Low-bitterness zanthoxylum bungeanum seed peptide freeze-dried flash-release tablet and preparation method thereof

Abstract

The application belongs to the technical field of functional polypeptides, and particularly relates to a low-bitterness Zanthoxylum bungeanum seed peptide freeze-drying flash-release tablet and a preparation method thereof. The Zanthoxylum bungeanum seed peptide freeze-drying flash-release tablet provided by the application comprises the following components in parts by weight: 0.1-0.5 parts of Zanthoxylum bungeanum seed peptide, 10-15 parts of mannitol and 4-6 parts of gelatin. The Zanthoxylum bungeanum seed peptide is at least one of Zanthoxylum bungeanum seed antioxidant peptide and Zanthoxylum bungeanum seed antihypertensive peptide. The preparation method of the Zanthoxylum bungeanum seed peptide freeze-drying flash-release tablet is as follows: Zanthoxylum bungeanum seed pretreatment, Zanthoxylum bungeanum seed protein extraction, Zanthoxylum bungeanum seed peptide preparation, Zanthoxylum bungeanum seed peptide debittering and combination with other excipients to prepare the Zanthoxylum bungeanum seed peptide freeze-drying flash-release tablet. The application prepares a low-bitterness Zanthoxylum bungeanum seed peptide freeze-drying flash-release tablet by using Zanthoxylum bungeanum seeds as raw materials, not only realizes full use of Zanthoxylum bungeanum seeds and improves the nutritional value of Zanthoxylum bungeanum seed protein, but also has the advantages that the prepared product is easy to store, has strong dispersing capacity, can be dissolved quickly in the mouth without swallowing or water dissolution, and has no unpleasant taste after dissolution.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of functional polypeptides, and particularly relates to a low-bitterness Zanthoxylum bungeanum seed peptide freeze-dried flash-release tablet and a preparation method thereof. BACKGROUND

[0002] Zanthoxylum bungeanum is originally from China and has a cultivation history of more than 2,000 years. The planting area and annual output of Zanthoxylum bungeanum in China rank first in the world. However, the main by-product of Zanthoxylum bungeanum pericarp, Zanthoxylum bungeanum seed, has not been fully utilized. Only a small part is used as fuel or as fertilizer, and most of the Zanthoxylum bungeanum seeds are discarded, causing resource waste and environmental pollution. The protein content in Zanthoxylum bungeanum seed is 9-15%, and various amino acid components are complete. Moreover, the polypeptide obtained by hydrolyzing Zanthoxylum bungeanum seed protein with protease has the functions of antibacterial, antioxidant and blood pressure reduction, and is a plant protein resource with great development potential. However, Zanthoxylum bungeanum has a unique aroma and numbness, is bitter and spicy, as mentioned in “Theory of Drug Properties” (the taste is bitter and spicy, and has a small amount of poison), and “Tang Bencao” (bitter, cold, and non-toxic). This characteristic limits the further utilization of Zanthoxylum bungeanum seed.

[0003] At present, most of the researches on the application of Zanthoxylum bungeanum seed are to prepare Zanthoxylum bungeanum polypeptide by treating Zanthoxylum bungeanum seed. For example, patent numbers CN111387338A and CN116143871A disclose a preparation method of Zanthoxylum bungeanum seed polypeptide. However, the above-mentioned technologies need to use expensive equipment, resulting in high production cost of the product. Moreover, in the process of enzymatic hydrolysis of Zanthoxylum bungeanum polypeptide by using enzyme preparation, the content of bitter and spicy substances in Zanthoxylum bungeanum seed is relatively high, and the bitter taste of the enzymatic hydrolysate of protein also exists due to the presence of hydrophobic polypeptide, so that the prepared Zanthoxylum bungeanum seed polypeptide has more or less bitter taste, which further affects the edible property.

[0004] At present, several methods for removing bitterness from active peptides have been reported, such as active carbon adsorption, embedding, biological enzymatic hydrolysis, and adding bitter taste treatment agent. However, the active carbon adsorption method for removing bitterness is not selective, and the nutritional components and bioactive components are also damaged simultaneously. The embedding method for removing bitterness is actually a certain masking of bitter components. Although the embedding method has high efficiency for removing bitterness and small influence on the activity of protein / polypeptide, the embedding method may affect the taste and texture of the product due to the outer embedding wall material, and the uneven embedding may also cause the problem of re-exposure of bitterness. The biological enzymatic hydrolysis method for removing bitterness has low cost, high peptide yield, and obvious de-bittering effect. However, the further enzymatic hydrolysis of amino acids and small peptides may also cause changes in biological activity.

[0005] And at present, there are few studies on the method for removing the bitterness of Zanthoxylum bungeanum seed polypeptide. Patent CN112210434A uses a bitter taste treatment agent composed of adenosine triphosphate and sodium ascorbate to treat the bitterness of Zanthoxylum bungeanum oil, but the treatment method has a high cost, and it is unknown whether it can be applied to the removal of the bitterness of Zanthoxylum bungeanum polypeptide.

[0006] In addition, Zanthoxylum bungeanum seed polypeptide on the market is mostly in the form of powder, and the powder-like polypeptide is prone to moisture absorption and deterioration, and is difficult to store and transport. If Zanthoxylum bungeanum seed polypeptide is prepared into common drug preparations such as capsule type, tablet type and granule type, it inevitably has the disadvantages of being difficult to swallow and being difficult to store for a long time. SUMMARY

[0007] In order to solve the above technical problems, the present application provides a low-bitterness Zanthoxylum bungeanum seed peptide freeze-dried flash release tablet and a preparation method thereof.

[0008] The low-bitterness Zanthoxylum bungeanum seed peptide freeze-dried flash release tablet provided by the present application comprises the following components in parts by weight: 0.1-0.5 parts of Zanthoxylum bungeanum seed peptide, 10-15 parts of mannitol and 4-6 parts of gelatin; the Zanthoxylum bungeanum seed peptide is at least one of Zanthoxylum bungeanum seed antioxidant peptide and Zanthoxylum bungeanum seed antihypertensive peptide.

[0009] The present application also provides a preparation method of the low-bitterness Zanthoxylum bungeanum seed peptide freeze-dried flash release tablet, comprising the following steps:

[0010] (1) Zanthoxylum bungeanum seed pretreatment: removing impurities from dried Zanthoxylum bungeanum seeds, crushing them with a flour mill, sieving and reserving;

[0011] (2) Extraction of Zanthoxylum bungeanum seed protein: dissolving the Zanthoxylum bungeanum seed powder in (1) in water to prepare a solution, the mass-volume ratio of Zanthoxylum bungeanum seed powder to water being 1g:20-30mL, adjusting the pH of the solution to 8.0-10.0, water bath heating for 3-6h, centrifugation, retaining the supernatant; then adjusting the pH of the supernatant to 3.0-5.0, centrifugation, discarding the supernatant and retaining the precipitate, washing the precipitate to neutral, obtaining a Zanthoxylum bungeanum seed protein solution, freezing and standing, and drying;

[0012] (3) Preparation of Zanthoxylum bungeanum seed peptide: preparing the Zanthoxylum bungeanum seed protein obtained in (2) into a solution, and performing enzymolysis with a protease; before adding the protease, the pH of the solution is adjusted to the optimal reaction pH of the protease; the pH of the solution is maintained stable within the range of the optimal reaction pH±0.05 during the enzymolysis process; after the enzymolysis reaction is completed, boiling water bath is performed for 10-20min to inactivate the enzyme, and then the solution is cooled to obtain Zanthoxylum bungeanum seed peptide;

[0013] (4) Bitterness removal: mixing the Zanthoxylum bungeanum seed peptide solution obtained in (3) with n-butanol to extract the bitter peptides therefrom, collecting the hydrolyzate layer to obtain a low-bitterness Zanthoxylum bungeanum seed peptide solution, freeze-drying the solution, and then freezing and storing the solution at-20℃ for standby use.

[0014] (5) dissolving the Zanthoxylum bungeanum seed peptide and mannitol prepared in (4) in the pre-dissolved gelatin solution, uniformly mixing the mixed solution, then constant volume with pure water, then performing injection molding and freeze-drying to obtain the Zanthoxylum bungeanum seed peptide freeze-dried flash release tablet.

[0015] In the preparation method of the Zanthoxylum bungeanum seed peptide freeze-dried flash release tablet with low bitterness mentioned above:

[0016] Preferably, in (2), the mass-volume ratio of the Zanthoxylum bungeanum seed powder to water is 1g: 22-26mL.

[0017] Preferably, in (2), the mass-volume ratio of the Zanthoxylum bungeanum seed powder to water is 1g: 25mL.

[0018] Preferably, in (2), the pH of the solution is adjusted to 8.0-10.0 by using NaOH, and the concentration of the NaOH solution is 0.8-1.2mol / L; the pH of the solution is adjusted to 3.0-5.0 by using HCl, and the concentration of the HCl solution is 1.0mol / L; the precipitate is washed to neutral by using 0.1-1.5mol / L NaOH solution and HCl solution.

[0019] Preferably, in (2), the temperature of the water bath heating is 45-55℃.

[0020] Preferably, in (2), the temperature of the water bath heating is 50℃.

[0021] Preferably, in (3), the protease is selected from any one of alkaline protease, complex protease, neutral protease, trypsin and pepsin; and the Zanthoxylum bungeanum seed peptide is any one of Zanthoxylum bungeanum seed antioxidant peptide and Zanthoxylum bungeanum seed antihypertensive peptide.

[0022] Preferably, in (3), when the Zanthoxylum bungeanum seed peptide is Zanthoxylum bungeanum seed antioxidant peptide, the protease is alkaline protease; and when the Zanthoxylum bungeanum seed peptide is Zanthoxylum bungeanum seed antihypertensive peptide, the protease is pepsin.

[0023] In the present application, different types of proteases are used to hydrolyze the protein solution of Zanthoxylum bungeanum seeds, and the physiological effects of the Zanthoxylum bungeanum seed polypeptides obtained show great differences.

[0024] Among them, the Zanthoxylum bungeanum seed peptide obtained after alkaline protease hydrolysis has significant DPPH free radical scavenging ability, Fe 2+ chelating ability and hydroxyl radical scavenging ability, and its antioxidant effect is even significantly better than Zanthoxylum bungeanum seed protein and the reducing agent glutathione, so the Zanthoxylum bungeanum seed peptide obtained after protease treatment in the present application is named "Zanthoxylum bungeanum seed antioxidant peptide".

[0025] In addition, the ACE inhibiting ability of the Zanthoxylum bungeanum seed peptide obtained by pepsin enzymolysis of Zanthoxylum bungeanum seed protein is significantly higher than that of Zanthoxylum bungeanum seed protein, and is even comparable to that of the antihypertensive drug captopril, therefore, the Zanthoxylum bungeanum seed peptide with strong ACE inhibiting ability obtained after pepsin action in the application is named as "Zanthoxylum bungeanum seed antihypertensive peptide".

[0026] Preferably, in (3), the concentration of the protein solution is 8-15 mg / mL, the added amount of the protease is 4000-8000 U / g of protein, and the hydrolysis time is 3-6 h.

[0027] More preferably, in (3), the concentration of the protein solution is 10 mg / mL, the added amount of the protease is 6000 U / g of protein, and the hydrolysis time is 4-5 h.

[0028] Preferably, in (4), the volume ratio of the hydrolysate to n-butanol is 1:1-4.

[0029] In the application, the raw material Zanthoxylum bungeanum seed contains more bitter and pungent components, and the Zanthoxylum bungeanum seed polypeptide needs to be enzymolyzed in the preparation process. Although the operation of enzymolysis can make the Zanthoxylum bungeanum seed protein enzymolyzed into small molecular polypeptides, this process also increases the exposure degree of bitter polypeptides, increases the content of bitter polypeptides, and makes the enzymolyzed Zanthoxylum bungeanum seed polypeptide have a certain bitter taste, which affects the eating experience.

[0030] The n-butanol, as an alcohol organic extractant, has hydrophobicity and hydrophilicity, can be used for the extraction of natural medicines, and can be used for removing bitter peptides from Zanthoxylum bungeanum seed, which is simple in operation, good in debittering effect, and conducive to industrialized production.

[0031] Preferably, in (5), the gelatin is dissolved in the following manner: after being soaked in cold water for 6-15 min, stirring is carried out in a water bath at 40-80 DEG C until complete dissolution.

[0032] Preferably, in (5), the solution needs to be emulsified before freeze-drying, the rotation speed of the emulsifying machine is 8000-12000 rpm, emulsification is carried out for 8-20 min, and in the freeze-drying, pre-freezing is first carried out at a temperature lower than -60 DEG C for 3-5 h, and then the solution is transferred into a vacuum freeze-drying machine, and freeze-drying is carried out for 12-48 h.

[0033] More preferably, in (5), the solution needs to be emulsified before freeze-drying, the rotation speed of the emulsifying machine is 10000 rpm, emulsification is carried out for 8-12 min, and in the freeze-drying, pre-freezing is first carried out at a temperature of -80 DEG C for 3-5 h, and then the solution is transferred into a vacuum freeze-drying machine, and freeze-drying is carried out for 24 h.

[0034] The application has the following beneficial effects:

[0035] (1) The present application respectively adopts different kinds of enzyme preparations to carry out enzymolysis on the Zanthoxylum bungeanum seed protein solution, wherein the Zanthoxylum bungeanum seed peptide obtained by adopting pepsin to carry out enzymolysis on the Zanthoxylum bungeanum seed protein has high ACE inhibiting capacity, and the Zanthoxylum bungeanum seed antioxidant peptide obtained by adopting alkaline protease to carry out hydrolysis exhibits significant antioxidant property, so that the Zanthoxylum bungeanum seed peptide with a certain specific efficacy can be directly applied or combined in actual production, which is beneficial to realize full utilization of the Zanthoxylum bungeanum seed and improve the nutritional value of the Zanthoxylum bungeanum seed protein.

[0036] (2) The present application reduces the bitterness of the Zanthoxylum bungeanum seed peptide product and improves the palatability of the product by simply treating the Zanthoxylum bungeanum seed peptide, i.e. removing bitter peptides and other components in the Zanthoxylum bungeanum seed peptide by adopting n-butanol.

[0037] (3) The present application improves the storage mode of the Zanthoxylum bungeanum seed peptide, i.e. preparing the Zanthoxylum bungeanum seed peptide into a freeze-dried flash release tablet, so that the prepared product is easy to store, and the freeze-dried flash release tablet has the advantages of strong dispersing capacity, no need for swallowing or water dissolution, rapid dissolution in the mouth, no unpalatable feeling after dissolution, etc. In addition, the freeze-dried flash release tablet in the present application is a small molecular peptide prepared at low temperature, the active components of the Zanthoxylum bungeanum seed peptide are well maintained, and the Zanthoxylum bungeanum seed peptide can be quickly absorbed by the human body after being taken. BRIEF DESCRIPTION OF DRAWINGS

[0038] Figure 1 Figure 1 is the DPPH free radical scavenging rate of the Zanthoxylum bungeanum seed peptide obtained under different protease enzymolysis conditions in Example 1 of the present application.

[0039] Figure 2 Figure 2 is the Fe2+chelating rate of the Zanthoxylum bungeanum seed peptide obtained under different protease enzymolysis conditions in Example 1 of the present application. 2+

[0040] Figure 3 Figure 3 is the hydroxyl free radical scavenging rate of the Zanthoxylum bungeanum seed peptide obtained under different protease enzymolysis conditions in Example 1 of the present application.

[0041] Figure 4 Figure 4 is the ACE inhibiting rate of the Zanthoxylum bungeanum seed peptide obtained under different protease enzymolysis conditions in Example 1 of the present application. DETAILED DESCRIPTION

[0042] In order to enable those skilled in the art to better understand the present application, the present application will be further described in conjunction with specific embodiments.

[0043] Example 1: Preparation and property evaluation of Zanthoxylum bungeanum seed peptide

[0044] The preparation steps of the Zanthoxylum bungeanum seed peptide are as follows:

[0045] (1) Zanthoxylum bungeanum seed pretreatment: remove impurities from the dried Zanthoxylum bungeanum seed, crush it with a flour mill, and pass it through a 200-mesh sieve for standby use;​

[0046] (2) Extraction of Zanthoxylum bungeanum seed protein: Zanthoxylum bungeanum seed powder was dissolved in water to prepare a solution, the mass-volume ratio of Zanthoxylum bungeanum seed powder to water was 1 g:25 mL, 1 mol / L NaOH solution was added to adjust the pH value to 10.0, and the solution was stirred in a water bath at 50°C for 4.5 h. After the reaction was completed, centrifugation was performed, the supernatant was taken, 1 mol / L HCl solution was added to adjust the pH value to 4.5, centrifugation was performed, and the precipitate was reserved; finally, the precipitate was washed to neutral with 0.1 mol / L NaOH solution and 0.1 mol / L HCl solution to obtain a Zanthoxylum bungeanum seed protein solution, which was freeze-dried to collect the protein, and the protein was stored at -20°C for standby use;

[0047] (3) Preparation of Zanthoxylum bungeanum seed peptide: the Zanthoxylum bungeanum seed protein prepared in (2) was prepared into a solution with a concentration of 10 mg / mL, and a total amount of 6000 U / g protein of protease (alkaline protease, complex protease, neutral protease, trypsin, pepsin) was added, and the solution was hydrolyzed under the optimum reaction conditions of the protease for 4 h. The hydrolysis conditions of different proteases are shown in Table 1 below, and 0.2 mol / L NaOH and HCl solutions were used to maintain the pH value of the solution within ±0.05 during the reaction. After the reaction was completed, a boiling water bath was immediately performed for 15 min, and then the solution was naturally cooled and freeze-dried. The freeze-dried Zanthoxylum bungeanum seed peptide was stored at -20°C for standby use.

[0048] Table 1 Optimum pH value and reaction temperature of different proteases

[0049] Protease type pH Temperature / °C Alkaline protease 8.0 60 Complex protease 6.5 50 Neutral protease 7.0 50 Trypsin 8.0 37 Pepsin 2.0 37

[0050] The DPPH free radical scavenging rate, Fe 2+ chelating ability, and hydroxyl radical scavenging rate of the Zanthoxylum bungeanum seed peptide prepared above were determined.

[0051] Test Example 1 Determination of DPPH free radical scavenging rate

[0052] The DPPH free radical scavenging rate of the Zanthoxylum bungeanum seed peptide was determined by the following method:

[0053] Sample solutions with different concentrations (0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL) were prepared, reagents were added to test tubes according to the reagent addition amount shown in Table 2 below, the test tubes were mixed and then placed in a dark environment at room temperature for 30 min, and then the absorbance value at 517 nm was determined. Zanthoxylum bungeanum seed protein and strong reducing agent glutathione were used as controls. The calculation formula of the DPPH free radical scavenging rate is as follows:

[0054]

[0055] In the formula:

[0056] OD0 - absorbance value of ethanol + sample;

[0057] OD1 - absorbance value of ethanol + DPPH;

[0058] OD2 - absorbance value of sample + DPPH.

[0059] Table 2 Sample addition amount of DPPH free radical scavenging rate

[0060] Test tube number 0 1 2 Sample (mL) 2 0 2 95% ethanol (mL) 2 2 0 DPPH-0.1 mmol / L ethanol solution (mL) 0 2 2

[0061] Test Example 2: Fe 2+ Chelating ability determination

[0062] The Fe of Zanthoxylum bungeanum seed peptide was determined using the following method 2+ Chelating ability:

[0063] Sample solutions of different concentrations (0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL) were prepared, and reagents were added to test tubes according to the reagent addition amount in Table 3 below, mixed well, and then allowed to stand for 15 min of reaction, and then the absorbance value at 562 nm was determined, and Zanthoxylum bungeanum seed protein and strong reducing agent glutathione were used as controls. 2+ The calculation formula of chelating ability is as follows:

[0064]

[0065] In the formula:

[0066] OD0 - absorbance value of water + FeCl2+ phenanthroline;

[0067] OD1 - absorbance value of sample + FeCl2+ phenanthroline;

[0068] OD2 - absorbance value of sample + water.

[0069] Table 3 Fe 2+ Chelating ability experiment of each reagent addition amount

[0070] Test tube number 0 1 2 Sample (mL) 0 1 1 Water (mL) 1 0 4 0.05 mmol / L FeCl2solution (mL) 2 2 0 0.5 mmol / L phlorrhizin solution (mL) 2 2 0

[0071] Test Example 3: Hydroxyl radical scavenging rate

[0072] The hydroxyl radical scavenging rate of Zanthoxylum bungeanum seed peptide was determined using the following method:

[0073] Prepare sample solutions with different concentrations (0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL), and add reagents to test tubes according to the reagent addition amount in Table 4 below, mix well, and then place in a 37°C water bath for 30 min, and then measure the absorbance value at 510 nm, while using Zanthoxylum bungeanum seed protein and strong reducing agent light glutathione as controls. The calculation formula of the hydroxyl radical scavenging rate is as follows:

[0074]

[0075] In the formula:

[0076] OD0 is the absorbance value of water + FeSO4 + salicylic acid-ethanol + H2O2;

[0077] OD1 is the absorbance value of sample + water + FeSO4 + salicylic acid-ethanol;

[0078] OD2 is the absorbance value of sample + FeSO4 + salicylic acid-ethanol + H2O2.

[0079] Table 4 Addition amount of each reagent in the determination experiment of the hydroxyl radical scavenging rate

[0080] Test tube number 0 1 2 Sample (mL) 0 1 1 Water (mL) 1 1 0 2 mmol / L FeSO4 solution (mL) 1 1 1 6 mmol / L salicylic acid-ethanol solution (mL) 1 1 1 6 mmol / L H2O2(mL) 1 0 1

[0081] Test Example 4: ACE inhibition rate

[0082] The ACE inhibition capacity is determined using the following method:

[0083] Prepare sample solutions with different concentrations (0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL), and add reagents to test tubes according to the reagent addition amount in Table 5 below, add 1 mol / L HCl to terminate the reaction, then add 2 mL of pre-cooled ethyl acetate, vortex for 20 s, then centrifuge at 6000 r / min for 10 min, take 1.5 mL of the upper layer solution to another clean test tube, evaporate the ethyl acetate in a boiling water bath, then add 2 mL of 60°C hot water to the test tube, shake to dissolve, mix well, and then measure the absorbance value at 195 nm, while using Zanthoxylum bungeanum seed protein and the antihypertensive drug captopril as controls. The ACE inhibition rate is calculated according to the following formula:

[0084]

[0085] In the formula:

[0086] OD0 is the absorbance value of hippurylhistidylleucine (HHL) + phosphate buffer + ACE;

[0087] OD1 is the absorbance value of HHL + phosphate buffer;

[0088] OD2—absorbance value of HHL+sample+ACE.

[0089] Table 5 ACE inhibition rate experiment The amount of each reagent added

[0090]

[0091] The test results of Test Examples 1 to 4 are shown in Tables 6 to 9 below. In order to more intuitively compare the antioxidant properties of Zanthoxylum bungeanum seed peptides obtained by enzymatic hydrolysis with different types of proteases, Tables 6 to 9 are plotted in the form of images, as shown in the attached figure. Figures 1-3 shown.

[0092] Table 6 DPPH free radical scavenging rate of Zanthoxylum bungeanum seed antioxidant peptides (%)

[0093] Protease type 0.1 mg / mL 0.2 mg / mL 0.3 mg / mL 0.4 mg / mL 0.5 mg / mL Complex protease 10.65 18.91 30.91 43.95 50.28 Alkaline protease 15.27 33.61 52.02 75.38 90.21 Neutral protease 10.76 26.06 38.20 53.09 70.75 Trypsin 14.62 25.48 36.99 48.48 59.05 Pepsin 10.67 15.16 21.51 24.43 25.22 Zanthoxylum bungeanum seed protein 11.36 17.28 21.37 25.90 27.99 Glutathione - - - - 47.79

[0094] Table 7 Fe of Zanthoxylum bungeanum seed peptide 2+ Chelation rate (%)

[0095] Protease type 0.5 mg / mL 1.0 mg / mL 1.5 mg / mL 2.0 mg / mL 2.5 mg / mL Complex protease 8.05 18.42 28.20 38.16 44.00 Alkaline protease 40.90 67.52 87.10 90.42 92.17 Neutral protease 1.95 3.33 7.36 16.96 24.95 Trypsin 11.46 30.66 36.80 44.82 56.89 Pepsin 2.11 11.30 17.53 21.77 28.95 Zanthoxylum bungeanum seed protein 1.74 6.59 11.54 13.76 29.11 Glutathione - - - - 15.13

[0096] Table 8 Hydroxyl radical scavenging rate of Zanthoxylum bungeanum seed peptide (%)

[0097]

[0098]

[0099] Table 9 ACE inhibition rate of Zanthoxylum bungeanum seeds (%)

[0100] Protease type 0.5 mg / mL 1.0 mg / mL 1.5 mg / mL 2.0 mg / mL 2.5 mg / mL Complex protease 45.55 77.93 80.98 86.59 87.36 Alkaline protease 33.07 61.00 78.78 92.82 93.24 Neutral protease 12.94 28.52 36.39 65.81 73.41 Trypsin 24.13 62.10 72.34 80.69 82.75 Pepsin 47.02 83.01 87.17 95.25 96.46 Zanthoxylum bungeanum seed protein 5.37 15.14 19.32 22.24 25.68 Captopril 65.79 83.64 91.12 97.98 98.99

[0101] From the above results, it can be seen that the Zanthoxylum bungeanum seed peptides obtained after hydrolysis by different types of proteases have certain DPPH free radical scavenging ability, Fe 2+ The chelating ability and hydroxyl radical scavenging ability of the pepper seed peptide obtained by alkaline protease hydrolysis are the strongest, and the antioxidant capacity is much higher than that of pepper seed protein and the reducing agent glutathione. Therefore, in the present invention, alkaline protease is used to hydrolyze pepper seed protein to prepare pepper seed antioxidant peptide.

[0102] In addition, it can be seen from the data on the inhibition rate of Zanthoxylum bungeanum seed peptide on ACE that the Zanthoxylum bungeanum seed peptide obtained by pepsin hydrolysis has an ACE inhibitory ability much higher than that of Zanthoxylum bungeanum seed protein, and is equivalent to the ACE inhibitory ability of the antihypertensive drug captopril. Therefore, in the present invention, pepsin is used to hydrolyze Zanthoxylum bungeanum seed protein to prepare Zanthoxylum bungeanum antihypertensive peptide.

[0103] Example 2

[0104] The application discloses a preparation method of a low-bitterness Zanthoxylum bungeanum seed antioxidant peptide freeze-drying flash release tablet.

[0105] The Zanthoxylum bungeanum seed antioxidant peptide is 0.25 parts, mannitol is 10 parts, gelatin is 4 parts, and the volume ratio of the hydrolysis liquid during debittering to n-butanol is 1:2.5.

[0106] The preparation method of the low-bitterness Zanthoxylum bungeanum seed antioxidant peptide freeze-drying flash release tablet is specifically as follows.

[0107] Firstly, the gelatin is soaked in cold water for 10 minutes, then stirred in a 60 DEG C water bath pot until completely dissolved, then the Zanthoxylum bungeanum seed antioxidant peptide and mannitol are dissolved in the gelatin solution, uniformly mixed, and then diluted with pure water to 100ml, emulsified at a speed of 10000rpm for 10 minutes; finally, the emulsified solution is injected into a mold, pre-frozen in a-80 DEG C refrigerator for 4 hours, and then transferred into a vacuum freeze-drying machine for freeze-drying for 24 hours, so as to obtain the Zanthoxylum bungeanum seed antioxidant peptide freeze-drying flash release tablet.

[0108] Example 3

[0109] The preparation method of the low-bitterness Zanthoxylum bungeanum seed antioxidant peptide freeze-drying flash release tablet is the same as that in Example 2, except that the proportion of each raw material is as follows: the Zanthoxylum bungeanum seed antioxidant peptide is 0.25 parts, the mannitol is 12 parts, the gelatin is 6 parts, and the volume ratio of the hydrolysis liquid during debittering to n-butanol is 1:2.5.

[0110] Example 4

[0111] The preparation method of the low-bitterness Zanthoxylum bungeanum seed antioxidant peptide freeze-drying flash release tablet is the same as that in Example 2, except that the proportion of each raw material is as follows:

[0112] The Zanthoxylum bungeanum seed antioxidant peptide is 0.5 parts, the mannitol is 14 parts, the gelatin is 4 parts, and the volume ratio of the hydrolysis liquid during debittering to n-butanol is 1:1.

[0113] Example 5

[0114] The preparation method of the low-bitterness Zanthoxylum bungeanum seed antioxidant peptide freeze-drying flash release tablet is the same as that in Example 2, except that the proportion of each raw material is as follows:

[0115] The Zanthoxylum bungeanum seed antioxidant peptide is 0.35 parts, the mannitol is 15 parts, the gelatin is 5 parts, and the volume ratio of the hydrolysis liquid during debittering to n-butanol is 1:4.

[0116] Example 6

[0117] A method for preparing a low-bitterness Zanthoxylum bungeanum seed antioxidant peptide freeze-dried flash-release tablet, the ratio of each raw material and the preparation method are the same as in Example 5, except that in the last step, after the emulsified solution is injected into a mold, it is pre-frozen for 4 hours in a -20°C refrigerator, and then transferred into a vacuum freeze dryer for freeze-drying for 24 hours, to obtain the Zanthoxylum bungeanum seed antioxidant peptide freeze-dried flash-release tablet.

[0118] Example 7

[0119] A method for preparing a low-bitterness Zanthoxylum bungeanum seed antioxidant peptide freeze-dried flash-release tablet is the same as in Example 2, except that the ratio of each raw material is as follows:

[0120] Zanthoxylum bungeanum seed antioxidant peptide 0.3 parts, mannitol 15 parts, gelatin 5 parts, and no n-butanol is added for debittering.

[0121] Example 8

[0122] A method for preparing a low-bitterness Zanthoxylum bungeanum seed antihypertensive peptide freeze-dried flash-release tablet is prepared according to the following weight component ratio:

[0123] Zanthoxylum bungeanum seed antihypertensive peptide 0.2 parts, mannitol 10 parts, gelatin 4 parts, and the volume ratio of hydrolysis liquid to n-butanol is 1:1 during debittering.

[0124] The method for preparing the low-bitterness Zanthoxylum bungeanum seed antihypertensive peptide freeze-dried flash-release tablet is the same as in Example 2.

[0125] Example 9

[0126] A method for preparing a low-bitterness Zanthoxylum bungeanum seed antihypertensive peptide freeze-dried flash-release tablet is the same as in Example 8, except that the weight fractions of each component are as follows:

[0127] Zanthoxylum bungeanum seed antihypertensive peptide 0.25 parts, mannitol 12 parts, gelatin 6 parts, and the volume ratio of hydrolysis liquid to n-butanol is 1:2.5 during debittering.

[0128] Example 10

[0129] A method for preparing a low-bitterness Zanthoxylum bungeanum seed antihypertensive peptide freeze-dried flash-release tablet is the same as in Example 8, except that the weight fractions of each component are as follows:

[0130] Zanthoxylum bungeanum seed antihypertensive peptide 0.25 parts, mannitol 15 parts, gelatin 5 parts, and the volume ratio of hydrolysis liquid to n-butanol is 1:3 during debittering.

[0131] Example 11

[0132] A method for preparing a low-bitterness Zanthoxylum bungeanum seed antihypertensive peptide freeze-dried flash-release tablet is the same as in Example 8, except that the weight fractions of each component are as follows:

[0133] Zanthoxylum bungeanum seed antihypertensive peptide 0.3 parts, mannitol 15 parts, gelatin 5 parts, volume ratio of butanol to water in the debittering solution: 1:4.

[0134] Example 12

[0135] A method for preparing a low-bitterness Zanthoxylum bungeanum seed antihypertensive peptide freeze-dried flash-release tablet is the same as in Example 11, except that in the final step, the emulsified solution is injected into a mold, pre-frozen for 4 h in a -20℃ refrigerator, and then transferred to a vacuum freeze-drying machine for freeze-drying for 24 h.

[0136] Example 13

[0137] A method for preparing a low-bitterness Zanthoxylum bungeanum seed antihypertensive peptide freeze-dried flash-release tablet is the same as in Example 8, except that the weight fractions of the components are as follows:

[0138] Zanthoxylum bungeanum seed antihypertensive peptide 0.2 parts, mannitol 15 parts, gelatin 5 parts, no butanol added for debittering.

[0139] The ratios of the raw materials in Examples 2-7 are shown in Table 10 below, and the ratios of the raw materials in Examples 8-13 are shown in Table 11 below.

[0140] Table 10 Ratios of raw materials in Examples 2-7

[0141] Example Zanthoxylum bungeanum seed antioxidant peptide Mannitol Gelatin Hydrolyzate to n-butanol ratio Freeze-drying temperature (°C) Example 2 0.25 10 4 1:2.5 -80 Example 3 0.25 12 6 1:2.5 -80 Example 4 0.5 14 4 1:1 -80 Example 5 0.35 15 5 1:4 -80 Example 6 0.35 15 5 1:4 -20 Example 7 0.3 15 5 1:0 -80

[0142] Table 11 Ratios of raw materials in Examples 8-13

[0143]

[0144]

[0145] Performance evaluation of Example 14

[0146] The Zanthoxylum bungeanum seed antioxidant peptide and Zanthoxylum bungeanum seed antihypertensive peptide freeze-dried flash-release tablets in Examples 2-13 were evaluated using a scoring system, and the evaluation score table is shown in Table 12 below.

[0147] Table 12 Evaluation indicators for Zanthoxylum bungeanum seed peptide freeze-dried flash-release tablets

[0148]

[0149] The sensory evaluation score results for the Zanthoxylum bungeanum seed antioxidant peptide freeze-dried flash-release tablets prepared in Examples 2-7 are shown in Table 13.

[0150] Table 13 Sensory evaluation score table for Zanthoxylum bungeanum seed antioxidant peptide freeze-dried flash-release tablets

[0151] Evaluation index Example 2 Example 3 Example 4 Example 5 Example 6 Example 7 Form (points) 3 5 5 5 1 5 Structure (points) 3 5 5 5 1 5 Dissolution speed (points) 5 5 5 5 3 5 Taste (points) 5 1 5 5 5 1 Flavor (points) 3 5 2 5 5 1 Total score (points) 19 21 22 25 15 17

[0152] As can be seen from the table:

[0153] The total score of the freeze-dried flash release tablet of Zanthoxylum bungeanum seed antioxidant peptide prepared by using the raw material formula of Example 5 is the highest, the best ratio is 0.35 parts of Zanthoxylum bungeanum seed antioxidant peptide, 15 parts of mannitol and 5 parts of gelatin, and the volume ratio of the hydrolysate to n-butanol during debittering is 1:4, the freeze-dried flash release tablet prepared has not only good surface morphology, but also neat cross section after being broken, fast dissolution speed, melting in mouth, no bitterness and strong palatability.

[0154] Compared with Example 5, the amount of Zanthoxylum bungeanum seed antioxidant peptide is reduced in Example 2, and the amount of mannitol and gelatin and other auxiliary materials is also greatly reduced. From the score, the appearance of the freeze-dried flash release tablet of Zanthoxylum bungeanum seed antioxidant peptide is slightly worse, there is a little grain feeling in mouth, and the product may have a little bitterness due to incomplete debittering because of the reduced amount of n-butanol during debittering.

[0155] In Example 3, the amount of mannitol and gelatin is adjusted on the basis of Example 2, 12 parts of mannitol and 6 parts of gelatin are added, but the content of auxiliary materials is slightly less than that of Example 5. Compared with Example 2, the structure and morphology of the freeze-dried flash release tablet of Zanthoxylum bungeanum seed antioxidant peptide are improved, and the taste of the product is slightly improved. It can be seen that the appropriate increase of the amount of gelatin and mannitol helps to disperse or embed the peculiar smell.

[0156] In Example 4, compared with Example 5, the amount of antioxidant peptide is increased, but the amount of auxiliary materials is slightly lower than that of Example 5. The most important thing is that the volume ratio of the hydrolysate to n-butanol during debittering is 1:1. Because Zanthoxylum bungeanum seed antioxidant peptide itself has special Zanthoxylum bungeanum smell and bitterness, and the amount of n-butanol is low, the freeze-dried flash release tablet of Zanthoxylum bungeanum seed antioxidant peptide prepared in Example 4 has heavy bitterness and peculiar smell, and the taste of the product is poor, but the overall morphology of the product is good.

[0157] In Example 6, the mixed solution of the flash release tablet of Zanthoxylum bungeanum seed antioxidant peptide is pre-frozen at-20℃, the tablet has poor morphology, rough surface, cracks and chipping, soft texture and slow dissolution speed.

[0158] In Example 7, n-butanol is not used for debittering of Zanthoxylum bungeanum seed antioxidant peptide, and the freeze-dried flash release tablet of Zanthoxylum bungeanum seed antioxidant peptide prepared has obvious bitterness, poor palatability and is difficult to take.

[0159] The sensory evaluation results of the freeze-dried flash release tablets of Zanthoxylum bungeanum seed antihypertensive peptide prepared in Examples 8-13 are shown in Table 14.

[0160] Table 14 Sensory evaluation score table of freeze-dried flash release tablets of Zanthoxylum bungeanum seed antihypertensive peptide

[0161] Evaluation index Example 8 Example 9 Example 10 Example 11 Example 12 Example 13 Form (points) 5 5 5 5 1 5 Structure (points) 3 5 5 5 1 5 Dissolution speed (points) 5 3 5 5 3 5 Taste (points) 5 5 5 5 5 1 Flavor (points) 2 3 5 3 5 1 Total score (points) 22 21 25 23 15 17

[0162] The above results can be seen: the use of raw materials ratio of example 10 obtained from the pepper seed antihypertensive peptide freeze-dried flash release tablet score the highest, wherein the pepper seed antihypertensive peptide 0.25 parts, mannitol 15 parts, gelatin 5 parts, pepper seed antihypertensive peptide debittering when the volume ratio of hydrolysate and n-butanol is 1:3, can be seen, the prepared product whether in the form, or in the taste, all show good.

[0163] And in example 13, the pepper seed antihypertensive peptide is not used in the preparation of n-butanol debittering, the final product prepared bitter, difficult to enter.

[0164] In addition, the amount of auxiliary material in examples 8-9 is adjusted respectively, the appearance of the prepared product is slightly lower than that of the product of example 10.

[0165] From the above test results, the auxiliary material mannitol, gelatin in the pepper seed peptide freeze-dried flash release tablet has a certain dispersion and embedding effect on the bitter taste of the product, relatively more auxiliary materials are beneficial to improve the overall form of the product, and the product has appropriate viscosity and moderate dissolution rate, while the content of auxiliary material is too low, which may lead to poor adhesion of the product, and thus cause poor fineness of the product, rough surface and even easy to fall off, causing the product to have a sand feeling when eating; In addition, the amount of n-butanol added in the debittering of pepper seed peptide plays a key role in eliminating the bitter and pungent taste of the product. If the content of n-butanol is too low, the prepared pepper seed peptide freeze-dried flash release tablet has obvious bitter taste and poor palatability, while the content of n-butanol is higher, since the debittering of pepper seed peptide is more thorough, thus the prepared product hardly feels any bitter taste or pungent taste of pepper seed.

Claims

1. A low-bitterness Zanthoxylum bungeanum seed peptide freeze-dried flash-release tablet, characterized in that, The composition comprises the following components in parts by weight: 0.25-0.35 parts of Zanthoxylum bungeanum seed peptide, 15 parts of mannitol, and 5 parts of gelatin; the Zanthoxylum bungeanum seed peptide is at least one of Zanthoxylum bungeanum seed antioxidant peptide and Zanthoxylum bungeanum seed antihypertensive peptide; The freeze-drying condition for preparing the freeze-dried flash release tablet is: pre-freezing for 4 hours in a -80°C refrigerator, then transferring into a vacuum freeze dryer, and freeze-drying for 24 hours. The Zanthoxylum bungeanum seed peptide is prepared by the following method: (1) Zanthoxylum bungeanum seed pretreatment: removing impurities from dried Zanthoxylum bungeanum seeds, crushing with a flour mill, sieving, and reserving; (2) Extraction of Zanthoxylum bungeanum seed protein: dissolving the Zanthoxylum bungeanum seed powder in water to prepare a solution, the mass-volume ratio of the Zanthoxylum bungeanum seed powder to water being 1 g:20-30 mL, adjusting the pH of the solution to 8.0-10.0, water bath heating for 3-6 hours, centrifugation, retaining the supernatant; then adjusting the pH of the supernatant to 3.0-5.0, centrifugation, discarding the supernatant, retaining the precipitate, washing the precipitate to neutral, obtaining a Zanthoxylum bungeanum seed protein solution, freezing and standing, and drying; (3) Preparation of Zanthoxylum bungeanum seed peptide: preparing the Zanthoxylum bungeanum seed protein obtained in (2) into a solution, and performing enzymolysis with a protease; before adding the protease, the pH of the solution is adjusted to the optimal reaction pH of the protease; during the enzymolysis, the pH of the solution is maintained within the optimal reaction pH±0.05 range; after the enzymolysis is completed, boiling water bath is performed for 10-20 minutes to inactivate the enzyme, and the solution is cooled to obtain the Zanthoxylum bungeanum seed peptide; When the Zanthoxylum bungeanum seed peptide is Zanthoxylum bungeanum seed antioxidant peptide, the protease is alkaline protease; when the Zanthoxylum bungeanum seed peptide is Zanthoxylum bungeanum seed antihypertensive peptide, the protease is pepsin; (4) Debittering: mixing the Zanthoxylum bungeanum seed peptide solution obtained in (3) with n-butanol to extract bitter peptides therefrom, collecting the hydrolysate layer to obtain a low-bitter Zanthoxylum bungeanum seed peptide solution, freeze-drying, and then freezing at -20°C for preservation, wherein the volume ratio of the hydrolysate to n-butanol is 1:

4.

2. The low-bitterness Zanthoxylum bungeanum seed peptide freeze-dried flash-release tablet of claim 1, characterized in that, In the hydrolysis of the Zanthoxylum bungeanum seed protein in (3), the concentration of the protein solution is 8-15 mg / mL, the addition amount of the protease is 4000-8000 U / g of protein, and the hydrolysis time is 3-6 hours.

3. The method for preparing the low-bitterness freeze-dried flash-release tablets of Zanthoxylum bungeanum seed peptide according to claim 1, characterized in that: The method comprises the following steps: The Zanthoxylum bungeanum seed peptide and mannitol are dissolved in a pre-dissolved gelatin solution, the mixed solution is uniformly mixed, then water is added to the solution to make the volume constant, and then the solution is subjected to injection molding and freeze-drying to obtain the Zanthoxylum bungeanum seed peptide freeze-dried flash release tablet.

4. The method for preparing the low-bitterness freeze-dried flash-release tablets of Zanthoxylum bungeanum seed peptide according to claim 3, characterized in that: The gelatin is dissolved by the following method: soaking in cold water for 6-15 minutes, and then stirring in a 40-80°C water bath until completely dissolved.

5. The method for preparing the low-bitterness freeze-dried flash-release tablets of Zanthoxylum bungeanum seed peptide according to claim 3, characterized in that: After the volume is made constant, emulsification is performed before injection molding, and the rotation speed of the emulsifier is 8000-12000 rpm, and the emulsification time is 8-20 minutes.

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