CN111317126A

CN111317126A - Papaya enzyme composition for recovering and maintaining digestive function, preparation method and composite nano preparation

Info

Publication number: CN111317126A
Application number: CN201811531043.7A
Authority: CN
Inventors: 解冰
Original assignee: Individual
Current assignee: Individual
Priority date: 2018-12-14
Filing date: 2018-12-14
Publication date: 2020-06-23

Abstract

The invention relates to a papaya enzyme composition for recovering and maintaining digestive function, a preparation method and a composite nano preparation in the field of medical health-care food. The papaya ferment composition for recovering and maintaining digestive function comprises the following components in percentage by weight: main materials: 0.5-5% of papain and 0.01-0.1% of nano tungsten ions; auxiliary materials: 30-60% of glucose, 35-60% of cane sugar, 0.5-3.5% of plant stearic acid, 0.01-0.1% of spice, 0.01-0.1% of edible pigment and 0.1-3% of magnesium silicate. The papaya enzyme composition can achieve the best digestion aiding effect, is prepared from natural muskmelon fruits, has good taste, has the effects of regulating blood fat, reducing blood pressure and the like, and does not have any adverse effect on a human body.

Description

Papaya enzyme composition for recovering and maintaining digestive function, preparation method and composite nano preparation

Technical Field

The invention relates to the field of medical health-care food, in particular to a papaya ferment composition for recovering and maintaining digestive function, a preparation method and a composite nano preparation.

Background

With the continuous development of science and technology and the continuous progress of human society, the dietary structure of people is changed, high-energy foods such as high protein and high fat are excessively taken, or the diet is extremely irregular, so that the metabolic disorder of human bodies and the secretion disorder of various enzymes in the bodies are caused, light people show the characteristics of halitosis, bad smell, fart and the like, and serious people give off a peculiar smell to the whole body. If the patient does not need to promote digestion, improve digestion and eliminate risks in time, various diseases are easily caused.

People usually choose chewing gum to relieve dyspepsia such as halitosis and flatus. However, it has been found that an important factor causing human dyspepsia is that due to the absence of the corresponding enzymes secreted by the human body, macromolecular substances ingested by the human body, such as proteins, starch polysaccharides, fats, etc., cannot be digested by the human body within an effective period of time, and thus are accumulated in the digestive system of the human body (mainly concentrated in the intestinal tract), causing the food in the human body to be digested heterogeneously, and generating odor which is then discharged through the oral cavity and the intestinal tract. The traditional chewing gum only covers the odor through the faint scent of the mint on the local oral cavity, and cannot eliminate the pathology forming the halitosis. Meanwhile, the chemical coating formed at the end of chewing the gum is difficult to degrade, which causes environmental pollution.

However, the method of treating with chemical drugs, yeast and Chinese herbal medicines is generally and mainly adopted in medicine, but adverse reactions such as abdominal distension and nausea generally exist, and the method cannot well function. The medicine is three-way toxic, long-term administration not only causes irritation reaction to gastric and intestinal mucosa and has toxic and side effects of different degrees, but also the antibacterial medicine can kill germs and destroy lactobacillus for maintaining the functions of the gastric and intestinal tracts, further damage is caused to a digestion system and an absorption system, and great harm is caused to human bodies.

Papaya is a tropical fruit abundant in south China, is rich in precious digestive elements, has a strong biocatalysis function because active protease is a main component for softening meat, can strengthen the catalytic functions of other enzymes, and starts to act on food before the food is delivered to the stomach; meanwhile, it can also regulate the endocrine function of the body and the like. The papain contained in the composition has high content, excellent stability and strong protease hydrolysis capability, and has good degradation effect on various proteins. The inventor researches the current dietary structure of Chinese people, the food composition with digestive difficulties ingested by human bodies, the mimicry digestive system environment (various enzyme components and the overall function under different proportions), and the digestive elements and functions of papaya enzyme from 2000 years; the effect of papaya proteinase on the function of other enzymes; product dosage form versus activity (function); the product tastes, and the like, and the digestive function is restored and improved by supplementing and improving the activity of the digestive enzyme of the human body, so that the internal balance is improved, the digestive capability is enhanced, and the immune function of the human body is adjusted, and the effect is obvious.

Disclosure of Invention

In order to prevent excessive intake of high-protein and high-fat foods from being a burden of absorption, particularly prevent stimulation and support of certain digestive components from causing excessive growth of certain bacteria and 'expelling' beneficial intestinal bacteria during diseases to cause imbalance of intestinal environment, the invention analyzes and proves that one form of tungsten (nano tungsten ions) can selectively prevent enterobacteriaceae from 'excessive reproduction' in the intestinal tract in an absorption system on the basis of solving the problems of the digestive system in the prior period, and beneficial bacteria nearby are not affected, so that the invention has positive and important significance for recovering and improving self-digestion function and avoiding damaging the intestinal absorption system.

In order to solve the above problems in the prior art, the present invention provides a papain enzyme composition for restoring and maintaining digestive functions. In particular to a papaya ferment composition for recovering and maintaining digestive function, a preparation method and a composite nano preparation.

One of the objectives of the present invention is to provide a papaya enzyme composition for recovering and maintaining digestive function, which comprises the following components by weight percent:

main materials:

0.5-5% of papain, preferably 1-2.5%;

0.01-0.1% of nano tungsten ions, preferably 0.02-0.05%;

auxiliary materials:

wherein,

the food pigment is selected from FD & yellow no6 lake;

the papain is 3000-8000 USP units/mg, preferably 5500-6500 USP units/mg. The papain is a proteolytic enzyme separated from papaya fruits, is the most common component of a plurality of digestive enzyme nutriments, is used for hydrolyzing proteins and softening meat, can activate the secretion of endogenous enzymes, supplements the deficiency of the endogenous enzymes, adjusts the endogenous enzymes, ensures the normal digestive absorption function, prevents diseases and promotes growth; can also eliminate anti-nutritional factors and promote the healthy growth of organisms; also has the functions of maintaining the complete shape of the villi of the small intestine and promoting the absorption of nutrient substances.

The nano tungsten ions are 2-10 USP units/mg, and preferably 3-8 USP units/mg.

The invention also aims to provide a preparation method of the papaya enzyme composition for recovering and maintaining digestive function, which specifically comprises the following steps:

preparing papain into papain nanocrystals; adding nanometer tungsten ion, binder, spice, edible pigment, magnesium silicate, and plant stearic acid, and making into tablet to obtain composite nanometer preparation.

The papain nanocrystal is prepared by a method comprising the following steps:

a medium grinding method: taking a proper amount of surfactant SDS with the weight concentration of 2-5% in a grinding tank, adding distilled water, performing ultrasonic treatment for 2-10 min, weighing the raw material papain, placing the papain in the grinding tank, performing ultrasonic treatment, fully dispersing the medicine, and adding 10-30 g of zirconium oxide (ZrO) with the diameter of 0.2-0.8 mm₂) Grinding the beads; installing a machine, adjusting the rotating speed of the machine to 200-600 r/min, operating the ball mill, stopping the machine for 15-40 min every 0.5-2 h, grinding for 6-9 h, stopping the machine, standing, and freeze-drying the obtained suspension for 2-12 h to obtain the papain nanocrystal;

the nano tungsten ions are prepared by the method comprising the following steps:

the method for preparing the nano tungstate by a direct precipitation-solvothermal method comprises the following steps:

mixing Na₂WO₄·2H₂O and Zn (NO)₃)₂·6H₂Dissolving O in distilled water respectively to form colorless solutions; under strong stirring, adding Na₂WO₄The solution was slowly added dropwise to Zn (NO)₃)₂Solutions ofForming white precipitate, adjusting the pH value to 6-8 by using an lmol/LNAOH solution, stirring for 0.5-2 h, transferring the white precipitate into a stainless steel reaction kettle with a Teflon liner, reacting for 20-60 h at 80-140 ℃, naturally cooling to room temperature, centrifugally separating, washing for several times by using ethylene glycol, and vacuum-drying for 4-8 h at 40-80 ℃ to obtain off-white particles, namely the nano tungsten ions;

wherein, the raw material Na₂WO₄·2H₂O and Zn (NO)₃)₂·6H₂The molar ratio of O is: (2-4) to (0.5-2).

The tablet is prepared by a method comprising the following steps:

sieving the papain nanocrystal with a 20-mesh sieve; preparing a binder from glucose and sucrose according to the using amount; measuring papain nanocrystal and nano tungsten ions according to the dosage, and adding an adhesive to prepare a soft material; granulating with a 16-mesh sieve, and drying at 40-60 ℃ until the water content is less than 3%; finishing the grains with a 14-mesh sieve; dissolving the perfume and the edible pigment in ethanol, and uniformly spraying the solution on the dried particles; adding magnesium silicate and plant stearic acid as lubricant; mixing, and tabletting. The tableting step may include: tabletting by using a punch with the diameter of 8 mm; the hardness of the tablet is controlled to be 1-4 kg^-2About 20-100 mg in tablet weight.

The third purpose of the invention is to provide a composite nano preparation prepared by the preparation method of the papaya ferment composition for recovering and maintaining digestive function; the composite nano-formulation may include various types including tablets such as chewable tablets.

The inventor researches the dietary structure of Chinese, the food composition of digestive difficulties ingested by human body, the synergistic function of various enzymes under different proportions, the digestive elements and functions of papaya fruits, the influence of papaya fruit protease on other enzyme functions, the relation between product dosage form and activity (function), the taste of products and other conditions, and prepares the papaya enzyme composition, which can effectively solve the problems of dyspepsia and difficult absorption without generating any side effect.

The papaya ferment composition can be used as a compound functional food. The invention relates to a formula product designed aiming at the dietary structure and consumption habits of Chinese (Asian) people, which takes active protease extracted from natural plant papaya fruits as a main body, and the papaya fruit protease can be used as a substrate of other enzymes, namely a catalyst, besides the self digestion function, so that the digestion function of the papaya fruit protease in the digestive tract can be efficiently and smoothly carried out. The papaya enzyme composition can achieve the best digestion promoting effect, has the effects of regulating blood fat, reducing blood pressure and the like, and has no adverse effect on a human body. The papaya ferment composition is prepared from natural muskmelon fruits, has good taste, and can digest the starch which is 2230 times of the weight of the papaya ferment composition.

The proportion of enzyme components of the product is designed according to various dietary structure characteristics of Chinese people, and particularly, the product aims at improving the dietary digestion problem of people with increasingly degraded digestive systems after the age of 40.

The papaya ferment composition can be used as a compound functional food, and has another effect of decomposing a settled layer on a blood vessel wall into particles and quickly discharging the particles out of a body; increase the permeability of sebocytes and tissues, which is beneficial for reducing inflammation and absorbing cellular debris; and effectively reduce atherosclerosis. The protein eaten by the human body can be converted into amino acid to the maximum extent, the secretion function of the organism is regulated, the immunity of the human body is enhanced, harmful free radicals in the organism are eliminated, the blood circulation is promoted, the blood stasis is removed, and the platelet aggregation is prevented; promoting metabolism and hormone secretion.

Detailed Description

The present invention will be further described with reference to the following examples. However, the present invention is not limited to these examples.

Example 1

Main materials:

2% of papain;

0.04% of nano tungsten ions;

auxiliary materials:

45% of glucose;

50% of sucrose;

2% of plant stearic acid;

0.03% of spice;

0.03% of edible pigment;

0.9 percent of magnesium silicate.

The preparation method comprises the following steps:

1) preparing main material papain nanocrystal:

a medium grinding method: preparing 5 wt% of surfactant SDS, putting the surfactant SDS into a 100mL grinding tank, adding 15mL of distilled water, and performing ultrasonic treatment for 2 min; weighing raw material medicine papain, placing in the grinding tank, performing ultrasonic treatment for several minutes, adding 30g of zirconium oxide (ZrO) with diameter of 0.8mm after the medicine is fully dispersed₂) Grinding the beads; installing a machine, adjusting the rotating speed of the machine to 600r/min, operating the ball mill, stopping the machine for 15min every 2h, grinding for 6h, stopping the machine, standing, and freeze-drying the obtained suspension for 8h to obtain the papain nanocrystal;

2) preparing nano tungsten ions: the method for preparing the nano tungstate by a direct precipitation-solvothermal method comprises the following steps: 2mmol of Na₂WO₄·2H₂O and 2mmol Zn (NO)₃)₂·6H₂O is respectively dissolved in 20mL of distilled water to form colorless solutions; under strong stirring, adding Na₂WO₄The solution was slowly added dropwise to Zn (NO)₃)₂And (3) forming white precipitate in the solution, adjusting the pH value to 8 by using 1mol/LNAOH solution, stirring for 0.5h, transferring the white precipitate into a stainless steel reaction kettle with a Teflon liner, reacting for 48h at 140 ℃, naturally cooling to room temperature, performing centrifugal separation, washing for several times by using ethylene glycol, and performing vacuum drying for 5h at 60 ℃ to obtain grey particles, namely the nano tungsten ions.

3) Preparing chewable tablet:

sieving papain nanocrystal with 20 mesh sieve; preparing a binder from glucose and sucrose according to the using amount; measuring papain nanocrystal and nano tungsten ions according to the dosage, and adding an adhesive to prepare a soft material; granulating with 16 mesh sieve, and drying at 50 deg.C; finishing the grains with a 14-mesh sieve; dissolving the perfume and the edible pigment in ethanol, and uniformly spraying the solution on the dried particles; adding magnesium silicate and plant stearic acid as lubricant; mixing, and tabletting to obtain nanometer tablet.

And (3) performance testing:

1) and (3) observing the shape and the particle size of the main material nanocrystal: the particle size and particle size distribution of the papain nanocrystals as a bulk material were measured using a dynamic light scattering nanometer particle size analyzer (PSSNicomp 380, usa) (table 1).

TABLE 1 nanocrystal particle size distribution

Name of a category	D10	D50	D90
				Nanocrystals	20.15	46.32	81.54

The test result shows that the prepared nano crystal has the grain diameter of about 100nm and is in a rod-shaped or needle-shaped crystal structure; the product performance and stability of the nano preparation are obviously improved while the specification is reduced.

2) And (3) determining the encapsulation efficiency and the drug loading capacity:

weighing a certain amount of the nano tablet, dissolving and diluting the nano tablet by using PBS buffer solution with pH of 6.8 to obtain external liquid, weighing the certain amount of PBS buffer solution, placing the PBS buffer solution in a dialysis bag soaked in the PBS buffer solution for 24 hours, dialyzing the solution for 8 hours under the condition of stirring at constant temperature of 4 ℃ for 1 time at intervals of 30min, measuring the absorbance of the obtained internal liquid, converting the concentration of the papain in the internal liquid corresponding to the absorbance according to a standard curve of the papain, calculating the amount of unencapsulated papain by knowing the volume of the internal liquid, and substituting the calculated amount into the following formula to obtain the encapsulation rate of a sample, wherein the encapsulation rate is × 100 percent (the total amount of the injected medicine-the mass of the unencapsulated medicine)/the total amount of the injected medicine).

The results show that the average encapsulation rate of the main component in the nano-tablets of the invention is higher than 90%, and the average drug loading is about 2% (see table 2 below). The indexes include illumination, temperature and the like, and the result is stable and is good for repeated use.

TABLE 2 mean entrapment efficiency and drug loading of the major component in the Nanoprostoles

Composition (I)	Encapsulation efficiency	Drug loading
			Papain	90.8％	2％

3) Dissolution test and enzyme activity determination

The in vitro dissolution rate of the nano tablet is determined by referring to the dissolution rate determination method of Chinese pharmacopoeia.

Papain can hydrolyze ethyl N-benzoyl-L-arginine (substrate) to release N-benzoyl-L-arginine, and the release amount can be titrated with sodium hydroxide solution to determine enzyme activity. The amount of enzyme capable of catalytically decomposing 1umol of substrate per minute at 25 ℃ is referred to as 1 unit. The activity P (units/mg) of papain was calculated as follows: p is C.DELTA.V.1000/W. In the formula: c: concentration of sodium hydroxide solution, mol/L; Δ V: the volume of sodium hydroxide solution consumed per minute minus the blank value; w: mass of papain in the reaction solution, mg. The results of measuring the activity of released protease according to the national standard assay (SB/T10317-1999 protease activity assay) are shown in Table 3 below.

TABLE 3

Sample (I)	Mass g	Dilution factor	Average absorbance	Diluent enzyme activity ug/mL	Average enzyme activity u/g
						Sample (I)	1.001	250	0.589	81.9	8102

The result shows that the dissolution rate of the nano tablet in dissolution media such as hydrochloric acid solution with pH 1.2 and phosphate buffer solution with pH 6.9 can reach more than 80% at 40min, and the nano tablet shows better solubility and dissolution rate; and the repeatability of the prescription process is good.

4) Effect of digestive enzyme composite nano preparation on intestinal function of rat

Constructing a rat liver injury model and a severe model.

100 to 150g Wistar rats 100 rats were divided into 50 rats in the normal group and 50 rats in the liver injury group. A rat liver injury model is constructed by using carbon tetrachloride, and particularly, 2ml/kg of carbon tetrachloride is injected into a rat intraperitoneally for 9 weeks at 2 weeks, so that cirrhosis accompanied with stem cell necrosis and obvious inflammation can be constructed.

The normal group and the liver injury group are divided into 5 groups, each group comprises 10 mice, each half of the mice are male and female, and the mice are randomly divided into a single medicine-papain group, a common tablet group (the common tablet refers to the main component distribution form produced by the traditional process on the preparation method according to the formula, namely mixing raw materials and auxiliary materials → preparing soft materials → granulating → drying → tabletting), a nano tablet group, a common rat food group and a blank control group. Feeding continuously for 4 weeks.

The specific feeding dosage method comprises the following steps: the feeding amount of the rats is supplied according to a rat feed formula (wherein the used common rat food is Schuckbebata SPF high-reproduction-rate rat food and Jiangsu synbiotics), and the dosage of the added drugs is calculated by the following method: 100-150g Wistar rat, tablet 1g, according to the average weight of 60kg per person, the administration amount of human is 1000mg/60kg, the administration amount of rat is 1000mg/60 kg/100 g, the daily food intake of rat is 30g, therefore the content of drug in the feed is 1000mg/60 kg/100 g/30. The amount of other single-component drugs is calculated according to the method and added into the rat food.

Weighing the weight of the rat every day, and measuring the effect of the test substance on the growth of the rat; rat feces are observed every day, feces samples are collected, the quantity of feces dominant flora and the change condition of short-chain fatty acid are detected, and the detection method refers to a method disclosed in Chinese patent with publication number CN101368154A (application number 200810048065.8).

The results show that the nano-tablet group can obviously improve the appetite of rats, increase the food consumption and promote the weight increase of rats compared with other groups. The results are given in Table 4 below. Compared with single-medicine preparation and common tablets, the nano-preparation group can obviously improve the digestion level of animals and promote the growth rate of the animals.

TABLE 4 Effect of Nanodiesents on growth force

Healthy rat group	Number of animals	Average weight gain%
			Blank control group	10	20
Common rat grain group-enzyme-free group	10	31
			Rat grain + papain group	10	38
Mouse grain and common tablet group	10	49
			Mouse grain and nano tablet group	10	58

The influence of the nano-tablet group on the flora in the digestive tract shows that the nano-tablet group can obviously improve the amount of beneficial flora (such as bifidobacterium) of rats with liver injury and maintain the intestinal health of the rats compared with other groups (see the following table 5).

TABLE 5

The influence of the nano-tablet group on the Short Chain Fatty Acid (SCFA) in the digestive tract was found to significantly improve the recovery of the metabolic balance of the short chain fatty acid (e.g., acetic acid) in the intestinal tract compared to the other groups (see table 6 below).

TABLE 6

Claims

1. A papaya ferment composition for recovering and maintaining digestive function comprises the following components in percentage by weight:

main materials:

0.5-5% of papain;

0.01-0.1% of nano tungsten ions;

auxiliary materials:

2. the papaya ferment composition for recovering and maintaining digestive function according to claim 1, characterized by comprising the following components in weight percentage:

main materials:

1-2.5% of papain;

0.02-0.05% of nano tungsten ions;

auxiliary materials:

3. the papaya ferment composition for restoring and maintaining digestive function according to claim 1 or 2, characterized in that:

the edible pigment is FD & yellow no6 lake;

the papain is 3000-8000 USP units/mg, preferably 5500-6500 USP units/mg;

the nano tungsten ions are 2-10 USP units/mg, and preferably 3-8 USP units/mg.

4. The method for preparing the papain composition for recovering and preserving digestive functions according to any one of claims 1 to 3, characterized by comprising the following steps:

preparing papain into papain nanocrystals; adding nanometer tungsten ion, binder, spice, edible pigment, magnesium silicate, and plant stearic acid, and making into tablet.

5. The method of preparing the papaya ferment composition for recovering and maintaining digestive function according to claim 4, wherein the papaya ferment composition comprises:

the papain nanocrystal is prepared by a method comprising the following steps:

a medium grinding method: placing a surfactant SDS with the weight concentration of 2-5% in a grinding tank, adding distilled water, and performing ultrasonic treatment; weighing raw material medicine papain, placing the papain in the grinding tank for ultrasonic treatment, and adding zirconia grinding beads after the medicine is fully dispersed; installing a machine, operating the ball mill, stopping the machine for 15-40 min every 0.5-2 h, grinding for 6-9 h, stopping the machine, standing, and freeze-drying the obtained suspension to obtain the papain nanocrystal.

6. The method of preparing the papaya ferment composition for recovering and maintaining digestive function according to claim 4, wherein the papaya ferment composition comprises:

mixing Na₂WO₄·2H₂O and Zn (NO)₃)₂·6H₂Dissolving O in distilled water respectively to form colorless solutions; under strong stirring, adding Na₂WO₄The solution was slowly added dropwise to Zn (NO)₃)₂Forming white precipitate in the solution, adjusting the pH value to 6-8, stirring, transferring the white precipitate to a stainless steel reaction kettle with a Teflon liner, reacting at 80-140 ℃, naturally cooling to room temperature, centrifugally separating, washing for several times, and vacuum drying at 40-80 ℃ to obtain off-white particles, namely the nano tungsten ions;

wherein, the Na₂WO₄·2H₂O and Zn (NO)₃)₂·6H₂The molar ratio of O is (2-4) to (0.5-2).

7. The method of preparing the papaya ferment composition for recovering and maintaining digestive function according to claim 4, wherein the papaya ferment composition comprises:

the tablet is prepared by a method comprising the following steps:

sieving the papain nanocrystal with a 20-mesh sieve; preparing a binder from glucose and sucrose according to the using amount; measuring papain nanocrystal and nano tungsten ions according to the dosage, and adding an adhesive to prepare a soft material; granulating with 16 mesh sieve, and drying; finishing the grains; dissolving the perfume and the edible pigment in ethanol, and uniformly spraying the solution on the particles; adding magnesium silicate and plant stearic acid as lubricant; mixing, and tabletting.

8. The method of preparing the papaya ferment composition for recovering and maintaining digestive function according to claim 7, wherein the papaya ferment composition comprises:

the drying is carried out at 40-60 ℃ until the moisture content is less than 3%.

9. The method of preparing the papaya ferment composition for recovering and maintaining digestive function according to claim 7, wherein the papaya ferment composition comprises:

the tabletting step comprises: tabletting by using a punch with the diameter of 8 mm; the hardness of the tablet is controlled to be 1-4 kg^-2The tablet weight is 20-100 mg.

10. The composite nano preparation prepared by the preparation method of the papain composition for recovering and maintaining digestive functions according to any one of claims 4 to 9.