LU101863B1 - Nano-liposome simultaneously containing lutein and cordyceps militaris alcohol extract, and preparation method therefor - Google Patents
Nano-liposome simultaneously containing lutein and cordyceps militaris alcohol extract, and preparation method therefor Download PDFInfo
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- LU101863B1 LU101863B1 LU101863A LU101863A LU101863B1 LU 101863 B1 LU101863 B1 LU 101863B1 LU 101863 A LU101863 A LU 101863A LU 101863 A LU101863 A LU 101863A LU 101863 B1 LU101863 B1 LU 101863B1
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- Luxembourg
- Prior art keywords
- lutein
- alcohol extract
- cordyceps militaris
- liposome
- nano
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title claims abstract description 78
- 235000012680 lutein Nutrition 0.000 title claims abstract description 64
- 229960005375 lutein Drugs 0.000 title claims abstract description 64
- 239000001656 lutein Substances 0.000 title claims abstract description 64
- KBPHJBAIARWVSC-RGZFRNHPSA-N lutein Chemical compound C([C@H](O)CC=1C)C(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\[C@H]1C(C)=C[C@H](O)CC1(C)C KBPHJBAIARWVSC-RGZFRNHPSA-N 0.000 title claims abstract description 64
- ORAKUVXRZWMARG-WZLJTJAWSA-N lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C ORAKUVXRZWMARG-WZLJTJAWSA-N 0.000 title claims abstract description 64
- KBPHJBAIARWVSC-XQIHNALSSA-N trans-lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C KBPHJBAIARWVSC-XQIHNALSSA-N 0.000 title claims abstract description 64
- FJHBOVDFOQMZRV-XQIHNALSSA-N xanthophyll Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C=C(C)C(O)CC2(C)C FJHBOVDFOQMZRV-XQIHNALSSA-N 0.000 title claims abstract description 64
- 239000002502 liposome Substances 0.000 title claims abstract description 62
- 241001264174 Cordyceps militaris Species 0.000 title claims abstract description 48
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 238000000605 extraction Methods 0.000 title 1
- 238000000034 method Methods 0.000 claims abstract description 13
- 235000019441 ethanol Nutrition 0.000 claims description 52
- 239000000725 suspension Substances 0.000 claims description 20
- 229920000053 polysorbate 80 Polymers 0.000 claims description 18
- 239000000243 solution Substances 0.000 claims description 17
- 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 claims description 16
- 239000000203 mixture Substances 0.000 claims description 13
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 12
- KQLDDLUWUFBQHP-UHFFFAOYSA-N Cordycepin Natural products C1=NC=2C(N)=NC=NC=2N1C1OCC(CO)C1O KQLDDLUWUFBQHP-UHFFFAOYSA-N 0.000 claims description 11
- OFEZSBMBBKLLBJ-BAJZRUMYSA-N cordycepin Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)C[C@H]1O OFEZSBMBBKLLBJ-BAJZRUMYSA-N 0.000 claims description 11
- OFEZSBMBBKLLBJ-UHFFFAOYSA-N cordycepine Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(CO)CC1O OFEZSBMBBKLLBJ-UHFFFAOYSA-N 0.000 claims description 11
- 235000010469 Glycine max Nutrition 0.000 claims description 10
- 244000068988 Glycine max Species 0.000 claims description 10
- 239000011259 mixed solution Substances 0.000 claims description 10
- 235000012000 cholesterol Nutrition 0.000 claims description 8
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 claims description 7
- 239000008347 soybean phospholipid Substances 0.000 claims description 7
- 230000000887 hydrating effect Effects 0.000 claims description 5
- 238000003760 magnetic stirring Methods 0.000 claims description 5
- 239000008055 phosphate buffer solution Substances 0.000 claims description 5
- 238000001291 vacuum drying Methods 0.000 claims description 5
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 claims description 4
- 239000002126 C01EB10 - Adenosine Substances 0.000 claims description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 2
- 229960005305 adenosine Drugs 0.000 claims description 2
- 230000001804 emulsifying effect Effects 0.000 claims description 2
- 238000005303 weighing Methods 0.000 claims description 2
- 238000011068 loading method Methods 0.000 abstract description 15
- 239000002245 particle Substances 0.000 abstract description 11
- 239000013543 active substance Substances 0.000 abstract description 8
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- 238000005516 engineering process Methods 0.000 abstract description 3
- 238000002347 injection Methods 0.000 abstract description 3
- 239000007924 injection Substances 0.000 abstract description 3
- 230000009286 beneficial effect Effects 0.000 abstract description 2
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- 230000035764 nutrition Effects 0.000 abstract description 2
- 238000009776 industrial production Methods 0.000 abstract 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 229940079593 drug Drugs 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 230000000259 anti-tumor effect Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 241000190633 Cordyceps Species 0.000 description 2
- UPYKUZBSLRQECL-UKMVMLAPSA-N Lycopene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1C(=C)CCCC1(C)C)C=CC=C(/C)C=CC2C(=C)CCCC2(C)C UPYKUZBSLRQECL-UKMVMLAPSA-N 0.000 description 2
- JEVVKJMRZMXFBT-XWDZUXABSA-N Lycophyll Natural products OC/C(=C/CC/C(=C\C=C\C(=C/C=C/C(=C\C=C\C=C(/C=C/C=C(\C=C\C=C(/CC/C=C(/CO)\C)\C)/C)\C)/C)\C)/C)/C JEVVKJMRZMXFBT-XWDZUXABSA-N 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
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- 238000005538 encapsulation Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 235000012661 lycopene Nutrition 0.000 description 2
- OAIJSZIZWZSQBC-GYZMGTAESA-N lycopene Chemical compound CC(C)=CCC\C(C)=C\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C=C(/C)CCC=C(C)C OAIJSZIZWZSQBC-GYZMGTAESA-N 0.000 description 2
- 229960004999 lycopene Drugs 0.000 description 2
- 239000001751 lycopene Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
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- 239000006228 supernatant Substances 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- ZCIHMQAPACOQHT-ZGMPDRQDSA-N trans-isorenieratene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/c1c(C)ccc(C)c1C)C=CC=C(/C)C=Cc2c(C)ccc(C)c2C ZCIHMQAPACOQHT-ZGMPDRQDSA-N 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- JQWAHKMIYCERGA-UHFFFAOYSA-N (2-nonanoyloxy-3-octadeca-9,12-dienoyloxypropoxy)-[2-(trimethylazaniumyl)ethyl]phosphinate Chemical compound CCCCCCCCC(=O)OC(COP([O-])(=O)CC[N+](C)(C)C)COC(=O)CCCCCCCC=CCC=CCCCCC JQWAHKMIYCERGA-UHFFFAOYSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 239000000232 Lipid Bilayer Substances 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
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- 235000021466 carotenoid Nutrition 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
- A23L31/10—Yeasts or derivatives thereof
- A23L31/15—Extracts
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/035—Organic compounds containing oxygen as heteroatom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/05—Organic compounds containing phosphorus as heteroatom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/10—Foods or foodstuffs containing additives; Preparation or treatment thereof containing emulsifiers
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
- A61K31/07—Retinol compounds, e.g. vitamin A
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/062—Ascomycota
- A61K36/066—Clavicipitaceae
- A61K36/068—Cordyceps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
Disclosed is a nano-liposome simultaneously containing lutein and a cordyceps militaris alcohol extract, which relates to the field of food nutrition and health. A mass percentage content of the cordyceps militaris alcohol extract in the nano-liposome of the invention is 4% to 8%, and a mass ratio of the lutein to the cordyceps militaris alcohol extract is 1:3 to 1:5. According to the invention, an ethanol injection method is combined with a microfluidization technology to jointly load two active substances including the lutein and the cordyceps militaris alcohol extract which have different hydrophobicities into the nano-liposome, so that the prepared liposome has a particle size less than 200 nm, a loading rate of the lutein greater than 90%, a loading rate of the cordyceps militaris alcohol extract greater than 85%, has a relatively good stability, a safe and simple preparation technology and a low cost, and is beneficial for industrial production.
Description
lu101863 NANO-LIPOSOME SIMULTANEOUSLY CONTAINING LUTEIN AND | CORDYCEPS MILITARIS ALCOHOL EXTRACT, AND PREPARATION METHOD | THEREFOR
TECHNICAL FIELD | 5 The present invention relates to the field of food nutrition and health, in particular to a nano- | liposome simultaneously containing lutein and a cordyceps militaris alcohol extract, and a preparation method therefor. | BACKGROUND | Study shows that intaking of a certain amount of lutein from foods is related to reducing the risks of AMD, cancers and cardiovascular diseases; and a cordyceps militaris alcohol extract may inhibit proliferation of a variety of human tumor cell lines, wherein cordycepin is known as a major anti-tumor component, and can more effectively induce tumor cell apoptosis in cooperation with other anti-tumor components. In order to improve an in-vivo stability, a solubility and a bioavailability of the two natural active substances so as to better exert a pharmacological effect thereof, a novel nano-drug delivery system has become a research hotspot and a research difficulty in pharmacy and food industry in recent years. Producers prefer a self-assembled colloid delivery system based on lipid due to a safety of lipid molecules, a capability of enhancing transcellular transportation and improvement of paracellular drug transportation. A liposome has a structure similar to a biological membrane, and can improve an in-vivo solubility of active substances, reduce a toxicity caused by the active substances, and realize continuous and specific targeted delivery; and the liposome is able to deliver lipophilic and hydrophilic compounds. At present, a total of 29 nano-liposome drugs in the world have been approved in the market, showing a broad application prospect. It has been reported in literatures that a lutein nano-liposome with a particle size less than 230 nm prepared by an ethanol injection method can improve a dispersity and a bioavailability of lutein (Preparation process optimization and oxidation stability of lutein nano-liposome [J]. Food Science, 2017, 38(18): 259-265). A mixture of lutein and lycopene is jointly loaded into the liposome to play a role of synergistic antioxidation (Carotenoid mixtures protect multilamellar liposomes against oxidative damage: synergistic effects of lycopene and lutein [J]. FEBS Letters, 1998, 427(2): 305-308). In an intellectual property aspect, Chinese patent (publication No.: CN105726482A, and publication date: July 6, 2016) discloses a lutein nano-liposome and a preparation method therefor, and the lutein nano-liposome is prepared by high-pressure homogenization, ultrasonic dispersion and hydration, and millipore filtration of lutein crystalline powder, egg yolk lecithin and cholesterol. Chinese patent (publication No.: CN106798725A, and publication date: June 6, 2017) discloses a method for preparing a cordycepin | 35 — nano-liposome, which combines a pH gradient method with a reverse evaporating method to prepare | the liposome, so that an in-vivo activity of the cordycepin is well protected and hepatoma cells|gre1863 ; obviously inhibited, and the liposome obtained by the method has a high encapsulation efficiency ‘ (290%), an even particle size and a good stability. | In conventional process and technical solutions, issues such as single encapsulation of the active | 5 substances, relatively poor pharmacology and drug efficacy, and the like in the case that two drugs | including the lutein and the cordyceps militaris alcohol extract which have different hydrophobicities | are synergistically loaded into the liposome are not considered. Development of many kinds of | healthy functional foods makes it necessary to design a carrier that simultaneously encapsulates a plurality of active substances. | 10 SUMMARY | An objective of the present invention is to provide a nano-liposome simultaneously containing | lutein and a cordyceps militaris alcohol extract, and a preparation method therefor, so as to jointly | load active substances which have different hydrophobicities into the nano-liposome, and improve a loading rate and a stability of the lutein and the cordyceps militaris alcohol extract. The objective of the present invention is achieved by the following ways. A method for preparing a nano-liposome simultaneously containing lutein and a cordyceps | militaris alcohol extract includes the following preparation steps: 1) weighing soybean phospholipid, cholesterol and Tween 80 according to a mass ratio of 4:1:1, and fully dissolving the soybean phospholipid, the cholesterol and the Tween 80 into absolute ethyl alcohol to obtain a soybean phospholipid-cholesterol-Tween 80 mixed solution; 2) slowly adding 5 mL to 10 mL of lutein-ethanol solution into 15 mL to 25 mL of the soybean | phospholipid-cholesterol-Tween 80 mixed solution in the step 1) under magnetic stirring of 400 rpm, and homogeneously emulsifying the mixture at 9000 rpm for 2 minutes to 4 minutes to obtain a lutein suspension; 3) performing vacuum concentration on the lutein suspension in the step 2) at 35°C, and forming a pale yellow and transparent film by vacuum drying protected from light; 4) adding 20 mL to 30 mL of phosphate buffer solution containing the cordyceps militaris alcohol extract into the dry film formed in the step 3), and performing vacuum rotating and hydrating on the mixture at 40 rpm at 35°C to obtain a crude liposome suspension containing the lutein and the cordyceps militaris alcohol extract; and 5) performing high-pressure microfluidization on the crude liposome suspension obtained in the step 4) twice at 9,000 psi to 12,000 psi to obtain a nano-liposome solution containing the lutein and the cordyceps militaris alcohol extract.
According to the present invention, a mass percentage content of the cordyceps militaris alcohol CT.
| 3 extract in the nano-liposome simultaneously containing the lutein and the cordyceps militaris alcqhpd1863 extract is 4% to 8%, and a mass ratio of the lutein to the cordyceps militaris alcohol extract is 1:3 to | 1:5. | The cordyceps militaris alcohol extract is extracted and obtained by assistance of ultrasonic | 5 wave with an ethanol solution with a volume ratio ranging from 50% to 75%, and the alcohol extract contains cordycepin, adenosine and cordycepic acid. | : Beneficial effects | An ethanol injection method is combined with a microfluidization technology to jointly load two | active substances including the lutein and the cordyceps militaris alcohol extract which have different hydrophobicities into the nano-liposome to an optimal degree according to a certain mass ratio, so that the prepared liposome has a particle size less than 200 nm, a loading rate of the lutein greater than 90%, a loading rate of the cordyceps militaris alcohol extract greater than 85%, and has a relatively good stability. Moreover, compared with the liposome containing a single active substance, a double-layer embedded liposome (FIG. 1) may synergistically exert stronger anti-tumor activity, and plays a variety of roles good to the health in the body.
BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a structure diagram of a nano-liposome simultaneously containing lutein and a cordyceps militaris alcohol extract according to the present invention; FIG. 2 is a particle size distribution diagram of the nano-liposome simultaneously containing the lutein and the cordyceps militaris alcohol extract according to the present invention; and FIG. 3 is a photo of a lutein-cordyceps militaris alcohol extract nano-liposome solution according to the present invention. In FIG. 1, 1 refers to cordyceps militaris alcohol extract; 2 refers to lutein; 3 refers to lipid bilayer; and 4 refers to Tween-80. | DETAILED DESCRIPTION The present invention provides a nano-liposome simultaneously containing lutein and a cordyceps militaris alcohol extract, and a preparation method therefor, and the present invention is | 30 further described hereinafter with reference to the specific embodiments. Embodiment 1 1) Soybean phospholipid, cholesterol and Tween 80 were weighed according to a mass ratio of 4:1:1, and fully dissolved into absolute ethyl alcohol to obtain a soybean phospholipid-cholesterol- Zn
! 4 Tween 80 mixed solution. 1u101863 | 2) 5 mL of lutein-ethanol solution was slowly added into 20 mL of the soybean phospholipid- | cholesterol-Tween 80 mixed solution in the step 1) under magnetic stirring of 400 rpm, and the mixture was homogeneously emulsified at 9,000 rpm for 2 minutes to obtain a lute suspension. 3) Vacuum concentration was performed on the lutein suspension in the step 2) at 35°C, and a pale yellow and transparent film was formed by vacuum drying protected from light.
4) 20 mL of phosphate buffer solution containing the cordyceps militaris alcohol extract was added into the dry film formed in the step 3), and vacuum rotating and hydrating were performed on the mixture at 40 rpm at 35°C to obtain a crude liposome suspension containing the lutein and the cordyceps militaris alcohol extract.
5) High-pressure microfluidization was performed on the crude liposome suspension obtained in the step 4) twice at 10,000 psi to obtain a nano-liposome solution containing the lutein and the cordyceps militaris alcohol extract (FIG. 3).
A PSS nano laser particle size analyzer was used to measure a particle size of the liposome by a measurement method as follows: a liposome solution sample was diluted to 100 times with deionized water, 4 mL of the diluted liposome solution sample was added into a polystyrene tank and balanced for 2 minutes, a measured temperature was 25°C, a nature of water was used as a measurement parameter, and a viscosity and a refractive index of the solution were 0.933 CP and 1.333 respectively. See FIG. 2 for a particle size distribution diagram.
500 uL of newly prepared lutein-cordyceps militaris extract nano-liposome was added into a 2 mL centrifuge tube and centrifuged at 1,000 rpm for 30 minutes; after centrifugation, 250 pL of supernatant was added into a 10 mL centrifuge tube, and then 2 mL of DCM (dichloromethane: | methyl alcohol=2:1, V:V) solvent was added. After vortex oscillation for 5 minutes, the mixture was | extracted for three times with 1 mL of normal hexane, extract liquids were mixed and filtered with a
0.45 um organic system filter head, an absorbance value was measured at a maximum absorption wavelength of 460 nm, and a content of the lutein was calculated, which was namely a content of free lutein. In addition, 500 uL of the liposome was added with methyl alcohol to a volume of 5 mL, and subjected to 100 W ultrasonic processing for 30 minutes till demulsification, and 250 pL of the demulsified liquid was added into a 10 mL centrifuge tube. The above method for extracting the lutein was repeated to obtain the content of the lutein by calculation, and the content was namely a total content of the lutein in 250 pL of the liquid added. A loading rate (%) of the lutein was calculated according to a formula as follows: Loading rate of lutein = Total content of lutein — Content of free lutein Total content of lutein The cordycepin was used as an index of an embedding rate of the cordyceps militaris alcohol | C—O
| [ extract, and 500 uL of newly prepared lutein-cordyceps militaris alcohol extract nano-liposome wa 863 | added into a 2 mL centrifuge tube, and centrifuged at 10,000 rpm for 30 minutes. After centrifugation, | a supernatant was firstly filtered through a 0.45 pm filter membrane, and then filtered through a 0.22 | um filter membrane to remove solid impurities. After analysis by a high performance liquid chromatography-ultraviolet detector, a wavelength was detected to be 260 nm. À content of free cordycepin was calculated. 500 pL of the liposome was additionally added with 0.1% Triton X-100 to a volume of 5 mL, and subjected to 100 W ultrasonic processing for 30 minutes till demulsification. The above filtering process was repeated, and a total content of the cordycepin in 500 pL of the liposome was analyzed by the high performance liquid chromatography-ultraviolet detector and calculated. A loading rate (%) of the cordyceps militaris alcohol extract was calculated according to a formula as follows: Loading rate of cordyceps ~~ Toal content of cordycepin — Content of free cordycepin militaris alcohol extract (%) — Toal content of cordycepin 100% An average particle size of a lutein-cordyceps militaris alcohol extract composite nano-liposome was measured to be 131 nm + 72 nm by the above method, the loading rate of the lutein was 98.2%, and the loading rate of the cordyceps militaris alcohol extract was 89.1%.
Embodiment 2 1) Soybean phospholipid, cholesterol and Tween 80 were weighed according to a mass ratio of | 4:1:1, and fully dissolved into absolute ethyl alcohol to obtain a soybean phospholipid-cholesterol- Tween 80 mixed solution.
© 2) 10 mL of lutein-ethanol solution was slowly added into 20 mL of the soybean phospholipid- cholesterol-Tween 80 mixed solution in the step 1) under magnetic stirring of 400 rpm, and the mixture was homogeneously emulsified at 9,000 rpm for 3 minutes to obtain a lutein suspension.
3) Vacuum concentration was performed on the lutein suspension in the step 2) at 35°C, and a pale yellow and transparent film was formed by vacuum drying protected from light.
4) 30 mL of phosphate buffer solution containing the cordyceps militaris alcohol extract was added into the dry film formed in the step 3), and vacuum rotating and hydrating were performed on the mixture at 40 rpm at 35°C to obtain a crude liposome suspension containing the lutein and the cordyceps militaris alcohol extract.
5) High-pressure microfluidization was performed on the crude liposome suspension obtained in the step 4) once at 12,000 psi to obtain a nano-liposome solution containing the lutein and the cordyceps militaris alcohol, wherein a particle size of the liposome was 163 nm = 69 nm, a loading rate of the lutein was 93.7%, and a loading rate of the cordyceps militaris alcohol extract was 88.4%.
Embodiment 3
1) Soybean phospholipid, cholesterol and Tween 80 were weighed according to a mass ratidwt01863 4:1:1, and fully dissolved into absolute ethyl alcohol to obtain a soybean phospholipid-cholesterol- Tween 80 mixed solution. 2) 8 mL of lutein-ethanol solution was slowly added into 15 mL of the soybean phospholipid- cholesterol-Tween 80 mixed solution in the step 1) under magnetic stirring of 400 rpm, and the mixture was homogeneously emulsified at 9,000 rpm for 4 minutes to obtain a lutein suspension. 3) Vacuum concentration was performed on the lutein suspension in the step 2) at 35°C, and a pale yellow and transparent film was formed by vacuum drying protected from light. 4) 25 mL of phosphate buffer solution containing the cordyceps militaris alcohol extract was added into the dry film formed in the step 3), and vacuum rotating and hydrating were performed on the mixture at 40 rpm at 35°C to obtain a crude liposome suspension containing the lutein and the cordyceps militaris alcohol extract. 5) High-pressure microfluidization was performed on the crude liposome suspension obtained in the step 4) for three times at 9,000 psi to obtain a nano-liposome solution containing the lutein and the cordyceps militaris alcohol, wherein a particle size of the liposome was 203 nm + 89 nm, a loading rate of the lutein was 92.5%, and a loading rate of the cordyceps militaris alcohol extract was 87.3%.
Claims (3)
1. A nano-liposome simultaneously containing lutein and a cordyceps militaris alcohol extract, wherein a mass percentage content of the cordyceps militaris alcohol extract in the nano-liposome is 4% to 8%, and a mass ratio of the lutein to the cordyceps militaris alcohol extract is 1:3 to 1:5.
2. The nano-liposome simultaneously containing the lutein and the cordyceps militaris alcohol extract according to claim 1, wherein the cordyceps militaris alcohol extract is extracted and obtained by assistance of ultrasonic wave with an ethanol solution with a volume ratio ranging from 50% to 75%, and the alcohol extract contains cordycepin, adenosine and cordycepic acid.
3. A method for preparing the nano-liposome simultaneously containing the lutein and the cordyceps militaris alcohol extract according to claim 1, comprising the following preparation steps: 1) weighing soybean phospholipid, cholesterol and Tween 80 according to a mass ratio of 4:1:1, and fully dissolving the soybean phospholipid, the cholesterol and the Tween 80 into absolute ethyl alcohol to obtain a soybean phospholipid-cholesterol- Tween 80 mixed solution; 2) slowly adding 5 mL to 10 mL of lutein-ethanol solution into 15 mL to 25 mL of the soybean phospholipid-cholesterol-Tween 80 mixed solution into the step 1) under magnetic stirring of 400 rpm, and homogeneously emulsifying the mixture at 9,000 rpm for 2 minutes to 4 minutes to obtain a lutein suspension; 3) performing vacuum concentration on the lutein suspension in the step 2) at 35°C, and forming a pale yellow and transparent film by vacuum drying protected from light; 4) adding 20 mL to 30 mL of phosphate buffer solution containing the cordyceps militaris alcohol extract into the dry film formed in the step 3), and performing vacuum rotating and hydrating on the mixture at 40 rpm at 35°C to obtain a crude liposome suspension containing the lutein and the cordyceps militaris alcohol extract; and 5) performing high-pressure microfluidization on the crude liposome suspension obtained in the step 4) twice at 9,000 psi to 12,000 psi to obtain a nano-liposome solution containing the lutein and the cordyceps militaris alcohol extract.
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| IT1269569B (en) * | 1994-04-22 | 1997-04-08 | Ugo Citernesi | PROCEDURE FOR THE PREPARATION OF COMPLEX BETWEEN PHOSPHOLIPIDS AND ACTIVE PRINCIPLES USEFUL FOR THE PRODUCTION OF LIPOSOMES AND ACTIVE PRINCIPLES AND LIPOSOMES OBTAINED WITH THE PROCEDURE |
| CN1294133A (en) * | 2000-11-06 | 2001-05-09 | 吉林东北虎药业股份有限公司 | Process for extracting cordycepin and its preparation |
| CN101816632B (en) * | 2010-04-23 | 2012-08-22 | 广东药学院 | Composite functional liposome and preparation method and application thereof |
| CN102793177B (en) * | 2012-09-07 | 2013-11-27 | 广州安德生物科技有限公司 | Biological adhesion type cordyceps militaris liposome and preparation method thereof |
| KR20150034517A (en) * | 2013-09-26 | 2015-04-03 | 삼성전자주식회사 | Liposome comprising a complex of a hydrophobic active ingredient and a polypeptide, and use thereof |
| CN103689554B (en) * | 2013-12-25 | 2016-02-10 | 广州安德健康科技有限公司 | A kind of preparation method of Cordyceps militaris liposome and application thereof |
| CN106798725A (en) * | 2015-11-26 | 2017-06-06 | 王春梅 | A kind of cordycepin nano liposomes and preparation method thereof and antitumor activity application |
| CN105726482A (en) * | 2016-01-27 | 2016-07-06 | 江苏省农业科学院 | Xanthophyll nano-liposome and preparation method thereof |
| CN107927739A (en) * | 2017-12-26 | 2018-04-20 | 北京牧笛生物科技有限公司 | A kind of food enrichment and preparation method thereof |
| CN108851072A (en) * | 2018-04-18 | 2018-11-23 | 江苏师范大学 | A kind of purple sweet potato cyanidin nano liposomes and preparation method thereof |
| CN110302161B (en) * | 2019-07-08 | 2021-10-26 | 大连理工大学 | Composite nano liposome and application thereof |
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