CN115444797B - Preparation method of Hibiscus sabdariffa liposome freeze-dried powder, hibiscus sabdariffa liposome freeze-dried powder and application of Hibiscus sabdariffa liposome freeze-dried powder - Google Patents
Preparation method of Hibiscus sabdariffa liposome freeze-dried powder, hibiscus sabdariffa liposome freeze-dried powder and application of Hibiscus sabdariffa liposome freeze-dried powder Download PDFInfo
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- CN115444797B CN115444797B CN202211145593.1A CN202211145593A CN115444797B CN 115444797 B CN115444797 B CN 115444797B CN 202211145593 A CN202211145593 A CN 202211145593A CN 115444797 B CN115444797 B CN 115444797B
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- 240000004153 Hibiscus sabdariffa Species 0.000 title claims abstract description 131
- 235000001018 Hibiscus sabdariffa Nutrition 0.000 title claims abstract description 131
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- 238000002360 preparation method Methods 0.000 title claims abstract description 23
- 239000011259 mixed solution Substances 0.000 claims abstract description 48
- 238000000034 method Methods 0.000 claims abstract description 29
- 239000003223 protective agent Substances 0.000 claims abstract description 24
- 239000000243 solution Substances 0.000 claims abstract description 23
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims abstract description 22
- 239000008101 lactose Substances 0.000 claims abstract description 22
- 238000009777 vacuum freeze-drying Methods 0.000 claims abstract description 22
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 claims abstract description 20
- 229910021642 ultra pure water Inorganic materials 0.000 claims abstract description 18
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- 238000000859 sublimation Methods 0.000 claims description 25
- 230000008022 sublimation Effects 0.000 claims description 25
- PUPZLCDOIYMWBV-UHFFFAOYSA-N (+/-)-1,3-Butanediol Chemical compound CC(O)CCO PUPZLCDOIYMWBV-UHFFFAOYSA-N 0.000 claims description 22
- 238000004108 freeze drying Methods 0.000 claims description 18
- 235000019437 butane-1,3-diol Nutrition 0.000 claims description 11
- 239000000284 extract Substances 0.000 claims description 10
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical group CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims description 7
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- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 claims description 5
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- 235000007185 Hibiscus lunariifolius Nutrition 0.000 description 11
- 239000008176 lyophilized powder Substances 0.000 description 10
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 9
- 239000000047 product Substances 0.000 description 7
- 241000451942 Abutilon sonneratianum Species 0.000 description 6
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- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
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- 239000001116 FEMA 4028 Substances 0.000 description 1
- NIPNSKYNPDTRPC-UHFFFAOYSA-N N-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 NIPNSKYNPDTRPC-UHFFFAOYSA-N 0.000 description 1
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- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 description 1
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- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
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- ODLHGICHYURWBS-LKONHMLTSA-N trappsol cyclo Chemical compound CC(O)COC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)COCC(O)C)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1COCC(C)O ODLHGICHYURWBS-LKONHMLTSA-N 0.000 description 1
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Classifications
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- 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
- A61K8/96—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
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
- A61K8/0216—Solid or semisolid forms
- A61K8/022—Powders; Compacted Powders
-
- 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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/345—Alcohols containing more than one hydroxy group
-
- 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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/60—Sugars; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- 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
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
-
- 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
Abstract
The present disclosure relates to a preparation method of a freeze-dried powder of Hibiscus sabdariffa liposome, the freeze-dried powder of Hibiscus sabdariffa liposome and application thereof, wherein the preparation method of the freeze-dried powder of Hibiscus sabdariffa liposome comprises the following steps: according to the mass fraction, mannose with the mass fraction of 5-10 percent and lactose with the mass fraction of 1-3 percent are mixed with the rest ultrapure water, heated, stirred and dissolved to obtain a protective agent solution; adding 3-7% of Hibiscus sabdariffa liposome into the protective agent solution, and stirring for dissolving to obtain a mixed solution; and (3) placing the mixed solution into a freeze dryer for vacuum freeze drying to obtain the freeze-dried powder of the Hibiscus sabdariffa liposome. The method is simple, the prepared freeze-dried powder of the Hibiscus sabdariffa liposome has good re-solubility and dispersion performance, and the particles are uniform, the particle size is small, aggregation or agglomeration phenomenon is avoided after the re-dissolution of the Hibiscus sabdariffa liposome is observed under a microscope, the damage to the structure of the freeze-dried Hibiscus sabdariffa liposome is small, and the transportation and the storage of skin care products or cosmetics prepared from the Hibiscus sabdariffa liposome are easy.
Description
Technical Field
The present disclosure relates to the technical field of cosmetics, and in particular relates to a preparation method of freeze-dried powder of Hibiscus sabdariffa liposome, freeze-dried powder of Hibiscus sabdariffa liposome and application thereof.
Background
Most of the active ingredients added into the cosmetics are animal and plant extracts, which have the problems of unstable light/heat, poor solubility, easy oxidation and the like, are not beneficial to skin absorption, are difficult to exert efficacy, and are required to improve the bioavailability by modifying or entrapment and other technologies. The liposome entrapment technique is a simple and efficient means. Liposomes are vesicles formed from one or more bilayer membranes, and are composed of generally human-inherent lecithin or cholesterol, and therefore have low cytotoxicity. The phospholipid molecules are composed of a hydrophilic head group and a hydrophobic hydrocarbon chain, can spontaneously assemble into liposome in aqueous solution, and encapsulate water-soluble or oil-soluble molecules. Because of the advantages of good biocompatibility, biodegradability, low cytotoxicity and the like, the liposome has wide application as a delivery carrier of an active substance in the fields of medicine, food, cosmetics and the like.
The liposome is encapsulated, so that percutaneous absorption of active molecules can be effectively promoted, and the effect of improving the efficacy is achieved, but the liposome has poor stability in aqueous medium, is easy to leak, is hydrolyzed in lecithin and the like, and is not beneficial to transportation and storage of products. The freeze-drying technology is the most effective and common method for maintaining the activity of various biological products at present, and the technology obtains a dried product by dehydrating a sample at a low temperature under vacuum, greatly prolongs the preservation time of the biological products, and has the advantages of small loss, easy renaturation and the like. Ice crystals generated in the process of freeze-drying the liposome easily damage the vesicle structure of the liposome, so that the physicochemical properties of the freeze-dried product are changed, and therefore, the need of adding a proper freeze-drying protective agent before the freeze-drying process is a difficulty to be solved in preparing the freeze-dried powder of the liposome.
The foregoing is provided merely for the purpose of facilitating understanding of the technical solutions of the present invention and is not intended to represent an admission that the foregoing is prior art.
Disclosure of Invention
In order to overcome the problems in the related art, the present disclosure provides a preparation method of a freeze-dried powder of wild hibiscus liposome, the freeze-dried powder of wild hibiscus liposome and application thereof.
According to a first aspect of embodiments of the present disclosure, there is provided a method for preparing a freeze-dried powder of hibiscus liposome, comprising the steps of: according to the mass fraction, mannose with the mass fraction of 5-10 percent and lactose with the mass fraction of 1-3 percent are mixed with the rest ultrapure water, heated, stirred and dissolved to obtain a protective agent solution; adding 3-7% of Hibiscus sabdariffa liposome into the protective agent solution, and stirring for dissolving to obtain a mixed solution; and (3) placing the mixed solution into a freeze dryer for vacuum freeze drying to obtain the freeze-dried powder of the Hibiscus sabdariffa liposome.
In one embodiment, the vacuum freeze drying comprises the steps of: prefreezing procedure, sublimation drying and analytical drying.
In one embodiment, the pre-freezing procedure comprises sequentially passing a temperature of-3+ -1deg.C, a pre-freezing time of 90min, a temperature of-45+ -5deg.C, a pre-freezing time of 120min, and a pre-freezing time of 90min at-15+ -1deg.C.
In one embodiment, the sublimation drying comprises sequentially passing a temperature of 0±5 ℃, a time of 180min, a temperature of 18±3 ℃, a time of 1080min, and a temperature of 25±2 ℃ for 120min.
In one embodiment, the analytical drying comprises sequentially passing a temperature of 30+ -3deg.C, a time of 120min, a temperature of 35+ -5deg.C, a time of 120min, a temperature of 40+ -5deg.C, a time of 120min, and a temperature of 45+ -2deg.C for 400min.
In one embodiment, the packaging material of the mixed solution is any one of a penicillin bottle, a freeze-drying ampoule and a freeze-drying bubble cap.
In one embodiment, the Hibiscus sabdariffa liposome is prepared from root extract or stem extract of Hibiscus sabdariffa and lecithin.
According to a second aspect of embodiments of the present disclosure, there is provided a hibiscus liposome lyophilized powder, comprising: the Hibiscus sabdariffa liposome freeze-dried powder is prepared by the preparation method of the Hibiscus sabdariffa liposome freeze-dried powder provided by the embodiment of the disclosure.
According to a third aspect of the embodiments of the present disclosure, an application of a freeze-dried powder of Hibiscus sabdariffa liposome is provided, wherein the freeze-dried powder of Hibiscus sabdariffa liposome is prepared by the preparation method of the freeze-dried powder of Hibiscus sabdariffa liposome provided by the embodiments of the present disclosure, and the freeze-dried powder of Hibiscus sabdariffa liposome is applied in cosmetics.
In one embodiment, the freeze-dried powder of the Hibiscus sabdariffa liposome is used by being compounded with cosmetic water, essence or essence, or the freeze-dried powder of the Hibiscus sabdariffa liposome is used by being compounded with an aqueous solution of 1, 3-butanediol with the mass fraction of 10%.
The technical scheme provided by the embodiment of the disclosure can comprise the following beneficial effects:
the method is simple, the prepared freeze-dried powder of the Hibiscus sabdariffa liposome has good re-solubility and dispersion performance, and the particles are uniform, the particle size is small, aggregation or agglomeration phenomenon is avoided after the re-dissolution of the Hibiscus sabdariffa liposome is observed under a microscope, and the structural damage of the freeze-dried Hibiscus sabdariffa liposome is small; the mannose and lactose are adopted as the freeze-drying protective agent, so that the protective effect on the Hibiscus sabdariffa liposome is improved, the stability of the Hibiscus sabdariffa liposome is improved, and the skin care product or cosmetic prepared from the Hibiscus sabdariffa liposome is easy to transport and store; the 1, 3-butanediol is used as a demulsifier to dissolve the Hibiscus sabdariffa liposome freeze-dried powder, so that the active ingredients can be released, and the corresponding effects can be achieved.
It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the disclosure.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the disclosure and together with the description, serve to explain the principles of the disclosure.
Fig. 1 is an external view of freeze-dried powder of Hibiscus sabdariffa liposome prepared in example 1 and comparative examples 1-3 according to the preparation method of freeze-dried powder of Hibiscus sabdariffa liposome provided in the embodiment of the disclosure.
Fig. 2 is a microscopic image of the freeze-dried powder of the Hibiscus sabdariffa liposome prepared in example 1 and comparative examples 1-3 according to the preparation method of the freeze-dried powder of the Hibiscus sabdariffa liposome provided in the embodiment of the disclosure after redissolution.
Fig. 3 is a schematic diagram showing the effect of the freeze-dried powder of wild hibiscus liposome prepared in example 1 and comparative examples 1-3 on the activity of RAW264.7 cells.
Fig. 4 is a comparative schematic diagram of the effect of the freeze-dried powder of wild lotus liposome prepared in example 1 and comparative examples 1-3 on LPS-induced NO release of RAW264.7 cells according to the preparation method of the freeze-dried powder of wild lotus liposome provided in the examples of the present disclosure.
Fig. 5 is a comparative diagram of the dissolution effect of 1, 3-butanediol on the active ingredients of the freeze-dried powder of hibiscus liposome in the preparation method of the freeze-dried powder of hibiscus liposome provided by the embodiment of the disclosure.
Detailed Description
Reference will now be made in detail to exemplary embodiments, examples of which are illustrated in the accompanying drawings. When the following description refers to the accompanying drawings, the same numbers in different drawings refer to the same or similar elements, unless otherwise indicated. The implementations described in the following exemplary examples are not representative of all implementations consistent with the present disclosure. Rather, they are merely examples of apparatus and methods consistent with some aspects of the present disclosure as detailed in the accompanying claims.
In order to solve the related technical problems, the embodiment of the disclosure provides a preparation method of Hibiscus sabdariffa liposome freeze-dried powder, hibiscus sabdariffa liposome freeze-dried powder and application thereof.
Examples
Example 1
The first aspect of the embodiments of the present disclosure provides a method for preparing freeze-dried powder of hibiscus liposome, wherein the method comprises the steps of weighing 8% of mannose, 2% of lactose, 85% of ultrapure water at 80 ℃ and 5% of hibiscus liposome according to mass fraction. Mannose and lactose are added into ultrapure water to be stirred and dissolved, and the protective agent solution is obtained. And adding the Hibiscus sabdariffa liposome into the protective agent solution, stirring and dissolving to obtain a mixed solution. And (3) placing the mixed solution into a freeze dryer for vacuum freeze drying to obtain the freeze-dried powder of the Hibiscus sabdariffa liposome. Specifically, the vacuum freeze-drying comprises the following steps: prefreezing procedure, sublimation drying and analytical drying. In the pre-freezing procedure step, the mixed solution sequentially passes through a pre-freezing time of 90min at-3 ℃, a pre-freezing time of 120min at-45 ℃ and a pre-freezing time of 90min at-15 ℃ and enters a sublimation drying step. In the sublimation drying step, the mixed solution sequentially passes through the temperature of 0 ℃, the time of 180min, the temperature of 20 ℃, the time of 1080min and the temperature of 25 ℃ and the time of 120min, and then enters the analytical drying step. In the analytical drying step, the mixed solution is subjected to freeze-drying at 30 ℃ for 120min, at 35 ℃ for 120min, at 40 ℃ for 120min and at 45 ℃ for 400min, and the freeze-dried powder of the Hibiscus manihot liposome No. 1 is obtained after being taken out of the box.
In the embodiment of the disclosure, the packaging material of the mixed solution is any one of a penicillin bottle, a freeze-dried ampoule and a freeze-dried bubble cap, and in the example of the penicillin bottle, the wild hibiscus liposome is added into the protective agent solution to be stirred and dissolved, and the obtained mixed solution is placed in the penicillin bottle.
In the embodiment of the disclosure, the hibiscus lipidosome is prepared from a hibiscus root extract or a hibiscus stem extract and lecithin.
Comparative example 1
Unlike example 1, beta-cyclodextrin accounting for 2% of the mass fraction is weighed to replace lactose accounting for 2% of the mass fraction, and the mixed solution is subjected to vacuum freeze drying to obtain No. 2 Hibiscus sabdariffa liposome freeze-dried powder.
Comparative example 2
Different from example 1, 2-hydroxypropyl-beta-cyclodextrin accounting for 2% of the mass fraction is weighed to replace lactose accounting for 2% of the mass fraction, and the mixed solution is subjected to vacuum freeze drying to obtain No. 3 Hibiscus sabdariffa liposome freeze-dried powder.
Comparative example 3
Unlike example 1, which did not contain lactose component, mannose in an amount of 8% by mass, ultrapure water at 80 ℃ in an amount of 87% by mass, and Hibiscus sabdariffa liposome in an amount of 5% by mass were weighed, respectively, and the mixed solution was subjected to vacuum freeze-drying to obtain No. 4 Hibiscus sabdariffa liposome lyophilized powder.
Fig. 1 is an external view of freeze-dried powder of Hibiscus sabdariffa liposome prepared in example 1 and comparative examples 1-3 according to the preparation method of freeze-dried powder of Hibiscus sabdariffa liposome provided in the embodiment of the disclosure. Fig. 2 is a microscopic image of the freeze-dried powder of the Hibiscus sabdariffa liposome prepared in example 1 and comparative examples 1-3 according to the preparation method of the freeze-dried powder of the Hibiscus sabdariffa liposome provided in the embodiment of the disclosure after redissolution.
The appearance of the freeze-dried powder of the Hibiscus sabdariffa liposome is evaluated: the freeze-dried powder of Hibiscus sabdariffa liposome prepared in example 1 and comparative example 1-3 has appearance shown in figure 1, and its appearance artificial evaluation results are shown in table 1 below,
table 1 liposome lyophilized score for appearance
As can be seen from Table 1, the Liposome of Hibiscus sabdariffa No. 1 and No. 4 has the most smooth surface morphology and more uniform thickness.
The optical microscope characterization result of the water solution of the freeze-dried powder of the Hibiscus sabdariffa liposome is shown in figure 2, the freeze-dried powder of the No. 2 Hibiscus sabdariffa liposome has serious local agglomeration phenomenon, poor dispersion performance and uneven size of the Hibiscus sabdariffa liposome particles with larger particle size. The freeze-dried powder of the Hibiscus sabdariffa liposome has the advantages of aggregation phenomenon, poor dispersion performance, uneven size of the Hibiscus sabdariffa liposome particles with larger particle size, but obviously weaker aggregation phenomenon compared with No. 2. Compared with No. 3, the freeze-dried powder particles of the Hibiscus sabdariffa liposome No. 4 are more uniform, but a small part of the particles are larger, and the local part is slightly aggregated. Compared with No. 4, the freeze-dried powder of the Hibiscus sabdariffa liposome has uniform particles and good dispersibility. From this, it can be seen that: after freeze-drying, the quality and the degree of dispersity, the uniformity of the particle size and the size of the wild hibiscus liposome are ordered as follows: no. 1 > 4 > 3 > 2.
Therefore, the preparation method of the freeze-dried powder of the Hibiscus sabdariffa liposome provided by the embodiment of the disclosure is simple, the prepared freeze-dried powder of the Hibiscus sabdariffa liposome has good re-solubility and good dispersion performance, and particles are uniform, the particle size is small, and aggregation or agglomeration phenomenon does not exist after the re-dissolution of the Hibiscus sabdariffa liposome is observed under a microscope.
Fig. 3 is a schematic diagram showing the effect of the freeze-dried powder of wild hibiscus liposome prepared in example 1 and comparative examples 1-3 on the activity of RAW264.7 cells. Fig. 4 is a comparative schematic diagram of the effect of the freeze-dried powder of wild lotus liposome prepared in example 1 and comparative examples 1-3 on LPS-induced NO release of RAW264.7 cells according to the preparation method of the freeze-dried powder of wild lotus liposome provided in the examples of the present disclosure.
RAW264.7 (mouse mononuclear macrophage leukemia cell) cell model detects the influence of lyoprotectant on the stability of the Hibiscus sabdariffa liposome lyophilized powder: W/O (water-in-oil emulsion) Hibiscus sabdariffa liposome is easy to break emulsion and has poor stability in alcohol substances. In the embodiment of the disclosure, the comparative example adopts 1, 3-butanediol to dissolve No. 1-4 Hibiscus sabdariffa liposome lyophilized powder, and the influence of the lyophilized powder on NO release of RAW264.7 cells is detected, so that the stability difference of the Hibiscus sabdariffa liposome lyophilized powder prepared by different lyoprotectants is indirectly illustrated.
Firstly, CCK-8 is utilized to detect the influence of No. 1-4 Hibiscus sabdariffa liposome freeze-dried powder on RAW264.7 cell activity, and as shown in figure 3, no significant influence on cell activity is confirmed. And then evaluating the soothing effect of the freeze-dried powder of the Hibiscus sabdariffa liposome by using an NO detection kit: RAW264.7 cells were plated into 48-well plates, samples were added, and LPS (final concentration 1. Mu.g/mL) was added. After 24h incubation of the cells, cell supernatants were collected and assayed for NO content using the NO assay kit. As shown in fig. 4, 1, 3-butanediol itself had NO significant effect on RAW264.7 cell NO release. NO content of the Hibiscus sabdariffa liposome lyophilized powder group 1-4 is ranked from large to small as follows: no. 1 > 4 > 3 > 2. The active ingredient is wrapped by the Hibiscus sabdariffa liposome, so that a slow release effect can be achieved, and the better the Hibiscus sabdariffa liposome wrapping effect is, the higher the NO content is. This is consistent with the results shown in fig. 2. Therefore, if mannose and lactose are used as protective agents, the protective effect on the Hibiscus sabdariffa liposome is optimal, the stability of the Hibiscus sabdariffa liposome can be effectively maintained, and the Hibiscus sabdariffa liposome is easier to store.
Example 2
According to mass fraction, 5% mannose, 1% lactose, 91% ultrapure water at 80 ℃ and 3% Hibiscus sabdariffa liposome are respectively weighed. Mannose and lactose are added into ultrapure water to be stirred and dissolved, and the protective agent solution is obtained. And adding the Hibiscus sabdariffa liposome into the protective agent solution, stirring and dissolving to obtain a mixed solution. And (3) placing the mixed solution into a freeze dryer for vacuum freeze drying to obtain the freeze-dried powder of the Hibiscus sabdariffa liposome. Specifically, the vacuum freeze-drying comprises the following steps: prefreezing procedure, sublimation drying and analytical drying. In the pre-freezing procedure step, the mixed solution sequentially passes through a pre-freezing time of 90min at-3 ℃, a pre-freezing time of 120min at-45 ℃ and a pre-freezing time of 90min at-15 ℃ and enters a sublimation drying step. In the sublimation drying step, the mixed solution sequentially passes through the temperature of 0 ℃, the time of 180min, the temperature of 20 ℃, the time of 1080min and the temperature of 25 ℃ and the time of 120min, and then enters the analytical drying step. In the analytical drying step, the mixed solution is subjected to freeze-drying at 30 ℃ for 120min, at 35 ℃ for 120min, at 40 ℃ for 120min and at 45 ℃ for 400min, and the freeze-dried powder of the Hibiscus manihot liposome No. 5 is obtained after being taken out of the box.
Example 3
According to mass fraction, respectively weighing 10% mannose, 3% lactose, 80% ultrapure water at 80 ℃ and 7% Hibiscus sabdariffa liposome. Mannose and lactose are added into ultrapure water to be stirred and dissolved, and the protective agent solution is obtained. And adding the Hibiscus sabdariffa liposome into the protective agent solution, stirring and dissolving to obtain a mixed solution. And (3) placing the mixed solution into a freeze dryer for vacuum freeze drying to obtain the freeze-dried powder of the Hibiscus sabdariffa liposome. Specifically, the vacuum freeze-drying comprises the following steps: prefreezing procedure, sublimation drying and analytical drying. In the pre-freezing procedure step, the mixed solution sequentially passes through a pre-freezing time of 90min at-3 ℃, a pre-freezing time of 120min at-45 ℃ and a pre-freezing time of 90min at-15 ℃ and enters a sublimation drying step. In the sublimation drying step, the mixed solution sequentially passes through the temperature of 0 ℃, the time of 180min, the temperature of 20 ℃, the time of 1080min and the temperature of 25 ℃ and the time of 120min, and then enters the analytical drying step. In the analytical drying step, the mixed solution is subjected to freeze-drying at 30 ℃ for 120min, at 35 ℃ for 120min, at 40 ℃ for 120min and at 45 ℃ for 400min, and the freeze-dried powder of the Hibiscus manihot liposome No. 6 is obtained after being taken out of the box.
Example 4
According to mass fraction, mannose accounting for 8% of mass fraction, lactose accounting for 2% of mass fraction, ultrapure water accounting for 85% of mass fraction at 80 ℃ and Hibiscus sabdariffa liposome accounting for 5% of mass fraction are respectively weighed. Mannose and lactose are added into ultrapure water to be stirred and dissolved, and the protective agent solution is obtained. And adding the Hibiscus sabdariffa liposome into the protective agent solution, stirring and dissolving to obtain a mixed solution. And (3) placing the mixed solution into a freeze dryer for vacuum freeze drying to obtain the freeze-dried powder of the Hibiscus sabdariffa liposome. Specifically, the vacuum freeze-drying comprises the following steps: prefreezing procedure, sublimation drying and analytical drying. In the pre-freezing procedure step, the mixed solution sequentially passes through a pre-freezing time of 90min at-2 ℃, a pre-freezing time of 120min at-40 ℃ and a pre-freezing time of 90min at-14 ℃ and enters a sublimation drying step. In the sublimation drying step, the mixed solution sequentially passes through a temperature of 5 ℃, a time of 180min, a temperature of 21 ℃, a time of 1080min, and a temperature of 27 ℃ for 120min, and then enters into the analytical drying step. In the analytical drying step, the mixed solution is subjected to freeze-drying at the temperature of 33 ℃ for 120min, at the temperature of 40 ℃ for 120min, at the temperature of 45 ℃ for 120min and at the temperature of 47 ℃ for 400min in sequence, and the freeze-dried powder of the No. 7 Hibiscus sabdariffa liposome is obtained after being taken out of the box.
Example 5
According to mass fraction, 6% mannose, 3% lactose, 84% ultrapure water at 80 ℃ and 7% Hibiscus sabdariffa liposome are respectively weighed. Mannose and lactose are added into ultrapure water to be stirred and dissolved, and the protective agent solution is obtained. And adding the Hibiscus sabdariffa liposome into the protective agent solution, stirring and dissolving to obtain a mixed solution. And (3) placing the mixed solution into a freeze dryer for vacuum freeze drying to obtain the freeze-dried powder of the Hibiscus sabdariffa liposome. Specifically, the vacuum freeze-drying comprises the following steps: prefreezing procedure, sublimation drying and analytical drying. In the pre-freezing procedure step, the mixed solution sequentially passes through a pre-freezing time of 90min at-4 ℃, a pre-freezing time of 120min at-50 ℃ and a pre-freezing time of 90min at-16 ℃ and enters a sublimation drying step. In the sublimation drying step, the mixed solution sequentially passes through a temperature of-5 ℃, a time of 180min, a temperature of 18 ℃, a time of 1080min, a temperature of 23 ℃ and a time of 120min, and then enters the analytical drying step. In the analytical drying step, the mixed solution is subjected to freeze-drying at the temperature of 27 ℃ for 120min, at the temperature of 30 ℃ for 120min, at the temperature of 35 ℃, at the temperature of 120min and at the temperature of 43 ℃ for 400min in sequence, and the freeze-dried powder of the No. 8 Hibiscus manihot liposome is obtained after being taken out of the box.
Example 6
According to mass fraction, mannose 9% by mass fraction, lactose 2% by mass fraction, ultrapure water at 80 ℃ 82% by mass fraction and Hibiscus sabdariffa liposome 7% by mass fraction are weighed respectively. Mannose and lactose are added into ultrapure water to be stirred and dissolved, and the protective agent solution is obtained. And adding the Hibiscus sabdariffa liposome into the protective agent solution, stirring and dissolving to obtain a mixed solution. And (3) placing the mixed solution into a freeze dryer for vacuum freeze drying to obtain the freeze-dried powder of the Hibiscus sabdariffa liposome. Specifically, the vacuum freeze-drying comprises the following steps: prefreezing procedure, sublimation drying and analytical drying. In the pre-freezing procedure step, the mixed solution sequentially passes through a pre-freezing time of 90min at-3 ℃, a pre-freezing time of 120min at-43 ℃ and a pre-freezing time of 90min at-15 ℃ and enters a sublimation drying step. In the sublimation drying step, the mixed solution sequentially passes through the temperature of 2 ℃, the time of 180min, the temperature of 19 ℃, the time of 1080min and the temperature of 24 ℃ and the time of 120min, and then enters the analytical drying step. In the analytical drying step, the mixed solution is subjected to freeze-drying at 28 ℃ for 120min, at 33 ℃ for 120min, at 38 ℃ for 120min and at 42 ℃ for 400min in sequence, and the No. 9 Hibiscus sabdariffa liposome freeze-dried powder is obtained after being taken out of the box.
Example 7
According to mass fraction, mannose 7%, lactose 2%, ultrapure water of 80deg.C 87% and Hibiscus sabdariffa liposome 4% are weighed respectively. Mannose and lactose are added into ultrapure water to be stirred and dissolved, and the protective agent solution is obtained. And adding the Hibiscus sabdariffa liposome into the protective agent solution, stirring and dissolving to obtain a mixed solution. And (3) placing the mixed solution into a freeze dryer for vacuum freeze drying to obtain the freeze-dried powder of the Hibiscus sabdariffa liposome. Specifically, the vacuum freeze-drying comprises the following steps: prefreezing procedure, sublimation drying and analytical drying. In the pre-freezing procedure step, the mixed solution sequentially passes through a pre-freezing time of 90min at-3 ℃, a pre-freezing time of 120min at-48 ℃ and a pre-freezing time of 90min at-15 ℃ and enters a sublimation drying step. In the sublimation drying step, the mixed solution sequentially passes through a temperature of-3 ℃, a time of 180min, a temperature of 21 ℃, a time of 1080min, a temperature of 26 ℃ and a time of 120min, and then enters the analytical drying step. In the analytical drying step, the mixed solution is subjected to freeze-drying at the temperature of 32 ℃ for 120min, at the temperature of 38 ℃ for 120min, at the temperature of 42 ℃, at the temperature of 120min and at the temperature of 46 ℃ for 400min in sequence, and the freeze-dried powder of the No. 10 Hibiscus manihot liposome is obtained after being taken out of the box.
In the examples of the disclosure, the freeze-dried powder of Hibiscus sabdariffa liposome prepared in examples 2, 3, 4, 5, 6 and 7 has a slightly different appearance form and re-solubility. Under a microscope, the aggregation phenomenon does not occur, the particles are uniform, the particle size is small, and the structure of the Hibiscus sabdariffa liposome can be effectively maintained.
Release effect of demulsifier 1, 3-butanediol on soothing activity of Hibiscus sabdariffa liposome lyophilized powder 1 prepared in example 1: the No. 1 Hibiscus sabdariffa liposome freeze-dried powder is dissolved and demulsified by 2mL of 10%1, 3-butanediol, and is added into RAW264.7 cells to detect the NO content and calculate the NO release inhibition rate, and the result is shown in FIG. 5. Compared with a sample of the Hibiscus sabdariffa extract which is not coated with lecithin, the NO inhibition rate of the Hibiscus sabdariffa liposome is obviously reduced, which shows that the lecithin coating limits the release of active ingredients, and the No. 1 Hibiscus sabdariffa liposome freeze-dried powder which is processed by a freeze-drying process and demulsified enables the NO inhibition rate to rise again, which shows that the 1, 3-butanediol is taken as a demulsifier to effectively release the active ingredients of the Hibiscus sabdariffa extract, and when the Hibiscus sabdariffa liposome freeze-dried powder is used, essence/water containing 10% of 1, 3-butanediol can be added to enable the components of the Hibiscus sabdariffa extract to be released from the Hibiscus sabdariffa liposome, so that the effects of relieving and the like are achieved.
Based on the same conception, the second aspect of the embodiment of the disclosure also provides a Hibiscus sabdariffa liposome lyophilized powder, which is prepared by the preparation method of the Hibiscus sabdariffa liposome lyophilized powder.
The third aspect of the embodiments of the present disclosure further provides an application of the freeze-dried powder of the hibiscus liposome, which is prepared by the preparation method of the freeze-dried powder of the hibiscus liposome provided by the embodiments of the present disclosure, and the freeze-dried powder of the hibiscus liposome is applied in cosmetics, for example, whitening skin care products, such as cream, emulsion, essence, mask, and the like. In the embodiment of the disclosure, the freeze-dried powder of the Hibiscus sabdariffa liposome is used by being compounded with the cosmetic water or essence, or the freeze-dried powder of the Hibiscus sabdariffa liposome is used by being compounded with a 1, 3-butanediol aqueous solution or essence with the mass fraction of 10%, so that the components of the Hibiscus sabdariffa extract are released from the Hibiscus sabdariffa liposome, the re-solubility of the Hibiscus sabdariffa liposome is improved, the efficacy stability of the cosmetic is maintained, and the transport and the storage are easy.
It is understood that the term "plurality" in this disclosure means two or more, and other adjectives are similar thereto. "and/or", describes an association relationship of an association object, and indicates that there may be three relationships, for example, a and/or B, and may indicate: a exists alone, A and B exist together, and B exists alone. The character "/" generally indicates that the context-dependent object is an "or" relationship. The singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.
It is further understood that the terms "first," "second," and the like are used to describe various information, but such information should not be limited to these terms. These terms are only used to distinguish one type of information from another and do not denote a particular order or importance. Indeed, the expressions "first", "second", etc. may be used entirely interchangeably. For example, first information may also be referred to as second information, and similarly, second information may also be referred to as first information, without departing from the scope of the present disclosure.
It will be further understood that the terms "center," "longitudinal," "transverse," "front," "rear," "upper," "lower," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like, as used herein, refer to an orientation or positional relationship based on that shown in the drawings, merely for convenience in describing the present embodiments and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed and operate in a particular orientation.
It will be further understood that "connected" includes both direct connection where no other member is present and indirect connection where other element is present, unless specifically stated otherwise.
It will be further understood that although operations are depicted in the drawings in a particular order, this should not be understood as requiring that such operations be performed in the particular order shown or in sequential order, or that all illustrated operations be performed, to achieve desirable results. In certain circumstances, multitasking and parallel processing may be advantageous.
Other embodiments of the disclosure will be apparent to those skilled in the art from consideration of the specification and practice of the disclosure disclosed herein. This application is intended to cover any adaptations, uses, or adaptations of the disclosure following, in general, the principles of the disclosure and including such departures from the present disclosure as come within known or customary practice within the art to which the disclosure pertains. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the disclosure being indicated by the following claims.
It is to be understood that the present disclosure is not limited to the precise arrangements and instrumentalities shown in the drawings, and that various modifications and changes may be effected without departing from the scope thereof. The scope of the present disclosure is limited only by the appended claims.
Claims (6)
1. The preparation method of the freeze-dried powder of the Hibiscus sabdariffa liposome is characterized by comprising the following steps:
according to the mass fraction, mannose with the mass fraction of 5-10 percent and lactose with the mass fraction of 1-3 percent are mixed with the rest ultrapure water, heated, stirred and dissolved to obtain a protective agent solution;
adding 3-7% of Hibiscus sabdariffa liposome into the protective agent solution, stirring and dissolving to obtain mixed solution, wherein the Hibiscus sabdariffa liposome is lecithin-coated root extract or stem extract of Hibiscus sabdariffa;
placing the mixed solution in a freeze dryer for vacuum freeze drying to obtain freeze-dried powder of the Hibiscus sabdariffa liposome, wherein the vacuum freeze drying comprises a pre-freezing procedure, sublimation drying and analytical drying;
the pre-freezing procedure comprises the steps of sequentially passing through a temperature of-3+/-1 ℃ and a pre-freezing time of 90min, a temperature of-45+/-5 ℃ and a pre-freezing time of 120min and a temperature of-15+/-1 ℃ and a pre-freezing time of 90 min;
the sublimation drying comprises the steps of sequentially passing through a temperature of 0+/-5 ℃ for 180min, a temperature of 18+/-3 ℃ for 1080min and a temperature of 25+/-2 ℃ for 120 min;
the analytical drying comprises the steps of sequentially passing through the temperature of 30+/-3 ℃ for 120min, the temperature of 35+/-5 ℃ for 120min, the temperature of 40+/-5 ℃ for 120min and the temperature of 45+/-2 ℃ for 400min.
2. The preparation method of the Hibiscus sabdariffa liposome freeze-dried powder according to claim 1, which is characterized in that:
the packaging material of the mixed solution is any one of a penicillin bottle, a freeze-drying ampoule and a freeze-drying bubble cap.
3. The freeze-dried powder of the Hibiscus sabdariffa liposome is characterized by comprising the following components: the freeze-dried powder of the Hibiscus sabdariffa liposome is prepared by the preparation method of the freeze-dried powder of the Hibiscus sabdariffa liposome according to any one of claims 1 to 2.
4. The application of the freeze-dried powder of the Hibiscus sabdariffa liposome is characterized in that: the freeze-dried powder of the Hibiscus sabdariffa liposome is described in claim 3, and the freeze-dried powder of the Hibiscus sabdariffa liposome is applied to the preparation of cosmetics.
5. The application of the Hibiscus sabdariffa liposome freeze-dried powder according to claim 4, which is characterized in that:
the freeze-dried powder of the Hibiscus sabdariffa liposome is compounded with cosmetic water or essence.
6. The application of the Hibiscus sabdariffa liposome freeze-dried powder according to claim 4, which is characterized in that:
the freeze-dried powder of the Hibiscus sabdariffa liposome is compounded with 10% of aqueous solution or essence of 1, 3-butanediol by mass fraction.
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| TWM526845U (en) * | 2016-01-20 | 2016-08-11 | Tenart Biotech Ltd | Conditioning membrane with Taiwan cotton-rose extract essence |
| CN112076126A (en) * | 2020-09-28 | 2020-12-15 | 嘉文丽（福建）化妆品有限公司 | Separation method of whitening functional components in hibiscus mutabilis and application of whitening functional components in cosmetics |
| TWM627072U (en) * | 2022-01-19 | 2022-05-11 | 百草禾生技有限公司 | Hibiscus taiwanensis S.Y. Hu extract embedding structure |
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| TWM526845U (en) * | 2016-01-20 | 2016-08-11 | Tenart Biotech Ltd | Conditioning membrane with Taiwan cotton-rose extract essence |
| CN112076126A (en) * | 2020-09-28 | 2020-12-15 | 嘉文丽（福建）化妆品有限公司 | Separation method of whitening functional components in hibiscus mutabilis and application of whitening functional components in cosmetics |
| TWM627072U (en) * | 2022-01-19 | 2022-05-11 | 百草禾生技有限公司 | Hibiscus taiwanensis S.Y. Hu extract embedding structure |
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