CN115735926A - Tea polyphenol, tea saponin and compound liquid thereof with antibacterial effect - Google Patents
Tea polyphenol, tea saponin and compound liquid thereof with antibacterial effect Download PDFInfo
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- CN115735926A CN115735926A CN202211174668.9A CN202211174668A CN115735926A CN 115735926 A CN115735926 A CN 115735926A CN 202211174668 A CN202211174668 A CN 202211174668A CN 115735926 A CN115735926 A CN 115735926A
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- 241001122767 Theaceae Species 0.000 title claims abstract description 236
- 150000008442 polyphenolic compounds Chemical class 0.000 title claims abstract description 120
- 235000013824 polyphenols Nutrition 0.000 title claims abstract description 120
- 239000001397 quillaja saponaria molina bark Substances 0.000 title claims abstract description 102
- 229930182490 saponin Natural products 0.000 title claims abstract description 102
- 150000007949 saponins Chemical class 0.000 title claims abstract description 102
- 239000007788 liquid Substances 0.000 title claims abstract description 43
- 150000001875 compounds Chemical class 0.000 title claims abstract description 35
- 230000000844 anti-bacterial effect Effects 0.000 title abstract description 25
- 238000002156 mixing Methods 0.000 claims abstract description 23
- 238000013329 compounding Methods 0.000 claims abstract description 7
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical group CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 36
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 27
- 239000003480 eluent Substances 0.000 claims description 25
- 230000003385 bacteriostatic effect Effects 0.000 claims description 23
- 230000001954 sterilising effect Effects 0.000 claims description 14
- 238000001035 drying Methods 0.000 claims description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 239000012153 distilled water Substances 0.000 claims description 9
- 239000003960 organic solvent Substances 0.000 claims description 9
- 238000000926 separation method Methods 0.000 claims description 9
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 6
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 6
- 239000000284 extract Substances 0.000 claims description 6
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- 239000000843 powder Substances 0.000 claims description 6
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 claims description 6
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- 238000010438 heat treatment Methods 0.000 claims description 3
- 235000012054 meals Nutrition 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 238000002791 soaking Methods 0.000 claims description 3
- 239000008223 sterile water Substances 0.000 claims description 3
- 238000000108 ultra-filtration Methods 0.000 claims description 3
- 241000894006 Bacteria Species 0.000 abstract description 16
- 230000002195 synergetic effect Effects 0.000 abstract description 11
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- 230000000694 effects Effects 0.000 description 24
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- 241000588724 Escherichia coli Species 0.000 description 15
- 241000191967 Staphylococcus aureus Species 0.000 description 12
- 239000001963 growth medium Substances 0.000 description 8
- 238000004659 sterilization and disinfection Methods 0.000 description 8
- 235000013305 food Nutrition 0.000 description 6
- 238000011160 research Methods 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
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- 230000000975 bioactive effect Effects 0.000 description 1
- 235000012970 cakes Nutrition 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000005536 corrosion prevention Methods 0.000 description 1
- 238000010227 cup method (microbiological evaluation) Methods 0.000 description 1
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- 238000004945 emulsification Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000005187 foaming Methods 0.000 description 1
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention relates to the technical field of biochemistry, and particularly discloses tea polyphenol, tea saponin and a compound liquid thereof with an antibacterial effect, wherein the compound liquid is prepared by compounding and mixing a tea polyphenol solution with the concentration of 2-4 mg/mL and a tea saponin solution with the concentration of 100-200 mg/mL in the same volume, the compound liquid enables the tea polyphenol and the tea saponin to achieve a complementary or synergistic effect, the antibacterial effect is enhanced, the surface area of the contact between the tea polyphenol and bacteria is increased by the surfactant property of the tea saponin, the effective action concentration of the tea polyphenol is increased, the oxidative damage of the tea saponin to the bacteria is caused, and the sensitivity of pathogenic bacteria to the tea polyphenol is enhanced.
Description
Technical Field
The application relates to the technical field of biochemistry, and particularly discloses tea polyphenol, tea saponin and compound liquid thereof with antibacterial effect.
Background
The tea contains various bioactive substances such as Tea Polyphenols (TP), tea Saponin (TS), amino acid and the like, and has high nutritive value for human bodies. The tea polyphenol is the main chemical component in the tea, and a large number of researches show that the tea polyphenol has strong effects of eliminating free radicals, resisting radiation, resisting cancer, resisting aging and the like. In addition, the tea polyphenol also has the bacteriostatic action, can prevent food from putrefaction and deterioration, prolongs the fresh-keeping period of food, and is a safe and ideal antioxidant and bacteriostatic agent. The preservative is widely applied to the corrosion prevention and the fresh keeping of fruits and vegetables, meat products, cakes and the like. The tea saponin is a chemical component with higher content in tea seeds, not only has good surface activity effects of emulsification, foaming, lubrication and the like, but also has obvious effects of resisting virus, resisting bacteria, diminishing inflammation, killing insects, expelling parasites and the like, also has strong inhibition effect on typical gram bacteria and microzyme, and has food preservation and fresh-keeping functions. Domestic and foreign researches show that the sterilization conditions of the food are milder by using a plurality of different natural preservatives in a matching way, and the complementary or synergistic effect is achieved. The research shows that the compounding of tea polyphenol and some plant extracts has a synergistic bacteriostasis effect. However, no report is found on the research of carrying out a mixed bacteriostasis experiment on the tea polyphenol and the tea saponin extracted from the tea leaves, so that the inventor provides the tea polyphenol, the tea saponin and the compound liquid thereof with bacteriostasis effects in view of the above, so as to solve the problems.
Disclosure of Invention
The invention aims to provide tea polyphenol, tea saponin and a compound liquid thereof with an antibacterial effect, so that the tea polyphenol and the tea saponin achieve a complementary or synergistic effect, and the antibacterial effect is enhanced.
In order to achieve the above object, the basic scheme of the present invention provides a tea polyphenol solution with bacteriostatic effect, comprising a tea polyphenol solution with a concentration of 2-4 mg/mL, which is prepared according to the following steps:
step S001, drying and crushing tea leaves to prepare tea leaf powder;
step S002, soaking the tea powder in hot water at 65-75 ℃ for 60-91 min, extracting the filtered filtrate, centrifuging the filtrate to obtain a separation solution, and ultrafiltering the separation solution through an ultrafiltration membrane to obtain an ultrafiltrate;
step S003, adding chloroform into the ultrafiltrate, stirring and mixing, standing and layering to obtain a water layer tea polyphenol extract, heating to 80-85 ℃, stirring for 2-4 min, and cooling to below 40 ℃;
step S004, adsorbing the cooled liquid by resin, and eluting by an eluant to obtain an eluent;
step S005, concentrating and drying the eluent to prepare crude tea polyphenol, and carrying out column chromatography treatment on the crude tea polyphenol to obtain refined tea polyphenol;
step S006, sterilizing the refined tea polyphenol, and mixing the sterilized tea polyphenol with sterile distilled water to form a tea polyphenol solution with the concentration of 2-4 mg/mL.
Further, in step S004, the eluent includes a solution prepared by combining water and an organic solvent.
Further, the eluent is an ethyl acetate solution, and the mass ratio of the tea polyphenol ultrafiltrate to the ethyl acetate solution is 1:4-7.
Further, in the step S004, the temperature of the system is kept between 25 and 28 ℃, and the elution flow rate is 4.2 to 5.3BV/h.
Based on the same invention concept, the invention provides a tea saponin solution with bacteriostatic effect, which is characterized by comprising the tea saponin solution with the concentration of 100-200 mg/mL prepared according to the following steps:
step A001, mixing tea seed meal with the extracting solution, and drying to prepare crude tea saponin;
step A002, mixing the crude tea saponin with sterile water, adsorbing the mixture by resin, and eluting the mixture by an eluant to obtain an eluent;
step A003, removing the organic solvent in the eluent, mixing the eluent with a flocculating agent, performing solid-liquid separation to obtain a clear liquid, and drying the clear liquid to obtain refined tea saponin;
step A004, sterilizing the refined tea saponin, and mixing the sterilized refined tea saponin with sterile distilled water to form a tea saponin solution with the concentration of 100-200 mg/mL.
Further, in the step S002, the eluent includes a solution prepared by combining water and an organic solvent.
Further, the eluent is an ethyl acetate solution, and the mass ratio of the tea polyphenol ultrafiltrate to the ethyl acetate solution is 1.5-4.
Further, in step S001, the extracting solution is one or more of methanol, ethanol, water, n-propanol, isopropanol and butanol.
Based on the same inventive concept, the invention discloses a compound liquid with an antibacterial effect, which is prepared by compounding and mixing the tea polyphenol solution and the tea saponin solution in the same volume.
Further, the concentration of the tea polyphenol solution is 3mg/mL, and the concentration of the tea saponin solution is 150mg/mL.
When the tea polyphenol and the tea saponin are independently used, the growth of escherichia coli and staphylococcus aureus is inhibited to a certain degree, and when the tea polyphenol and the tea saponin are compounded according to a certain proportion, the antibacterial effect is enhanced along with the increase of the concentration proportion of the compound liquid of the tea polyphenol and the tea saponin, the antibacterial effect is better than that of the independent use of the tea polyphenol and the tea saponin with the same concentration, the antibacterial effect of the compound tea saponin and the tea polyphenol on the escherichia coli is stronger than that of the independent use of the tea saponin and the tea saponin, namely, the compound tea polyphenol and the tea saponin with proper concentration have certain synergistic effect, and the synergistic effect is positively correlated with the compound concentration of the tea polyphenol and the tea saponin.
Compared with the prior art, the compound liquid disclosed by the invention has the advantages that the tea polyphenol and the tea saponin achieve complementary or synergistic interaction, the antibacterial effect is enhanced, the surface area of the tea polyphenol in contact with bacteria is increased due to the surfactant property of the tea saponin, the effective action concentration of the tea polyphenol is increased, the tea saponin can cause oxidative damage to the bacteria, and the sensitivity of pathogenic bacteria to the tea polyphenol is enhanced.
Detailed Description
To further illustrate the technical means and effects of the present invention adopted to achieve the predetermined objects, the embodiments, structures, features and effects according to the present invention are described in detail as follows.
A tea polyphenol solution with bacteriostatic effect is prepared according to the following steps:
step S001, drying and crushing tea leaves to prepare tea leaf powder;
step S002, soaking the tea powder in hot water at 65-75 ℃ for 60-90 min, extracting and filtering to obtain filtrate, centrifuging and filtering the filtrate to obtain separation liquid, and ultrafiltering the separation liquid through an ultrafiltration membrane to obtain ultrafiltrate;
step S003, adding chloroform into the ultrafiltrate, stirring and mixing, standing and layering to obtain a water layer tea polyphenol extract, heating to 80-85 ℃, stirring for 2-4 min, and cooling to below 40 ℃;
and step S004, adsorbing the cooled liquid by resin, and eluting by an eluent to obtain an eluent, wherein the eluent comprises a solution prepared by combining water and an organic solvent, an ethyl acetate solution is preferably adopted, the mass ratio of the tea polyphenol ultrafiltrate to the ethyl acetate solution is 1:4-7, and in the process, the system temperature is kept at 25-28 ℃, and the elution flow rate is 4.2-5.3 BV/h.
Step S005, concentrating and drying the eluent to prepare crude tea polyphenol, and carrying out column chromatography treatment on the crude tea polyphenol to obtain refined tea polyphenol;
step S006, sterilizing the refined tea polyphenol, and mixing the sterilized tea polyphenol with sterile distilled water to form a tea polyphenol solution with the concentration of 1-2 mg/mL.
A tea saponin solution with antibacterial effect is prepared according to the following steps:
step A001, mixing the tea seed meal with an extracting solution, and drying to prepare crude tea saponin, wherein the extracting solution is one or more of methanol, ethanol, water, n-propanol, isopropanol and butanol;
step A002, mixing the crude tea saponin with sterile water, adsorbing by resin, and eluting by an eluent to obtain an eluent, wherein the eluent comprises a solution prepared by combining water and an organic solvent, preferably an ethyl acetate solution, and the mass ratio of the tea polyphenol ultrafiltrate to the ethyl acetate solution is 1.5-4;
step A003, removing the organic solvent in the eluent, mixing the mixture with a flocculating agent, carrying out solid-liquid separation to obtain a clarified liquid, and drying the clarified liquid to obtain the refined tea saponin;
step A004, sterilizing the refined tea saponin, and mixing the sterilized refined tea saponin with sterile distilled water to form a tea saponin solution with the concentration of 25-100 mg/mL.
A compound liquid with an antibacterial effect is prepared by compounding and mixing a tea polyphenol solution with the concentration of 1-2 mg/mL and a tea saponin solution with the concentration of 25-100 mg/mL in the same volume.
In order to compare the sterilization effects of the tea polyphenol solution, the tea saponin solution and the compound liquid with different concentrations in the embodiment, the inhibition effects of the tea polyphenol and the tea saponin which are used independently and the tea polyphenol and the tea saponin which are used in a compound mode according to a certain proportion on two common food contamination bacteria of escherichia coli and staphylococcus aureus are observed through an oxford cup method.
Firstly, respectively inoculating escherichia coli and staphylococcus aureus to a beef extract peptone agar slant culture medium on a sterile operating platform, and placing the beef extract peptone agar slant culture medium in a constant-temperature incubator at 37 ℃ for 24 hours. Then respectively picking out bacterial colonies of the tested strains from the slant agar culture medium, streaking and inoculating the bacterial colonies to a proper plate culture medium, placing the plate culture medium in a constant-temperature incubator at 37 ℃ for 24 hours, and then carrying out bacterial counting according to a conventional plate counting method. According to the plate counting result, the bacteria to be tested are selected and diluted to 1X 106CFU/mL by using sterile physiological saline for the next experiment.
Taking the prepared 2.0mg/mL tea polyphenol solution, fixing the volume to the scale with distilled water, and sequentially diluting into 1.6 and 1.0mg/mL tea polyphenol solutions as 1, 2 and 3 treatment groups respectively; obtaining prepared 100mg/mL tea saponin solution, fixing the volume to scale with distilled water, and sequentially diluting into 50mg/mL and 20mg/mL tea saponin solutions as 4, 5 and 6 treatment groups respectively;
preparing a compound liquid: the prepared tea saponin and tea polyphenol solution are compounded in the same volume, the concentration of the compounded solution is respectively (2.0 + 100.0), (2.0 + 50.0), (2.0 + 25.0), (1.60 + 100.0), (1.60 + 50.0), (1.60 + 25.0), (1.0 + 100.0), (1.0 + 50.0) and (1.0 + 25.0) mg/mL, the former figure is the concentration of the tea saponin solution, and the latter figure is the concentration of the tea polyphenol solution.
A double-layer flat plate transparent ring method is adopted, namely 5mL of autoclaved agar (lower layer) is poured into a sterile culture dish, and after solidification, 4 sterilized oxford cups are quickly placed on a flat plate in a cross shape. 15mL of the culture medium mixed with the test bacterium was added under aseptic conditions (1 mL of the test bacterium suspension was added to the upper layer at a concentration of 1X 106CFU/mL per 100mL of the culture medium). Taking out the Oxford cup after the culture medium is solidified, respectively adding the prepared tea saponin solution, the prepared tea polyphenol solution and the prepared compound solution of the tea saponin solution and the tea polyphenol solution into each hole by 100 mu L, and taking a blank control as sterile distilled water with the same volume. The plate was placed in a 37 ℃ light incubator for culture, after 20h the diameter of the zone of inhibition was observed and measured, 3 times for each zone of inhibition, 3 replicates for each concentration, and the average was taken.
The experimental results are as follows:
TABLE 1 bacteriostatic effect of tea polyphenol solution, tea saponin solution and its compound liquid on Escherichia coli
And (4) note that: - "negative in table; the same letters in the same column indicate no significant difference (p > 0.05), the upper case letters indicate very significant difference (p < 0.01), and the lower case letters in parentheses indicate significant difference (p < 0.05).
As can be seen from Table 1, the tea polyphenol solution and the tea saponin solution both show certain inhibition effect on the growth of escherichia coli when being used independently, and the inhibition effect is enhanced along with the increase of the concentration, wherein when the concentration of the tea polyphenol solution is 2.00mg/mL, the inhibition zone on the escherichia coli is extremely obviously larger than the inhibition zones of the tea polyphenol solutions with the concentrations of 1.6mg/mL and 1.00mg/mL respectively (p is less than 0.01); when the concentration of the tea saponin solution is 100.00mg/mL, the inhibition zone to Escherichia coli is remarkably larger than that of other two low concentrations (p is less than 0.01). The antibacterial effect of the tea polyphenol solution and the tea saponin solution on escherichia coli is related to the concentration of the escherichia coli. When the tea polyphenol solution and the tea saponin solution are compounded according to a certain proportion, the bacteriostatic effect is enhanced along with the increase of the concentration proportion of the compound solution of the tea polyphenol solution and the tea saponin solution, and the bacteriostatic effect is superior to that of the bacteriostatic effect of the two solutions with the same concentration when the two solutions are used independently. When the concentration of the tea polyphenol solution is 2.0mg/mL and the concentration of the tea saponin solution is 100mg/mL, the bacteriostatic action is strongest, the diameter of a bacteriostatic ring is 18.10mm, and the bacteriostatic ring is remarkably larger than that of a compound solution with other concentrations (p is less than 0.01). The antibacterial effect of the compounded tea saponin solution and tea polyphenol solution on escherichia coli is higher than that of the two solutions used independently, namely the compounding of the tea polyphenol solution and the tea saponin solution with proper concentration has a certain synergistic effect, and the synergistic effect is in positive correlation with the concentration of the compounded tea saponin solution and the tea polyphenol solution.
TABLE 2 bacteriostatic effect of tea polyphenol solution, tea saponin solution and its compound liquid on staphylococcus aureus
Note that: - "negative in table; the same letters in the same column indicate no significant difference (p > 0.05), the upper case letters indicate very significant difference (p < 0.01), and the lower case letters in parentheses indicate significant difference (p < 0.05).
As can be seen from Table 2, the tea polyphenol solution and the tea saponin solution both have inhibitory effects on Staphylococcus aureus when used alone, and the inhibitory effects are enhanced as the concentrations of the two increase. Wherein when the concentration of the tea polyphenol solution is 2.00mg/mL, the inhibition zone for staphylococcus aureus is obviously greater than that of the tea polyphenol solution with the concentrations of 1.60mg/mL and 1.00mg/mL (p is less than 0.05). When the concentration of the tea saponin solution is 100.00mg/mL, the inhibition zone on staphylococcus aureus is remarkably greater than that of the other two groups (p is less than 0.01). And when the inhibition effect on staphylococcus aureus after the tea polyphenol solution and the tea saponin solution are compounded according to a certain proportion is increased along with the increase of the compound concentration of the two, the antibacterial effect is more obvious, wherein when the tea polyphenol solution with the concentration of 2.0mg/mL and the tea saponin solution with the concentration of 100mg/mL are compounded, the antibacterial effect is strongest, the diameter of an antibacterial ring is 13.89mm, and the antibacterial effect is extremely obviously greater than that of the compound solution with other concentrations (p is less than 0.01). The antibacterial activity of the tea polyphenol solution and the tea saponin solution which are jointly used is higher than that of the tea polyphenol with the same concentration which is singly used, the two solutions are compounded to show a certain synergistic interaction effect, and the high-concentration compounding effect of the two solutions is most obvious.
The results show that the tea polyphenol solution, the tea saponin solution and the compound liquid thereof have different degrees of inhibition effects on two food pathogenic bacteria, the inhibition effect of the tea polyphenol solution, the tea saponin solution and the compound liquid thereof with the same concentration on escherichia coli is stronger than that on staphylococcus aureus, and the reason is presumed to be related to the species, the growth pH, the selectivity of the tea polyphenol solution and the tea saponin solution on the bacteriostasis of microorganisms and the like. In addition, the tea polyphenol solution and the tea saponin solution have a synergistic antibacterial effect, the antibacterial effect is enhanced along with the increase of the concentration ratio of the two compound solutions, particularly, the inhibition effect of the tea polyphenol solution compounded with the tea saponin solution with a certain concentration on two bacteria is obviously stronger than the antibacterial activity of the tea saponin solution used alone, the effect of the tea saponin solution on the synergism of the tea polyphenol is probably because the surface area of the tea polyphenol solution in contact with the bacteria is increased due to the surfactant property of the tea saponin solution, the effective action concentration of the tea polyphenol solution is improved, or the oxidative damage of the tea saponin solution on the bacteria is caused, and the sensitivity of pathogenic bacteria on the tea polyphenol solution is enhanced.
In conclusion, the tea polyphenol solution, the tea saponin solution and the compound thereof have certain inhibition effect on common food pollution bacteria, namely staphylococcus aureus and escherichia coli, the stronger the inhibition effect is along with the increase of the concentration, and the two have synergistic inhibition effect when used together, and are positively correlated with the concentrations of the two. The tea saponin solution compounded by the tea polyphenol solution with a certain concentration has a good bacteriostatic effect, namely the tea saponin solution has a synergistic effect on the tea polyphenol solution. Meanwhile, the tea polyphenol solution, the tea saponin solution and the compound solution thereof with the same concentration have stronger inhibition effect on escherichia coli than staphylococcus aureus.
Taking the refined tea polyphenol prepared in step S005 again, preparing tea polyphenol solutions with concentrations of 1mg/mL, 3mg/mL and 5mg/mL according to step S006, taking the refined tea saponin prepared in step A003, preparing tea saponin solutions with concentrations of 25mg/mL, 150mg/mL and 275mg/mL according to step A004, mixing the prepared tea polyphenol solutions and the tea saponin solutions according to the proportion of 1:1 to form a reconstituted liquid, wherein the reconstituted liquid concentrations are respectively (5.0 + 275.0), (5.0 + 150.0), (5.0 + 25.0), (3.0 + 275.0), (3.0 + 150.0), (3.0 + 25.0), (1.0 + 275.0), (1.0 + 150.0), (1.0 + 25.0) mg/mL, the former number is the tea polyphenol solution, and the latter number is the tea polyphenol solution concentration. The 24h escherichia coli and staphylococcus aureus sterilization experiment is carried out in a closed environment, 3 experimental mice are added in the experimental environment to observe the influence of the concentration of the reconstituted liquid on the 3 experimental mice, experimental comparison is carried out again according to the above experimental steps, the experimental results show that the concentrations of the reconstituted liquid (5.0 + 275.0), (5.0 + 150.0), (5.0 + 25.0), (3.0 + 275.0), (3.0 + 150.0), (1.0 + 275.0) and (1.0 + 150.0) can achieve better sterilization efficiency, even (5.0 + 275.0), (5.0 + 150.0), (3.0 + 275.0) and (3.0 + 150.0), but with the increase of the concentration of the reconstituted liquid, the experimental mice show phenomena of continuous excitation, reduction of sleep time and the like, and the conditions of diarrhea and the like appear in subsequent feeding, the research shows that the initial influence of the reconstituted tea on the experimental mice can be the sterilization solution, and the concentration of the reconstituted liquid can be considered as 0, the sterilization error of the stomach sterilization is the optimal sterilization experiment is 0.0.
In consideration of highest cost performance and safe use of the disinfectant, the concentration of the tea saponin in the compound liquid should be 2-4 mg/mL, and the concentration of the tea polyphenol should be 100-200 mg/mL.
Compared with the prior art, the compound liquid disclosed by the invention has the advantages that the tea polyphenol and the tea saponin achieve complementary or synergistic interaction, the antibacterial effect is enhanced, the surface area of the tea polyphenol in contact with bacteria is increased due to the surfactant property of the tea saponin, the effective action concentration of the tea polyphenol is increased, the tea saponin can cause oxidative damage to the bacteria, and the sensitivity of pathogenic bacteria to the tea polyphenol is enhanced.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. A tea polyphenol solution with bacteriostatic effect is characterized by comprising a tea polyphenol solution with the concentration of 2-4 mg/mL, which is prepared according to the following steps:
step S001, drying and crushing tea leaves to prepare tea leaf powder;
step S002, soaking the tea powder in hot water at 65-75 ℃ for 60-91 min, extracting the filtered filtrate, centrifuging the filtrate to obtain a separation solution, and ultrafiltering the separation solution through an ultrafiltration membrane to obtain an ultrafiltrate;
step S003, adding chloroform into the ultrafiltrate, stirring and mixing, standing and layering to obtain a water layer tea polyphenol extract, heating to 80-85 ℃, stirring for 2-4 min, and cooling to below 40 ℃;
step S004, adsorbing the cooled liquid by resin, and eluting by an eluant to obtain an eluent;
step S005, concentrating and drying the eluent to prepare crude tea polyphenol, and carrying out column chromatography treatment on the crude tea polyphenol to obtain refined tea polyphenol;
step S006, sterilizing the refined tea polyphenol, and mixing the sterilized tea polyphenol with sterile distilled water to form a tea polyphenol solution with the concentration of 2-4 mg/mL.
2. The tea polyphenol solution with bacteriostatic effect as claimed in claim 1, wherein in step S004, the eluent comprises a solution prepared by combining water and an organic solvent.
3. The tea polyphenol solution with bacteriostatic effect as claimed in claim 2, wherein the eluent is ethyl acetate solution and the mass ratio of the tea polyphenol ultrafiltrate to the ethyl acetate solution is 1:4-7.
4. The tea polyphenol solution with bacteriostatic effect according to claim 3, wherein in the step S004, the temperature of the system is kept at 25-28 ℃, and the elution flow rate is 4.2-5.3 BV/h.
5. The tea saponin solution with the bacteriostatic effect is characterized by comprising the tea saponin solution with the concentration of 100-200 mg/mL prepared by the following steps:
step A001, mixing tea seed meal with the extracting solution, and drying to prepare crude tea saponin;
step A002, mixing the crude tea saponin with sterile water, adsorbing by resin, and eluting by an eluant to obtain an eluent;
step A003, removing the organic solvent in the eluent, mixing the mixture with a flocculating agent, carrying out solid-liquid separation to obtain a clarified liquid, and drying the clarified liquid to obtain the refined tea saponin;
step A004, sterilizing the refined tea saponin, and mixing the sterilized refined tea saponin with sterile distilled water to form a tea saponin solution with the concentration of 100-200 mg/mL.
6. The tea saponin solution with bacteriostatic effect of claim 5, wherein in step S002, the eluent comprises a solution prepared by combining water and an organic solvent.
7. The tea saponin solution with bacteriostatic effect according to claim 6, wherein the eluent is ethyl acetate solution and the mass ratio of the tea polyphenol ultrafiltrate to the ethyl acetate solution is 1.5-4.
8. The tea saponin solution with bacteriostatic effect of claim 7, wherein in step S001, the extract is one or more selected from methanol, ethanol, water, n-propanol, isopropanol and butanol.
9. The compound liquid with the bacteriostatic effect is characterized in that: the compound liquid is prepared by compounding and mixing the tea polyphenol solution as claimed in any one of claims 1 to 4 and the tea saponin solution as claimed in any one of claims 5 to 8 in the same volume.
10. The compound liquid with bacteriostatic effect according to claim 9, wherein the concentration of the tea polyphenol solution is 3mg/mL, and the concentration of the tea saponin solution is 150mg/mL.
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| CN101874518A (en) * | 2010-06-11 | 2010-11-03 | 华中农业大学 | Natural preservative for nectarine and application thereof |
| CN108653713A (en) * | 2018-06-12 | 2018-10-16 | 广西庆翠堂生物科技有限责任公司 | A kind of natural composite antibacterial composition and preparation method thereof containing Tea Saponin |
| CN110590861A (en) * | 2019-09-18 | 2019-12-20 | 紫云自治县新山农业发展有限公司 | Method for extracting tea polyphenol from fresh tea leaves |
| CN112110979A (en) * | 2020-10-26 | 2020-12-22 | 长沙中战茶油集团有限公司 | Method for extracting tea saponin |
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| CN101874518A (en) * | 2010-06-11 | 2010-11-03 | 华中农业大学 | Natural preservative for nectarine and application thereof |
| CN108653713A (en) * | 2018-06-12 | 2018-10-16 | 广西庆翠堂生物科技有限责任公司 | A kind of natural composite antibacterial composition and preparation method thereof containing Tea Saponin |
| CN110590861A (en) * | 2019-09-18 | 2019-12-20 | 紫云自治县新山农业发展有限公司 | Method for extracting tea polyphenol from fresh tea leaves |
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