阅读说明：本技术 一种采用动态高压微射流预处理提取鱼鳞多糖的方法 (Method for extracting scale polysaccharide by adopting dynamic high-pressure microjet pretreatment ) 是由 赵岩岩 张�浩 崔震昆 王书彦 莫海珍 赵圣明 梁新红 周威 于 2020-07-15 设计创作，主要内容包括：本发明公开一种采用动态高压微射流预处理提取鱼鳞多糖的方法：将新鲜鱼鳞洗净,依次进行脱钙、烘干、粉碎,得鱼鳞粉末；将鱼鳞粉末与水混合均匀,使用动态高压纳米均质仪进行处理,得鱼鳞粉末与水的混合物；将所得鱼鳞粉末与水的混合物进行水浴浸提、离心,收集上清液,得到含鱼鳞多糖的产品。本发明动态高压微射流预处理可有效降低鱼鳞粉末的平均粒度,提高鱼鳞多糖的提取得率,随着处理压力的提高,多糖提取液对DPPH·、·O<Sub>2</Sub><Sup>-</Sup>、ABTS·<Sup>+</Sup>和·OH的清除率以及Fe<Sup>3+</Sup>的还原能力逐渐增强,均显著高于空白对照组,本发明采用动态高压微射流预处理提取鱼鳞多糖的方法可以有效提高鱼鳞多糖的提取得率和体外抗氧化能力。(The invention discloses a method for extracting scale polysaccharide by adopting dynamic high-pressure microjet pretreatment, which comprises the following steps: cleaning fresh fish scales, sequentially decalcifying, drying and crushing to obtain fish scale powder; mixing the fish scale powder and water uniformly, and treating by using a dynamic high-pressure nano homogenizer to obtain a mixture of the fish scale powder and water; and (3) carrying out water bath leaching and centrifugation on the mixture of the fish scale powder and water, and collecting supernatant to obtain a product containing the fish scale polysaccharide. The dynamic high-pressure microjet pretreatment can effectively reduce the average particle size of the fish scale powder and improve the extraction yield of the fish scale polysaccharide, and the polysaccharide extract is used for treating DPPH and O with the improvement of the treatment pressure 2 ‑ 、ABTS· + And OH clearance and Fe 3+ The reduction capability of the fish scale polysaccharide is gradually enhanced and is obviously higher than that of a blank control group, and the method for extracting the fish scale polysaccharide by adopting the dynamic high-pressure microjet pretreatment can effectively improve the effect of the fish scale polysaccharideExtraction yield and in vitro antioxidant ability.)

1. A method for extracting fish scale polysaccharide by adopting dynamic high-pressure microjet pretreatment is characterized by comprising the following steps:

(1) cleaning fresh fish scales, sequentially decalcifying, drying and crushing to obtain fish scale powder for later use;

(2) mixing the fish scale powder obtained in the step (1) with water, uniformly stirring, and then treating by using a dynamic high-pressure nano homogenizer to obtain a mixture of the fish scale powder and the water for later use;

(3) and (3) carrying out water bath leaching and centrifugation on the mixture of the fish scale powder obtained in the step (2) and water, and collecting supernatant to obtain a product containing the fish scale polysaccharide.

2. The method for extracting fish scale polysaccharide by dynamic high-pressure microjet pretreatment of claim 1, wherein the decalcification in step (1) is carried out by the following specific methods: adding the fish scales into phosphoric acid solution, stirring at the rotating speed of 60-100r/min for 40-60min, and filtering through 100-mesh and 200-mesh filter cloth to obtain the decalcified fish scales.

3. The method for extracting fish scale polysaccharide by using dynamic high-pressure microjet pretreatment of claim 2, wherein the concentration of the phosphoric acid solution is 0.6-1.2mol/L, and the mass-to-volume ratio of the fish scales to the phosphoric acid solution is 1 g: 30-50 mL.

4. The method for extracting fish scale polysaccharide by using dynamic high-pressure microjet pretreatment of claim 1, wherein the fish scale drying temperature in step (1) is 48-53 ℃, and the drying time is 2-2.5 h.

5. The method for extracting fish scale polysaccharide by using dynamic high-pressure micro-jet pretreatment of claim 1, wherein the fish scale in step (1) is pulverized into particles with a size of 80-120 meshes.

6. The method for extracting fish scale polysaccharide by using dynamic high-pressure micro-jet pretreatment as claimed in claim 1, wherein the stirring speed in step (2) is 200-300r/min, and the stirring time is 1-2 min.

7. The method for extracting fish scale polysaccharide by using dynamic high-pressure microjet pretreatment of claim 1, wherein the mass volume ratio of the fish scale powder to water in the step (2) is 1 g: 20-40 ml.

8. The method for extracting fish scale polysaccharide by dynamic high-pressure microjet pretreatment of claim 1, wherein the step (2) uses a dynamic high-pressure nano homogenizer with a processing pressure of 50-150MPa, a processing temperature of 30-50 ℃ and a processing time of 30-60 s.

9. The method for extracting fish scale polysaccharide by dynamic high-pressure micro-jet pretreatment of claim 1, wherein the temperature of the water bath leaching in the step (3) is 75-85 ℃, and the time of the water bath leaching is 1.5-6 h.

10. The method for extracting fish scale polysaccharide by using dynamic high-pressure microjet pretreatment as claimed in claim 1, wherein the centrifugation speed in step (3) is 4000-6000r/min, and the centrifugation time is 10-15 min.

Technical Field

The invention belongs to the technical field of polysaccharide extraction, and particularly relates to a method for extracting fish scale polysaccharide by adopting dynamic high-pressure microjet pretreatment.

Background

Carp is widely distributed in continental europe and asia, and is one of the main aquaculture species in the world, China is the largest carp breeding country, and the annual output is 302 ten thousand tons, which accounts for 80% of the total world output. Each l00g carp meat contains protein 17.6g, fat 4.1g, phosphorus 204mg, calcium 50mg and vitamins; in addition, most of the fat in the carp is unsaturated fatty acid, and the carp has the functions of reducing cholesterol of a human body, preventing and treating arteriosclerosis, coronary heart disease and the like, so the carp is considered to be a healthy food.

About 30% of byproducts are generated in the carp processing process, wherein the fish scales account for about 5%, and are generally treated as waste, thereby causing environmental pollution. The fish scales are rich in protein, vitamins, polysaccharide, unsaturated fatty acid and the like, wherein the content of the polysaccharide reaches 9.5%.

At present, the extraction of the phosphate polysaccharide of the freshwater fish is mainly carried out by adopting an alkaline extraction method, but the waste liquid extracted by the alkaline method pollutes the environment and does not accord with the concept of environmental protection.

The dynamic high-pressure micro-jet is a new ultramicro homogenizing technology, and the materials are crushed through the actions of high-speed impact, high-frequency oscillation, instantaneous high pressure, strong shearing and the like, so that the better superfine and homogenizing effects are achieved, and active components cannot be damaged.

At present, the dynamic high-pressure micro-jet technology is widely applied to sterilization, protein modification, polysaccharide modification and extraction of natural active ingredients, and can effectively improve the extraction rate of lentinan, sweet potato leaf polyphenol and the like due to the advantages of reducing the granularity of materials, increasing the penetrating capacity of solvents and the like. However, the application of the dynamic high-pressure micro-jet technology to the extraction of fish scale polysaccharide is not reported.

Therefore, the research and development of a method for extracting the fish scale polysaccharide by adopting dynamic high-pressure microjet pretreatment, which can improve the extraction yield and the in vitro antioxidant capacity of the carp fish scale polysaccharide, are problems to be solved by technical personnel in the field.

Disclosure of Invention

In view of the above, the present invention provides a method for extracting fish scale polysaccharide by using dynamic high-pressure microjet pretreatment.

The technical scheme is as follows:

a method for extracting fish scale polysaccharide by adopting dynamic high-pressure microjet pretreatment comprises the following steps:

(1) cleaning fresh fish scales, sequentially decalcifying, drying and crushing to obtain fish scale powder for later use;

(2) mixing the fish scale powder obtained in the step (1) with water, uniformly stirring, and then treating by using a dynamic high-pressure nano homogenizer to obtain a mixture of the fish scale powder and the water for later use;

(3) and (3) carrying out water bath leaching and centrifugation on the mixture of the fish scale powder obtained in the step (2) and water, and collecting supernatant to obtain a product containing the fish scale polysaccharide.

Further, the specific method for decalcification in the step (1) comprises the following steps: adding the fish scales into phosphoric acid solution, stirring at the rotating speed of 60-100r/min for 40-60min, and filtering through 100-mesh and 200-mesh filter cloth to obtain the decalcified fish scales.

The adoption of the further beneficial effects is as follows: can effectively promote the dissolution of calcium in the fish scales and improve the decalcification efficiency of the fish scales.

Further, the concentration of the phosphoric acid solution is 0.6-1.2mol/L, and the mass volume ratio of the fish scales to the phosphoric acid solution is 1 g: 30-50 mL.

The adoption of the further beneficial effects is as follows: phosphoric acid belongs to medium and strong acid, does not cause serious environmental pollution like strong acid, and is easy to damage the structure of polysaccharide in fish scales while decalcifying by strong acid.

Further, the drying temperature of the fish scales in the step (1) is 48-53 ℃, and the drying time is 2-2.5 h.

The adoption of the further beneficial effects is as follows: improve drying efficiency, be convenient for further smash.

Further, the fish scales in the step (1) are crushed to reach the grain diameter of 80-120 meshes.

The adoption of the further beneficial effects is as follows: after the fish scales are crushed into powder, the dynamic high-pressure micro-flow treatment is further conveniently carried out.

Further, the stirring speed in the step (2) is 200-300r/min, and the stirring time is 1-2 min.

The adoption of the further beneficial effects is as follows: prevent the scale powder from gathering into blocks and blocking the feeding pipe of the dynamic high-pressure nano homogenizer.

Further, the mass volume ratio of the fish scale powder to water in the step (2) is 1 g: 20-40 ml.

The adoption of the further beneficial effects is as follows: the concentration of the fish scale powder solution is moderate, so that the particle size distribution of a sample processed by a dynamic high-pressure nano homogenizer is more uniform.

Further, the step (2) uses a dynamic high-pressure nano homogenizer to process for 30-60s at the temperature of 30-50 ℃ and under the pressure of 50-150 MPa.

The adoption of the further beneficial effects is as follows: the grain size of the fish scale powder processed by the dynamic high-pressure nano homogenizer is smaller.

Furthermore, the dynamic high-pressure nano homogenizer is used for 2-4 times in the step (2), and after each time of treatment, the temperature of the dynamic high-pressure nano homogenizer is cooled to 20-30 ℃ for next treatment.

The further beneficial effects of the adoption are as follows: prevent the machine from overheating to cause the damage of partial functional activity of the fish scale polysaccharide due to the overlong continuous processing time.

Further, the temperature of the water bath leaching in the step (3) is 75-85 ℃, and the time of the water bath leaching is 1.5-6 h.

The adoption of the further beneficial effects is as follows: the leaching of the fish scale polysaccharide is facilitated, and the functional activity of the fish scale polysaccharide is not damaged when the temperature is not higher than 90 ℃.

Further, the centrifugal rotation speed in the step (3) is 4000-.

The adoption of the further beneficial effects is as follows: the fish scale polysaccharide is obtained through centrifugal collection, and the structure of the fish scale polysaccharide cannot be damaged through proper centrifugal rotating speed.

Preferably, the fish scales are carp scales and/or grass carp scales.

The preferable beneficial effects are as follows: carp and grass carp belong to freshwater fish belonging to the family Cyprinaceae, the scale structures are similar, and the scale polysaccharide can be extracted by the same method.

The invention has the beneficial effects that: the dynamic high-pressure microjet pretreatment can effectively reduce the average particle size of the fish scale powder from unprocessed 499.8nm to 192.57nm, so that the extraction yield of carp scale polysaccharide is improved, and when the treatment pressure is 130MPa, the extraction yield reaches 7.81%. The polysaccharide extracts are subjected to DPPH and O with increasing treatment pressure₂ ^-、ABTS·⁺And OH clearance and Fe³⁺The reducing capability of the composition is gradually enhanced, and the highest values respectively reach 86.04%, 61.23%, 89.58%, 90.59% and 0.64(OD values), which are all significantly higher than that of a blank control group (P)<0.05), the method for extracting the fish scale polysaccharide by adopting the dynamic high-pressure microjet pretreatment can effectively improve the extraction yield and the in-vitro antioxidant capacity of the carp fish scale polysaccharide.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.

FIG. 1 is a graph showing the influence of dynamic high-pressure microjet pretreatment on the yield of carp scale polysaccharide;

FIG. 2 is a graph showing the effect of dynamic high-pressure microjet pretreatment on DPPH (DPPH-removing potential) of carp scale polysaccharide;

FIG. 3 shows the pretreatment of carp scale polysaccharide O by dynamic high-pressure microjet^2-A graph of impact of cleaning ability;

FIG. 4 is a drawing showingDynamic high-pressure micro-jet pretreatment on carp scale polysaccharide ABTS⁺A graph of impact of cleaning ability;

FIG. 5 is a graph showing the effect of dynamic high-pressure microjet pretreatment on carp fish scale OH removal capacity;

FIG. 6 shows the pretreatment of carp scale polysaccharide Fe by dynamic high-pressure microjet³⁺Influence graph of reducing power;

FIG. 7 is a graph showing the effect of dynamic high-pressure microjet pretreatment on the particle size distribution of carp scale polysaccharides.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments, not all embodiments, of the present invention 1. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.