Preparation method and application of abalone nutritional seasoning

Abstract

The invention discloses a preparation method and application of an abalone nutritional seasoning, and the preparation method comprises the following steps: (1) removing solid impurities from an abalone cooking waste liquid by using a centrifugal machine, and then removing heavy metals by using ion exchange resin; (2) adding the material obtained in the step (1) into specific protease of which the weight is 2-3% of the weight of the solid in the material obtained in the step (1), carrying out enzymolysis reaction, then removing macromolecular impurities by using a nanofiltration membrane, and then carrying out spray drying to obtain a mixture; and (3) dissolving the mixture obtained in the step (2), adding reducing sugar to carry out Maillard reaction, and then carrying out spray drying to obtain the abalone nutritional seasoning. The prepared abalone nutritional seasoning is high in biological activity for improving immunity, high in safety and low in heavy metal content; meanwhile, the seasoning is rich in pyrazine and ketone flavor substances, has rich seafood flavor, barbecue flavor and meat flavor, and can be widely applied to the field of functional nutritional seasonings.

Description

Technical Field

[0001] The invention belongs to the technical field of food seasonings, and particularly relates to a preparation method and application of an abalone nutritional seasoning. Background Art

[0002] Abalone is a precious marine delicacy with extremely high nutritional value. It is known as the first of the "eight marine delicacies". It is not only nutritious, but also has high edible value and health value. China is the world's largest abalone farming country, with an annual output of 180,300 tons and an annual output value of about 20 billion yuan. Abalone farming in my country is mainly concentrated in Fujian, Shandong, Liaoning and other places. Among them, the annual output in Fujian is 143,900 tons, accounting for 79.8% of the national total output, and a large-scale farming industry cluster has been formed. However, in the processing of abalone, a large amount of cooking waste liquid is generated, which seriously pollutes the environment. Although these waste liquids are rich in polypeptides, water-soluble proteins, polysaccharides and trace elements, they are high-quality raw materials for the development of high-end nutritional condiments, but the problem of excessive heavy metals in abalone cooking waste liquid has hindered its industrial production and promotion. Therefore, it is urgent to develop a process technology that can effectively remove heavy metals from abalone cooking waste liquid.

[0003] In recent years, with the increasing awareness of nutrition and health care, natural, healthy and nutritious condiments are becoming more and more popular among consumers and have become a hot topic of research. There are many types of these nutritious condiments on the market in China, but they have not yet formed an industrial scale. This is mainly because the raw material cost and production cost of nutritious condiments are high, which leads to the high price of condiments, limiting their large-scale promotion. In addition, although these condiments are rich in nutrition, not all of them have the effect of improving immunity. Therefore, it is particularly important to develop abalone nutritious seasonings with the effect of improving immunity and preparation methods thereof. Summary of the invention

[0004] The invention aims to overcome the defects of the prior art and provide a method for preparing an abalone nutritional seasoning.

[0005] Another object of the present invention is to provide the application of the abalone nutritional seasoning prepared by the above preparation method.

[0006] The technical solution of the present invention is as follows:

[0007] A method for preparing an abalone nutritional seasoning comprises the following steps:

[0008] (1) after the abalone cooking waste liquid is removed solid impurities with a centrifuge, ion exchange resin is utilized to remove heavy metals;

[0009] (2) placing the material obtained in step (1) in a reaction kettle, adding 2-3% of the weight of the solid content in the material obtained in step (1) of a specific protease to carry out enzymatic hydrolysis reaction, then removing macromolecular impurities with a nanofiltration membrane, and then spray drying to obtain a solid mixture; the specific protease is selected from at least one of A3 animal hydrolyzing protease, bromelain, papain and flavor protease;

[0010] (3) After dissolving the solid mixture obtained in step (2) with water, adding reducing sugar to carry out Maillard reaction, and then spray drying to obtain the abalone nutritional seasoning.

[0011] In a preferred embodiment of the present invention, in step (1), the flow rate of the material using the ion exchange resin to remove heavy metals is 10-50m 3 / h.

[0012] In a preferred embodiment of the present invention, the specific protease is A3 animal hydrolyzing protease, A3 animal hydrolyzing protease + neutral protease or A3 animal hydrolyzing protease + neutral protease + flavor protease.

[0013] In a preferred embodiment of the present invention, the temperature of the enzymatic hydrolysis reaction is 40-55°C, the pH value is 6-7, and the time is 1-3 hours.

[0014] In a preferred embodiment of the present invention, the molecular weight cut-off of the nanofiltration membrane is 500-2000 Da.

[0015] In a preferred embodiment of the present invention, the reducing sugar is selected from the group consisting of arabinose, fructose and lactose.

[0016] Further preferably, the mass ratio of the solid mixture to the reducing sugar is 1-5:1.

[0017] More preferably, the Maillard reaction is carried out at a temperature of 50-80°C and a time of 1-3 hours.

[0018] The abalone nutritional seasoning prepared by the preparation method is used in preparing an immunity enhancing composition.

[0019] An immunity enhancing composition, the raw material of which contains the abalone nutritional seasoning prepared by the preparation method.

[0020] The beneficial effects of the present invention are:

[0021] 1. The present invention takes abalone cooking waste liquid as the object, adopts centrifuge separation and column chromatography technology to remove heavy metals in the waste liquid, and develops a new source of low-cost abalone functional nutritional seasoning raw material; this innovation realizes "turning waste into treasure", solves the problem of environmental pollution caused by cooking waste liquid generated in the processing of abalone products, and is suitable for large-scale production.

[0022] 2. The abalone nutritional seasoning prepared by the present invention not only has high biological activity for improving immunity, high safety, but also low heavy metal content; at the same time, it is rich in pyrazine and ketone flavor substances, has strong seafood flavor, barbecue flavor and meat flavor, and can be widely used in the field of functional nutritional seasonings. DETAILED DESCRIPTION

[0023] The technical solution of the present invention is further illustrated and described below through specific implementation methods.

[0024] Example 1

[0025] (1) 1000L of abalone cooking wastewater was centrifuged to remove solid impurities, and then heavy metals were removed using ion exchange resin. The flow rate was 10m 3 / h.

[0026] (2) The material obtained in step (1) is placed in a reactor, and 2% of the weight of the solid calculated by the solid content in the material obtained in the above step (1) is added to perform enzymatic hydrolysis, and the pH value of the reaction system is controlled to be 7, the temperature of the enzymatic hydrolysis reaction is 40° C., and the time of the enzymatic hydrolysis reaction is 3 hours. After the enzymatic hydrolysis reaction is completed, a nanofiltration membrane with a cutoff relative molecular mass of 2000Da is used to remove macromolecular impurities, and the material solution after membrane separation is spray-dried to obtain a solid mixture (mixed by abalone peptide, polysaccharide and amino acid).

[0027] (3) After dissolving the solid mixture obtained in step (2) with water, arabinose is added at a mass ratio of 5:1 between the solid mixture and reducing sugar, and the Maillard reaction is carried out at a temperature of 50° C. and a time of 3 hours. After the Maillard reaction is completed, the material is dried by spray drying to obtain abalone nutritional seasoning.

[0028] [Identification of flavor substances]

[0029] The abalone nutritional seasoning prepared in this embodiment was detected by SPME-GC-MS. After the sample was equilibrated in a water bath at 60°C for 30 minutes, the CAR / PDMS extraction fiber was used, and the headspace sampling was performed for 45 minutes for gas chromatography-mass spectrometry analysis. The gas chromatography conditions were: TG-1701MS column (30m×0.25mm, 0.25μm); electron bombardment (electron ionization, EI) ion source; mass scanning range m / z 33~450. As shown in Table 1, the test results show that the abalone nutritional seasoning prepared in this embodiment is rich in pyrazine flavor substances (29.17%) and ketone flavor substances (28.92%), and has a strong seafood flavor, barbecue flavor and meat flavor.

[0030] Table 1. Content of flavor substances in abalone nutritional seasoning prepared in Example 1

[0031]

[0032]

[0033] [Heavy metal content detection]

[0034] GB 500.11-2014 detects the content of inorganic arsenic in the abalone nutritional seasoning prepared in this embodiment, GB500.12-2017 detects the content of lead in the product, and GB 500.17-2021 detects the content of methylmercury in the product. The experimental results show that the contents of inorganic arsenic, lead and methylmercury in the abalone nutritional seasoning prepared in this embodiment are all less than 0.1, indicating that the abalone nutritional seasoning prepared in this embodiment is highly safe.

[0035] [Safety Evaluation]

[0036] Different doses of the abalone nutritional seasoning prepared in this embodiment were gavaged into Kunming mice, and the dose groups were 1 g / kg, 5 g / kg, 10 g / kg, and 15 g / kg, respectively. There was no death in the animals in each dose group, and no abnormal behavior, and no obvious weight gain, indicating that the abalone nutritional seasoning prepared in this embodiment has no toxic side effects.

[0037] [Evaluation of biological activity to improve immunity]

[0038] After sensitization of Kunming mice with 5% (v / v) sheep red blood cells (SRBC) intraperitoneal immunization, different doses of abalone nutritional seasonings prepared in the present embodiment were administered to mice (different doses were 5mg / kg / day, 10mg / kg / day and 50mg / kg / day), and each was injected with 0.2mL. After immunization for 4 days, the thickness of the left hind foot plantar was measured, and then 20% (V / V) SRBC was injected subcutaneously at the measurement site, and 20uL of each mouse was injected. The thickness of the left hind foot plantar was measured at 24h after injection, and the same position was measured three times, and the average value was taken. The abalone nutritional seasoning prepared in the present embodiment showed (table 1) to the thickness increase experiment of normal mouse plantar, compared with the normal control group, the thickness increase of the mouse plantar in each dosage group was significantly improved (p < 0.05), indicating that the abalone nutritional seasoning prepared in the present embodiment can promote the specific cellular immunity of mice, and belongs to the functional nutritional seasoning that improves immunity.

[0039] Table 2. Effect of the abalone nutritional seasoning prepared in Example 1 on the thickness of the paw of normal mice

[0040] Grouping Plantar thickness (mm) Normal control group 0.048±0.004 5mg / kg / day <![CDATA[0.103±0.005 ** ]]> 10 mg / kg / day <![CDATA[0.132±0.002 *** ]]> 50mg / kg / day <![CDATA[0.156±0.003 *** ]]>

[0041] Note: Compared with blank, * p<0.05, ** p<0.01, *** p<0.001.

[0042] Example 2

[0043] (1) 3000L of abalone cooking wastewater, after the solid impurities are removed by centrifuge, heavy metals are removed by ion exchange resin, flow rate 30m 3 / h.

[0044] (2) The material obtained in step (1) is placed in a reactor, and 1% of the weight of the solid calculated by the solid content in the material obtained in the above step (1) is added with A3 animal hydrolyzing protease and 1% of neutral protease for enzymolysis reaction, the pH value of the reaction system is controlled to be 6, the temperature of the enzymolysis reaction is 45° C., and the time of the enzymolysis reaction is 2 hours. After the enzymolysis reaction is completed, a nanofiltration membrane with a cutoff relative molecular mass of 1000Da is used to remove macromolecular impurities, and the material solution after membrane separation is spray-dried to obtain a solid mixture (mixed by abalone peptide, polysaccharide and amino acid).

[0045] (3) After dissolving the solid mixture obtained in step (2) with water, fructose is added in a mass ratio of 3:1 between the solid mixture and reducing sugar to carry out a Maillard reaction at a temperature of 70° C. for 2 h. After the Maillard reaction is completed, the material is dried by spray drying to obtain an abalone nutritional seasoning.

[0046] [Identification of flavor substances]

[0047] The abalone nutritional seasoning prepared in this embodiment was detected by SPME-GC-MS. After the sample was equilibrated in a water bath at 60°C for 30 minutes, the CAR / PDMS extraction fiber was used, and the headspace sampling was performed for 45 minutes for gas chromatography-mass spectrometry analysis. The gas chromatography conditions were: TG-1701MS column (30m×0.25mm, 0.25μm); electron bombardment (electron ionization, EI) ion source; mass scanning range m / z 33~450. As shown in Table 3, the experimental results show that the abalone nutritional seasoning is rich in pyrazine flavor substances (32.52%) and ketone flavor substances (24.55%), and has a strong seafood flavor, barbecue flavor and meat flavor.

[0048] Table 3. Content of flavor substances in abalone nutritional seasoning prepared in Example 2

[0049]

[0050]

[0051] [Heavy metal content detection]

[0052] GB 500.11-2014 detects the content of inorganic arsenic in the abalone nutritional seasoning prepared in this embodiment, GB500.12-2017 detects the content of lead in the product, and GB 500.17-2021 detects the content of methylmercury in the product. The experimental results show that the contents of inorganic arsenic, lead and methylmercury in the abalone nutritional seasoning prepared in this embodiment are all less than 0.1, indicating that the abalone nutritional seasoning prepared in this embodiment is highly safe.

[0053] [Safety Evaluation]

[0054] Different doses of the abalone nutritional seasoning prepared in this embodiment were gavaged into Kunming mice, and the dose groups were 1 g / kg, 5 g / kg, 10 g / kg, and 15 g / kg, respectively. There was no death in the animals in each dose group, and no abnormal behavior, and no obvious weight gain, indicating that the abalone nutritional seasoning prepared in this embodiment has no toxic side effects.

[0055] [Evaluation of biological activity to improve immunity]

[0056] After sensitization of Kunming mice with 5% (v / v) sheep red blood cells (SRBC) intraperitoneal immunization, different doses of abalone nutritional seasonings prepared in the present embodiment were administered to mice (different doses were 5mg / kg / day, 10mg / kg / day and 50mg / kg / day), and each was injected with 0.2mL. After immunization for 4 days, the thickness of the left hind foot plantar was measured, and then 20% (V / V) SRBC was injected subcutaneously at the measurement site, and 20uL of each mouse was injected. The thickness of the left hind foot plantar was measured at 24h after injection, and the same position was measured three times, and the average value was taken. The abalone nutritional seasoning prepared in the present embodiment showed (table 1) to the thickness increase experiment of normal mouse plantar, compared with the normal control group, the thickness increase of the mouse plantar in each dosage group was significantly improved (p < 0.05), indicating that the abalone nutritional seasoning prepared in the present embodiment can promote the specific cellular immunity of mice, and belongs to the functional nutritional seasoning that improves immunity.

[0057] Table 4. Effect of the abalone nutritional seasoning prepared in Example 2 on the thickness of the paw of normal mice

[0058] Grouping Plantar thickness (mm) Normal control group 0.046±0.003 5mg / kg / day <![CDATA[0.114±0.002 ** ]]> 10mg / kg / day <![CDATA[0.136±0.001 *** ]]> 50mg / kg / day <![CDATA[0.141±0.002 *** ]]>

[0059] Note: Compared with blank, * p<0.05, ** p<0.01, *** p<0.001.

[0060] Example 3

[0061] (1) 5000L of abalone cooking wastewater, after the solid impurities are removed by centrifuge, heavy metals are removed by ion exchange resin, flow rate 50m 3 / h.

[0062] (2) The material obtained in step (1) is placed in a reaction kettle, and 1% of the weight of the solid content in the material obtained in step (1) is added with A3 animal hydrolyzing protease, 1% of the neutral protease and 0.5% of the flavor protease to carry out enzymatic hydrolysis reaction, and the pH value of the reaction system is controlled to be 7, the temperature of the enzymatic hydrolysis reaction is 55° C., and the time of the enzymatic hydrolysis reaction is 1 hour. After the enzymatic hydrolysis reaction is completed, a nanofiltration membrane with a cutoff relative molecular mass of 500Da is used to remove macromolecular impurities, and the material solution after membrane separation is spray-dried to obtain a solid mixture (mixed with abalone peptides, polysaccharides and amino acids).

[0063] (3) After dissolving the solid mixture obtained in step (2) with water, lactose is added in a mass ratio of 1:1 between the solid mixture and reducing sugar to carry out a Maillard reaction at a temperature of 80° C. for 1 h. After the Maillard reaction is completed, the material is dried by spray drying to obtain an abalone nutritional seasoning.

[0064] [Identification of flavor substances]

[0065] The abalone nutritional seasoning prepared in this embodiment was detected by SPME-GC-MS. After the sample was equilibrated in a water bath at 60°C for 30 minutes, the CAR / PDMS extraction fiber was used, and the headspace sampling was performed for 45 minutes for gas chromatography-mass spectrometry analysis. The gas chromatography conditions were: TG-1701MS column (30m×0.25mm, 0.25μm); electron bombardment (electron ionization, EI) ion source; mass scanning range m / z 33~450. As shown in Table 5, the experimental results show that the abalone nutritional seasoning is rich in pyrazine flavor substances (32.24%) and ketone flavor substances (21.18%), and has a strong seafood flavor, barbecue flavor and meat flavor.

[0066] Table 5. Content of flavor substances in abalone nutritional seasoning prepared in Example 3

[0067]

[0068]

[0069] [Heavy metal content detection]

[0070] GB 500.11-2014 detects the content of inorganic arsenic in the abalone nutritional seasoning prepared in this embodiment, GB500.12-2017 detects the content of lead in the product, and GB 500.17-2021 detects the content of methylmercury in the product. The experimental results show that the contents of inorganic arsenic, lead and methylmercury in the abalone nutritional seasoning prepared in this embodiment are all less than 0.1, indicating that the abalone nutritional seasoning prepared in this embodiment is highly safe.

[0071] [Safety Evaluation]

[0072] Different doses of the abalone nutritional seasoning prepared in this embodiment were gavaged into Kunming mice, and the dose groups were 1 g / kg, 5 g / kg, 10 g / kg, and 15 g / kg, respectively. There was no death in the animals in each dose group, and no abnormal behavior, and no obvious weight gain, indicating that the abalone nutritional seasoning prepared in this embodiment has no toxic side effects.

[0073] [Evaluation of biological activity to improve immunity]

[0074] After sensitization of Kunming mice with 5% (v / v) sheep red blood cells (SRBC) intraperitoneal immunization, different doses of abalone nutritional seasonings prepared in the present embodiment were administered to mice (different doses were 5mg / kg / day, 10mg / kg / day and 50mg / kg / day), and each was injected with 0.2mL. After immunization for 4 days, the thickness of the left hind foot plantar was measured, and then 20% (V / V) SRBC was injected subcutaneously at the measurement site, and 20uL of each mouse was injected. The thickness of the left hind foot plantar was measured at 24h after injection, and the same position was measured three times, and the average value was taken. The abalone nutritional seasoning prepared in the present embodiment showed (table 1) to the thickness increase experiment of normal mouse plantar, compared with the normal control group, the thickness increase of the mouse plantar in each dosage group was significantly improved (p < 0.05), indicating that the abalone nutritional seasoning prepared in the present embodiment can promote the specific cellular immunity of mice, and belongs to the functional nutritional seasoning that improves immunity.

[0075] Table 6. Effect of the abalone nutritional seasoning prepared in Example 3 on the thickness of the plantar fossa of normal mice

[0076] Grouping Plantar thickness (mm) Normal control group 0.041±0.003 5mg / kg / day <![CDATA[0.117±0.002 ** ]]> 10 mg / kg / day <![CDATA[0.139±0.001 *** ]]> 50mg / kg / day <![CDATA[0.142±0.002 *** ]]>

[0077] Note: Compared with blank, * p<0.05, ** p<0.01, *** p<0.001.

[0078] The above description is only a preferred embodiment of the present invention, and therefore cannot be used to limit the scope of the present invention. That is, equivalent changes and modifications made according to the patent 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 an abalone nutritional seasoning, characterized in that: The steps include: (1) after the abalone cooking waste liquid is removed solid impurities with a centrifuge, ion exchange resin is utilized to remove heavy metals; (2) placing the material obtained in step (1) in a reaction kettle, adding 2-3% of the weight of the solid content in the material obtained in step (1) of a specific protease to carry out enzymatic hydrolysis reaction, then removing macromolecular impurities with a nanofiltration membrane, and then spray drying to obtain a solid mixture; the specific protease is selected from at least one of A3 animal hydrolyzing protease, bromelain, papain and flavor protease; (3) After dissolving the solid mixture obtained in step (2) with water, adding reducing sugar to carry out Maillard reaction, and then spray drying to obtain the abalone nutritional seasoning.

2. The preparation method according to claim 1, characterized in that: In the step (1), the flow rate of the material using ion exchange resin to remove heavy metals is 10-50m 3 / h.

3. The preparation method according to claim 1, characterized in that: The specific protease is A3 animal hydrolyzing protease, A3 animal hydrolyzing protease+neutral protease or A3 animal hydrolyzing protease+neutral protease+flavor protease.

4. The preparation method according to claim 1, characterized in that: The temperature of the enzymatic hydrolysis reaction is 40-55° C., the pH value is 6-7, and the time is 1-3 hours.

5. The preparation method according to claim 1, characterized in that: The molecular weight cut-off of the nanofiltration membrane is 500-2000Da.

6. The preparation method according to claim 1, characterized in that: The reducing sugar is selected from the group consisting of arabinose, fructose and lactose.

7. The preparation method according to claim 6, characterized in that: The mass ratio of the solid mixture to the reducing sugar is 1-5:

1.

8. The preparation method according to claim 7, characterized in that: The Maillard reaction has a temperature of 50-80°C and a reaction time of 1-3 hours.

9. Use of the abalone nutritional 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 raw material contains abalone nutritional seasoning prepared by the preparation method according to any one of claims 1 to 8.

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