CN105559074B - A kind of tealeaves volatile essential oil is preparing the application in cancer-preventing health product or anticancer drug - Google Patents

A kind of tealeaves volatile essential oil is preparing the application in cancer-preventing health product or anticancer drug Download PDF

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CN105559074B
CN105559074B CN201610077038.8A CN201610077038A CN105559074B CN 105559074 B CN105559074 B CN 105559074B CN 201610077038 A CN201610077038 A CN 201610077038A CN 105559074 B CN105559074 B CN 105559074B
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tealeaves
tea
essential oil
cancer
volatile essential
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CN105559074A (en
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陈忠正
李斌
张媛媛
林晓蓉
燕妮
高雄
李晓飞
李晓玲
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South China Agricultural University
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/82Theaceae (Tea family), e.g. camellia
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/0012Galenical forms characterised by the site of application
    • A61K9/007Pulmonary tract; Aromatherapy
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation 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
    • A61K2236/30Extraction of the material
    • A61K2236/35Extraction with lipophilic solvents, e.g. Hexane or petrol ether

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  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
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  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
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  • Biotechnology (AREA)
  • Engineering & Computer Science (AREA)
  • Pulmonology (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention belongs to tea processing field, discloses a kind of tealeaves volatile essential oil and preparing the application in cancer-preventing health product or anticancer drug.The cancer-preventing health product or anticancer drug refer to that prevention and treatment or treatment have the health care product or drug of the cancer of quinone reductase inductive effect.Tealeaves volatile essential oil is to be extracted using tealeaves as raw material by Simultaneous distillation.The tealeaves volatile essential oil of acquisition not only has the smell of pleasant, can releive anxiety, but also there is certain anticancer effect, 6 big teas include that the volatile essential oil concentration extracted of black tea, green tea, green tea, yellow tea, white tea and dark green tea can induce quinone reductase multiplication in the range of 3~30 μ g/ml and play antitumaous effect.Result of study shows that tealeaves volatile essential oil by inducing quinone reductase to play antitumaous effect, is a kind of potential anti-cancer active matter, can prepare cancer-preventing health product and anticancer drug.

Description

A kind of tealeaves volatile essential oil is preparing the application in cancer-preventing health product or anticancer drug
Technical field
The invention belongs to tealeaves intensive processing technical field, in particular to a kind of tealeaves volatile essential oil is protected in preparation anti-cancer Application in strong product or anticancer drug.
Background technique
Chinese tea drinking is with a long history, and Resources of Tea Plant is abundant, this provides advantageous condition for the person of drinking tea.Tea be most by One of the healthy beverage that people welcome, contains a variety of function such as tea albumen, tea polyphenols, tea polysaccharide, caffeine, tea saponin in tealeaves Ingredient is imitated, there are the physiological functions such as antitumor, hypoglycemic, reducing blood lipid, blood pressure lowering, anti-oxidant and enhancing immunity of organisms, as Health food and drug have wide development prospect.
Tea aroma substance classes are abundant, and content is atomic, typically constitutes from the 0.01~0.05% of dry matter of tea content, green Tea is about 0.005~0.02%, and black tea is about 0.01~0.03%, and oolong tea is about 0.05%.Tea aroma substance is by organic Chemical classification can be divided into hydrocarbon, alcohols, aldehydes, ketone, acids, esters, lactone, phenols, oxa- class compound, Sulfur-bearing class compound, containing nitrogen compound etc., wherein based on alcohols, ketone, esters, aldehydes and hydrocarbon.Fragrance contains Highest oolong tea is measured, fragrance is because of home environment, tea tree breed, fresh leaf, manufacture craft, quality grade, storage method Respective distinctive odor characteristic is shown Deng different, is affected especially with manufacture craft: attenuation degree less serious case, fragrance component Middle nerolidol, jasmine lactone, indoles equal size are higher;Attenuation degree severe one, linalool and its oxidation product, geraniol, benzene The component contents such as methanol are higher.In terms of tea comprehensive processing, Aqua Folium Camelliae sinensis extraction prepares instant tea, concentrated tea juice is processed for tea beverage It is used widely, the tea polyphenols separated in tealeaves include that catechin therein is widely used in food additives or production health care food Product, anticancer research even for field of medicaments such as anti-inflammatory drug and is done in part, but volatile essential oil, that is, aroma substance of tealeaves, protects Health-care function is paid attention to using seldom work in-process.There is patent using hydro carbons and its derivative (it is derivative to be originated from grease) to 6 big tea The volatile essential oil of class tealeaves has carried out the extraction of selectivity, prepares high fragrance after having patent to utilize correlation down enzymatic treatment tealeaves Tea extract, there are also patents, and microwave coupled ion liquid to be utilized to carry out the extraction of tea aroma and for analyzing.
Summary of the invention
In order to solve the disadvantage that the prior art and shortcoming, the purpose of the present invention is to provide a kind of tealeaves volatility essences Oil is preparing the application in cancer-preventing health product or anticancer drug.This method is simple, quickly.
The purpose of the present invention is achieved through the following technical solutions:
A kind of tealeaves volatile essential oil is preparing the application in cancer-preventing health product or anticancer drug.
The cancer-preventing health product or anticancer drug refer to that prevention and treatment or treatment have the cancer of quinone reductase (QR) inductive effect The health care product or drug of disease.
The tealeaves volatile essential oil is preferably extracted by Simultaneous distillation using tealeaves as raw material.? It can be prepared using other methods.
The Simultaneous distillation is specifically according to the following steps: tea raw material being ground, according to every gram of tealeaves original The ratio of material addition 5~20ml water adds water, obtains tea mixture;Using methylene chloride as extractant, using distilling simultaneously The methods of extraction equipment extracts, and extraction time is 60min~180min;Obtained solution is transferred to nitrogen in sample bottle to blow It is dry, obtain tealeaves essential oil.
Described to be extracted specifically according to the following steps using SDE method: tea mixture is placed in round bottom burning In bottle, it is connected to one end of distilling and extracting device, and heated with electric jacket, kept slightly boiled;The other end of distilling and extracting device connects Tipping has the round-bottomed flask of methylene chloride, is heated with water-bath, temperature is maintained at 55 DEG C.
When tealeaves dosage≤50 gram, the dosage of the methylene chloride is 50mL;When 50 grams of tealeaves dosage >, described two The dosage of chloromethanes is ml of tealeaves grams corresponding number.
The tealeaves is black tea, green tea, green tea, yellow tea, white tea and dark green tea.
The present invention induces its quinone reductase to by representative of murine hepatocarcinoma cell Hepa1c1c7 using tealeaves volatile essential oil (QR) activity rises to anticancer activity evaluation, and tealeaves volatile essential oil there is preferable inhibition to make Hepa1c1c7 cancer cell line With being a kind of potential cancer-resisting active material.
The present invention have the following advantages compared with prior art and the utility model has the advantages that
(1) present invention, which extracts obtained tealeaves volatile essential oil, has stronger cancer-preventing health and anticancer effect, and this hair Operation is simple for Simultaneous distillation involved in bright and correlation method, at low cost, and the rate of recovery of essential oil is high;Therefore, have Higher practical application and promotional value.
(2) the obtained tealeaves essential oil of the present invention can be applied in aromatotherapy, can not only relieving mood, eliminate burnt Consider, and also there is the health-care efficacy for inhibiting growth of cancer cells, have a good application prospect, can be applied to medicine, health care Product and daily use chemicals industry.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail, and embodiments of the present invention are not limited thereto.
The preparation method of 1 tealeaves volatile essential oil of embodiment
(1) it weighs 25g and grinds tea sample in 1L round-bottomed flask, according to the ratio of every gram of tea raw material addition 5~20ml water Level-one water is added, is connected to one end of SDE method, and heated with electric jacket, is kept slightly boiled;Other end connection dress There is the round-bottomed flask of 50mL methylene chloride, heated with water-bath, temperature is maintained at 55 DEG C;Extraction time is 60~180min.
(2) it collects distillate after extracting to be transferred in sample bottle, nitrogen, which is blown, volatilizes methylene chloride, obtains tealeaves volatility essence Oil samples.
The research of 2 tealeaves volatile essential oil antitumor cell efficacy in vitro of embodiment, comprising the following steps:
(1) Hepa1c1c7 murine hepatocarcinoma cell suspension, logarithmic growth phase, cell in good condition, with 0.25% are prepared Trypsase (mass percent) digestion, is configured to 5 × 10 with culture medium3The cell suspension of a cell/mL is inoculated in the training of 96 holes It supports in plate, every 200 μ L of hole, is inoculated with two boards.37 DEG C, 5% (percentage by volume) CO2Cell is cultivated for 24 hours in incubator.
(2) tealeaves volatile essential oil is used to the a-MEM culture solution (volume basis for containing 10% fetal calf serum according to doses Number) it is configured to certain concentration solution with sample, 150 μ L solution are added in every hole, will be added without murine hepatocarcinoma cell group as sky White control group, control group add equivalent fresh medium.37 DEG C, 5%CO2Continue to cultivate cell in incubator (percentage by volume).
(3) after culture for 24 hours, two boards are divided into A plate and B plate.
The measurement of cell survival rate
The crystal violet of A plate 0.2% (mass percent) of addition, every 100 μ L of hole, (20-25 DEG C) placement 10-15min of room temperature, Paper suck dry moisture is used after rinsing in water.0.5%SDS (mass percent) solution, the 100rpm on shaking table, 37 DEG C of oscillations are added After 1h, absorbance value (OD value) is surveyed at 610nm with microplate reader.
The measurement of quinone reductase (QR)
The digitonin (mass percent) of 50 μ L 0.08% is added in B plate, on 100rpm shaking table, 37 DEG C of oscillations > 0.5h.Then by reaction system: being poured into loading slot after enzyme solution, MTT, menadione mixing, it is anti-to be rapidly added 200 μ L/well enzymes Answer liquid.Using microplate reader using KINETIC METHOD in 490nm, 1min measurement is primary.
(4) calculation method
The opposite QR activity of sample indicates with CD value, and being defined as opposite QR activity, to reach corresponding sample when multiplication dense Degree, when cell viability is greater than 0.5, and opposite QR activity is greater than 2, it is believed that there is sample the activity of induction quinone reductase to have Anticancer activity.
The anticancer activity of 3 tealeaves essential oil of embodiment evaluates (one)
Black tea, green tea, dark green tea, white tea, green tea, the yellow tea method that totally 6 big teas representatives use embodiment 1 are obtained into tealeaves Volatile essential oil obtains the tool that induction quinone reductase generates multiplication by linear fit curve using method as described in example 2 Bulk concentration.
The big teas tealeaves essential oil of table 1. 6 induces quinone reductase to generate the concentration (μ g/ml) to double
Teas Teas is tested to represent QR multiplication concentration
Black tea Big-leaf species in yunnan black tea 5.32
Green tea Big-leaf species in yunnan green tea 12.32
Dark green tea Big-leaf species in yunnan Pu'er tea 3.03
White tea Fuding white tea 8.96
Green tea Phoenix is single from (honey is blue fragrant) 23.08
Yellow tea Hunan Junshan Silver Needle Tea 7.39
Result above is the average value of parallel laboratory test three times
According to table 1 the result shows that: the tea of different teas volatilization oil concentration can induce quinone also in the range of 3~30 μ g/mL Antitumaous effect is played in protoenzyme multiplication.Illustrate that tealeaves volatile essential oil is a kind of potential cancer-resisting active material.
The anticancer activity of 4 tealeaves essential oil of embodiment evaluates (two)
Different flavor Various Seasonal oolong tea (green tea) is obtained into tealeaves volatile essential oil using the method for embodiment 1, is made With method as described in example 2, the specific concentration that induction quinone reductase generates multiplication is obtained by linear fit curve.
2. different flavor Various Seasonal phoenix Dan Congcha (green tea) of table etc. induces QR to generate the concentration (μ g/mL) to double
Result above is the average value of parallel laboratory test three times
According to table 2 the result shows that: the phoenix of different flavor Various Seasonal it is single from oolong tea volatilization oil concentration in 3~30 μ g/ It equally can induce quinone reductase multiplication in the range of mL and play antitumaous effect.Illustrate that tealeaves volatile essential oil is a kind of potential Cancer-resisting active material.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention, It should be equivalent substitute mode, be included within the scope of the present invention.

Claims (4)

1. a kind of tealeaves volatile essential oil is preparing the application in anticancer drug;The anticancer drug refers to that treatment has quinone reduction The drug of the liver cancer of enzyme induction effect;The tealeaves volatile essential oil is to be mentioned using tealeaves as raw material by Simultaneous distillation It obtains;The Simultaneous distillation is specifically according to the following steps: tea raw material being ground, according to every gram of tealeaves original The ratio of material addition 5~20ml water adds water, obtains tea mixture;Using methylene chloride as extractant, using distilling simultaneously Extraction equipment method extracts, and extraction time is 60min~180min;Obtained solution is transferred to nitrogen in sample bottle to dry up It is dry, obtain tealeaves essential oil.
2. application according to claim 1, it is characterised in that: described to be extracted specifically using SDE method According to the following steps: tea mixture sets the one end for being connected to distilling and extracting device in a round bottom flask, and is added with electric jacket Heat is kept slightly boiled;The other end connection of distilling and extracting device is equipped with the round-bottomed flask of methylene chloride, is heated with water-bath, temperature It is maintained at 55 DEG C.
3. application according to claim 1, it is characterised in that: when tealeaves dosage≤50 gram, the use of the methylene chloride Amount is 50mL;When 50 grams of tealeaves dosage >, the dosage of the methylene chloride is ml of tealeaves grams corresponding number.
4. application according to claim 1, it is characterised in that: the tealeaves be black tea, green tea, green tea, yellow tea, white tea or Dark green tea.
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