Method for extracting codonopsis pilosula concentrated juice through combination of ultrahigh pressure and enzymolysis

Abstract

The invention relates to the technical field of health-care drinks, in particular to a method for extracting codonopsis pilosula concentrated juice by ultrahigh pressure combined with enzymolysis, which comprises the following steps: selecting mildewproof and insect-free codonopsis pilosula, cleaning soil impurities on the surface, drying water on the surface, and slicing after drying for later use; weighing 100g of codonopsis pilosula slices, putting the codonopsis pilosula slices into a crusher, crushing, and sieving with a 100-mesh sieve to obtain codonopsis pilosula powder for later use; taking codonopsis pilosula powder, adding purified water according to a material-liquid ratio of 1: 15 (g / mL), and fully and uniformly stirring to obtain a codonopsis pilosula stock solution; heating the codonopsis pilosula stock solution to 45 DEG C, adding a compound enzyme for enzymolysis, and heating for enzyme deactivation after 1 h to obtain a primary codonopsis pilosula extracting solution; transferring the enzyme-deactivated primary codonopsis pilosula extracting solution into a polyethylene plastic bag, sealing the polyethylene plastic bag, putting the polyethylene plastic bag into ultrahigh-pressure equipment for ultrahigh-pressure treatment, and centrifuging the codonopsis pilosula extracting solution subjected to ultrahigh-pressure treatment at 8000 rpm for 15 minutes to obtain a secondary codonopsis pilosula extracting solution; and performing vacuum concentration on the secondary codonopsis pilosula extracting solution for 15 minutes to obtain codonopsis pilosula concentrated juice, and finally sterilizing the concentrated juice and performing sterile filling.

Description

Technical Field

[0001] This invention relates to the field of health beverage technology, specifically a method for extracting concentrated Codonopsis pilosula juice using ultra-high pressure combined with enzymatic hydrolysis. Background Technology

[0002] Codonopsis pilosula is derived from the dried root of Codonopsis pilosula, Codonopsis lanceolata, or Codonopsis chuanxiong, all belonging to the Campanulaceae family. It has a sweet taste and neutral properties. It is used to invigorate the spleen and lungs, replenish qi, quench thirst, and promote blood circulation and fluid production. As a traditional tonic Chinese medicinal herb, Codonopsis pilosula is rich in nutrients. Its active ingredient, codonopsis glycoside, has anti-ulcer activity; Codonopsis pilosula polysaccharides have protective and immunomodulatory effects and improve inflammation. The extraction efficiency and stability of the core active ingredients directly affect the efficacy of the product.

[0003] Currently, conventional water or alcohol extraction techniques for preparing concentrated juice result in the breakage of Codonopsis pilosula polysaccharide molecular chains, loss of heat-sensitive flavonoids through oxidation, and loss of volatile aroma components. Enzymatic extraction requires prolonged enzymatic hydrolysis, and residual enzymes affect the stability of the concentrated juice. Therefore, there is a need to find a simple, efficient method for preparing Codonopsis pilosula concentrated juice that retains more of the active ingredients of Codonopsis pilosula. Summary of the Invention

[0004] The present invention aims to provide a method for extracting concentrated Codonopsis pilosula juice under ultra-high pressure combined with enzymatic hydrolysis, which can improve the retention of flavonoid active ingredients and solve the problem of low extraction rate of active ingredients such as flavonoids and polysaccharides caused by traditional water decoction method.

[0005] To achieve the above objectives, the following technical solution is provided: A method for extracting concentrated Codonopsis pilosula juice under ultra-high pressure combined with enzymatic hydrolysis, characterized by comprising the following steps: Step 1: First, select Codonopsis pilosula that is free from mold and pests, wash off the dirt and impurities on the surface, dry the surface moisture, and then slice it for later use. Step 2: Weigh 100 g of Codonopsis pilosula slices, put them into a grinder to grind them, and then pass them through a 100-mesh sieve to obtain Codonopsis pilosula powder for later use; take the Codonopsis pilosula powder, add purified water at a material-to-liquid ratio of 1:15 (g / mL), and stir thoroughly to obtain Codonopsis pilosula stock solution; heat the Codonopsis pilosula stock solution to 45 ℃, add a compound enzyme for enzymatic hydrolysis, and heat to inactivate the enzyme after 1 h to obtain a first Codonopsis pilosula extract; Step 3: Transfer the enzyme-inactivated primary Codonopsis pilosula extract into a polyethylene plastic bag, seal it, and place it in an ultra-high pressure device for ultra-high pressure treatment. After ultra-high pressure treatment, centrifuge the Codonopsis pilosula extract at 8000 rpm for 15 min to obtain the secondary Codonopsis pilosula extract. Step four involves vacuum concentrating the secondary Codonopsis pilosula extract for 15 minutes to obtain concentrated Codonopsis pilosula juice. Finally, the concentrated juice is sterilized and aseptically bottled.

[0006] Furthermore, in step two, the composite enzyme is cellulase and papain, the ratio of cellulase to papain is 1:3, the enzymatic hydrolysis temperature is 45 ℃, cellulase is added first for 30 min of enzymatic hydrolysis, and then papain is added for 30 min of enzymatic hydrolysis.

[0007] Furthermore, in step three, in order to obtain the optimal ultra-high pressure, the primary Codonopsis pilosula extract is treated for 10 min at 0 MPa, 100 MPa, 200 MPa, 300 MPa, and 400 MPa.

[0008] Furthermore, in step three, in order to obtain the optimal ultra-high pressure time, the Codonopsis pilosula extract is treated at 300 MPa for 5 min, 10 min, 15 min, and 20 min.

[0009] The beneficial effects of this invention are as follows: 1. Ultra-high pressure instantly destroys the cell wall structure, and combined with cellulase and papain for efficient enzymatic hydrolysis, it significantly shortens the extraction time and improves the dissolution rate and yield of the target components.

[0010] 2. Ultra-high pressure physical force causes the cell wall to rupture instantly, combined with enzymatic hydrolysis to specifically degrade cell wall polysaccharides, the dual effect significantly improves the cell wall rupture rate and extraction rate.

[0011] 3. Ultra-high pressure technology can achieve physical cell wall disruption at room temperature or low temperature, avoiding greater thermal damage and maintaining the natural structure and biological activity of active ingredients. Attached Figure Description

[0012] Figure 1 This is a process flow diagram of the present invention; Figure 2 This illustrates the effect of different pressures on the polysaccharide content of Codonopsis pilosula in this invention. Figure 3 This illustrates the effect of different pressures on the total flavonoid content in this embodiment of the invention. Figure 4 This illustrates the effect of different pressures on antioxidant activity in the embodiments of the present invention. Figure 5 This illustrates the effect of different time points on the polysaccharide content of Codonopsis pilosula in this embodiment of the invention. Figure 6 This illustrates the effect of different time points on the total flavonoid content in the embodiments of the present invention; Figure 7 This illustrates the effect of different time points on antioxidant activity in the embodiments of the present invention. Detailed Implementation

[0013] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention. Example

[0014] A method for extracting concentrated Codonopsis pilosula juice using ultra-high pressure combined with enzymatic hydrolysis, such as... Figure 1 As shown, it includes the following steps: Step 1: First, select Codonopsis pilosula that is free from mold and pests, wash off the dirt and impurities on the surface, dry the surface moisture, slice it after drying, and set aside. Step 2: Weigh 100 g of Codonopsis pilosula slices, put them into a grinder to grind them, and then pass them through a 100-mesh sieve to obtain Codonopsis pilosula powder for later use; take the Codonopsis pilosula powder, add purified water at a material-to-liquid ratio of 1:15 (g / mL), and stir thoroughly to obtain Codonopsis pilosula stock solution; heat the Codonopsis pilosula stock solution to 45 ℃, add a compound enzyme for enzymatic hydrolysis, and heat to inactivate the enzyme after 1 h to obtain a first Codonopsis pilosula extract; Step 3: Transfer the enzyme-inactivated primary Codonopsis pilosula extract into a polyethylene plastic bag, seal it, and place it in an ultra-high pressure device for ultra-high pressure treatment. After ultra-high pressure treatment, centrifuge the Codonopsis pilosula extract at 8000 rpm for 15 min to obtain secondary Codonopsis pilosula extract. Vacuum concentrate the secondary Codonopsis pilosula extract for 15 min to obtain Codonopsis pilosula concentrate. Finally, sterilize the concentrate and aseptically fill it.

[0015] Furthermore, in step two, the composite enzyme is cellulase and papain, the ratio of cellulase to papain is 1:3, the enzymatic hydrolysis temperature is 45 ℃, cellulase is added first for 30 min of enzymatic hydrolysis, and then papain is added for 30 min of enzymatic hydrolysis.

[0016] Furthermore, in step three, to obtain the optimal ultra-high pressure, the primary Codonopsis pilosula extract was treated for 10 min at 0 MPa, 100 MPa, 200 MPa, 300 MPa, and 400 MPa. Finally, the following samples were obtained: control sample (without ultra-high pressure treatment), sample 1 (0 MPa, 10 min), sample 2 (100 MPa, 10 min), sample 3 (200 MPa, 10 min), sample 4 (300 MPa, 10 min), and sample 5 (400 MPa, 10 min).

[0017] Furthermore, in step three, in order to obtain the optimal ultra-high pressure time, the Codonopsis pilosula extract was treated at 300 MPa for 5 min, 10 min, 15 min, and 20 min. Samples 6 (300 MPa, 5 min), 7 (400 MPa, 10 min), 8 (300 MPa, 15 min), and 9 (300 MPa, 20 min) were obtained.

[0018] Example 2: The polysaccharides, total flavonoids, and antioxidant properties of Codonopsis pilosula concentrate were tested.

[0019] (1) Detection of Codonopsis pilosula polysaccharide content Preparation of the linear regression equation: Prepare a 5 mg / mL glucose standard solution. Take 0 mL, 1 mL, 2 mL, 3 mL, 4 mL, and 5 mL of the 5 mg / mL glucose standard solution and dilute to volume in 10 mL volumetric flasks. Take 1 mL of each concentration of glucose standard solution in a test tube, add 2 mL of 5% phenol solution, shake well, and quickly add 2 mL of concentrated sulfuric acid. Heat in boiling water for 20 min, then remove and cool to room temperature. Measure the absorbance at a wavelength of 490 nm. Plot a linear regression equation with the glucose standard solution concentration on the x-axis and absorbance on the y-axis.

[0020] Take 1 mL of the control sample and the concentrated juice of samples 1-9 into a test tube, add 2 mL of 5% phenol solution, shake well, quickly add 2 mL of concentrated sulfuric acid, heat in a boiling water bath for 20 min, and then cool to room temperature. Measure the absorbance at a wavelength of 490 nm, and calculate the content of Codonopsis pilosula polysaccharides according to the linear regression equation. Each sample was measured in triplicate, and the average value was taken.

[0021] , In the formula: C - concentration calculated from the regression equation, mg / mL; V - sample volume, mL; N - dilution factor; M - sample volume, mL. For example... Figure 2 and Figure 4 As shown.

[0022] (2) Detection of total flavonoid content in Codonopsis pilosula Preparation of the standard curve: Weigh 5.2 mg of rutin standard, add 90% ethanol solution to make up to 25 mL and shake well. Pipette 0 mL, 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1.0 mL of the standard solution into 10 mL volumetric flasks, add 0.4 mL of 5% sodium nitrite solution and let stand for 6 min; then add 0.4 mL of 10% aluminum nitrate solution and let stand for 6 min; add 4 mL of 4% sodium hydroxide solution, then add 90% ethanol solution to make up to the mark, shake well and let stand for 15 min. Measure the absorbance at the maximum absorption peak of 510 nm. Plot the standard curve with absorbance value on the ordinate and rutin concentration on the x-axis.

[0023] Take 1 mL of the sample solution, the control sample, and the concentrated juice of samples 1-9, and place them in a 10 mL volumetric flask. Add 0.4 mL of 5% sodium nitrite solution and let stand for 6 min. Then add 0.4 mL of 10% aluminum nitrate solution and let stand for 6 min. Add 4 mL of 4% sodium hydroxide solution, and then add 90% ethanol solution to make up to the mark. Shake well and let stand for 15 min. Measure the absorbance at the maximum absorption peak of 510 nm. Calculate the total flavonoid content according to the standard curve. Each sample was measured in triplicate, and the average value was taken.

[0024] , Where: N - dilution factor; C - concentration calculated from the standard curve, mg / mL; V 测 - Volume of liquid to be tested, mL; V 样 - Sample volume, mL. For example... Figure 3 and Figure 5 As shown.

[0025] (3) DPPH scavenging rate determination: The DPPH free radical scavenging rate kit was used for testing. 1 mL of the control sample and the concentrated juice of samples 1-9 were added to the kit. Figure 4 and Figure 7 As shown.

[0026] , The analytical results of the concentrated Codonopsis pilosula juice obtained under different processing methods are as follows. Figure 2 , Figure 3 and Figure 4As shown, the Codonopsis pilosula concentrate obtained under ultra-high pressure of 300 MPa had the highest content of Codonopsis pilosula polysaccharides and total flavonoids, and the best antioxidant capacity. This is because during ultra-high pressure extraction, pressure affects the degree of cell wall rupture and the dissolution efficiency of active ingredients. When the pressure is too low (such as 100 MPa or 200 MPa), the cell wall is not sufficiently destroyed, resulting in a low extraction rate of polysaccharides and flavonoids. When the pressure is too high (such as 400 MPa), some heat-sensitive components (such as certain flavonoids) may denature or degrade, which will reduce the extraction efficiency. When the pressure is 300 MPa, the plant cell wall can be effectively destroyed, promoting the release of active ingredients (polysaccharides and flavonoids) from the cells.

[0027] like Figure 5 , Figure 6 , Figure 7 As shown, the Codonopsis pilosula concentrate obtained by ultra-high pressure treatment for 15 minutes contained the highest content of Codonopsis pilosula polysaccharides and total flavonoids, and had the best antioxidant properties. This is because too short a time (such as 5 minutes or 10 minutes) will lead to incomplete cell wall rupture and insufficient dissolution of active ingredients, resulting in lower polysaccharide and flavonoid contents. Too long a time (such as 20 minutes) may cause some active ingredients (such as certain flavonoids) to degrade due to prolonged high pressure, reducing the bioactivity of the extract.

[0028] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

Claims

1. A method for extracting concentrated juice of radix codonopsis by superhigh pressure combined with enzymolysis, characterized in that, The method comprises the following steps: Step one, select the no mildew no insect party, wash off the surface of the soil impurities, dry the surface moisture, dry and cut into slices for use; Step two, take 100 g of party slice, put it into the crusher and crush it through a 100 mesh sieve to obtain party powder for use; take party powder, add purified water according to the ratio of 1:15, fully stir and mix, obtain party original liquid; heat the party original liquid to 45 ℃, add compound enzyme for enzymolysis, 1 h after heating to kill enzyme, obtain the first party extract; Step three, transfer the first party extract after enzyme inactivation to a polyethylene plastic bag, seal, place in an ultra-high pressure device for ultra-high pressure treatment, centrifuge the party extract after ultra-high pressure treatment at 8000 rpm for 15 min to obtain the second party extract; Step four, vacuum concentrate the second party extract for 15 min to obtain party concentrated juice, and finally sterilize the concentrated juice and aseptically fill it.

2. The method of claim 1, wherein the method is characterized by: In step two, the compound enzyme is cellulase and papain, the ratio of cellulase and papain is 1:3, the enzymolysis temperature is 45 ℃, cellulase is added first for enzymolysis for 30 min, and then papain is added for enzymolysis for 30 min.

3. The method for extracting concentrated Codonopsis pilosula juice under ultra-high pressure combined with enzymatic hydrolysis according to claim 1, characterized in that: In step three, the first party extract is treated under the conditions of 0 MPa, 100 MPa, 200 MPa, 300 MPa and 400 MPa for 10 min.

4. The method for extracting concentrated Codonopsis pilosula juice under ultra-high pressure combined with enzymatic hydrolysis according to claim 1, characterized in that: In step three, the first party extract is treated under the condition of 300 MPa for 5 min, 10 min, 15 min and 20 min.

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