Preparation method and application of squid melanin seasoning
The preparation of squid melanin seasoning through ultrasound synergistic and Maillard reactions solved the problem of poor solubility of squid ink melanin, achieved functional seasonings with high biological activity and rich flavor, and promoted the sustainable utilization and market promotion of squid processing waste.
Patent Information
- Application Number
- CN202510749354.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-06
- Publication Date
- 2025-08-12
AI Technical Summary
The poor melanin solubility of squid ink has caused its biological activity and flavor to be insufficient, limiting its application and commercialization process in food.
Squid melanin seasoning was prepared by ultrasonic synergistic and Maillard reaction combined with spray drying, and enzymatically dissolved using acidic, alkaline and neutral proteases, followed by Maillard reaction with reduced sugar, and finally spray-drying the finished product.
It significantly improves the water solubility and biological activity of squid melanin, enhances its application potential in food, and provides seasonings with rich flavors, which have the function of enhancing immunity, is cost-effective, safe and non-toxic side effects.
Smart Images
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of functional seasonings, and particularly relates to a preparation method and application of a squid melanin seasoning. Background Art
[0002] Squid (Loligo japonica) is a key marine economic species in China, highly valued for its tender meat, high protein content, and short growth cycle. According to statistics, China's annual squid production exceeds one million tons, and processing produces approximately 25% of its total weight as byproducts, including squid ink, hyaline cartilage, and testes. Squid ink sacs are rich in melanin, proteins, and polysaccharides, exhibiting diverse biological activities such as immune enhancement, antioxidant properties, and tumor inhibition. Furthermore, squid ink's unique flavor makes it a popular ingredient for food innovation, with widespread use in products such as squid ink noodles, squid ink steamed buns, and squid ink biscuits. However, existing squid ink products have low added value and have yet to be produced on an industrial scale. A major limitation is the poor solubility of squid melanin, preventing direct absorption by the human body. This insolubility not only reduces the bioavailability of the melanin's biological activity but also complicates the formulation of squid ink-based products, hindering their large-scale production and commercialization. For example, traditional squid ink food processing methods typically involve simply adding the ink directly to dough or fillings, failing to fully utilize its nutritional value and functional properties. Furthermore, while squid ink's unique flavor is appealing, it hasn't been optimized to meet the complex flavor demands of modern consumers.
[0003] In recent years, with improving living standards, consumer demand for natural, healthy, and functional seasonings has increased significantly. Functional seasonings must not only provide basic flavor enhancement but also offer additional health benefits. China is the largest consumer and producer of seasonings and spices in the Asia-Pacific region. Its large population, diverse culinary culture, and growing food processing industry make it a major processing and export center for seasonings and spices worldwide. The Asia-Pacific region will account for approximately 78% of the global seasoning market revenue in 2023. However, existing functional seasonings often face high raw material costs and complex production processes, resulting in high final product prices and limited consumer acceptance. This cost barrier restricts the market penetration and promotion of functional seasonings.
[0004] Existing technologies primarily utilize squid ink by adding it directly to foods, such as pasta, but this approach fails to fully exploit its bioactivity and flavor potential. The low solubility of squid melanin is a major bottleneck in its application, necessitating a novel approach to improve its solubility while preserving and enhancing its bioactivity and flavor. Summary of the Invention
[0005] The present invention aims to overcome the defects of the prior art and provide a method for preparing a squid melanin seasoning.
[0006] Another object of the present invention is to provide an application of the squid melanin seasoning prepared by the above preparation method.
[0007] The technical solutions of the present invention are as follows:
[0008] A method for preparing a squid melanin seasoning comprises the following steps:
[0009] (1) After removing impurities from the squid ink sac, squid melanin is prepared by using ultrasound-assisted enzymatic hydrolysis, the ultrasound power is 1000-3000W, and the enzyme used in the enzymatic hydrolysis is selected from acidic protease, alkaline protease and neutral protease;
[0010] (2) Adding reducing sugar to the material obtained in step (1) to carry out a Maillard reaction, followed by spray drying to obtain the squid melanin seasoning, wherein the mass ratio of the reducing sugar to the material obtained in step (1) is 1:1-3.
[0011] In a preferred embodiment of the present invention, during the enzymatic hydrolysis, the temperature is 50-60°C.
[0012] More preferably, during the enzymatic hydrolysis, the pH is 1-9.
[0013] More preferably, the enzymatic hydrolysis time is 3-4 hours.
[0014] In a preferred embodiment of the present invention, the reducing sugar is selected from the group consisting of arabinose, lactose and fructose.
[0015] In a preferred embodiment of the present invention, the temperature of the Maillard reaction is 50-80°C.
[0016] More preferably, the Maillard reaction time is 2-3 hours.
[0017] In a preferred embodiment of the present invention, during the enzymatic hydrolysis, the temperature is 50-60°C, the pH is 1-9, and the time is 3-4 hours; the reducing sugar is selected from arabinose, lactose and fructose; and the temperature of the Maillard reaction is 50-80°C, and the time is 2-3 hours.
[0018] The squid melanin seasoning prepared by the preparation method is used in preparing an immunity enhancing composition.
[0019] An immunity enhancing composition, the effective component of which includes the squid melanin seasoning prepared by the above preparation method.
[0020] The beneficial effects of the present invention are:
[0021] By converting squid ink, a processing byproduct, into a functional seasoning, this invention not only solves waste disposal issues but also creates a new value-added product. This method helps reduce environmental pollution associated with squid processing waste and promotes the sustainable use of marine resources.
[0022] 2. The ultrasound-assisted enzymatic hydrolysis process in this invention significantly increases the water solubility of squid melanin, overcoming a major limitation of its direct application in food. Experiments have shown that the seasoning produced by this invention has good solubility in water (10-30% w / v solution) and is free of visible impurities, making it easy to use in a variety of foods.
[0023] 3. The squid melanin seasoning produced by the present invention exhibits high biological activity in enhancing immunity. Animal experiments have shown that it can significantly enhance specific cellular immunity. Safety assessments have shown that no toxicity or adverse reactions were observed at high doses (up to 15g / kg), confirming its safety for human consumption.
[0024] 4. The present invention provides a cost-effective alternative, overcoming the high cost barrier of existing functional seasonings. This cost-effectiveness makes functional seasonings more accessible to consumers, thereby expanding the market. DETAILED DESCRIPTION
[0025] The technical solution of the present invention is further illustrated and described below through specific implementation methods.
[0026] Example 1
[0027] (1) After removing the capsule, 500 L of squid ink was added to an ultrasonic reactor, and acidic protease was added for enzymatic hydrolysis. The pH value of the reaction system was controlled to be 1, the enzymatic hydrolysis temperature was 50 ° C, the ultrasonic power was 1000 W, and the reaction time was 3 h.
[0028] (2) After the ultrasonic enzymatic hydrolysis reaction, the enzymatic hydrolyzate obtained was subjected to a Maillard reaction by adding arabinose in a mass ratio of 1:1 between the solid mixture and the reducing sugar. The reaction temperature was 50°C and the reaction time was 3 hours. After the Maillard reaction, the material was dried by spray drying to obtain a squid melanin seasoning.
[0029] Flavor Compound Identification: The squid melanin seasoning prepared in this example was analyzed using SPME-GC-MS. After equilibration in a water bath at 60°C for 30 minutes, the sample was extracted with a CAR / PDMS fiber and headspace sampling was performed for 45 minutes before gas chromatography-mass spectrometry (GC-MS). The GC conditions were: a TG-1701MS column (30 m × 0.25 mm, 0.25 μm); an electron ionization (EI) source; and a mass scan range of m / z 33-450. As shown in Table 1, the results indicate that the squid melanin seasoning prepared in this example is rich in ester flavor compounds (27.49%) and pyridine flavor compounds (17.13%), exhibiting rich floral, fruity, roasted, and meaty aromas.
[0030] Table 1. Content of flavor substances in the squid melanin seasoning prepared in Example 1
[0031]
[0032]
[0033] Solubility evaluation: The squid melanin seasoning powder prepared in this example was dissolved in water to prepare solutions with a mass volume ratio of 10%, 20%, and 30%. No particulate impurities were visible to the naked eye.
[0034] Safety evaluation: Kunming mice were gavaged with different doses of the squid melanin seasoning prepared in this example, with the dose groups being 1 g / kg, 5 g / kg, 10 g / kg, and 15 g / kg, respectively. No animals in each dose group died, had abnormal behavior, or showed significant weight gain, indicating that the squid melanin seasoning prepared in this example had no toxic side effects.
[0035] Immunity Enhancement Bioactivity Evaluation: Sensitized Kunming mice were immunized intraperitoneally with 5% (v / v) sheep red blood cells (SRBC). Different doses of the squid melanin seasoning prepared in this example (5 mg / kg / day, 10 mg / kg / day, and 50 mg / kg / day) were administered to the mice, with each mouse receiving an injection of 0.2 mL. Four days after immunization, the plantar thickness of the left hind paw was measured. Then, 20 μL of 20% (v / v) SRBC was subcutaneously injected at the measurement site. The plantar thickness of the left hind paw was measured 24 hours after injection. Three measurements were taken at the same site, and the average value was calculated. The squid melanin seasoning prepared in this example showed an effect on the thickening of the paws of normal mice (Table 2). Compared with the normal control group, the paw thickening of mice in each dosage group was significantly increased (p < 0.05), indicating that the squid melanin seasoning prepared in this example can promote the specific cellular immunity of mice and is a functional nutritional seasoning for enhancing immunity.
[0036] Table 2. Effect of the squid melanin seasoning prepared in Example 1 on the thickness of the paw of normal mice
[0037] Grouping Plantar thickness (mm) Normal control group 0.036±0.003 5mg / kg / day <![CDATA[0.95±0.004 ** ]]> 10mg / kg / day <![CDATA[0.128±0.004 *** ]]> 50mg / kg / day <![CDATA[0.147±0.005 *** ]]>
[0038] Note: Compared with blank, * p<0.05, ** p<0.01, *** p<0.001.
[0039] Example 2
[0040] (1) After removing the capsule, 500 L of squid ink was added to an ultrasonic reactor, and alkaline protease was added for enzymatic hydrolysis. The pH value of the reaction system was controlled to be 9, the enzymatic hydrolysis temperature was 55 ° C, the ultrasonic power was 3000 W, and the reaction time was 4 h.
[0041] (3) After the ultrasonic enzymatic hydrolysis reaction, lactose was added to the resulting enzymatic hydrolyzate at a mass ratio of 2:1 between the solid mixture and the reducing sugar, and a Maillard reaction was performed at a temperature of 70°C for 4 hours. After the Maillard reaction, the material was dried by spray drying to obtain a squid melanin seasoning.
[0042] Flavor Compound Identification: The squid melanin seasoning prepared in this example was analyzed using SPME-GC-MS. After equilibration in a water bath at 60°C for 30 minutes, the sample was extracted using a CAR / PDMS fiber, and headspace sampling was performed for 45 minutes before GC-MS analysis. GC conditions included a TG-1701MS column (30 m × 0.25 mm, 0.25 μm), an electron ionization (EI) source, and a mass scan range of m / z 33-450. As shown in Table 3, the experimental results indicate that the squid melanin seasoning is rich in ester flavor compounds (24.41%) and ketone flavor compounds (17.77%), as well as pyrazine flavor compounds (0.81%). It exhibits a rich floral, fruity, roasted, and meaty aroma.
[0043] Table 3. Content of flavor substances in the squid melanin seasoning prepared in Example 2
[0044]
[0045]
[0046] Solubility evaluation: The squid melanin seasoning powder prepared in this example was dissolved in water to prepare solutions with a mass volume ratio of 10%, 20%, and 30%. No particulate impurities were visible to the naked eye.
[0047] Safety evaluation: Kunming mice were gavaged with different doses of the squid melanin seasoning prepared in this example, with the dose groups being 1 g / kg, 5 g / kg, 10 g / kg, and 15 g / kg, respectively. No animals in each dose group died, had abnormal behavior, or showed significant weight gain, indicating that the squid melanin seasoning prepared in this example had no toxic side effects.
[0048] Immunity Enhancement Bioactivity Evaluation: Sensitized Kunming mice were immunized intraperitoneally with 5% (v / v) sheep red blood cells (SRBC). Different doses of the squid melanin seasoning prepared in this example (5 mg / kg / day, 10 mg / kg / day, and 50 mg / kg / day) were administered to the mice, with each mouse receiving an injection of 0.2 mL. Four days after immunization, the plantar thickness of the left hind paw was measured. Then, 20 μL of 20% (v / v) SRBC was subcutaneously injected at the measurement site. The plantar thickness of the left hind paw was measured 24 hours after injection. Three measurements were taken at the same site, and the average value was calculated. The squid melanin seasoning prepared in this example showed an effect on the thickening of the paws of normal mice (Table 4). Compared with the normal control group, the paw thickening of mice in each dosage group was significantly increased (p < 0.05). This indicates that the squid melanin seasoning prepared in this example can promote specific cellular immunity in mice and is a functional nutritional seasoning for enhancing immunity.
[0049] Table 4. Effect of the squid melanin seasoning prepared in Example 2 on the thickness of the paw of normal mice
[0050] Grouping Plantar thickness (mm) Normal control group 0.033±0.002 5mg / kg / day <![CDATA[0.101±0.003 ** ]]> 10mg / kg / day <![CDATA[0.125±0.002 *** ]]> 50mg / kg / day <![CDATA[0.132±0.003 *** ]]>
[0051] Note: Compared with blank, * p<0.05, ** p<0.01, *** p<0.001.
[0052] Example 3
[0053] (1) After removing the capsule, 500 L of squid ink was added to an ultrasonic reactor, and neutral protease was added for enzymatic hydrolysis. The pH value of the reaction system was controlled to be 7, the temperature of the enzymatic hydrolysis reaction was 60°C, the ultrasonic power was 2000 W, and the reaction time was 3 h.
[0054] After the ultrasonic enzymatic hydrolysis reaction, fructose was added to the resulting enzymatic hydrolyzate at a mass ratio of 3:1 between the solid mixture and reducing sugar, and a Maillard reaction was performed at 80°C for 3 hours. After the Maillard reaction, the material was spray dried to obtain a squid melanin seasoning.
[0055] Flavor Compound Identification: The squid melanin seasoning prepared in this example was analyzed using SPME-GC-MS. After equilibration in a water bath at 60°C for 30 minutes, the sample was extracted using a CAR / PDMS fiber, and headspace sampling was performed for 45 minutes before GC-MS analysis. GC conditions included a TG-1701MS column (30 m × 0.25 mm, 0.25 μm), an electron ionization (EI) source, and a mass scan range of m / z 33-450. As shown in Table 5, the squid melanin seasoning was rich in ester flavor compounds (22.34%) and ketone flavor compounds (23.36%), as well as pyrrole flavor compounds (3.97%). It exhibited a rich floral, fruity, roasted, and meaty aroma.
[0056] Table 5. Content of flavor substances in the squid melanin seasoning prepared in Example 3
[0057]
[0058] Solubility evaluation: The squid melanin seasoning powder prepared in this example was dissolved in water to prepare solutions with a mass volume ratio of 10%, 20%, and 30%. No particulate impurities were visible to the naked eye.
[0059] Safety evaluation: Kunming mice were gavaged with different doses of the squid melanin seasoning prepared in this example, with the dose groups being 1 g / kg, 5 g / kg, 10 g / kg, and 15 g / kg, respectively. No animals in each dose group died, had abnormal behavior, or showed significant weight gain, indicating that the squid melanin seasoning prepared in this example had no toxic side effects.
[0060] Immunity Enhancement Bioactivity Evaluation: Sensitized Kunming mice were immunized intraperitoneally with 5% (v / v) sheep red blood cells (SRBC). Different doses of the squid melanin seasoning prepared in this example (5 mg / kg / day, 10 mg / kg / day, and 50 mg / kg / day) were administered to the mice, with each mouse receiving an injection of 0.2 mL. Four days after immunization, the plantar thickness of the left hind paw was measured. Then, 20 μL of 20% (v / v) SRBC was subcutaneously injected at the measurement site. The plantar thickness of the left hind paw was measured 24 hours after injection. Three measurements were taken at the same site, and the average value was calculated. The squid melanin seasoning prepared in this example showed an effect on the thickening of the paws of normal mice (Table 6). Compared with the normal control group, the paw thickening of mice in each dosage group was significantly increased (p < 0.05). This indicates that the squid melanin seasoning prepared in this example can promote specific cellular immunity in mice and is a functional nutritional seasoning for enhancing immunity.
[0061] Table 6. Effect of the squid melanin seasoning prepared in Example 3 on the thickening of the paw of normal mice
[0062] Grouping Plantar thickness (mm) Normal control group 0.029±0.004 5mg / kg / day <![CDATA[0.105±0.004 ** ]]> 10mg / kg / day <![CDATA[0.128±0.003 *** ]]> 50mg / kg / day <![CDATA[0.131±0.005 *** ]]>
[0063] Note: Compared with blank, * p<0.05, ** p<0.01, *** p<0.001.
[0064] The above description is merely a preferred embodiment of the present invention and therefore cannot be used to limit the scope of the present invention. In other words, equivalent changes and modifications made within the scope of the present invention and the contents of the specification should still fall within the scope of the present invention.
Claims
1. A method for preparing a squid melanin seasoning, characterized in that: The following steps are involved: (1) After removing impurities from the squid ink sac, squid melanin is prepared by using ultrasound-assisted enzymatic hydrolysis, the ultrasound power is 1000-3000W, and the enzyme used in the enzymatic hydrolysis is selected from acidic protease, alkaline protease and neutral protease; (2) Adding reducing sugar to the material obtained in step (1) to carry out a Maillard reaction, followed by spray drying to obtain the squid melanin seasoning, wherein the mass ratio of the reducing sugar to the material obtained in step (1) is 1:1-3.
2. The preparation method according to claim 1, wherein: During the enzymatic hydrolysis, the temperature is 50-60°C.
3. The preparation method according to claim 2, wherein: During the enzymatic hydrolysis, the pH is 1-9.
4. The preparation method according to claim 3, wherein: The enzymatic hydrolysis time is 3-4 hours.
5. The preparation method according to claim 1, wherein: The reducing sugar is selected from the group consisting of arabinose, lactose and fructose.
6. The preparation method according to claim 1, wherein: The temperature of the Maillard reaction is 50-80°C.
7. The preparation method according to claim 6, wherein: The Maillard reaction time is 2-3 hours.
8. The preparation method according to claim 1, wherein: During the enzymatic hydrolysis, the temperature is 50-60° C., the pH is 1-9, and the time is 3-4 hours; the reducing sugar is selected from arabinose, lactose, and fructose; and the temperature of the Maillard reaction is 50-80° C., and the time is 2-3 hours.
9. Use of the squid melanin seasoning prepared by the preparation method according to any one of claims 1 to 8 in preparing an immunity enhancing composition.
10. An immunity-enhancing composition, characterized in that: The effective component thereof comprises the squid melanin seasoning prepared by the preparation method according to any one of claims 1 to 8.