenization is performed a total of 3-5 times.

7. The method according to claim 1 or 2, wherein, In step (1), the first contact is carried out under stirring conditions, and at least meets the following requirements: a rotation speed of 800-1200 rpm and a time of 0.8-1.5 h; and / or In step (3), the dissolution is carried out under stirring conditions, and at least meets the following requirements: temperature 58-62℃, rotation speed 800-1200 rpm, time 30-45 min; and / or In step (3), the gelatinization conditions must at least satisfy: a temperature of 80-90°C and a time of 1-2.5 h; and / or In step (3), the third contact is carried out under stirring conditions, and at least meets the following requirements: temperature 32-38℃, rotation speed 800-1200rpm, and time 30-45min; and / or In step (4), the fourth contact is carried out under stirring conditions, and at least the following conditions must be met: the rotation speed is 1300-1700 rpm and the time is 15-30 min.

8. The method according to claim 1 or 2, wherein, In step (4), the casting amount of the film is 0.2-0.35 mL / cm. 2 .

9. The chitosan membrane loaded with kaempferia oil prepared by the method according to any one of claims 1-8.

10. The application of the chitosan membrane loaded with kaempferol essential oil as described in claim 9 in the field of food safety control.

Images