CN113750126A - Artemisia annua essential oil and white tea essential oil for treating skin herpes and composition thereof - Google Patents
Artemisia annua essential oil and white tea essential oil for treating skin herpes and composition thereof Download PDFInfo
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- CN113750126A CN113750126A CN202111163300.8A CN202111163300A CN113750126A CN 113750126 A CN113750126 A CN 113750126A CN 202111163300 A CN202111163300 A CN 202111163300A CN 113750126 A CN113750126 A CN 113750126A
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- 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
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/282—Artemisia, e.g. wormwood or sagebrush
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- 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
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/82—Theaceae (Tea family), e.g. camellia
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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
- A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
- A61K8/922—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof of vegetable origin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
- A61P31/22—Antivirals for DNA viruses for herpes viruses
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- 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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
-
- 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
- A61K2800/10—General cosmetic use
Abstract
The invention relates to sweet wormwood essential oil and white tea essential oil for treating skin herpes and a composition thereof. The sweet wormwood essential oil is prepared by the following method: collecting fresh herba Artemisiae Annuae tender seedling, collecting leaves, drying, cutting into pieces, soaking, and distilling at a material-liquid ratio of 1 (0.5-2) for 60-90 min. The white tea essential oil is prepared by adopting the following methodPreparing: collecting fresh white tea tender shoots, withering in a backlight room at 30-35 deg.C and 50-60% humidity until weight loss is 40-60%, drying, and subjecting to supercritical CO2And (4) fluid extraction. The invention further analyzes the components of the obtained essential oil by GC-MS. The effects of the sweet wormwood essential oil, the white tea essential oil and the composition thereof on the skin herpes viruses are also detected, and the two have obvious inhibition effects on the skin herpes viruses and have certain synergistic effect when combined. The invention provides a method for preparing a medicament or a cosmetic for inhibiting skin herpes by using the sweet wormwood essential oil, the white tea essential oil and the composition thereof.
Description
Technical Field
The invention relates to extraction and application of plant essential oil, in particular to sweet wormwood essential oil and white tea essential oil for treating skin herpes, a composition of the sweet wormwood essential oil and the white tea essential oil, and application of the sweet wormwood essential oil and the white tea essential oil in resisting skin herpes viruses and treating skin herpes.
Background
Skin herpes (cutaneous skin) is a skin lesion caused by herpes simplex virus infection and can occur anywhere in the body, especially at the skin mucosal junction, such as the lips and corners of the mouth. Clinically, the rice grains are clustered into blisters, which are not fused, have thin blister walls and clear liquid. The ulcer, erosion and scab are broken in a short period, and no scar is generally left. The total course of disease is 2-3 weeks. The attack may be repeated. Herpes simplex virus type 1 (HSV-1) causes herpes labialis, gingivitis, keratitis, encephalitis, and the like.
The plant essential oil belongs to a safe, natural and pollution-free excellent plant source product, and has various effects of sterilization, disinsection, moistening and the like. Journal articles (topical treatment of viral infections such as herpes labialis with mixtures of essential oils [ J ] foreign medicine: botanical drug Manual, 2004(2):82-82.) disclose mixtures consisting of 2 or more of the following plant essential oils or their major components: the mint, cajeput, eucalyptus, juniper and holly have the lowest concentration of 30 percent when being applied, are externally used for treating skin or mucosal virus infection, in particular to I-type herpes labialis infection, have the advantages of quick response and high curative effect compared with the traditional antiviral drug (penciclovir), can reduce the recurrence rate of the herpes labialis during medium-term and long-term treatment, have no side effect, and have the functions of relieving pain and local anesthesia. Patent document CN108498378A discloses a compound plant essential oil for repelling mosquitoes, relieving itching and reducing swelling, which comprises the following raw materials: folium artemisiae argyi essential oil, lemon eucalyptus essential oil, clove essential oil, camellia seed essential oil, catnip essential oil, thyme essential oil, geranium essential oil, rosemary essential oil, castor oil, citronella oil, litsea cubeba essential oil, lavender essential oil, lithospermum essential oil, clary sage essential oil, spearmint essential oil, cypress essential oil, platycladus orientalis essential oil, calendula officinalis essential oil, cedar essential oil, juniper berry essential oil, wild chrysanthemum essential oil, motherwort essential oil, eucommia ulmoides essential oil, jasmine essential oil, eucalyptus essential oil, valerian essential oil, rose essential oil, sweet osmanthus essential oil and lemon grass essential oil. The plant essential oil has obvious mosquito repellent effect, high itching relieving and swelling eliminating speed, obvious improvement effect on eczema, suppuration, herpes, whelk, furuncle, swelling and skin pruritus, and obvious antibacterial and anti-inflammatory effects.
The herba Artemisiae Annuae essential oil is volatile oil extracted from herba Artemisiae Annuae (Artemisia annua L.). Modern researches have proved that the volatile oil of sweet wormwood has the activities of antibiosis, anticancer, antivirus, anti-inflammatory, mosquito repellent, insect killing, antioxidation and the like. The main uses are acne treatment, influenza virus inhibition, inflammation diminishing, ulcer resistance, local stimulation and heart strengthening, fever relieving, cough relieving, phlegm eliminating, asthma relieving and pain easing, and can be used as a health food and can also be used for mosquito and insect repelling.
The white tea is a special treasure in Chinese tea, belongs to micro-fermented tea, and is prepared by withering leaves and buds of white tea tree in tea (Camellia sinensis (L.) O.Ktze.), baking (or drying in the shade), picking out, and annealing through a special process. The white tea essential oil is volatile oil extracted from tea leaves and buds of white tea. At present, the research on the efficacy of white tea essential oil is less.
At present, no study and report on the treatment of skin herpes by using artemisia apiacea essential oil or white tea essential oil is seen.
Disclosure of Invention
The invention aims to provide essential oil of sweet wormwood herb and white tea, a composition thereof, a preparation method and application thereof, which are used for resisting herpes viruses and aims to overcome the defects in the prior art.
In a first aspect, the invention provides an application of sweet wormwood essential oil in preparation of an anti-herpes virus product, wherein the sweet wormwood essential oil is prepared by the following method: collecting fresh herba Artemisiae Annuae young seedling, collecting leaves, drying, cutting into pieces, placing into distillation equipment, adding distilled water at a material-liquid ratio of 1 (0.5-2), soaking, distilling for 60-90min to obtain pure distillate mixture of essential oil, and separating with separator to obtain herba Artemisiae Annuae essential oil.
Preferably, the drying mode is drying at 60 ℃ or below for 2-5 hours, the size of the fragments is 4-6mm, and the soaking time is 1-2 hours.
Preferably, the parameters of the distillation are: the extraction temperature is 90-96 ℃, and the temperature of the condensation pipe is 15-25 ℃.
Preferably, the sweet wormwood herb is produced in Henan.
Preferably, the herpesvirus is a varicella-zoster virus or a herpes simplex virus.
Preferably, the product is a cosmetic, a pharmaceutical or a daily article or clothing having an anti-herpes virus effect.
In a second aspect, the invention provides the use of white tea essential oil in the manufacture of a product for combating herpes viruses, the white tea essential oil being prepared by the method comprising: collecting tender shoots of fresh white tea, regularly spreading in a backlight room until the thickness is 3-5cm, controlling the temperature at 30-35 ℃, controlling the humidity at 50-60%, withering until the weight loss rate is 50-60%, and then placing in a dryer for drying at the temperature of 100 ℃ and 120 ℃ for 20-40 min; pulverizing dried leaves of white tea, and supercritical CO2Fluid extraction is carried out to obtain the white tea essential oil.
Preferably, the supercritical CO2The parameters of the fluid extraction were: CO 22The flow rate of the fluid is 0.5-2L/min, the extraction temperature is 42-50 ℃, the extraction pressure is 20-25 MPa, the separation temperature is 50-60 ℃, the separation pressure is 4.5-5.5 MPa, and the extraction time is 1.5-2.5 h.
Preferably, the supercritical CO2The fluid extraction is pre-soaked for 5-20min at the beginning.
Preferably, the dried white tea leaves are crushed into 10-20 meshes of coarse powder.
Preferably, the herpesvirus is a varicella-zoster virus or a herpes simplex virus.
Preferably, the product is a cosmetic, a pharmaceutical or a daily article or clothing having an anti-herpes virus effect.
In a third aspect, the invention provides the use of a plant essential oil composition containing the sweet wormwood essential oil and the white tea essential oil in any one of the above in the preparation of an anti-herpesvirus product.
Preferably, the ratio of the sweet wormwood essential oil to the white tea essential oil is 1 (0.01-20).
Preferably, the herpesvirus is a varicella-zoster virus or a herpes simplex virus.
Preferably, the product is a cosmetic, a pharmaceutical or a daily article or clothing having an anti-herpes virus effect.
The sweet wormwood essential oil is extracted from Artemisia annua L of Compositae. The white tea essential oil is extracted from Camellia sinensis (L.) Siemens of Theaceae.
The invention has the advantages that:
1. the sweet wormwood essential oil and the white tea essential oil prepared by the invention have the effect of inhibiting herpes viruses.
2. The invention optimizes the preparation method of the sweet wormwood essential oil and the white tea essential oil, and obtains the product with remarkably improved effect of inhibiting the herpes virus.
3. The invention discovers that the composition of the sweet wormwood essential oil and the white tea essential oil has the synergistic effect on the aspect of inhibiting the herpesvirus or neuralgia caused by the herpesvirus.
Drawings
FIG. 1: and (3) carrying out a total ion flow diagram of GC-MS analysis on the sweet wormwood herb essential oil.
FIG. 2: and (3) performing GC-MS analysis on the white tea essential oil to obtain a total ion flow diagram.
Detailed Description
The following detailed description of the present invention will be made with reference to the accompanying drawings.
Example 1 Artemisia apiacea essential oil of the invention and preparation thereof
Tender seedlings of sweet wormwood in Henan province are taken as materials.
The instrument comprises the following steps: the water seal type distillation still comprises the following equipment:
a distillation kettle: the whole appearance is cylindrical, the bottom of the spherical crown type kettle is provided with an opening at the upper part as a charging hole;
a gooseneck: a conical cover with an upper opening of the distillation kettle is connected with a condenser;
a condenser: an aluminum tube-in-tube condenser to condense the steam and cool the distillate;
an oil-water separator: is made of aluminum material, and is a container for receiving distillate and a separator for essential oil and water.
The process flow for extracting the essential oil comprises the following steps: batch charging → water adding → distillation → cooling → oil-water separation → essential oil → packaging.
The method comprises the following specific steps:
collecting fresh herba Artemisiae Annuae young seedling of 5 months in Henan, removing stem and useless parts, cleaning leaves, draining, and drying in an electrothermal constant temperature drying oven at temperature below 60 deg.C for 3 hr. Cutting dried herba Artemisiae Annuae into 4-6mm pieces with a cutting machine to facilitate volatilization of essential oil molecules. Placing the smashed sweet wormwood leaves into a distillation inner tank, adding equal volume of Drech distilled water, injecting into an outer distillation tank, placing the distillation tank on a water tank, sealing the outer distillation tank, soaking for 1.5h, and distilling. Extracting at 90 deg.C and 15-25 deg.C in a condenser tube for 75min to obtain mixed liquid of essential oil and hydrosol, and separating with a separator to obtain herba Artemisiae Annuae essential oil.
Example 2 Artemisia apiacea essential oil of the invention and preparation thereof
The materials and apparatus were the same as in example 1.
The method comprises the following specific steps:
collecting fresh herba Artemisiae Annuae young seedling of 5 months in Henan, removing stem and useless parts, cleaning leaves, draining, and drying in an electrothermal constant temperature drying oven at temperature below 60 deg.C for 3 hr. Cutting dried herba Artemisiae Annuae into 4-6mm pieces with a cutting machine to facilitate volatilization of essential oil molecules. Placing the smashed sweet wormwood leaves into a distillation inner tank, adding 0.8-time volume of Drech distilled water, injecting into an outer distillation tank, placing the distillation tank on a water tank, sealing the outer distillation tank, soaking for 1.5h, and then distilling. Extracting at 90 deg.C and 15-25 deg.C in a condenser tube for 75min to obtain mixed liquid of essential oil and hydrosol, and separating with a separator to obtain herba Artemisiae Annuae essential oil.
Example 3 Artemisia annua essential oil of the invention and preparation thereof
The materials and apparatus were the same as in example 1.
The method comprises the following specific steps:
collecting fresh herba Artemisiae Annuae young seedling of 5 months in Henan, removing stem and useless parts, cleaning leaves, draining, and drying in an electrothermal constant temperature drying oven at temperature below 60 deg.C for 3 hr. Cutting dried herba Artemisiae Annuae into 4-6mm pieces with a cutting machine to facilitate volatilization of essential oil molecules. Placing the smashed sweet wormwood leaves into a distillation inner tank, adding 2 times of volume of Drech distilled water, injecting into an outer distillation tank, placing the distillation tank on a water tank, sealing the outer distillation tank, soaking for 1.5h, and then distilling. Extracting at 90 deg.C and 15-25 deg.C in a condenser tube for 75min to obtain mixed liquid of essential oil and hydrosol, and separating with a separator to obtain herba Artemisiae Annuae essential oil.
Example 4 Artemisia apiacea essential oil of the invention and preparation thereof (IV)
The materials and apparatus were the same as in example 1.
The method comprises the following specific steps:
collecting fresh herba Artemisiae Annuae young seedling of 5 months in Henan, removing stem and useless parts, cleaning leaves, draining, and drying in an electrothermal constant temperature drying oven at temperature below 60 deg.C for 3 hr. Cutting dried herba Artemisiae Annuae into 4-6mm pieces with a cutting machine to facilitate volatilization of essential oil molecules. Placing the smashed sweet wormwood leaves into a distillation inner tank, adding equal volume of Drech distilled water, injecting into an outer distillation tank, placing the distillation tank on a water tank, sealing the outer distillation tank, soaking for 1.5h, and distilling. Extracting at 90 deg.C, condensing at 15-25 deg.C for 60min to obtain pure distillate, and separating with separator to obtain herba Artemisiae Annuae essential oil.
Example 5 Artemisia apiacea essential oil of the invention and preparation thereof (V)
The materials and apparatus were the same as in example 1.
The method comprises the following specific steps:
collecting fresh herba Artemisiae Annuae young seedling of 5 months in Henan, removing stem and useless parts, cleaning leaves, draining, and drying in an electrothermal constant temperature drying oven at temperature below 60 deg.C for 3 hr. Cutting dried herba Artemisiae Annuae into 4-6mm pieces with a cutting machine to facilitate volatilization of essential oil molecules. Placing the smashed sweet wormwood leaves into a distillation inner tank, adding equal volume of Drech distilled water, injecting into an outer distillation tank, placing the distillation tank on a water tank, sealing the outer distillation tank, soaking for 1.5h, and distilling. Extracting at 90 deg.C, cooling at 15-25 deg.C in a condenser tube, distilling for 90min to obtain pure distillate of essential oil, and separating with a separator to obtain herba Artemisiae Annuae essential oil.
Example 6 Artemisia annua essential oil of the invention and preparation thereof (VI)
The materials and apparatus were the same as in example 1.
The method comprises the following specific steps:
collecting fresh herba Artemisiae Annuae young seedling of 5 months in Henan, removing stem and useless parts, cleaning leaves, draining, and drying in an electrothermal constant temperature drying oven at temperature below 60 deg.C for 5 hr. Cutting dried herba Artemisiae Annuae into 4-6mm pieces with a cutting machine to facilitate volatilization of essential oil molecules. Placing the smashed sweet wormwood leaves into a distillation inner tank, adding equal volume of Drech distilled water, injecting into an outer distillation tank, placing the distillation tank on a water tank, sealing the outer distillation tank, soaking for 2h, and distilling. Extracting at 90 deg.C and 15-25 deg.C in a condenser tube for 75min to obtain mixed liquid of essential oil and hydrosol, and separating with a separator to obtain herba Artemisiae Annuae essential oil.
Example 7 Artemisia apiacea essential oil of the invention and preparation thereof (seven)
The materials and apparatus were the same as in example 1.
The method comprises the following specific steps:
collecting fresh herba Artemisiae Annuae young seedling of 5 months in Henan, removing stem and useless parts, cleaning leaves, draining, and drying in electrothermal constant temperature drying oven at temperature below 60 deg.C for 2 hr. Cutting dried herba Artemisiae Annuae into 4-6mm pieces with a cutting machine to facilitate volatilization of essential oil molecules. Placing the smashed sweet wormwood leaves into a distillation inner tank, adding equal volume of Drech distilled water, injecting into an outer distillation tank, placing the distillation tank on a water tank, sealing the outer distillation tank, soaking for 1h, and distilling. Extracting at 96 deg.C and 15-25 deg.C in a condenser tube for 75min to obtain mixed liquid of essential oil and hydrosol, and separating with a separator to obtain herba Artemisiae Annuae essential oil.
Example 8 white tea essential oil of the invention and its preparation
The large-leaf white tea in Pu' er tea area of Yunnan is used as a material.
The instrument comprises the following steps: the main equipment is an extractor consisting of
The stainless steel tube is made, the pressurizing buffer is of a double-layer structure, the inner cylinder is made of stainless steel, and the outer cylinder is made of brass and is tightly matched with the inner cylinder and the outer cylinder. The equipment comprises: CO 22A steel cylinder, a dryer, a condensed ice water bath, a plunger pump, a pressurizing buffer bottle, a thermostatic bath, a preheating pipe, an extractor, an ice water beaker, a sedimentation separator, a buffer and a wet flowmeter.
The method comprises the following specific steps:
picking tender leaves of the same size of the first heart and the second heart by hand at 6 to 9 points in the morning, neatly spreading the tender leaves in a backlight room until the thickness is 3-5cm, controlling the temperature at 35 ℃, the humidity at 55-60%, withering until the weight loss rate is 50-55%, and then placing the tender leaves in a dryer for drying at 110 ℃ for 30 min. Then crushing the white tea leaves, sieving the crushed white tea leaves by a sieve of 10 meshes to obtain coarse powder, bagging the weighed coarse tea powder by abrasive cloth, putting the coarse tea powder into an extractor, and sealing a top cover. And starting a power supply of the thermostatic bath. The carbon dioxide cylinder valve was opened and the gas was passed through the dryer along the line, the cooling condenser (immersed in an ice-water bath), the plunger pump, and into the booster buffer bottle. Opening valve V1 when the pressure of the pressurizing buffer bottle greatly exceeds the experimental pressure, increasing the pressure of the extractor to the experimental pressure, closing valve V1, pre-soaking the tea for 10min, and performing supercritical CO2And (4) fluid extraction. CO 22The flow rate of the fluid is 1L/min, the extraction temperature is 50 ℃, the extraction pressure is 25MPa, the separation temperature is 50 ℃, the separation pressure is 5.0MPa, and the extraction time is 2h, so that the white tea essential oil is obtained.
Example 9 white tea essential oil of the invention and its preparation (II)
The materials and apparatus were the same as in example 8.
The method comprises the following specific steps:
picking tender leaves of the same size of the first heart and the second heart by hand at 6 to 9 points in the morning, neatly spreading the tender leaves in a backlight room until the thickness is 3-5cm, controlling the temperature at 35 ℃, the humidity at 55-60%, withering until the weight loss rate is 55-60%, and then placing the tender leaves in a dryer for drying at 110 ℃ for 30 min. Then crushing the white tea leaves, sieving the crushed white tea leaves by a sieve of 10 meshes to obtain coarse powder, bagging the weighed coarse tea powder by abrasive cloth,and loaded into an extractor and sealed with a top cover. And starting a power supply of the thermostatic bath. The carbon dioxide cylinder valve was opened and the gas was passed through the dryer along the line, the cooling condenser (immersed in an ice-water bath), the plunger pump, and into the booster buffer bottle. Opening valve V1 when the pressure of the pressurizing buffer bottle greatly exceeds the experimental pressure, increasing the pressure of the extractor to the experimental pressure, closing valve V1, pre-soaking the tea for 10min, and performing supercritical CO2And (4) fluid extraction. CO 22The flow rate of the fluid is 1L/min, the extraction temperature is 50 ℃, the extraction pressure is 25MPa, the separation temperature is 50 ℃, the separation pressure is 5.0MPa, and the extraction time is 2h, so that the white tea essential oil is obtained.
Example 10 white tea essential oil of the invention and its preparation (III)
The materials and apparatus were the same as in example 8.
The method comprises the following specific steps:
picking tender leaves of the same size of the first heart and the second heart by hand at 6 to 9 points in the morning, neatly spreading the tender leaves in a backlight room until the thickness is 3-5cm, controlling the temperature at 30 ℃, the humidity at 50-55%, withering until the weight loss rate is 50-55%, and then placing the tender leaves in a dryer for drying at 120 ℃ for 20 min. Then crushing the white tea leaves, sieving the crushed white tea leaves by a sieve of 10 meshes to obtain coarse powder, bagging the weighed coarse tea powder by abrasive cloth, putting the coarse tea powder into an extractor, and sealing a top cover. And starting a power supply of the thermostatic bath. The carbon dioxide cylinder valve was opened and the gas was passed through the dryer along the line, the cooling condenser (immersed in an ice-water bath), the plunger pump, and into the booster buffer bottle. Opening valve V1 when the pressure of the pressurizing buffer bottle greatly exceeds the experimental pressure, increasing the pressure of the extractor to the experimental pressure, closing valve V1, pre-infiltrating the tea for 5min, and performing supercritical CO2And (4) fluid extraction. CO 22The flow rate of the fluid is 2L/min, the extraction temperature is 42 ℃, the extraction pressure is 20MPa, the separation temperature is 60 ℃, the separation pressure is 5.5MPa, and the extraction time is 1.5h, so that the white tea essential oil is obtained.
Example 11 white tea essential oil of the invention and its preparation (IV)
The materials and apparatus were the same as in example 8.
The method comprises the following specific steps:
picking tender leaves of one heart and two hearts with the same size by hand at 6-9 am, spreading in a backlight room to 3-5cm thickness, and controlling temperatureAt the temperature of 32 ℃, the humidity is 50% -55%, and the withering is carried out until the weight loss rate is 50% -55%, and then the mixture is placed into a dryer for drying for 40min at the temperature of 110 ℃. Then crushing the white tea leaves, sieving the crushed white tea leaves by a sieve of 10 meshes to obtain coarse powder, bagging the weighed coarse tea powder by abrasive cloth, putting the coarse tea powder into an extractor, and sealing a top cover. And starting a power supply of the thermostatic bath. The carbon dioxide cylinder valve was opened and the gas was passed through the dryer along the line, the cooling condenser (immersed in an ice-water bath), the plunger pump, and into the booster buffer bottle. Opening valve V1 when the pressure of the pressurizing buffer bottle greatly exceeds the experimental pressure, increasing the pressure of the extractor to the experimental pressure, closing valve V1, pre-soaking the tea for 20min, and performing supercritical CO2And (4) fluid extraction. CO 22The flow rate of the fluid is 1.5L/min, the extraction temperature is 42 ℃, the extraction pressure is 20MPa, the separation temperature is 60 ℃, the separation pressure is 5.5MPa, and the extraction time is 1.5h, so that the white tea essential oil is obtained.
Comparative example 1 Artemisia apiacea essential oil control I and preparation thereof
The plant of southernwood in bud stage in Henan province is used as the material.
The apparatus is as in example 1.
The method comprises the following specific steps:
collecting fresh herba Artemisiae Annuae whole plant of 7 months in Henan, removing stem and useless parts, cleaning leaves, filtering to remove water, and drying in an electrothermal constant temperature drying oven at temperature below 60 deg.C for 3 hr. Cutting dried herba Artemisiae Annuae into 4-6mm pieces with a cutting machine to facilitate volatilization of essential oil molecules. Placing the smashed sweet wormwood leaves into a distillation inner tank, adding equal volume of Drech distilled water, injecting into an outer distillation tank, placing the distillation tank on a water tank, sealing the outer distillation tank, soaking for 1.5h, and distilling. Extracting at 90 deg.C and 15-25 deg.C in a condenser tube for 75min to obtain mixed liquid of essential oil and hydrosol, and separating with a separator to obtain herba Artemisiae Annuae essential oil.
Comparative example 2 Artemisia apiacea essential oil control two and preparation thereof
The materials and apparatus were the same as in example 1.
The method comprises the following specific steps:
collecting fresh herba Artemisiae Annuae young seedling of 5 months in Henan, removing stem and useless parts, cleaning leaves, draining, and drying in an electrothermal constant temperature drying oven at temperature below 60 deg.C for 3 hr. Cutting dried herba Artemisiae Annuae into 4-6mm pieces with a cutting machine to facilitate volatilization of essential oil molecules. Placing the smashed sweet wormwood leaves into a distillation inner tank, adding 3 times of volume of Drech distilled water, injecting into an outer distillation tank, placing the distillation tank on a water tank, sealing the outer distillation tank, soaking for 1.5h, and then distilling. Extracting at 90 deg.C and 15-25 deg.C in a condenser tube for 75min to obtain mixed liquid of essential oil and hydrosol, and separating with a separator to obtain herba Artemisiae Annuae essential oil.
Comparative example 3 Artemisia apiacea essential oil control III and preparation thereof
The materials and apparatus were the same as in example 1.
The method comprises the following specific steps:
collecting fresh herba Artemisiae Annuae young seedling of 5 months in Henan, removing stem and useless parts, cleaning leaves, draining, and drying in an electrothermal constant temperature drying oven at temperature below 60 deg.C for 3 hr. Cutting dried herba Artemisiae Annuae into 4-6mm pieces with a cutting machine to facilitate volatilization of essential oil molecules. Placing the smashed sweet wormwood leaves into a distillation inner tank, adding equal volume of Drech distilled water, injecting into an outer distillation tank, placing the distillation tank on a water tank, sealing the outer distillation tank, soaking for 1.5h, and distilling. Extracting at 90 deg.C, cooling at 15-25 deg.C in a condenser tube, distilling for 120min to obtain pure distillate of essential oil, and separating with a separator to obtain herba Artemisiae Annuae essential oil.
Comparative example 4 white tea essential oil control one and preparation thereof
The materials and apparatus were the same as in example 8.
The method comprises the following specific steps:
picking tender leaves of the same size as the first heart and the second heart by hand at 6 to 9 o' clock in the morning, airing the tender leaves in the daylight locally in the daytime, airing the tender leaves in a shady and cool place indoors at night for 3 days continuously, and then placing the tender leaves into a dryer to dry the tender leaves for 30min at 110 ℃. Then crushing the white tea leaves, sieving the crushed white tea leaves by a sieve of 10 meshes to obtain coarse powder, bagging the weighed coarse tea powder by abrasive cloth, putting the coarse tea powder into an extractor, and sealing a top cover. And starting a power supply of the thermostatic bath. The carbon dioxide cylinder valve was opened and the gas was passed along the line through the dryer, cooling condenser (immersed in an ice-water bath), plunger pump, and intoAnd (4) pressurizing the buffer bottle. Opening valve V1 when the pressure of the pressurizing buffer bottle greatly exceeds the experimental pressure, increasing the pressure of the extractor to the experimental pressure, closing valve V1, pre-soaking the tea for 10min, and performing supercritical CO2And (4) fluid extraction. CO 22The flow rate of the fluid is 1L/min, the extraction temperature is 50 ℃, the extraction pressure is 25MPa, the separation temperature is 50 ℃, the separation pressure is 5.0MPa, and the extraction time is 2h, so that the white tea essential oil is obtained.
Comparative example 5 white tea essential oil control two and preparation thereof
The materials and apparatus were the same as in example 8.
The method comprises the following specific steps:
picking tender leaves of the same size of the first heart and the second heart by hand at 6 to 9 points in the morning, neatly spreading the tender leaves in a backlight room until the thickness is 3-5cm, controlling the temperature at 35 ℃, the humidity at 55% -60%, withering until the weight loss rate is 65% -70%, and then placing the tender leaves in a dryer for drying at 110 ℃ for 30 min. Then crushing the white tea leaves, sieving the crushed white tea leaves by a sieve of 10 meshes to obtain coarse powder, bagging the weighed coarse tea powder by abrasive cloth, putting the coarse tea powder into an extractor, and sealing a top cover. And starting a power supply of the thermostatic bath. The carbon dioxide cylinder valve was opened and the gas was passed through the dryer along the line, the cooling condenser (immersed in an ice-water bath), the plunger pump, and into the booster buffer bottle. Opening valve V1 when the pressure of the pressurizing buffer bottle greatly exceeds the experimental pressure, increasing the pressure of the extractor to the experimental pressure, closing valve V1, pre-soaking the tea for 10min, and performing supercritical CO2And (4) fluid extraction. CO 22The flow rate of the fluid is 1L/min, the extraction temperature is 50 ℃, the extraction pressure is 25MPa, the separation temperature is 50 ℃, the separation pressure is 5.0MPa, and the extraction time is 2h, so that the white tea essential oil is obtained.
Example 12 GC-MS analysis of Artemisia annua essential oil
Reagent and material
Essential oil of sweet wormwood prepared in example 1.
Diethyl ether, analytically pure.
Second, experimental instrument
Agilent 7000B triple quadrupole gas phase mass spectrometer (Agilent).
Analysis of three components
1GC-MS analysis conditions
(1) Chromatographic stripA chromatographic column: HP-5ms (30m 0.25mm 0.25 μm); carrier gas: high-purity He; flow rate: 1 ml. min-1(ii) a Sample inlet temperature: 250 ℃; the split ratio is as follows: 10: 1; temperature rising procedure: the initial temperature is 60 deg.C, and the temperature is maintained for 3min, and then the temperature is raised to 300 deg.C at the speed of 5 deg.C/min, and the temperature is maintained for 20 min.
(2) Mass spectrum condition mode: full scanning; the temperature of the adapter: 250 ℃; ion source temperature: 230 ℃; electron energy: -70 ev; scanning range: 30-400 m/e; solvent retardation: and 4 min.
2 preparation of test solution
Precisely sucking 1 μ l of essential oil sample, adding diethyl ether to desired volume of 1ml, and making into sample solution.
3 assay method
Precisely sucking 1 μ l of the sample solution, injecting into a gas phase mass spectrometer, and measuring.
4 data analysis
Analyzing the obtained GC/MS original file of the sample by MassHunter Workstation Software (Version B.04.00) to obtain the corresponding retention time and peak area percentage of each component, and comparing by a database (NIST11.0) to obtain a qualitative identification result.
5 results of measurement
The total ion flow pattern is shown in figure 1. The results of the main component analysis are shown in Table 1.
TABLE 1GC-MS analysis results of the chemical components of Artemisia annua L.oil
Example 13 GC-MS analysis of white tea essential oil
Reagent and material
White tea essential oil prepared in example 8.
Diethyl ether (analytical grade).
Second, experimental instrument
Agilent 7000B triple quadrupole gas phase mass spectrometer (Agilent).
Analysis of three components
1GC-MS analysis conditions
(1) Chromatographic conditions chromatographic column: HP-5ms (30m 0.25mm 0.25 μm); carrier gas: high-purity He; flow rate: 1 ml. min-1(ii) a Sample inlet temperature: 250 ℃; the split ratio is as follows: 10: 1; temperature rising procedure: the initial temperature is 60 deg.C, and the temperature is maintained for 3min, and then the temperature is raised to 300 deg.C at the speed of 5 deg.C/min, and the temperature is maintained for 20 min.
(2) Mass spectrum condition mode: full scanning; the temperature of the adapter: 250 ℃; ion source temperature: 230 ℃; electron energy: -70 ev; scanning range: 30-400 m/e; solvent retardation: and 4 min.
2 preparation of test solution
Precisely sucking 1 μ l of essential oil sample, adding diethyl ether to desired volume of 1ml, and making into sample solution.
3 assay method
Precisely sucking 1 μ l of the sample solution, injecting into a gas phase mass spectrometer, and measuring.
4 data analysis
Analyzing the obtained GC/MS original file of the sample by MassHunter Workstation Software (Version B.04.00) to obtain the corresponding retention time and peak area percentage of each component, and comparing by a database (NIST11.0) to obtain a qualitative identification result.
5 results of measurement
The total ion flow pattern is shown in figure 2. The results of the main component analysis are shown in Table 2.
TABLE 2 GC-MS analysis results of the chemical components of the white tea essential oil
Example 14 in vitro experiment of anti-varicella-zoster Virus compositions and extracts of Artemisia annua essential oil, white tea essential oil and compositions of the present invention
1 materials of the experiment
Virus: varicella Zoster Virus (VZV) was given by the shanghai biologies institute.
Cell: vero cells were given by the shanghai biologicals institute.
Reagent testing: the compositions of the sweet wormwood essential oil prepared in examples 1-5 and comparative examples 1-3, the white tea essential oil prepared in examples 8-9 and comparative examples 4-5, the sweet wormwood essential oil prepared in example 1 and the white tea essential oil prepared in example 8.
2 method of experiment
2.1 pharmaceutical formulation
Each single essential oil was dissolved in a small amount of DMSO and diluted to 250. mu.g/mL with DMEM medium.
The sweet wormwood essential oil and the white tea essential oil are contained in every 1mL of the compound preparation, wherein each 1mL of the compound preparation contains 400 mu L of the sweet wormwood essential oil with 250 mu g/mL and 100 mu L of the white tea essential oil with 250 mu g/mL, and the rest is supplemented with DMEM medium.
Acyclovir: dissolved with a small amount of DMSO and diluted to 0.2mg/ml with DMEM medium.
2.2 Virus inhibition experiments
Will be 1 × 105The Vero cell suspension/mL was inoculated in 96-well plates at 37 ℃ with 5% CO2Culturing in incubator, spreading the culture well with monolayer, removing culture solution, adding VZV virus 100 times of TCID50, standing at 37 deg.C and 5% CO2Allowing virus to adsorb cells in the incubator, adding DMEM medium containing essential oil 2 hr later, 37 deg.C, and 5% CO2Incubate in incubator for 72 hours, then determine the rate of virus inhibition using MTT method: adding 5mg/mL MTT solution into each well, culturing for 4 hr, adding 150 μ L dimethyl sulfoxide into each well, shaking on a shaker at low speed for 10min to dissolve the crystal, and measuring OD with enzyme-labeling instrument490And (4) calculating the inhibition rate of the essential oil on the virus. Meanwhile, a normal control group (no virus and no medicine) and a virus control group (virus and no medicine) and a blank control group (no cell, no virus and no medicine) are set, and acyclovir (0.2mg/ml) is used as a positive control. The virus inhibition ratio (%) was (essential oil group OD value-virus control group OD value)/(normal control group OD value-virus control group OD value) × 100%. Each treatment was set up in 6 replicate wells and the experiment was repeated 3 times.
2.3 statistical analysis
Statistical analysis was performed on the data using SPSS 20.0 software, with data expressed as mean ± standard deviation, and comparisons between groups were analyzed using One-way ANOVA test in combination with LSD pairwise comparisons. The difference is statistically significant when P is less than 0.05.
3 results of the experiment
Analysis results show that the sweet wormwood essential oil has a remarkable inhibitory effect on VZV virus, and has the same antiviral effect as that of acyclovir serving as a positive medicine.
TABLE 3 inhibition of VZV virus by Artemisia annua essential oil
Note: p <0.05, P <0.01 compared to positive group; compared with the artemisia apiacea essential oil group of the comparative example 1, the # P is less than 0.01; compared with the artemisia apiacea essential oil group of the comparative example 2, the delta P is less than 0.05, and the delta P is less than 0.01; compared with the refined sweet wormwood oil in the comparative example 3, P is less than 0.05, P is less than 0.01.
Analysis results show that the white tea essential oil has a remarkable inhibitory effect on VZV virus, and has an antiviral effect equivalent to or even superior to that of acyclovir serving as a positive drug.
TABLE 4 inhibition of VZV virus by white tea essential oil
Note: p <0.01 compared to positive group; compared with the white tea essential oil group of the comparative example 4, # # P is less than 0.01; compared with the white tea essential oil group of the comparative example 5, the delta P is less than 0.01.
Analysis results show that the formula essential oil has a remarkable inhibitory effect on VZV virus, and is superior to the positive drug acyclovir.
TABLE 5 comparison of the inhibitory effects of the single essential oil and the formulated essential oil on dermatophytes
Note: p <0.01 compared to positive group.
Example 15 animal experiments on the treatment of postherpetic neuralgia caused by herpes simplex virus type I with the Artemisia apiacea essential oil, the white tea essential oil and the composition of the invention
1 materials of the experiment
1.1 Experimental animals
8 weeks old C57 mice, male, 18-22 g, purchased from Shanghai Jessie laboratory animals, Inc., license number: the production permission is SCXK (Shanghai) 2018 and 0004.
1.2 reagents and reagents
A combination of the essential oil of artemisia annua of example 1, the essential oil of white tea of example 8, the essential oil of artemisia annua of example 1 and the essential oil of white tea of example 8. Pregabalin capsules, zilu pharmaceuticals (hainan) limited, national drug standard H20203040. IL-6 and TNF-alpha enzyme linked immunosorbent assay (ELISA) assay kit.
1.3 instruments and devices
A biological safety cabinet, a full-automatic enzyme marking instrument, a centrifuge, a water bath kettle, a refrigerator with 20 degrees, a refrigerator with 4 degrees, guns and gun heads with different specifications such as 1000 mul, 200 mul, 10 mul and the like, a thermal pain stimulation instrument and a timer.
2 method of experiment
2.1 pharmaceutical formulation
The formula of the essential oil is as follows: the sweet wormwood essential oil and the white tea essential oil are added into 400 mu L of 250 mu g/mL sweet wormwood essential oil and 100 mu L of 250 mu g/mL white tea essential oil.
2.2 establishment of animal models
Mice were injected intraperitoneally with 1% sodium pentobarbital solution 50After mg/kg anesthesia, the left lower limb was depilated and the depilated area was rinsed with clear water. A microsyringe was used to subcutaneously inoculate 10. mu.l (1X 10 concentration) of herpes simplex virus type I (HSV-1) to the tibia of the left lower limb of each mouse6/ml)。
2.3 animal grouping and administration
8 mice were randomly selected as a normal group, and an equal volume of physiological saline was injected instead of the virus solution. The rest mice are used for establishing a postherpetic neuralgia animal model. The mice after modeling are randomly divided into 5 groups, including a model group, a southernwood essential oil treatment group, a white tea essential oil treatment group, a formula essential oil treatment group and a positive drug treatment group, wherein each group comprises 8 mice. The drug is administered on the 2 nd day of molding, 200 mul of essential oil diluted by 2 times of DMSO is respectively administered to the sweet wormwood herb essential oil treatment group, the white tea essential oil treatment group and the formula essential oil treatment group to smear the injection part of the tibia of the left lower limb of the mouse, and the pregabalin suspension is administered to the positive drug treatment group to achieve 27 mg/kg.d intragastric administration. The model and normal groups were given DMSO smeared in the left lower limb. The administration was continued for 14 days.
2.4 thermal pain threshold determination
The mice are placed in a hot pain threshold detection box, after the mice are adapted for 5 minutes, a radiation light source is turned on, 3 irradiation points are sequentially selected on toes and soles of the mice, the distance between every two irradiation points is 0.2cm, and the interval time between every two irradiations is 3 min. The time required for irradiation was recorded when the mouse developed either a hoarse roar, licking, flicking the tail, or flicking the leg. The mean of 3 irradiation spots was taken as the mouse final hot pain Threshold (TWL).
2.5 measurement of serum IL-6, TNF-a expression level
And (3) collecting mouse blood by measuring the vein of the auricle after the completion, separating serum, and detecting the expression levels of IL-6 and TNF-alpha in the serum by an ELISA method.
2.6 statistical analysis
Statistical analysis is carried out on data by adopting SPSS 20.0 software, the data is represented by mean +/-standard deviation, One-way ANOVA method is adopted for comparing multiple groups of means, an LSD method is selected when the variances are equal in pairwise comparison between the groups, a Dunnett's T3 method is selected when the variances are not equal, and the statistical significance is achieved by taking P <0.05 as the difference.
3 results of the experiment
3.1 thermal pain threshold assay results
The thermal pain threshold of mice in each group is measured before and after administration, and comparative analysis shows that the thermal pain threshold of each model group is remarkably reduced compared with that of a normal group before administration, and has a remarkable difference (P <0.05), which indicates that the model is successful, and the thermal pain threshold between the model group and each administration group does not have a remarkable difference (P > 0.05). Compared with the model group, the thermal pain threshold of each administration group is obviously improved, wherein the thermal pain threshold of the essential oil treatment group of the formula reaches the level of a normal group.
TABLE 6 comparison of the thermal pain threshold for each group of mice
Note: a to c, P <0.05 in the pre-administration group, or P <0.05 in the post-administration group.
3.2 mouse serum IL-6, TNF-a expression level determination results
Compared with the expression levels of IL-6 and TNF-a in serum of mice in each group after treatment, the IL-6 and TNF-a levels of each administration group are obviously lower than those of a model group (P is less than 0.05), which indicates that the inflammation progression can be obviously inhibited and reduced.
TABLE 7 comparison of IL-6 and TNF-a expression levels in the sera of various groups of mice
Note: a to c, which indicate a comparison between groups P < 0.05.
The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications and additions can be made without departing from the method of the present invention, and these modifications and additions should also be regarded as the protection scope of the present invention.
Claims (16)
1. The application of the sweet wormwood essential oil in preparing the anti-herpes virus product is characterized in that the sweet wormwood essential oil is prepared by the following method: collecting fresh herba Artemisiae Annuae young seedling, collecting leaves, drying, cutting into pieces, placing into distillation equipment, adding distilled water at a material-liquid ratio of 1 (0.5-2), soaking, distilling for 60-90min to obtain pure distillate mixture of essential oil, and separating with separator to obtain herba Artemisiae Annuae essential oil.
2. The use according to claim 1, wherein the drying is carried out at a temperature below 60 ℃ for 2-5 hours, the size of the fragments is 4-6mm, and the soaking time is 1-2 hours.
3. Use according to claim 1, wherein the parameters of the distillation are: the extraction temperature is 90-96 ℃, and the temperature of the condensation pipe is 15-25 ℃.
4. The use as claimed in claim 1, wherein the southernwood is produced from Henan.
5. The use according to claim 1, wherein the herpes virus is varicella-zoster virus or herpes simplex virus.
6. Use according to claim 1, characterized in that the product is a cosmetic, pharmaceutical or daily article or clothing with an anti-herpes virus effect.
7. The application of the white tea essential oil in preparing the anti-herpes virus product is characterized in that the white tea essential oil is prepared by adopting the following method: collecting tender shoot of fresh white tea, and putting on backThe light room is regularly spread to the thickness of 3-5cm, the temperature is controlled at 30-35 ℃, the humidity is 50-60%, the withering is carried out until the weight loss rate is 50-60%, and then the mixture is placed into a dryer for drying for 20-40min at the temperature of 100 ℃ and 120 ℃; pulverizing dried leaves of white tea, and supercritical CO2Fluid extraction is carried out to obtain the white tea essential oil.
8. Use according to claim 7, wherein the supercritical CO is2The parameters of the fluid extraction were: CO 22The flow rate of the fluid is 0.5-2L/min, the extraction temperature is 42-50 ℃, the extraction pressure is 20-25 MPa, the separation temperature is 50-60 ℃, the separation pressure is 4.5-5.5 MPa, and the extraction time is 1.5-2.5 h.
9. Use according to claim 7, wherein the supercritical CO is2The fluid extraction is pre-soaked for 5-20min at the beginning.
10. The use according to claim 7, wherein the dried leaves of white tea are ground to a 10-20 mesh coarse powder.
11. The use according to claim 7, wherein the herpes virus is varicella-zoster virus or herpes simplex virus.
12. Use according to claim 7, characterized in that the product is a cosmetic, pharmaceutical or daily article or clothing with an anti-herpes virus effect.
13. Use of a plant essential oil composition comprising the essential oil of artemisia annua as claimed in any one of claims 1 to 6 and the essential oil of white tea as claimed in any one of claims 7 to 12 in the manufacture of an anti-herpesvirus product.
14. The use as claimed in claim 13, wherein the ratio of the sweet wormwood essential oil and the white tea essential oil is 1 (0.01-20).
15. The use according to claim 13, wherein the herpes virus is varicella-zoster virus or herpes simplex virus.
16. Use according to claim 13, characterized in that the product is a cosmetic, pharmaceutical or daily article or clothing with an anti-herpes virus effect.
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