CN114711426A - Preparation method of allium chinensis extract, inclusion compound and application thereof - Google Patents

Preparation method of allium chinensis extract, inclusion compound and application thereof Download PDF

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Publication number
CN114711426A
CN114711426A CN202210433868.5A CN202210433868A CN114711426A CN 114711426 A CN114711426 A CN 114711426A CN 202210433868 A CN202210433868 A CN 202210433868A CN 114711426 A CN114711426 A CN 114711426A
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allium
extract
allium chinense
chinense
preparation
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王艳
高鹏
陈谦
何江
徐攀
黄敏
伏毅
陈浩
吴舸洋
邱娅璐
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SICHUAN INSTITUTE OF ATOMIC ENERGY
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SICHUAN INSTITUTE OF ATOMIC ENERGY
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
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    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
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    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
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    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
    • A61K36/8962—Allium, e.g. garden onion, leek, garlic or chives
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
    • A61K47/6951—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes using cyclodextrin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73—Polysaccharides
    • A61K8/738—Cyclodextrins
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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    • A61K2800/524—Preservatives
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Abstract

The invention discloses a preparation method of an allium chinense extract, which comprises the following steps: a. cleaning fresh Allium chinense with root and bulb with clear water, removing impurities, and draining water at normal temperature; b. taking the drained allium chinense in the step a, and mixing the allium chinense with the feed liquid ratio of 1: 2-5, adding distilled water, and stirring and crushing by using a crusher to obtain an allium chinense homogenate; c. c, homogenizing the allium chinense in the step b, adding an acidic reagent to adjust the pH value to be 4.0-5.0, adding food-grade enzyme, and extracting for 60-180 min at a constant temperature of 40-50 ℃ under the stirring condition to obtain an extracting solution; d. c, filtering the extracting solution in the step c, centrifuging, and taking supernate; e. and d, treating the supernatant obtained in the step d by using a filter membrane, and performing vacuum freeze drying to obtain solid powder to obtain allium chinensis extract powder, thereby solving the problems that the allium chinensis and products thereof in the prior art are narrow in applicable range, complex in preparation process of extracting and processing of the allium chinensis and low in economic benefit.

Description

Preparation method of allium chinensis extract, inclusion compound and application thereof
Technical Field
The invention relates to the technical field of extraction methods and application of plant active ingredients, and particularly relates to preparation methods and application of allium chinense extract and a cyclodextrin inclusion compound thereof.
Background
Allium chinense (A) and (B)Allium chinenseG. Don), also called Xie, also called Jie Zi and Qiao Tou, belonging to perennial lilyThe plant of the family Allium has the characteristic of strong tillering capability, and more than 20-30 bulbs can be generated after one bulb is planted. The allium chinense has already been planted in China for 3700 years, the planting technology is simple, the field management is easy, the requirement on the land is not high, and the yield per mu can reach 1500 kg-1800 kg. The meat quality of the Allium chinense is pure white, tender and crisp, and is rich in sugar, protein, calcium, iron, carotene, vitamin C, 16 amino acids and various pharmacological components, according to the record of the compendium of materia Medica, the allium chinense is pungent, warm, smooth and nontoxic, mainly treats the golden sore and detoxication, has the effects of light weight, no hunger and fatigue resistance, can also prevent various diseases, and is a green health-care food known as the lucid ganoderma in vegetables. The allium chinense has special spicy taste, is well loved by people, and has the functions of appetizing, tonifying spleen, stimulating appetite, regulating intestines and stomach, sterilizing, checking diarrhea and the like. The components contained in the allium chinense have good treatment or adjuvant treatment effects on coronary heart disease, angina pectoris, gastroneurosis, gastroenteritis, retch, chronic bronchitis, asthma, cough, chest pain, back induction, chronic dysentery, and cold diarrhea.
Research shows that the allium chinense has obvious inhibition effect on various pathogenic bacteria and has the potential of developing antibacterial drugs and food preservatives. Antibacterial active substance in Allium chinense to Escherichia coli (E:)Escherichia coli) Candida albicans (C.albicans) ((C.albicans))Candida albicans) Bacillus subtilis preparation (B)Bacillus subtilis) Shigella dysenteriae (Shigella dysenteriae) (II)Shigella dysenteriae) Staphylococcus aureus (1)Staphylococcus aureus) Salmonella typhi (A), (B)Salmonella typhimurium) And brewers' yeast: (Saccharomyces cerevisiae) The growth of bacteria and fungi has obvious inhibiting effect, and simultaneously, active substances of the allium chinense are found to have inhibiting effect on liver cancer (HepG-2) cells and Hela (Hela) cells. The allium chinense has obvious antibacterial and antiseptic effects, is a medicinal and edible food, has the advantages of high yield, low price, wide plantation and the like, and can ensure food safety and obtain greater economic benefit if being developed into a natural food antiseptic bacteriostatic agent.
At present, the research on the allium chinense at home and abroad mostly focuses on the extraction and purification identification of active ingredients, and organic solvents are mostly adopted as media, so that certain potential safety hazards exist when the allium chinense is applied to food. For example, the invention patent application named as 'preparation method of allium chinense polysaccharide' which is published by the national intellectual property office in 2013, 1 month and 2 days is 'CN 201210366841.5', and an alcohol precipitation technology is adopted; in the technical scheme disclosed by the invention patent with the application number of "CN 201210161354.5" and the name of "a method for extracting total saponins from Allium chinense" disclosed by the national intellectual property office on 9/19/2012, an alcohol extraction technology is also adopted, and an organic solvent is introduced to extract active substances of the Allium chinense, so that on one hand, the production cost of the whole production process is increased, and the popularization to batch production is limited; on the other hand, the organic solvent is adopted for processing, so that the requirement on equipment is high after the organic solvent is popularized to mass production; in addition, the organic solvent is adopted for processing, and the obtained extract has certain potential safety hazard when being applied to the food industry and the application range is limited.
In addition, the bacteriostatic effect of the allium chinense is the combined action of multiple components, the single component is not suitable as the index of extraction and optimization of the bacteriostatic active ingredients of the allium chinense, and the bacteriostatic effect is directly used as the evaluation index of the allium chinense and is more reliable. The allium chinense as a natural bacteriostatic agent can improve the quality of food, but has special spicy flavor, and limits the application range of the allium chinense to a certain extent. For the spicy taste of the allium chinense, a plurality of removal methods are reported by scholars, but researches on allium chinense deodorization, and the influence of different deodorization methods and deodorization degrees on the antibacterial activity of the allium chinense are fresh. In order to remove the special odor of the allium chinense, ensure the bacteriostatic effect as much as possible, improve the practical application value of the allium chinense, and develop the related research of the odor removing process of the allium chinense, which is necessary. At present, the allium chinense extract deodorization process research considering both the antibacterial activity and the deodorization effect does not exist.
Therefore, the searching for an effective and safer extraction method or treatment method of allium chinense, and the wider application of the extract thereof still needs continuous research and innovation by people in the industry.
Disclosure of Invention
The invention aims to provide a preparation method of an allium chinensis extract, an allium chinensis extract inclusion compound prepared by using the allium chinensis extract, a preparation method of the allium chinensis extract inclusion compound and application of a product, which solve the problems of narrow applicable range and low economic benefit of the allium chinensis extract and the allium chinensis extract product in the prior art, and also solve the problems of high production cost and high popularization difficulty caused by complex preparation processes of extraction and processing of the allium chinensis in the prior art.
The invention is realized by the following technical scheme:
a preparation method of an allium chinense extract comprises the following steps:
a. cleaning fresh Allium chinense with root and bulb with clear water, removing impurities, and draining water at normal temperature;
b. taking the drained allium chinense in the step a, and mixing the allium chinense with the feed liquid ratio of 1: 2-5, adding distilled water, and stirring and crushing by using a crusher to obtain an allium chinense homogenate;
c. c, homogenizing the allium chinense obtained in the step b, adding an acidic reagent to adjust the pH value to be 4.0-5.0, adding food grade enzyme, and extracting for 60-180 min at the constant temperature of 40-50 ℃ under the stirring condition to obtain an extracting solution;
d. c, filtering the extracting solution in the step c, centrifuging, and taking supernate;
e. and d, treating the supernatant obtained in the step d by using a filter membrane, and then carrying out vacuum freeze drying to obtain solid powder, thus obtaining the allium chinense extract powder.
Further, in the step c, the food grade enzyme is one or any combination of pectinase, cellulase, neutral protease and xylanase.
Further, in the step c, the addition amounts of the pectinase, the cellulase, the neutral protease and the xylanase in the fresh allium chinense are as follows: 0.1-0.7% of pectinase, 0.1-0.5% of cellulase, 0.1-0.5% of neutral protease and 0.5-0.9% of xylanase.
Further, in the step d, the extracting solution is filtered by a filter with the precision of 200-300 meshes; the centrifugation condition is 8000-10000 r/min for 5-10 min.
Further, in the step e, after the supernatant is subjected to suction filtration treatment by a 0.22-0.45 mu m aqueous mixed cellulose microporous filter membrane, the supernatant is subjected to vacuum freeze drying at-60 to-85 ℃ and under the pressure of 5-30 Pa to form solid powder.
An allium chinensis extract clathrate compound is prepared from allium chinensis extract prepared by the preparation scheme and embedding material.
An allium chinensis extract inclusion compound, wherein the embedding material is cyclodextrin.
A preparation method of an allium chinense extract clathrate compound comprises the following steps:
A. under the constant temperature condition of 40-50 ℃, according to the mass ratio of cyclodextrin to distilled water of 1:2.5 to 3.5 taking cyclodextrin, dissolving the cyclodextrin in distilled water, dripping the allium chinense extract at the speed of 10m to 15mL/min under the stirring condition, keeping out of the sun, and stirring under the stirring condition of 200 plus 400r/min until no precipitate exists;
C. and D, refrigerating the mixture obtained in the step A in a refrigerator at 4 ℃ for 24 hours, and then carrying out vacuum freeze drying at low temperature to obtain the allium chinense extract clathrate compound.
Further, the allium chinense extract prepared by the preparation method is applied to preparing additives and/or bacteriostatic agents and/or antibacterial agents of foods, health products, cosmetics, pet foods or medicines.
Further, the allium chinensis extract clathrate prepared by the preparation method is applied to preparing bacteriostatic agents and/or antibacterial agents of foods, health products, cosmetics, pet foods or medicines.
Compared with the prior art, the invention has the following advantages and beneficial effects:
1. the invention uses natural bacteriostatic components of allium chinense to treat escherichia coli (E.coli) The method has the advantages that the antibacterial effect of the allium chinense is used as an evaluation index, the extraction condition is optimized, the optimal extraction process of the allium chinense antibacterial active ingredients is obtained through response surface design, the safe and efficient extraction process of the allium chinense natural antibacterial ingredients is established, the odor removal effect and the antibacterial activity of the allium chinense extract are considered, the deodorization process is optimized, the application range of the allium chinense extract in the fields of food, health care products, cosmetics or medicines and the like is expanded, the use of chemical preservative additives is reduced, and the safety and the nutritional value of the food are improved. Development and utilization of natural bacteriostatic component of allium chinense for inhibiting bacteria and fungiThe bacterial property promotes the development of allium chinense industry, increases the added value of allium chinense agricultural products, and lays a foundation for the deep processing of allium chinense and the large-scale development and utilization of the natural bacteriostatic agent derived from allium chinense in the field of foods.
2. According to the invention, the fresh allium chinense with roots and bulbs is adopted, a large number of experiments show that the fresh allium chinense roots also contain antibacterial ingredients, the allium chinense with roots and bulbs is selected, the available parts of raw materials are enlarged, and after the method is popularized to batch production, the production cost can be obviously reduced, and resources are effectively utilized.
3. According to the invention, fresh allium chinense is used as a raw material, impurities are removed after cleaning, the content of the impurities is generally not more than 5%, the next step of treatment is carried out after water is drained at normal temperature, operations such as air drying are not carried out, and the antibacterial activity of the final extract is ensured.
4. According to the invention, food-grade enzymes such as pectinase, cellulase, neutral protease and xylanase are adopted to treat the allium chinensis homogenate, so that the extraction time can be obviously shortened, the requirement on the extraction temperature is reduced, the food-grade enzymes can be recycled until the food-grade enzymes lose activity, the production cost can be obviously reduced when the allium chinensis homogenate is applied to large-scale production, and the energy loss is reduced. In the invention, the preferable ranges of the addition amounts (%, w/w) of the enzymes are as follows: 0.1-0.7% of pectinase, 0.1-0.5% of cellulase, 0.5-0.9% of xylanase and 0.1-0.5% of neutral protease.
5. According to the invention, the inclusion compound of the allium chinense extract is prepared from the lyophilized powder of the allium chinense extract by adopting an embedding technology, so that the peculiar spicy smell of the allium chinense can be effectively removed, the inclusion compound is colorless and easy to dissolve in water, has the performances of radiation resistance and temperature resistance (below 120 ℃), has a good antibacterial effect, can be added into food to play an antibacterial role, reduces the use of chemical additives, improves the safety and the nutritional value of the food, and can be popularized to the application of additives and/or antibacterial agents of food, health products, cosmetics, pet food or medicines because the special spicy smell of the allium chinense extract inclusion compound is removed.
Drawings
Figure 1 is the picture of the inhibition zone experiment of 34 th group obtained allium chinense extract in example 1.
Figure 2 is a total ion flow diagram of allium chinense crude extract.
Fig. 3 is an infrared absorption spectrum of the allium chinensis extract freeze-dried powder, hydroxypropyl-beta-cyclodextrin, 1:1 mixed allium chinensis extract dry powder, hydroxypropyl-beta-cyclodextrin and hydroxypropyl-beta-cyclodextrin inclusion compound in example 2.
FIG. 4 is an image taken with a scanning electron microscope in example 2 (a-d: hydroxypropyl-. beta. -cyclodextrin, e-h: inclusion compound).
Fig. 5 is a graph of the electronic nose odor test results of hydroxypropyl- β -cyclodextrin inclusion compound of example 2.
FIG. 6 is a graph showing the results of measuring the bacteriostatic activity of hydroxypropyl-. beta. -cyclodextrin inclusion compound of example 2 with respect to radiation resistance.
Figure 7 is a graph of the temperature resistance results of the hydroxypropyl-beta-cyclodextrin inclusion compound bacteriostatic activity of example 2.
Fig. 8 is a graph of fermentation status of kimchi added with allium chinense extract and control group.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
The materials and equipment used in the following examples are as follows:
materials:
fresh allium chinense is purchased from the city farmer market in Sichuan province;
the enzyme activity of the pectinase is more than or equal to 10 ten thousand U/g, the enzyme amount of 1g of enzyme powder for catalyzing pectin hydrolysis to generate 1mg of galacturonic acid in 1 hour is 1 pectinase activity unit under the conditions of 50 ℃ and pH value of 3.5;
the enzyme activity of the cellulase is more than or equal to 10 ten thousand U/g, the enzyme amount of 1 mu g of glucose generated by hydrolyzing a substrate (filter paper, CMC, absorbent cotton or saligenin) for 1min is 1 enzyme activity unit under the conditions that 1g of enzyme powder is at 50 ℃ and the pH value is 5.0;
the enzyme activity of xylanase is more than or equal to 10 ten thousand U/g, the enzyme amount of 1g of xylose generated by hydrolyzing 1% xylan solution in 1min is 1 xylanase activity unit under the conditions that 1g of enzyme powder is at 50 ℃ and the pH value is 5.0;
the enzyme activity of neutral protease is more than or equal to 10 wumu/g, 1g of solid enzyme powder hydrolyzes casein for 1min under the conditions of 40 ℃ and pH =7.5 to generate 1 mu g of tyrosine which is an enzyme activity unit, and the four enzymes are purchased from Ningxia and Heishi wall biotechnology limited company;
LB, NA/NB and Sa's nutrient medium were purchased from Qingdao Haibo Biotech, Inc.;
all other reagents commonly used were in-house analytical grade.
The instrument comprises the following steps:
HH digital display constant temperature water bath kettle, Jiangsu province, jin Tan city, Jincheng Guosheng test instrument factory;
BSA224S-CW electronic balance Saedodes scientific instruments (Beijing) Inc.;
eppendorf centrifuge 5810R high speed centrifuge, Eppendorf Python, Inc.;
ELELAFDU-2110 lyophilizer ELELA, Tokyo, Japan, physiochemical Co., Ltd;
Mettler-Toledo-S210 pH Meter Mettler-Torledo instruments (Shanghai) Co., Ltd;
DNP-9272 electric heating constant temperature incubator Shanghai sperm macro experimental facilities Co., Ltd;
sample breaker U.S. blender PB 12.
Example 1:
a. cleaning fresh Allium chinense with root and bulb with clear water, removing impurities, draining water at normal temperature, and collecting 34 parts of crushed Allium chinense 1kg per crushed material;
b. b, adding double distilled water into the drained allium chinense in the step a according to the feed liquid ratio shown in the table 1, and stirring and crushing the allium chinense by using a crusher to obtain an allium chinense homogenate;
c. b, homogenizing the allium chinense in the step b, adding 2 mol/L hydrochloric acid or 2 mol/L sodium hydroxide reagent into each group to adjust the pH value, adding the types and the adding amount of the enzymes according to the table 1, continuously stirring, and extracting for 50 ℃ under the heat preservation condition according to the table 1, wherein the heat preservation condition in the step can be adjusted within 40-50 ℃, and the activity of the enzymes can be damaged when the temperature is too high;
d. c, filtering the extracting solution in the step c, centrifuging, taking supernate, and referring to the table 1 for the filtering precision and the centrifugation condition;
e. and d, treating the supernatant obtained in the step d by using a water-based mixed cellulose microporous filter membrane, and performing vacuum freeze drying to obtain solid powder to obtain the allium chinense extract powder, wherein the precision and the vacuum conditions of the filter membrane refer to table 1, and the rest conditions are the same.
In this embodiment, the double distilled water added to the allium chinense crushed aggregates in step b can be selected from distilled water, double distilled water, deionized water and tap water according to actual production requirements, and the double distilled water is taken as an example in this embodiment, so that other interference factors are eliminated, and the extraction conditions are convenient to optimize.
The comparative example in table 1 is selected from the same batch of fresh allium chinense with roots and bulbs, which is cleaned and dried in the sun, and the other treatment conditions are the same as those in group 3, so as to obtain the allium chinense extract powder.
The other conditions are the same as those in a certain group,
TABLE 1
Ratio of material to liquid (mass ratio) Conditioning agents pH value Enzyme (ratio of fresh Chinese onion) During extraction Interval (min) Filter fineness
Group 1 1:2 2 mol/L HcL 4 0.7% of pectinase 60 200 mesh filtration, 8000r/min separation Treating with heart machine for 5min to obtain 0.22 μm holes Filtering with 0.1MPa filter membrane
Group 2 1:4 2 mol/L HcL 5 0.7% of pectinase 75 Same as above
Group 3 1:3 2 mol/L HcL 5 0.7% of pectase 60 Same as above
Group 4 1:4 2 mol/L HcL 4 0.7% of pectinase 60 Same as above
Group 5 1:5 2 mol/L HcL 5 0.7% of pectinase 60 Same as above
Group 6 1:3 2 mol/L HcL 4 0.7% of pectase 75 Same as above
Group 7 1:5 2 mol/L HcL 4 The addition amount of the pectinase accounts for 0.7 percent of the allium chinense 45 Same as above
Group 8 1:3 2 mol/L HcL 3 0.7% of pectinase 60 Same as above
Group 9 1:5 2 mol/L HcL 3 0.7% of pectase 60 Same as above
Group 10 1:5 2 mol/L HcL 4 0.7% of pectinase 75 Same as above
Group 11 1:4 2 mol/L HcL 3 0.7% of pectinase 75 Same as above
Group 12 1:4 2 mol/L HcL 5 0.7% of pectinase 45 Same as above
Group 13 1:4 2 mol/L HcL 4 0.7% of pectinase 60 Same as above
Group 14 1:3 2 mol/L HcL 4 0.7% of pectinase 45 Same as above
Group 15 1:3 2 mol/L HcL 4.5 0.5% cellulase 60 Same as above
Group 16 1:3 2 mol/L HcL 4.5 0.5% xylanase 60 Same as above
Group 17 1:3 2 mol/LNaOH 7 0.5% of neutral protease 60 Same as above
Group 18 1:3 2 mol/L HcL 4.5 0.3% of pectinase and 0.3% of cellulase Adding 0.3% of xylanase, and adding 0.7% of xylanase 60 300 mesh filtration, 8000r/min separation Treating with heart machine for 10min, 0.22 μm Filtering with pore diameter microporous filter membrane of 0.1MPa
Group 19 1:3 2 mol/L HcL 4.5 0.5% of pectinase and 0.5% of cellulase Adding 0.1% of xylanase, and adding 0.9% of xylanase 60 Same as above
Group 20 1:3 2 mol/L HcL 4.5 0.3% of pectinase and 0.3% of cellulase Adding 0.3% of xylanase, adding 0.364% of xylanase 60 Same as above
Group 21 1:3 2 mol/L HcL 4.5 0.3% pectinase and 0.3% cellulase Adding 0.3% of xylanase, and adding 1.036% of xylanase 60 Same as above
Group 22 1:3 2 mol/L HcL 4.5 0.636 percent of pectinase and cellulase 0.3% of xylanase and 0.7% of xylanase are added 60 Same as above
Group 23 1:3 2 mol/L HcL 4.5 0% of pectinase and 0% of cellulase 0.3 percent of xylanase, and 0.7 percent of xylanase 60 Same as above
Group 24 1:3 2 mol/L HcL 4.5 0.5% pectinase and 0.5% cellulase Adding 0.5% of xylanase, and adding 0.9% of xylanase 60 Same as above
Group 25 1:3 2 mol/L HcL 4.5 0.1% of pectinase and 0.1% of cellulase Adding 0.5% of xylanase, and adding 0.9% of xylanase 60 Same as above
Group 26 1:3 2 mol/L HcL 4.5 0.1% of pectinase and 0.1% of cellulase 0.1 percent of xylanase and 0.9 percent of xylanase 60 Same as above
Group 27 1:3 2 mol/L HcL 4.5 0.5% of pectinase and 0.5% of cellulase Adding 0.1% of xylanase, and adding 0.5% of xylanase 60 Same as above
Group 28 1:3 2 mol/L HcL 4.5 0.3% of pectinase and fiberVitamin enzyme additive 0% of xylanase is added, and 0.7% of xylanase is added 60 Same as above
Group 29 1:3 2 mol/L HcL 4.5 0.5% of pectinase and 0.5% of cellulase Adding 0.5% of xylanase, and adding 0.5% of xylanase 60 Same as above
Group 30 1:3 2 mol/L HcL 4.5 0.3% of pectinase and 0.3% of cellulase Adding 0.636 percent of xylanase, and adding 0.7 percent of xylanase 60 Same as above
Group 31 1:3 2 mol/L HcL 4.5 0.1% of pectinase and 0.1% of cellulase Adding 0.5% of xylanase, and adding 0.5% of xylanase 60 Same as above
Group 32 1:3 2 mol/L HcL 4.5 0.1% pectinase and 0.1% cellulase Adding 0.1% of xylanase, and adding 0.5% of xylanase 120 Same as above
Group 33 1:3 2 mol/L HcL 4.5 0.1% of pectinase and 0.1% of cellulase Adding 0.1% of xylanase, and adding 0.5% of xylanase 180 Same as above
Group 34 1:3 2 mol/L HcL 4.5 0.1% of pectinase and 0.1% of cellulase 0.1 percent of xylanase and 0.5 percent of xylanase 60 Same as above
Comparative example 1 1:3 2 mol/L HcL 5 0.7% of pectinase 60 200 mesh filtration, 8000r/min separation Treating with heart machine for 5min to obtain 0.22 μm holes Filtering with 0.1MPa microporous filter membrane
According to the conditions of each group in the table 1, the Allium chinense extract powder is obtained by extraction and detected, the weight, the color and the smell of the powder are recorded, sterilized double distilled water is used for preparing 10% (w/v) antibacterial solution from the Allium chinense extract powder of each experimental group according to the mass volume ratio, and antibacterial experiments are carried out on escherichia coli, staphylococcus aureus and candida albicans. The experimental method comprises the following steps: diluting the activated bacteria with physiological saline to 106And (3) coating 100 muL of the bacterial suspension of CFU/mL on a corresponding culture medium flat plate, uniformly placing Oxford cups at intervals on the bacterial suspension, respectively adding 0.2mL of Allium chinense extract into the Oxford cups, taking sterile water as a control, repeating each sample for 6 times, culturing at the constant temperature of 37 ℃ for 24 hours, measuring the diameter of a bacteriostatic circle, and taking the diameter of the bacteriostatic circle of each group of bacteria as an evaluation index of bacteriostatic activity.
Through tests, the allium chinense extract powder obtained by 34 experimental groups and the allium chinense extract powder obtained by the comparative example group are all milky white solids and have special taste of the allium chinense, and other results are shown in table 2.
TABLE 2
Weight of powder (unit: g) Diameter of escherichia coli inhibition zone (mm) Staphylococcus aureus bacteriostasis ringDiameter (mm) Candida albicans zone diameter (mm)
Group 1 29.43 20.78±0.376 22.14±0.479 22.81±0.461
Group 2 31.56 19.95±0.317 21.75±0.407 23.75±0.613
Group 3 32.71 18.55±0.249 21.05±0.268 22.53±0.204
Group 4 31.12 22.11±0.827 24.52±0.45 25.15±0.32
Group 5 32.06 23.52±0.308 25.25±0.45 23.56±0.096
Group 6 32.57 19.85±0.28 23.54±0.234 23.95±0.187
Group 7 30.76 23.45±0.236 25.75±0.13 23.65±0.066
Group 8 30.61 22.45±0.51 23.12±0.191 24.75±0.186
Group 9 32.48 25.16±0.249 27.75±0.182 25.25±0.215
Group 10 30.49 25.25±0.544 27.21±0.442 25.57±0.495
Group 11 32.44 24.95±0.267 22.15±0.45 26.25±0.508
Group 12 30.91 22.84±0.473 18.25±0.242 23.54±0.132
Group 13 31.94 22.35±0.513 24.25±0.528 24.85±0.235
Group 14 32.16 21.16±0.609 19.25±0.537 23.75±0.344
Group 15 28.91 25.99±0.731 18.12±0.37 25±0.736
Group 16 28.13 24.92±0.521 17.31±0.352 24.45±0.374
Group 17 27.81 22.71±0.513 16.81±0.611 22.45±0.613
Group 18 36.78 26.57 ±0.46 20.08±0.18 28.92±0.62
Group 19 37.91 23.77 ±0.31 20.86±0.03 27.41±0.25
Group 20 34.75 26.89 ±0.6 21.9±0.66 29.96±0.87
Group 21 35.24 26.12 ±1.01 20±0.87 28.69±0.31
Group 22 39.34 24.18 ±0.27 21.03±0.55 25.28±0.28
Group 23 33.89 29.86 ±0.15 20.43±0.74 27.81±0.24
Group 24 39.26 26.10 ±0.22 19.91±0.82 26.93±0.51
Group 25 39.65 26.88 ±0.48 19.84±0.81 30.8±0.22
Group 26 39.25 29.49 ±0.5 20.22±0.39 27±0.54
Group 27 38.49 22.61 ±0.51 21.87±0.77 27.63±1.27
Group 28 34.41 26.28 ±0.28 21.47±0.44 26.95±0.77
Group 29 36.75 25.87 ±0.6 18.9±0.68 26.44±0.51
Group 30 36.8 26.82 ±0.61 20.99±0.48 27.6±0.61
Group 31 38.41 28.80 ±1.35 21.55±0.61 29.8±1.27
Group 32 40.25 29.29±0.43 20.82±0.36 29.16±0.62
Group 33 40.75 28.09±0.75 20.08±0.63 28.34±0.51
Group 34 41.28 30.09 ±0.83 22.18±0.6 30.01±1.08
Comparative example 1 30.17 10.55±0.25 9.05±0.27 10.13±0.22
Description of the drawings: inhibition cycles are expressed as mean ± standard deviation, n = 6.
As can be seen from Table 2, the effect of single-enzyme assisted extraction is not as good as that of complex-enzyme assisted extraction, and the ratio of 1: 3, the pH value is 4.5, and at the constant temperature of 50 ℃, 0.1 percent of pectinase, 0.1 percent of cellulase and 0.5 percent of xylanase are added, so that the allium chinense extracted according to the proportion is better in bacteriostatic activity and higher in yield. Compared with the experiment groups 31 to 34, the 34 th group has the highest yield (the yield is 16.5 percent), the weight of the powder is 41.28g, and under the condition of consistent other conditions, the extraction time of 60min is better than the bacteriostatic performance of the allium chinense extract obtained by the experiment groups of 120min and 180 min.
In addition, fig. 1 is a picture of a zone of inhibition experiment of the allium chinense extract obtained in group 3, and the experimental result can be visually understood. Various test bacteria and fungi are inoculated on a flat plate in advance, a small metal oxford cup is filled with bacteriostatic agents and can slowly diffuse into a culture medium below, and bacteria and fungi cannot grow on the culture medium with allium chinense extract, so that a transparent circle without bacteria growth can appear.
The diameter of the transparent inhibition zone is used to evaluate the bacteriostasis of the sample. The water in the oxford cup in the middle of each plate is used as a control group, and the oxford cup on which water is discharged can not inhibit the growth of bacteria and fungi, which indicates that the whole experimental process is reliable.
It can be known from table 2 that the antibacterial ring of the freeze-dried powder extracted from the dried allium chinense in the comparative example 1 is obviously smaller than that of the experiment of the group 3, and it can be known that the dried allium chinense is not beneficial to the retention of antibacterial substances of the allium chinense.
Analysis of components of Allium chinense extract
Taking 1g of allium chinense extract, and carrying out component analysis on the allium chinense extract by GC-MS.
GC conditions were as follows: an HP-5MS elastic quartz column with a size of 30m multiplied by 0.25mm multiplied by 0.25 mu m is selected, the initial temperature is 50 ℃, the initial temperature is kept for 1min, the temperature is increased to 280 ℃ at a speed of 5 ℃/min, the initial temperature is kept for 15 min, and the operation is carried out for 62 min. The injection port temperature is 250 ℃ and the carrier gas He.
MS conditions: EI ion source, electron energy 70 eV. Scan range 35-450u, electron multiplication voltage 2226.7. Transmission line temperature: 280 ℃ and the ion source temperature 250 ℃.
Finally, the map shown in fig. 2 is obtained, the map is searched by a computer and manually analyzed, and the analysis result is shown in the following table 3.
Table 3: chemical components and percentage content of allium chinense crude extract
Figure 322811DEST_PATH_IMAGE002
As can be seen from fig. 2 and table 3, 17 components can be detected from the allium chinense crude extract, wherein 14 are sulfur-containing compounds and are main components thereof. The components with higher content mainly comprise 24.18 percent of dimethyl disulfide and 54.51 percent of dimethyl trisulfide, which account for about 80 percent of the total content, and are basically the same as the products obtained by the prior art, but the yield is obviously improved (in the prior art, the yield of the dimethyl disulfide is generally 10 to 20 percent, and the yield of the dimethyl trisulfide is generally 25 to 40 percent).
Example 2
A preparation method of an allium chinensis extract clathrate comprises the following steps:
A. adopting the allium chinense extract prepared by the preparation method of the allium chinense extract 34 in the embodiment 1, taking 6 parts of the allium chinense extract, wherein each part is 3 g;
B. under the condition of constant temperature, proper inclusion materials are dissolved in distilled water according to a certain proportion, and under the condition of stirring, the allium chinense extract is slowly dripped at a constant speed and stirred in a dark place until no precipitate exists. In the scheme, the inclusion compound with the antibacterial effect can be obtained under the condition of no light shielding, but the antibacterial effect of the finished product obtained under the condition of light shielding is better;
C. and D, naturally cooling the mixture obtained in the step A, putting the mixture into a refrigerator with the temperature of 4 ℃ for refrigeration for 24 hours, freezing, and carrying out low-temperature vacuum freeze drying to obtain the allium chinense extract inclusion compound.
Specific conditions of each group are shown in Table 4, and the other conditions are the same.
TABLE 4
Inclusion material Material and handle Ratio of taking out Temperature of Time Filter fineness Method of operation
Group
35 Sodium alginate 3:25 60℃ Curing for 30min Is free of 12mL of 30g/L sodium alginate solution is prepared at constant temperature of 60 DEG C Mixing 6mL of 50% Allium chinense extract at 50 deg.C 30g/L of sodium alginate in a ratio of 1:2, uniformly mixing the mixture according to the proportion of 2, dropping the mixed solution into CaCl of 20g/L at constant speed2Formation in solution And (4) microcapsules. Solidifying the microcapsule in coagulating bath for 30min, and separating Discharging the wet microcapsule, rinsing with clear water, and removing the microcapsule Residual CaCl on the surface2Then drying at room temperature
Group 36 Beta-cyclodextrin 1:3 At normal temperature Grinding for 60min 0.22 mu m Pore diameter microporous filter Film 0.1MPa drawer Filter element 30mL of 10 percent Allium chinense extracting solution is added with 1g of beta-ring Grinding for 1h by dextrin, and filtering with a 0.22 mu m pore size microporous filter membrane Filtering under 0.1MPa, and vacuum freeze drying the filtrate
Group 37 Beta-cyclodextrin 8:3 Prepared at a constant temperature of 60 DEG C Beta-cyclodextrin saturated solution Cooling the solution to 50 deg.C Then, undissolved matter was filtered off Precipitation of beta-cyclodextrin Stirring for 5 hours 0.22 mu m Pore diameter microporous filter Film 0.1MPa drawer Filter element Beta-cyclodextrin saturated solution is dripped at constant temperature of 50 ℃ at uniform speed Stirring 50% allium chinense extract 6mL at the rotating speed of 1200r/min Placing in a refrigerator at 4 deg.C for 24 hr for 5 hr, refrigerating, and filtering0.22 mu m filter membrane Suction filtering, washing filter cake with distilled water, vacuum filtering Freeze drying to obtain solid clathrate
Group 38 Hydroxypropyl- Beta-cyclodextrin 1:1 50℃ Stirring for 2 hours Is free of Stirring 3g of hydroxypropyl-beta-cyclodextrin at the constant temperature of 50 DEG C Fully dissolved in 8mL of distilled water, and slowly dropwise added with 50 percent of allium chinense at constant speed 6mL of head extract (3 g of allium chinense extract) and 400r & lt/EN & gt protected from light Stirring for 2h at min speed. Naturally cooling, and cooling in a refrigerator at 4 deg.C Storing for 24h, and vacuum freeze-drying at low temperature for 24h to obtain the clathrate.
Group 39 Hydroxypropyl- Beta-cyclodextrin 5:1 50℃ Stirring for 2 hours Is free of Stirring 15g of hydroxypropyl-beta-cyclodextrin at a constant temperature of 50 DEG C Completely dissolved in 40mL of distilled water, slowly dropped at uniform speed 50% Allium chinense extract 6mL (3 g Allium chinense extract), and avoiding light Stirring at the rotating speed of 400r/min for 2 h. Naturally cooling, and adding into 4 deg.C Refrigerating for 24h in refrigerator, and vacuum freeze-drying at low temperature for 24h to obtain And (3) inclusion compound.
Group 40 Hydroxypropyl- Beta-cyclodextrin 3:1 50℃ Stirring for 2 hours Is free of Stirring 9g of hydroxypropyl-beta-cyclodextrin at the constant temperature of 50 DEG C Dissolving in 27mL distilled water, and slowly adding 50% dropwise at uniform speed Allium chinense extract 6mL (3 g Allium chinense extract), and light-proof 400 r- Stirring at min speed for 2 h. Naturally cooling, and cooling in a refrigerator at 4 deg.C Storing for 24h, and vacuum freeze-drying at low temperature for 24h to obtain the clathrate.
Extracting to obtain the inclusion compound of the allium chinense extract (hereinafter referred to as the inclusion compound) according to the conditions of each group in the table 4, weighing the solid weight of the inclusion compound, and calculating the inclusion yield (%) of each group of the inclusion compound. Wherein, the formula for calculating the inclusion yield is as follows:
the inclusion yield (%) = the total mass of the inclusion compound/(the mass of the inclusion material + the total mass of the added allium chinense extract) × 100%.
The color and morphology of each set of compositions was recorded. The odor of each group of the compositions is graded through sensory evaluation, the sensory evaluation is carried out on the odor of each group of the compositions, the odor removing effect of each group of the compositions is judged, the grading method is shown in the following table 5, the total score is 9, the score is increased or decreased by 0.1, the higher the score is, the better each sensory quality index of the sample is, and the score is reserved to one position behind the decimal point. Meanwhile, the odor of the allium chinense extract which is not deodorized is used as a control. 8 Ginseng and sensory evaluation, the evaluation members were individually conducted without contact and communication with each other, and the results of the sensory evaluation were expressed as mean ± standard deviation.
TABLE 5
Figure 771110DEST_PATH_IMAGE004
And preparing the inclusion compounds of 35-40 groups of the experimental groups into solution by using sterilized double distilled water, and performing bacteriostatic experiments on escherichia coli, staphylococcus aureus and candida albicans. The composition in group 35 was not readily soluble in water at room temperature, and 1g of the composition was mixed with 8mL of sterile distilled water to prepare a suspension of the inclusion compound (containing about 10% extract) for use in bacteriostatic experiments. The compositions in the 36 th group and the 37 th group are not easy to dissolve in water at normal temperature, and sterilized distilled water is prepared into a clathrate compound saturated solution for bacteriostasis experiments. And weighing 2g of the inclusion compound of the composition of the group 38, dissolving the inclusion compound with 8mL of distilled water to prepare a solution with the final concentration of the allium chinensis extract being about 10%, and taking 200 mu L of the solution to perform a zone experiment of inhibiting bacteria of escherichia coli in an Oxford cup. Weighing 2g of the inclusion compound in the 39 th group, dissolving the inclusion compound with 2mL of distilled water to prepare a solution with the final concentration of the allium chinensis extract being about 8%, and taking 200 mu L of the solution to perform a zone of inhibition experiment on escherichia coli in an Oxford cup. Weighing 5g of the inclusion compound in the 40 th group of compositions, dissolving the inclusion compound with 7.5mL of distilled water to prepare a solution with the final concentration of the allium chinensis extract being about 10%, and taking 200 mu L of the solution to be used as a bacteriostatic circle of escherichia coli in an Oxford cupAnd (5) carrying out experiments. The experimental method of the zone of inhibition: diluting the activated bacteria with physiological saline to 106And (3) coating 100 muL of the bacterial suspension of CFU/mL on a corresponding culture medium flat plate, uniformly placing Oxford cups at intervals on the Oxford cups, respectively adding 0.2mL of Allium chinense extract into the Oxford cups, taking sterile water as a control, repeating each sample for 6 times, culturing at constant temperature of 37 ℃ overnight, measuring the diameter of a bacteriostatic circle, and taking the diameter of the bacteriostatic circle of each group of bacteria as an evaluation index of bacteriostatic activity.
The evaluation results are shown in Table 6.
TABLE 6
Inclusion compound Solid stem Heavy (g) Is included to obtain Rate of change Colour(s) Sensory evaluation Color scoring Smell(s) Sensory evaluation Scoring of odor Sensory evaluation Total Receptivity Escherichia coli inhibitor Diameter of fungus ring （mm） Golden yellow grape Antibacterial zone for cocci Diameter (mm) Candida albicans Diameter of zone of inhibition （mm）
Group 35 2.874 85.54% Half light yellow Transparent solid 8.4±0.93 With a small amount of Chinese onion The taste of the tea is improved, 7.4±1.16 7.1±0.96 7.36±0.6 is free of 9.31±0.74
Group 36 0.462 11.55% White powder 8.5±1.21 With a small amount of Chinese onion Taste (I) 8.0±0.98 8.4±0.73 9.7±0.68 Is free of 10.68±0.57
Group 37 9.85 89.55% White powder Solid body 8.5±0.71 Is nearly odorless 8.4±0.45 8.5±0.85 12.42±0.45 8.72±0.53 14.97±0.61
Group 38 5.23 87.17% White powder Solid body 8.5±0.74 Is nearly odorless 85±0.65 8.5±0.81 25.57±0.43 15.33±0.48 26.78±0.41
Group 39 16.83 93.52% White powder Solid body 8.5±0.51 Is nearly odorless 8.5±0.42 8.4±0.49 21.32±0.47 11.81±0.67 22.15±0.49
Group 40 10.93 91.08% White powder Solid body 8.5±0.49 Low smell of odor 8.5±0.57 8.5±0.64 28.94±0.73 18.17±0.43 29.15±0.67
Description of the drawings: sensory evaluation scoring data are expressed as mean ± standard deviation, n = 8; inhibition cycles are expressed as mean ± standard deviation, n = 6.
As can be seen from table 6, the composition of group 40 had good odor removal effect and minimal loss of antibacterial activity, and the preferred inclusion conditions of allium chinense extract were: the proportion of the hydroxypropyl-beta-cyclodextrin to the extract is 3:1, the mixture is stirred for 2 hours at the constant temperature of 50 ℃ and the rotating speed of 400r/min, the mixture is naturally cooled and then put into a refrigerator at 4 ℃ for refrigeration for 24 hours, and the low-temperature vacuum freeze drying is carried out for 24 hours to obtain the inclusion compound.
The hydroxypropyl-beta-cyclodextrin allium chinense extract inclusion compound obtained in the 40 th group in the example is further identified.
Infrared spectra of allium chinense extract freeze-dried powder (example 1, group 34), hydroxypropyl- β -cyclodextrin, a mixture of allium chinense extract freeze-dried powder (example 1, group 34) mixed with hydroxypropyl- β -cyclodextrin 1:1, and an allium chinense extract inclusion compound (example 2, group 40) were respectively examined, and the results are shown in fig. 3.
As can be seen from fig. 3, the allium chinense extract freeze-dried powder and the hydroxypropyl-beta-cyclodextrin have different infrared spectrograms, the spectrogram of the mixture is basically the superposition of the allium chinense extract freeze-dried powder and the hydroxypropyl-beta-cyclodextrin, and the position and the intensity of a main absorption peak are not changed greatly. And the characteristic absorption peak of the inclusion compound of hydroxypropyl-beta-cyclodextrin allium chinense extract is 3428.60cm-1、1632.61cm-1Obviously diminishes or disappears, a plurality of new characteristic absorption peaks appear in the inclusion compound, and the analysis verifies the allium chinense extract in 40 th group of example 2The dry powder does form an inclusion complex with hydroxypropyl-beta-cyclodextrin.
The surface characteristics of the hydroxypropyl-beta-cyclodextrin and the hydroxypropyl-beta-cyclodextrin allium chinense extract inclusion compound obtained in group 40 of this example were observed by a scanning electron microscope, and the result is shown in fig. 4, in which the hydroxypropyl-beta-cyclodextrin is spherical particles with porous surfaces under an electron microscope, and the appearance of the hydroxypropyl-beta-cyclodextrin allium chinense extract inclusion compound is not spherical, and the surfaces of the hydroxypropyl-beta-cyclodextrin allium chinense extract inclusion compound are not porous any more, and the hydroxypropyl-beta-cyclodextrin allium chinense extract inclusion compound is changed into blocks. From the electron microscope photos, the appearance shape of the inclusion compound is greatly changed, and the implementation proves that the allium chinense extract freeze-dried powder and the hydroxypropyl-beta-cyclodextrin really form the inclusion compound.
Further investigate each performance, characteristic of allium chinensis extract freeze-dried powder, allium chinensis extract freeze-dried powder clathrate compound, analyze its stability, can provide data support for the popularization and application of allium chinensis extract freeze-dried powder and clathrate compound, promote the development of allium chinensis industry, for the intensive processing of allium chinensis, provide new way.
Taking the lyophilized powder of allium chinensis extract in the 34 th group of the example 1 and the lyophilized powder of allium chinensis extract in the 40 th group of the example 2, namely hydroxypropyl-beta-cyclodextrin inclusion compound, and investigating odor, color, solubility, bacteriostatic effect, radiation resistance and high temperature resistance.
Sensory evaluation is carried out on the allium chinense extract freeze-dried powder and the allium chinense extract hydroxypropyl-beta-cyclodextrin inclusion compound from three aspects of smell, color and total acceptance, and the results are shown in table 7. The odor removing effect of the hydroxypropyl-beta-cyclodextrin inclusion compound is judged through sensory evaluation results, and the results show that the peculiar pungent odor of the allium chinense contained in the allium chinense extract embedded by the hydroxypropyl-beta-cyclodextrin is removed, the pungent odor is difficult to smell, and the taste is close to odorless.
Table 7: odor evaluation of Allium chinense extract and hydroxypropyl-beta-cyclodextrin inclusion compound
Experimental group Smell(s) Colour(s) Total degree of acceptance
Hydroxypropyl-beta-cyclodextrin inclusion compounds 8.5±0.57 8.5±0.49 8.5±0.64
Allium chinense extract 3.5±1.44 8.0±0.75 5.1±0.92
Description of the drawings: data are presented as mean ± standard deviation, n = 8.
Further, the odor of the lyophilized powder of allium chinense extract and the hydroxypropyl-beta-cyclodextrin inclusion compound of allium chinense extract was measured by an electronic nose, and the data of the electronic nose at the 8 th second of the test process was compared, and the result is shown in fig. 5 (the left column in fig. 5 represents the inclusion compound, and the right column represents the extract). From the odor measurement results of the electronic nose of fig. 5, it was found that the odor of the inclusion compound was significantly less than that of the allium chinense extract which was not embedded.
The method for investigating the bacteriostatic effect comprises the following steps: preparing a solution (10% of allium chinense extract freeze-dried powder and 40% of hydroxypropyl-beta-cyclodextrin inclusion compound) containing 10% of allium chinense extract, inhibiting the growth of escherichia coli, and detecting the inhibition zone.
Radiation resistance test method: subjecting the object to be examined to 4kGy, 6kGy and 8kGy, respectively60After Co gamma ray irradiation, distilled water is respectively prepared into solutions (10% of Allium chinense extract freeze-dried powder and 40% of hydroxypropyl-beta-cyclodextrin inclusion compound) containing 10% of Allium chinense extract final concentration, growth inhibition is carried out on escherichia coli, and the radiation resistance of the hydroxypropyl-beta-cyclodextrin inclusion compound is deduced through the diameter of a bacteriostatic zone.
High temperature resistance test method: preparing a 10% extract solution of an object to be tested by using distilled water, heating the solution at different high temperatures of 60 ℃, 80 ℃, 100 ℃ and 121 ℃ for 25 minutes, and performing an escherichia coli inhibition experiment by using the extract heated at the high temperature.
The results of the examination are shown in FIGS. 6 and 7.
Fig. 6 is a graph of the results of the detection of bacteriostatic radiation resistance of the allium chinensis extract of example 1 and the hydroxypropyl- β -cyclodextrin inclusion compound of example 2. In fig. 6, A represents the antibacterial activity of the allium chinense extract after irradiation, and B represents the antibacterial activity of the inclusion after irradiation. Data are expressed as mean ± standard deviation, n = 6; different lower case letters indicate significant differences at the 5% level from the other groups.
Figure 7 is a graph of temperature resistance results for bacteriostatic activity of the allium chinense extract of example 1 and the hydroxypropyl- β -cyclodextrin inclusion compound of example 2. In fig. 7, A is the antibacterial activity of the heated allium chinense extract, and B is the antibacterial activity of the heated inclusion. Data are expressed as mean ± standard deviation, n = 6; different lower case letters indicate significant differences at the 5% level from the other groups.
As can be seen from fig. 6, after the allium chinense extract which is not included is irradiated, the antibacterial activity is obviously reduced, along with the increase of the irradiation dose, the antibacterial activity is reduced more, and the antibacterial effect of the allium chinense extract irradiated with the dose of 8kGy on escherichia coli can be maintained to 65%. The bacteriostatic property of the allium chinense extract can be more stable and more durable in the inclusion process60And (5) irradiating by Co gamma rays. The antibacterial effect of the hydroxypropyl-beta-cyclodextrin inclusion compound on escherichia coli is hardly influenced after the hydroxypropyl-beta-cyclodextrin inclusion compound is irradiated by 4kGy dose, and the diameter of an antibacterial ring on the escherichia coli after the hydroxypropyl-beta-cyclodextrin inclusion compound is irradiated by 6kGy and 8kGy doses can be maintained to be more than 85% of that of an unirradiated control group. The dosage of 4kGy-8kGy is the common dosage range for killing putrefying bacteria in food by irradiation. The inclusion compound can better keep the bacteriostatic activity in the dosage range, so the inclusion compound is very suitable to be used as a bacteriostatic additive for irradiation sterilized food.
As can be seen from fig. 7, both the allium chinense extract and the hydroxypropyl- β -cyclodextrin inclusion compound have good temperature resistance, and can tolerate temperatures below 100 ℃. If the allium chinense extract and the hydroxypropyl-beta-cyclodextrin inclusion compound are added into food, even if high-temperature treatment is adopted in the subsequent food processing process, the high-temperature treatment conditions do not influence the antibacterial activity, and the allium chinense extract and the hydroxypropyl-beta-cyclodextrin inclusion compound still have good antibacterial effect.
Further, the influence of pH change on the antibacterial performance of the allium chinensis extract and the hydroxypropyl-beta-cyclodextrin inclusion compound is investigated. The results are shown in Table 8.
TABLE 8
Figure 347585DEST_PATH_IMAGE006
Description of the drawings: inhibition cycles are expressed as mean ± standard deviation, n = 6.
As can be seen from table 8, the antibacterial performance of the allium chinense extract is basically unchanged under a neutral condition, but the antibacterial zone of the allium chinense extract is obviously reduced under an alkaline condition, and the allium chinense extract has no inhibitory effect on staphylococcus aureus. Therefore allium chinensis extract is not suitable for use in foods that are alkaline. Under alkaline conditions, the hydroxypropyl-beta-cyclodextrin inclusion compound is more stable than the extract, the antibacterial performance of the hydroxypropyl-beta-cyclodextrin inclusion compound is superior to that of the allium chinense extract, and the result provides data reference for the application field of the allium chinense extract.
Example 3
This example examines the fresh keeping effect of allium chinense extract and hydroxypropyl-beta-cyclodextrin allium chinense extract inclusion compound (hereinafter referred to as hydroxypropyl-beta-cyclodextrin inclusion compound) in food.
Taking the allium chinensis extract of the 34 th group in the example 1, taking the allium chinensis extract clathrate of the 40 th group in the example 2, and taking the bean curd for standby.
The operation method comprises the following steps: dicing bean curd, weighing → adding 9 times volume of phosphate buffer solution (PBS solution, 1 mol/L Na)2HPO4And 1 mol/L NaH2PO4) Homogenizing → split charging 20 mL/bottle → sterilizing → adding 0.5g Allium chinense extract (final concentration of 2.5% in the whole system), 2.2g hydroxypropyl-beta-cyclodextrin inclusion compound (final concentration of 2.5% in the whole system), and no adding in the control group → adding 3 kinds of test bacteria (10) such as Escherichia coli, Staphylococcus aureus and Candida albicans in all samples5CFU/mL) 2.5mL → 37 ℃ shake culture (150 r/min) → total number of colonies measured after 1, 2 and 3 days, respectively.
At room temperature, the fresh keeping effect of the allium chinense extract and the hydroxypropyl-beta-cyclodextrin inclusion compound on the tofu is examined, and the results are shown in tables 9 to 11.
Inhibitory effect of Allium chinense extract of Table 9 on Escherichia coli in bean curd sample
Figure 24685DEST_PATH_IMAGE008
TABLE 10 inhibitory effect of Allium chinense extract on Staphylococcus aureus in tofu samples
Figure 635795DEST_PATH_IMAGE010
Inhibitory effect of Allium chinense extract of Table 11 on Candida albicans in bean curd sample
Figure 989416DEST_PATH_IMAGE012
Description of the drawings: the minimum dilution on day one was 10-1The dilution was < 100 since it was not detected, and < 10 since the lowest dilution was the stock solution on days 2 and 3.
As can be seen from table 9: the initial bacterial count of the bean curd sample was 104Under the condition of CFU/mL, the content of coliform bacteria can reach 10 after the control culture is carried out for 1 day9CFU/mL, and the bacteria content of day 2 and 3 is maintained at 108CFU/mL, while the content of coliform bacteria in the tofu sample containing 2.5% of allium chinense extract is less than 100CFU/mL on day 1, and no coliform bacteria are detected on days 2 and 3, which shows that the allium chinense extract and the hydroxypropyl-beta-cyclodextrin inclusion compound have good inhibition effect on the coliform bacteria under the condition.
As can be seen from Table 10, the picked-up amounts of Staphylococcus aureus were 10 after 1, 2 and 3 days of culture of the bean curd control samples7CFU/mL, bean curd sample containing 2.5% Allium chinense extract, with CFU/mL < 100 on day 1, and no detection on days 2 and 3. The allium chinense extract and the hydroxypropyl-beta-cyclodextrin inclusion compound have good inhibitory effect on staphylococcus aureus.
As can be seen from Table 11, Candida albicans in the bean curd control samples were detected at 10 days in 1, 2 and 3 days7CFU/mL, the allium chinense extract content is 2.5% bean curd sample, 1 day < 100CFU/mL, 2, 3 days have not detected, show under this condition, allium chinense extract and hydroxypropyl-beta-cyclodextrin inclusionThe product has good inhibitory effect on Candida albicans.
From the results, the allium chinense extract with low dosage can play a good antibacterial role in the actual food preservation.
Example 4
This example examines the application effects of allium chinense extract and allium chinense extract hydroxypropyl-beta-cyclodextrin inclusion compound in pickles.
Taking the allium chinense extract of the 34 th group in the example 1, taking the allium chinense extract hydroxypropyl-beta-cyclodextrin inclusion compound of the 40 th group in the example 2, and taking fresh carrots as pickle for standby.
The operation method comprises the following steps: the ratio of the pickled vegetable carrots to the fermentation liquor in each experimental group is 2: 3. First blank group: no allium chinense extract is added; second allium chinense extract group: adding allium chinense extract with the final concentration of 2.5%; the third group of allium chinense extract inclusion compound group: adding clathrate of Allium chinensis extract with final concentration of 10% (Allium chinensis extract with final concentration of about 2.5%). Fermenting conventionally, and observing the fermentation condition of the pickle after 3 d.
The results are shown in FIG. 8.
The early research result of the project shows that the allium chinense extract has obvious inhibition effect on escherichia coli, staphylococcus aureus, candida albicans and staphylococcus albus, but hardly has inhibition effect on lactic acid bacteria. The experiment utilizes the characteristic of the allium chinense extract to add the allium chinense extract into the fermentation process of the pickle, thereby inhibiting the growth of yeast, escherichia coli and other miscellaneous bacteria, and improving the sanitary quality of the pickle under the condition of not influencing the normal fermentation of the pickle. As can be seen from fig. 8, in the fermentation process of the carrot pickle, the surfaces of the control group samples are full of bacteria such as yeast and the like to form bacterial films, which seriously affect the sensory and safety quality of the pickle, while the pickle fermentation liquor added with the allium chinense extract/allium chinense extract inclusion compound is clear and transparent, which indicates that the addition of a certain amount of allium chinense extract/allium chinense extract inclusion compound in the pickle is beneficial to inhibiting the growth and reproduction of the bacteria therein and greatly improving the quality of the pickle. Combining the research results, the Allium chinense extract is feasible and has outstanding effect in the pickle fermentation process, and has application potential.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, and all simple modifications and equivalent variations of the above embodiments according to the technical spirit of the present invention are included in the scope of the present invention.

Claims (10)

1. The preparation method of the allium chinense extract is characterized by comprising the following steps:
a. cleaning fresh Allium chinense with root and bulb with clear water, removing impurities, and draining water at normal temperature;
b. taking the drained allium chinense in the step a, and mixing the allium chinense with the feed liquid ratio of 1: 2-5, adding distilled water, and stirring and crushing by using a crusher to obtain an allium chinense homogenate;
c. c, homogenizing the allium chinense in the step b, adding an acidic reagent or an alkaline reagent to adjust the pH value to be 4.0-5.0, adding food-grade enzyme, and extracting for 60-180 min at a constant temperature of 40-50 ℃ under the stirring condition to obtain an extracting solution;
d. c, filtering the extracting solution in the step c, centrifuging, and taking supernate;
e. and d, treating the supernatant obtained in the step d by using a filter membrane, and then carrying out vacuum freeze drying to obtain solid powder, thus obtaining the allium chinense extract powder.
2. The preparation method of the allium chinense extract as claimed in claim 1, which is characterized in that: in the step c, the food-grade enzyme is one or any combination of pectinase, cellulase, neutral protease and xylanase.
3. The preparation method of the allium chinense extract as claimed in claim 2, which is characterized in that: in the step c, the addition amounts of the pectinase, the cellulase, the neutral protease and the xylanase account for the following mass percentages of the fresh allium chinense: 0.1-0.7% of pectinase, 0.1-0.5% of cellulase, 0.1-0.5% of neutral protease and 0.5-0.9% of xylanase.
4. The preparation method of the allium chinense extract as claimed in claim 1, which is characterized in that: in the step d, the extracting solution is filtered by a filter with the precision of 200-300 meshes; the centrifugation condition is 8000-10000 r/min for 5-10 min.
5. The preparation method of the allium chinense extract as claimed in claim 1, which is characterized in that: and e, performing suction filtration treatment on the supernatant through a 0.22-0.45 mu m aqueous mixed cellulose microporous filter membrane, and performing vacuum freeze drying at the temperature of-60 to-85 ℃ and under the pressure of 5-30 Pa to obtain solid powder.
6. An allium chinensis extract clathrate compound, which is prepared from allium chinensis extract prepared by the preparation scheme of any one of claims 1 to 5 and embedding material.
7. The allium chinensis extract clathrate compound of claim 6, which is characterized in that: the embedding material is cyclodextrin.
8. The preparation method of the allium chinensis extract clathrate compound of claim 7, comprising the following steps:
A. under the constant temperature condition of 40-50 ℃, taking cyclodextrin according to the mass ratio of the cyclodextrin to distilled water of 1:2.5-3.5, dissolving the cyclodextrin in the distilled water, dripping the allium chinense extract at the speed of 10m-15mL/min under the stirring condition, keeping out of the sun, and stirring under the stirring condition of 200-400 r/min until no precipitate exists;
C. and D, refrigerating the mixture obtained in the step A in a refrigerator at 4 ℃ for 24 hours, and then carrying out vacuum freeze drying at low temperature to obtain the allium chinense extract clathrate compound.
9. Use of allium chinense extract prepared by the preparation method of any one of claims 1 to 5 in the preparation of additives and/or bacteriostatic and/or antibacterial agents for foods, health products, cosmetics, pet foods or medicines.
10. Use of the allium chinensis extract clathrate prepared by the preparation method of any one of claims 6 to 8 in preparation of bacteriostatic and/or antibacterial agents for foods, health products, cosmetics, pet foods or medicines.
CN202210433868.5A 2022-04-24 2022-04-24 Preparation method of allium chinensis extract, inclusion compound and application thereof Pending CN114711426A (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1456150A (en) * 2003-06-04 2003-11-19 王景成 Garlicin injection preparation
CN104770571A (en) * 2015-04-22 2015-07-15 徐州三农生物科技有限公司 Production method of garlic powder-beta-cyclodextrin or derivative inclusion compound of garlic powder-beta-cyclodextrin
CN105639120A (en) * 2015-12-29 2016-06-08 湖南晶天科技实业有限公司 Cyclodextrin inclusion compound containing antibiotic components, a compounding method thereof and feed additive

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1456150A (en) * 2003-06-04 2003-11-19 王景成 Garlicin injection preparation
CN104770571A (en) * 2015-04-22 2015-07-15 徐州三农生物科技有限公司 Production method of garlic powder-beta-cyclodextrin or derivative inclusion compound of garlic powder-beta-cyclodextrin
CN105639120A (en) * 2015-12-29 2016-06-08 湖南晶天科技实业有限公司 Cyclodextrin inclusion compound containing antibiotic components, a compounding method thereof and feed additive

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
吴琴，等: ""响应面法优化藠头抑菌成分提取工艺"，吴琴，等，第209-213页，江苏农业科学，第47卷，第24期", 江苏农业科学, vol. 47, no. 24, pages 2 - 6 *

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