CN110133140A - A kind of shaggy-fruited dittany aerial parts volatile oil Components identification and its bacteriostatic activity research - Google Patents
A kind of shaggy-fruited dittany aerial parts volatile oil Components identification and its bacteriostatic activity research Download PDFInfo
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Abstract
This application involves a kind of shaggy-fruited dittany aerial parts volatile oil constituent analyses and its bacteriostatic activity to study.Shaggy-fruited dittany is a kind of economical and practical plant of China's wild cultivating, and dry root skin is commonly known as cortex dictamni, and for treating various diseases associated with inflammation, in order to improve the utilization rate of shaggy-fruited dittany whole plant, the aerial part discarded to shaggy-fruited dittany is developed.The volatile oil of shaggy-fruited dittany aerial part is obtained using steam distillation, yield 0.14% is identified using GC-MS method and contains 39 kinds of compounds, the 94.3% of the total ingredient of Zhan.Main component includes germacrene D(18.04%), terpinolene (12.97%), ocimenum (10.42%), β-carypohyllene (7.74%), phytol (5.20%) and viridiflorol (3.16%), the 57.5% of the total component content of Zhan.Shaggy-fruited dittany aerial parts volatile oil has bacteriostasis to staphylococcus aureus, Candida albicans, Escherichia coli, bacillus pumilus, bacillus subtilis, it is respectively wherein 1.56 mg/mL and 3.23 mg/mL to staphylococcus aureus and Candida albicans MIC, the exploitation for shaggy-fruited dittany aerial part provides scientific basis.
Description
Technical field
The present invention relates to shaggy-fruited dittany aerial parts volatile oils, and in particular to a kind of extraction of shaggy-fruited dittany aerial parts volatile oil, at
It analyzes and antibacterial.
Background technique
Since ancient times, plant is exactly the natural treasure-house of antibacterial substance, is widely used to food fresh keeping and medicine is auxiliary
Material, solves the problems, such as many in human production life, promotes the development and utilization of plant resources；Shaggy-fruited dittany belongs to Rutaceae man
Race, main chemical compositions include alkaloid, volatile oil etc.；Existing research shows that cortex dictamni volatile oil extracting rate is 0.09%,
Middle volatile oil is mainly made of sesquiterpenoids and long-chain fat sour component, sesquiterpenoids be its chief active at
Point, the main component of cortex dictamni is that bornylene, diformazan basic ring last of the ten Heavenly stems triolefin and green branch alkene etc. are removed along 7- hydroxyl -7- fennel；Cortex dictamni
With stronger antibacterial activity, wherein cortex dictamni volatile oil is to staphylococcus aureus and methicillin-resistant staphylococcus aureus
Bactericidal activity it is most strong, and have the effects that antitumor, anti-inflammatory, pest-resistant, antiallergy, hemostasis, anti-oxidant；Cortex dictamni is as a kind of
Conventional Chinese medicine is that history tree and " Chinese Pharmacopoeia " are collected, and there is heat-clearing and damp-drying drug, wind-dispelling to detoxify and other effects, cure mainly damp and hot sore,
The diseases such as yellow water is dripping, eczema, rubeola, scabies sore leprosy, beriberoid pyretic arthralgia, jaundice and reddish urine.
Volatile oil is Secondary metabolites, is made of the lesser simple compounds of relative molecular mass, is had certain
Aromatic odor；According to chemical structure difference, aliphatic compound, aromatic compounds and terpenoid can be divided into；This
A little compounds have low boiling point and good volatility.
There are many terpenoids in volatile oil, terpenes is a kind of alkene, and many terpenoids are to Gram-negative
Bacterium and gram-positive bacteria have good inhibiting effect；Germacrene D belongs to sesquiterpenoids according to the literature, accounts for
The 18.9% of artemisia annua volatile oil total amount its be respectively to the minimal inhibitory concentration of hay bacillus and staphylococcus aureus
0.00781 and 10 mg/mL；Same germacrene D is also the main component of Liang Zhong Turkey kuh-seng volatile oil, to large intestine
The minimal inhibitory concentration of bacillus is 11.0mg/mL；In pistacia chinensis bunge essential oil contain (Z)-β-ocimenum, up to 44.85%, it is right
The minimal inhibitory concentration of staphylococcus aureus is 0.032 mg/mL；β-carypohyllene content in aristolochia indica is higher, right
Staphylococcus aureus, Escherichia coli, bacillus subtilis have inhibiting effect；Phytol ratio shared in black nightshade volatile oil
It is 27.25%, the minimal inhibitory concentration to bacillus subtilis and Escherichia coli is respectively 62.5 ug/mL and 125 ug/mL.
Cortex dictamni is widely used in inflammation, microorganism infection, pruritus vulvue, eczema, cancer, allergic dermatitis, psoriasis etc.
Treating for skin disease；There are about five verieties for shaggy-fruited dittany, are distributed widely in the ground such as Europe and Asia, and there are two types of China, widely distributed,
The ground such as Jilin, Heilungkiang, Hebei, Sichuan, Jiangsu, and there is business to cultivate.
With the increase to shaggy-fruited dittany demand, the cultivated area of shaggy-fruited dittany also increases year by year, and aerial part causes large area
Waste；Cortex dictamni is expensive, and plantation is difficult, at high cost, and leaf has a unique smell, and shaggy-fruited dittany aerial parts volatile oil from
It is not studied to develop.
With essential oil product extensive use and demand it is vigorous, this research to Tonghua, Jilin Province city (42 ° 4 ' 52 " N,
125 ° 38 ' 8 " E) the shaggy-fruited dittany aerial parts volatile oil of growth carried out component analysis and bacteriostatic experiment, turns waste into wealth, exploitation is new
Type resource, and the exploitation for shaggy-fruited dittany aerial parts volatile oil product from now on provides scientific basis.
Summary of the invention
It is an object of the invention to the carry out GC-MS characterizations to a kind of shaggy-fruited dittany aerial parts volatile oil, in conjunction with retention index
Qualitative, the effective component of first identified volatile oil is carried out to volatile oil, and the bacteriostasis of the volatile oil is studied.
To achieve the above object the present invention adopts the following technical scheme:
The first aspect of the present invention provides a kind of extracting method of shaggy-fruited dittany aerial parts volatile oil, and steps are as follows:
Shaggy-fruited dittany aerial part is shredded, is added water and stirred, is extracted with volatile oil extractor, the volatilization of shaggy-fruited dittany aerial part is obtained
Oil；The mass ratio of above step shaggy-fruited dittany leaf aerial part and water is 1:5 ~ 8, preferably 1:6, steam distillation 5h；By extract
Being placed in -20 DEG C makes its precipitation Crystallization Separation go out volatile oil, will volatilize oil and is stored in air-tight bottle, is placed in refrigerator freezing layer.
The second aspect of the present invention provides a kind of analysis method of shaggy-fruited dittany aerial parts volatile oil, and steps are as follows:
Analysis detection is carried out to shaggy-fruited dittany aerial parts volatile oil using gas chromatography-mass spectrography.
The condition of analysis detection are as follows: Rxi-5sil chromatographic column (30m × 0.25mm × 0.25 μm), carrier gas are helium, flow velocity
For 1.0 mL/min, solvent delay 3min；Temperature program: initial temperature is 40 DEG C, keeps 4min, is warming up to 60 with 5 DEG C/min
DEG C, 2min is kept, 110 DEG C is warming up to 15 DEG C/min, keeps 4min, is warming up to 180 DEG C with 5 DEG C/min, keeps 5min, then with
25 DEG C/min is warming up to 280 DEG C, keeps 5min；230 DEG C of ion source temperature, electron impact ion source (EI) energy is 70eV, is passed
Defeated line temperature is 280 DEG C, scanning range (m/z): 40-450；Mass spectrogram is obtained after carrying out scanning of the mass spectrum to all chromatographic peaks, uses peak
Area normalization method determines relative amount of each composition in volatile oil, composes library Nist05 normal data library searching by retrieval
It is chemically defined, calculates each component relative amount using area normalization method, then utilize the retention index value and number calculated
It is compared according to retention index in library, and the type and content of shaggy-fruited dittany aerial parts volatile oil ingredient is analyzed.
The third aspect of the present invention provides and obtains shaggy-fruited dittany aerial parts volatile oil by said extracted method.Through analysis detection,
The chemical component of shaggy-fruited dittany aerial parts volatile oil of the invention include: germacrene D, terpinolene, (Z)-β-ocimenum,
β-carypohyllene, phytol, δ-dodecanol lactone, cedrol, linolenyl alcohol, δ-cadinene, α-thujene, β-phellandrene, laurene
, terpinene, linalool, 1,6,6- trimethyl cyclohexene -3- ketone, Α-cloth Chenopodiaceae alkene, bisabolence, carypohyllene, α-muurolene,
Trans--nerolidol, viridiflorol, eucalyptus oil enol, ledol, positive pentadecane aldehyde, ethyl linolenate, 1- (vinyl
Oxygroup) hexadecane, acetic acid octadecyl ester, hexadecanoyl, N, N- dimethyl lauramide, n-dotriacontane etc..
The fourth aspect of the present invention, provides above-mentioned shaggy-fruited dittany aerial parts volatile oil as the application in bacteriostatic agent, described
Bacteriostatic agent has staphylococcus aureus, Candida albicans, Escherichia coli, bacillus pumilus, bacillus subtilis preferable
Fungistatic effect, can be used for the fields such as cosmetics, health care product, food.
Specific embodiment:
The ingredient of volatile oil is sufficiently complex, and source is different, at present also not about to the extraction of shaggy-fruited dittany aerial parts volatile oil, analysis
With the report used；Based on this grinding present applicant proposes a kind of Components identification of shaggy-fruited dittany aerial parts volatile oil and bacteriostatic activity
Study carefully.
Example 1: in a kind of embodiment of the application, a kind of extracting method of shaggy-fruited dittany aerial parts volatile oil, step
It is as follows:
The shaggy-fruited dittany aerial part for taking weight 100g fresh is cleaned, and is crushed, and is stirred, is impregnated after addition distilled water, is filled with volatile oil extracting
Set and carry out steam distillation extraction, solid-liquid ratio: 1:6(g/mL), extraction time 5h, obtaining shaggy-fruited dittany aerial parts volatile oil is 0.14%,
Will volatilize oil be placed in -20 DEG C make its be precipitated crystallization, by water-oil separating, sample is obtained, at gas chromatography-mass spectrography (GC-MS)
Before analysis, it will volatilize oil and be stored in air-tight bottle, be placed in refrigerator freezing layer.
Example 2: in specific embodiment, a kind of shaggy-fruited dittany aerial part being prepared by said extracted method is provided and is waved
The discriminatory analysis method of hair oil, steps are as follows:
The detection of volatile oil includes: by species volatile oil to be measured using gas chromatography-mass spectrography (GC-MS) analysis detection, detection
Condition are as follows: Rxi-5sil chromatographic column (30m × 0.25mm × 0.25 μm), carrier gas are helium, and flow velocity is 1.0 mL/min, solvent
Postpone 3min；Temperature program: initial temperature is 40 DEG C, keeps 4min, is warming up to 60 DEG C with 5 DEG C/min, 2min is kept, with 15
DEG C/min is warming up to 110 DEG C, 4min is kept, 180 DEG C is warming up to 5 DEG C/min, keeps 5min, then be warming up to 25 DEG C/min
280 DEG C, keep 5min.230 DEG C of ion source temperature, electron impact ion source (EI) energy is 70eV, and transmission line temperature is 280
DEG C, scanning range (m/z): 40-450, mass spectrum retrieve picture library: Nist05.
By comparing the related retention index with standard alkane series (C8-C40), and by its mass spectrum and device databases (NIST
05) reference data or in document is compared, and is determined the characteristic of each component, be the results are shown in Table 1；
1 shaggy-fruited dittany aerial parts volatile oil ingredient GC-MS of table analyzes result
Shaggy-fruited dittany aerial part aerial part distills to obtain 0.14% (w/w) volatile oil through water, with fragrance striking the nose in faint yellow.Using
Gas chromatographyMass spectrometry identifies about 39 kinds of volatile compounds, and the 94.3% of the total ingredient of Zhan.Shaggy-fruited dittany aerial part
The chemical component of upper part volatile oil includes that qualitative and quantitative analysis is shown in Table 1.Sesquiterpenoids in shaggy-fruited dittany aerial part aerial part
Compounds content is higher, is 37.3%, is secondly monoterpene (29.4%).The main volatile chemical combination of shaggy-fruited dittany aerial part aerial part
Object germacrene D accounts for 18.0%, followed by terpinolene (13.0%), (Z)-β-ocimenum (10.4%), β-carypohyllene
(7.74%), phytol (5.20%) and viridiflorol (3.16%).
The testing result of shaggy-fruited dittany aerial parts volatile oil and Jiachuan Lei etc. 2008 to cortex dictamni volatile oil at
Point research be compared, in the research of cortex dictamni volatile oil, shaggy-fruited dittany aerial part and cortex dictamni volatile oil share 13 kinds at
Divide unanimously, but content is less, comprising: β-phellandrene, laurene, γ-terpinene, terpinolene, (Z)-β-ocimenum, fragrant camphor tree
Alcohol, terpinene, β-carypohyllene, δ-cadinene, germacrene D, α-muurolene, viridiflorol, but main component is not
Unanimously, the main component of cortex dictamni volatile oil has according to the literature: cis- -7- hydroxyl -7- fennel norbornene (29.4%),
Diformazan basic ring last of the ten Heavenly stems triolefin (15.5%) He Qingzhi alkene (11.4%)；These the result shows that shaggy-fruited dittany upper and lower part at point it
Between difference, it may be possible to caused by a lot of reasons, such as sunlight, soil, water and temperature, influence growth and the metabolite of plant
Biosynthesis etc.；Shaggy-fruited dittany aerial part aerial part is a kind of new natural resources rich in sesquiterpene essential oil.
Example 3: the bacteriostatic activity of present invention research shaggy-fruited dittany aerial parts volatile oil；
By detect shaggy-fruited dittany aerial part aerial part minimal inhibitory concentration (MIC), bacterial strain uses therefor include staphylococcus aureus,
Candida albicans, Escherichia coli, bacillus pumilus bacterium, hay bacillus；Shaggy-fruited dittany aerial part aerial part dilutes in 96 orifice plates
0.6 times, gradient is 0.78-200 mg/mL；A kind of standard suspension of bacterial strain (108 cfu/mL) is added, finally uses concentration
Volatile oil is dissolved for the dimethyl sulfoxide (DMSO) of 20% (v/v)；With chloramphenicol (0.78-200 mg/mL) for positive control, with
20% DMSO solution without volatile oil is negative control；All testing results are repeated 3 times；Experimental result is shown in Table 2.
2 shaggy-fruited dittany aerial parts volatile oil minimum bacteriostatic activity of table
| Staphylococcus aureus | Candida albicans | Escherichia coli | Bacillus pumilus | Bacillus subtilis | |
| Shaggy-fruited dittany aerial part aerial part (mg/mL) | 1.56 | 3.23 | 10 | 100 | 100 |
| Chloramphenicol (mg/mL) | < 0.78 | 12.5 | 3.12 | < 0.78 | < 0.78 |
Shaggy-fruited dittany aerial part aerial part is determined to 5 kinds of pathogens (including 4 kinds of gram-positive bacterias and a kind of Gram-negative
Bacterium)；Shaggy-fruited dittany aerial part aerial part is shown in Table 2 to the MIC value of above-mentioned strain；It can be seen that shaggy-fruited dittany aerial part aerial part is to gold
Staphylococcus aureus and Candida albicans show apparent antibacterial ability, and MIC is respectively 1.56 mg/mL and 3.23 mg/
mL；Especially to the antibacterial activity of Candida albicans, MIC is almost 4 times lower than a kind of antibacterial agent chloramphenicol on the market；Shaggy-fruited dittany
Some volatile components in aerial part aerial part, especially main component may play strong antibacterial activity, there is document table
Bright: germacrene D, terpinolene, (Z)-β-ocimenum, β-carypohyllene, phytol and viridiflorol are in natural volatilization
There is inhibiting effect mostly to the bacteriums such as staphylococcus aureus and Candida albicans in oil；These volatile compounds are shaggy-fruited dittanies
Main bioactive ingredients in aerial part aerial part, total content 57.5%.
Candida albicans is a kind of Common Pathogenic fungi widely distributed in nature, constitutes serious prestige to human health
The side of body, causes a series of infectious diseases；With complete and/or semisynthetic antibiotics abuse, more and more and infection by Candida albicans
Relevant disease also occurs therewith, and causes great concern, and shaggy-fruited dittany aerial part aerial part is as a kind of natural products
With good antifungal activity.
Claims (5)
1. a kind of extracting method of shaggy-fruited dittany aerial parts volatile oil, which is characterized in that step are as follows:
Shaggy-fruited dittany aerial part is shredded, is added water and stirred, is extracted with volatile oil extractor, the volatilization of shaggy-fruited dittany aerial part is obtained
Oil；
The mass ratio of above step shaggy-fruited dittany leaf aerial part and water is 1:5 ~ 8, preferably 1:6, steam distillation 5h；By extract
Being placed in -20 DEG C makes its precipitation Crystallization Separation go out volatile oil, will volatilize oil and is stored in air-tight bottle, be placed in refrigerator freezing layer, extracts
Rate is 0.14%.
2. a kind of detection method of shaggy-fruited dittany aerial parts volatile oil, which is characterized in that step are as follows:
Analysis detection is carried out to shaggy-fruited dittany aerial parts volatile oil using GC-MS.
3. detection method as claimed in claim 2, is characterized in that, in above step 2, analysis detection condition are as follows: Rxi-5sil
Chromatographic column (30m × 0.25mm × 0.25 μm), carrier gas are helium, and flow velocity is 1.0 mL/min, solvent delay 3min；Heat up journey
Sequence: initial temperature is 40 DEG C, keeps 4min, is warming up to 60 DEG C with 5 DEG C/min, keeps 2min, be warming up to 110 with 15 DEG C/min
DEG C, 4min is kept, is warming up to 180 DEG C with 5 DEG C/min, 5min is kept, then be warming up to 280 DEG C with 25 DEG C/min, keeps 5min；From
230 DEG C of source temperature, electron impact ion source (EI) energy is 70eV, and transmission line temperature is 280 DEG C, scanning range (m/z):
40-450；Mass spectrogram is obtained after carrying out scanning of the mass spectrum to all chromatographic peaks, determines each composition in volatile oil with areas of peak normalization method
In relative amount, by retrieval spectrum library Nist11 normal data library searching be chemically defined, utilize area normalization method meter
Each component relative amount is calculated, is then compared using the retention index value of calculating with retention index in database, and to shaggy-fruited dittany
The type and content of aerial parts volatile oil ingredient are analyzed.
4. as obtained in claim 3, shaggy-fruited dittany aerial parts volatile oil identifies 39 kinds of chemical combination by retention index using GC-MS method
Object, the 94.3% of the total component content of Zhan；Main component is germacrene D(18.0%), terpinolene (13.0%), (Z)-β-sieve
Alkene (10.4%), β-carypohyllene (7.74%), phytol (5.20%) and viridiflorol (3.16%) are strangled, the total component content of Zhan
57.5%。
5. a kind of bacteriostatic activity of shaggy-fruited dittany aerial parts volatile oil, it is characterised in that:
Shaggy-fruited dittany aerial parts volatile oil is to staphylococcus aureus, Candida albicans, Escherichia coli, bacillus pumilus, withered grass
The minimal inhibitory concentration of bacillus is respectively 1.56,3.23,10.0,100 and 100mg/mL, wherein to golden yellow grape
Coccus and Candida albicans show apparent antibacterial ability.
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| CN115745829B (en) * | 2022-11-08 | 2023-06-09 | 齐齐哈尔医学院 | Dictamni-acyl base A and preparation method and application thereof |
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| CN110133140B (en) | 2022-07-12 |
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