CN1789395A - Tibetan herb dracocephalum heterophyllum essential oil with bacteria inhibiting and/or killing functions - Google Patents

Tibetan herb dracocephalum heterophyllum essential oil with bacteria inhibiting and/or killing functions Download PDF

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CN1789395A
CN1789395A CNA2005102007913A CN200510200791A CN1789395A CN 1789395 A CN1789395 A CN 1789395A CN A2005102007913 A CNA2005102007913 A CN A2005102007913A CN 200510200791 A CN200510200791 A CN 200510200791A CN 1789395 A CN1789395 A CN 1789395A
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volatile oil
herba ajugae
bacterium
streptococcus
extract
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李红玉
张春江
允田
傅永红
支德娟
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Lanzhou University
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Lanzhou University
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    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract

The invention relates herba dracocephalum heterophylli volatile oil possessing the bacteriostatic and germicidal action and its application. The method adopts steam distillation to extract the herba dracocephalum heterophylli volatile oil and adopts gas-chromatography/mass spectra to analysis the volatile oil, identifying the 83 components and 89.83% total content. The volatile oil can kill and suppress the staphylococcus, streptococcus, gonococcus, enterobacter, Escherichia, salmonella, shigella, pseudomonas, bacillus, oidiomycin, cryptococcus, aspergillus, penicillium, mucor, epidermophytin, micromonospora, trichophyton and other bacteriums. The herba dracocephalum heterophylli volatile oil is used for foodstuffs, drugs, health products, cosmetic products and other industries.

Description

Tibetan medicine Herba Ajugae volatile oil with antibacterial and/or germicidal action
Technical field
The present invention relates to have the Tibetan medicine Herba Ajugae volatile oil of antibacterial and/or germicidal action and had application in the product of antibacterial and/or germicidal action in preparation.
Background technology
The infectious diseases serious harm human health that pathogenic micro-organism causes causes huge social and financial loss.Wherein bacterial infectious diseases is effectively used microbiotic, but a large amount of at present continuous rising of bacterial drug resistance of using microbiotic to cause has become one of common difficult problem of paying close attention in the whole world.Especially the resistant rate of streptococcus aureus has reached more than 90%, the intestinal microflora disorder that causes simultaneously, also increase the weight of primary infection, from natural drug, seek the new drug that exploitation has anti-microbial activity and receive publicity day by day, the generation that reduces bacterial drug resistance is had active operation significance.At present, people place on natural drug seeking the approach that upgrades better antibacterials.The natural drug wide material sources have applicating history in several thousand, and especially the herbal medicine of China comprises Chinese medicine, ethnic drug and herbal medicine, and these are treasure-houses of motherland's medicine, the exploitation antibacterials are well worth doing from this class natural drug.
From special geographical naturally resource, seek new antibacterial plant, and the separation effective active becomes to be divided into the active trend of present research natural drug restraining and sterilizing bacteria.As Suad.Khamis.S etc., Journal of Ethnopharmacology, 96:107～12 (2005) reports extract the composition that bacteriostatic activity is arranged from Root-bark of Coconut, and monomeric separation, evaluation.RamziA.A etc., Journalof Ethnopharmacology, 96:177～81 (2005) reports screening from the natural plant extracts that gather on island of Yemen has the sample of bacteriostatic activity.J.Yu etc., Phytochemistry 65:881-884 (2004) has reported that Portulaca grandiflora Hook volatile oil has antibacterial and germicidal action.
Chinese patent (ZL03132166.6) discloses the preparation method and the pharmacodynamic action of a kind of brambles volatile oil and preparation thereof.With brambles volatile oil is the compound pharmaceutical preparation with antimicrobial and anti-inflammation efficacy that effective constituent is made.In view of present natural drug antibacterial research present situation, promptly all concentrate on the anti-microbial activity research of herbal medicine or medicinal plant, and ethnic drug is rarely had research and report, therefore to China's abundant and the research of unique Tibetan medicine resource anti-microbial activity has great application value.
Tibetan medicine is a wonderful work of China's natural drug, and the Tibetan medicine material is grown in height above sea level more than 3000 meters, the habitat uniqueness, and physiologically active is strong, is the treasure-house of new drug research.Tibetan medicine dracocephalum heterophyllum (Dracocephalum heterophyllum Benth.) is a labiate, be distributed in ground such as Qinghai, Gansu, Xinjiang, Tibet and Sichuan, with all herbal medicine, Tibetan medicine lucky diligent blue or green guarantor by name, dracocephalum heterophyllum belongs to the antipyretic and antidotal type medicine, is used for icterohepatitis, swelling and aching of gum, hemorrhage, stomatocace, illnesss such as poradenolymphitis, cough due to lung-heat, influenza.
At present to the existing report of composition analysis research of Herba Ajugae volatile oil, for example Zhang Guobin, Wang Mingkui isolated main component from oil is isopinccamphone (account for total oil mass 38.71%) (Ningxia Medicine College's journal, 1995,17 (4): 325-326); Lu Man, Tian Xuan extracts Herba Ajugae volatile oil with steam distillation and by the gas chromatography/mass spectrometry technology it is analyzed, identify 86 kinds of compositions, contain a large amount of terpenoids in the volatile oil, wherein geraniol content is up to 50.17%, secondly be phantol (21.2%), geranial (0.19%), eucalyptol (0.19%), caryophyllene (0.18%), peaceful alkene (0.13%), α 2 Terpineol 350s (0.129%), Karvon (0.14%) etc., these medicinal ingredientss account for the 60%[Acta Pharmaceutica Sinica in the Herba Ajugae volatile oil total content, 1999,34 (12): 925～927]; Employings such as Qin Bo capillary gas chromatography-mass spectrometry method is studied the dracocephalum heterophyllum volatile chemical component, isolate more than 184 peak through capillary chromatography, wherein 154 kinds of compositions have been confirmed altogether, the content of institute's authenticating compound account for full oil 83.52%, measured the relative percentage composition of each composition with the gas-chromatography area normalization method, its main chemical compositions is a 4-methylthiazol, 6,6-dimethyl-two ring [3,3,1]-hept-2-ene"-2-methyl alcohol, the 4[1-methylethyl]-phenylcarbinol, ethyl palmitate [research and development of natural products, 2000 (1): 4-11].Shi Wei, Chai Erfeng adopt capillary gas chromatography-mass spectrometry method that the low polarity chemical ingredients of dracocephalum heterophyllum is analyzed and researched, the 12 kinds of chemical ingredientss of reflecting out altogether. measured the relative percentage composition of each composition with the gas-chromatography area normalization method, its main chemical compositions is a geraniol, secondly be phantol, geranial, eucalyptol, caryophyllene etc., [Gansu College of Education journal (natural science edition), 2000,14 (4): 50-52].
Above investigator is behind the composition of having analyzed Herba Ajugae volatile oil, pass through inspection information, all the possible effect of its main component has been made forecast analysis, they think that geraniol, geranial etc. have the bacteriostatic effect, eucalyptol, caryophyllene etc. have relieving asthma, the effect of antibechic, but these predictions all lack experimental evidence.
Because the difference of technology, the different experiments chamber has nothing in common with each other to the analytical results of Herba Ajugae volatile oil composition.Therefore, also be difficult to the main component of Herba Ajugae volatile oil is made clear and definite definition at present, also just can't make evaluation the fungistatic effect of each composition.In view of this reason, develop the pharmaceutical use of Herba Ajugae volatile oil, the work that can do at present is exactly the fungistatic effect of at first finding out dracocephalum heterophyllum general extractive and total volatile oil, after the main component for the treatment of Herba Ajugae volatile oil is made clear and definite definition, and fungistatic effect that could clear and definite each composition.Up to the present, also nobody does systematic study to the fungistatic effect of this valuable Tibetan medicine resource general extractive of dracocephalum heterophyllum and volatile oil specially, comprises at Herba Ajugae volatile oil in the concrete application aspect sterilization, the antibacterial series products but regrettably.
We adopt steam distillation to extract Herba Ajugae volatile oil and by the gas chromatography/mass spectrometry technology it are analyzed, identify 83 kinds of compositions, main chemical compositions is Terpane (14.89%), instead-and nerolidol (7.10%), hydrogenation-1,1,4,7-tetramethyl--1H-cyclopropane [e] azulene-4a-alcohol (4.94%), gima ethylenic D (4.84%), palmitic acid (4.42%), 4-methyl isophthalic acid-different methyl-3-tetrahydrobenzene-1-alcohol, acetic ester (4.34%), 4-methyl isophthalic acid-sec.-propyl-3-hexanaphthene-1-alcohol (4.10%), 2-methyl-5-sec.-propyl two rings [3.1.0] oneself-2-alcohol (3.31%), 1-sec.-propyl-4,7-dimethyl-naphthane-4a-alcohol (2.83%), 2,6,6-trimethylammonium-two ring [3.1.1] oneself-2-alkene-4-alcohol, acetic ester (2.87%) etc., total content accounts for 89.83% of full oil, and chemical constitution sees Table 1 in detail.This result shows that there is the difference of essence in Herba Ajugae volatile oil provided by the present invention with report in the past.Data retrieval shows, the Herba Ajugae volatile oil mixture is playing a part antibacterial or germicidal action but not single certain composition among the present invention.And the fungistatic effect of the wherein separate constituent that the fungistatic effect of Herba Ajugae volatile oil mixture is better than retrieving among the present invention.In addition, it is simple to extract wherein arbitrary single component technology for the extraction of Herba Ajugae volatile oil mixture, conservation.
Table 1: Herba Ajugae volatile oil gas chromatography/mass spectrometry analytical results.
Numbering Chemistry examination and title Retention time Molecular weight Area％
1 4-hydroxy-4-methyl-2 pentanone 3.53 116 0.04
2 Eicosane 55.55 282 1.31
3 Phenylethane 3.91 106 0.02
4 Heneicosane 54.40 296 0.29
5 P-Xylol 4.06 106 0.19
6 O-Xylol 4.50 106 0.10
7 1-sec.-propyl-4-methyl bicyclic [3.1.0] oneself-2-alkene 5.23 136 0.62
8 (9E, 12E) 9,12 octadecadienoic acid ethyl ester 50.11 308 0.06
9 2,6,6-trimethylammonium-two ring [3.1.1] oneself-2-alkene (a-firpene) 5.43 136 2.63
10 (3Z, 13Z)-2-methyl-3,13-18 carbon dienols 49.45 280 0.17
11 Trans verbenol 5.96 152 0.04
12 (11E, 14E)-11,14-eicosadienoic acid methyl esters 49.25 322 0.11
13 The a-phellandrene 6.47 136 0.93
14 2,6,10,15-tetramethyl--heptadecane 48.42 296 0.15
15 6,6-dimethyl-2-methylene radical-two ring [3.1.1] hexane (a-firpene) 6.64 136 0.98
16 Ethyl palmitate 45.10 284 0.25
17 1-methyl-4-isopropylidene-1-tetrahydrobenzene 7.82 136 0.44
18 Palmitic acid 44.07 256 1.08
19 1-methyl-2-isopropyl benzene 8.09 134 4.42
20 Dibutyl phthalate 43.68 278 0.29
21 D-limonene (limonene) 8.23 136 0.13
22 Methyl palmitate 42.89 270 0.04
23 Terpane 8.37 154 14.89
24 1-methyl-4-sec.-propyl-1 (terpinene) 9.24 136 1.89
25 6,10,14-trimethylammonium pentadecane-2-ketone 40.06 268 0.55
26 2-methyl-5-sec.-propyl two rings [3.1.0] oneself-2-alcohol 9.71 154 3.31
27 4-methyl isophthalic acid-different methyl-3-tetrahydrobenzene-1-alcohol, acetic ester 10.24 196 4.34
28 Retinene 35.27 284 0.10
29 (1E)-and the 1-octenol, acetic ester 11.08 170 0.82
30 2,7, the 10-trimethyldodecane 35.14 212 0.07
31 Chrysanthenone 11.67 150 0.91
32 2-methylene radical-5-cholane-3-alcohol 34.89 400 0.05
33 The oxidation ledene 34.61 220 0.54
34 2-(2,2,3-front three basic ring penta-2-alkene) acetaldehyde 11.89 152 0.28
35 4-sec.-propyl-1,6-dimethyl-1,2,3,4,4a, 7,8,8a-octalin-1-alcohol 33.32 222 0.21
36 Instead-beta pinene-4-alcohol 12.46 152 0.84
37 2-industry methyl-6,8,8-trimethylammonium three ring [5.2.2.0 (1,6)] undecanes-3-alcohol 31.89 220 0.17
38 (s)-suitable-verbenol 12.66 152 1.27
39 (3E, 7Z)-1,5,5,8-tetramethyl--12 oxabicyclo [9.1.0] 12-3,7-diene 31.59 220 0.97
40 5-sec.-propyl two ring [3.1.0] oneself-2-ketone 13.13 138 0.10
41 Green colored Herba Carpesii Cernui alcohol 31.39 222 1.11
42 2 (10)-firpenes-3-ketone 13.33 150 0.48
43 Pinocamphone 13.90 152 0.11
44 1-sec.-propyl-4,7-dimethyl-naphthane-4a-alcohol 30.94 222 2.83
45 4-methyl isophthalic acid-sec.-propyl-3-hexanaphthene-1-alcohol 14.10 154 4.10
46 Different oxidation aromadendrene 30.70 220 0.17
47 2-is to toluene propane-2-alcohol 14.37 150 1.03
48 The oxidation caryophyllene 30.51 220 1.29
49 5-methyl-3-sec.-propyl-1-tetrahydrobenzene 14.71 136 0.56
50 (-)-spainulenol 30.34 220 1.74
51 Beta pinene-4-ketone 15.19 150 0.58
52 Decahydroization-1,1,4,7-tetramethyl--1H-cyclopropane [e] azulene-4a-alcohol 30.08 220 4.94
53 Instead-nerolidol 29.85 222 7.10
54 2-methyl-5-pseudoallyl-1-tetrahydrobenzene-2-alcohol 15.72 152 0.11
55 2-Methyl Butyric Acid-(3Z)-3-hexene ester 16.20 184 0.12
56 Bergamol 17.00 196 0.92
57 2-methyl-3-phenylpropionaldehyde 16.68 148 0.30
58 4,4,7-trimethylammonium-5,6,7,7a-tetrahydro benzo furans-2-ketone 28.35 180 0.49
59 2,6,6-trimethylammonium-two ring [3.1.1] oneself-2-alkene-4-alcohol, acetic ester 17.29 194 2.87
60 1-sec.-propyl-4,7-dimethyl-1,2,3,5,6,8a hexahydro-naphthalene 28.07 204 0.35
61 (+)-trans elecampane oil cyclic alcohol acetic ester 18.29 194 0.12
62 5-(2-propenyl)-1，3-benzodioxole 18.68 162 0.39
63 Cananga oil alkene 27.86 204 0.41
64 (-)-trans pinocarveol acetic ester 18.94 194 1.30
65 1-(6,6-dimethyl two ring [3.1.0] oneself-2-alkene-2-yl) ethanol 19.17 150 0.04
66 1-methyl-4-(6-methyl heptan-1,5-diene-2-yl) hexamethylene-1 alkene 27.71 204 0.15
67 2,6-dimethyl-1,3,5-heptantriene 19.79 122 0.31
68 Pentadecane 27.51 212 0.12
69 (-)-Stenocalyx micheli acetic ester 20.09 194 0.09
70 1-sec.-propyl-4,7-dimethyl-1,2,4a, 5,6,8a hexahydro-naphthalene 27.29 204 0.08
71 Copaene 22.26 204 0.30
72 1-methyl isophthalic acid-vinyl-2-pseudoallyl-4-isopropylidene hexanaphthene 27.15 204 0.42
73 Basic ring butane [1,2:3,4] in decahydroization-3a-methyl-6-methylene radical-1-is different 22.59 204 1.43
74 Folium eucalypti 4 (14), the 11-diene 26.87 204 0.10
75 1-methyl isophthalic acid-ethene and-2,4-di-isopropyl hexanaphthene 22.86 204 0.28
76 Gima ethylenic D 26.57 204 4.84
77 1,1,4,7-tetramethyl--1a, 2,3,4a, 5,6,7b-octahydroization-1H-pentamethylene [e] azulene 23.55 204 0.18
78 2,6-dimethyl-6-(4-methyl-3-amylene)-two ring [3.1.1] oneself-2-alkene 23.85 204 0.04
79 Caryophyllene 24.06 204 0.82
80 4,11,11-trimethylammonium-8-industry methyl bicyclic [7.2.0] 11-3-alkene 25.70 204 0.08
80 7-methyl-3-methylene radical-4-sec.-propyl-1H-pentamethylene [1,3] cyclopropane [1,2] benzene 24.48 204 0.15
82 1,5,5,9-tetramethyl--ring 11-1,4,7-triolefin 25.52 204 1.02
83 3-methyl-2-isopropylidene-3, the 5-sorbic aldehyde 25.26 150 0.36
Add up to 89.83
The present invention relates to have the Tibetan medicine Herba Ajugae volatile oil of antibacterial and/or germicidal action and had application in the product of antibacterial and/or germicidal action, filled up the barren that the Tibetan medicine dracocephalum heterophyllum is used in this field in preparation.Volatile oil of the present invention has the antibacterial and germicidal action of wide spectrum to gram-positive microorganism, Gram-negative bacteria, superficial part fungal infection, deep infection fungi.This broad-spectrum antibacterial and germicidal action have positive application in practice and are worth.Such as to the antibacterial and sterilization effect of staphylococcus epidermidis de persister, present serious day by day antibiotics resistance had positive practice significance; To Candida albicans, gonococcal antibacterial and sterilization effect, to prevention and treatment gynecological inflammation, venereal disease has positive effect.
The Tibetan medicine dracocephalum heterophyllum is the kind that national ministerial standard Tibetan medicine is included, and its Tibetan medicine lucky diligent blue or green guarantor by name is that the researcher in this field can understand.Herba Ajugae volatile oil among the present invention can derive from any part or the combination of several sections or the whole strain plant of dracocephalum heterophyllum plant.
Goal of the invention: a kind of Tibetan medicine Herba Ajugae volatile oil of antibacterial and/or germicidal action is provided and has application in the product of antibacterial and/or germicidal action in preparation.
Summary of the invention:
1 Herba Ajugae volatile oil preparation method of the present invention is as follows:
The preparation of Tibetan medicine Herba Ajugae volatile oil: get the dry herb of dracocephalum heterophyllum, pulverize, with 95% extraction using alcohol, reclaim ethanol, it is outstanding molten with water to extract medicinal extract, uses petroleum ether extraction, reclaim sherwood oil, petroleum ether extract adds ether dissolution, adds distilled water and carries out steam distillation, collect distillate, use extracted with diethyl ether, merge ether extraction liquid, reclaim ether, use anhydrous sodium sulfate dehydration, get yellow-green colour oily matter, peat-reek is arranged, standby.Being to be example with the wet distillation in this technology, can also be CO 2Supercritical extraction, this be those skilled in the art can understand.
2 Herba Ajugae volatile oil chemical ingredientss of the present invention
Gas-chromatography-mass spectrometric hyphenated technique gained mass spectrum machine mass-spectrometric data library searching has as calculated been determined the chemical composition of part in the Herba Ajugae volatile oil.Gas phase phase chromatographic condition: quartz capillary column HP25MS, 30m * 0.25mm, thickness 0.25Lm.Heating schedule: be raised to 210 ℃ since 60 ℃ with 215 ℃/min, be raised to 280 ℃ with 10 ℃/min again, carrier gas is He, post flow 1.2m1/min, injector temperature: 280 ℃.Mass spectrum condition: EI source; Ionization voltage: 70eV; Ion source temperature: 230 ℃, sweep limit: 33～550aum, sample size: 014L1, splitting ratio: 20: 1.Analysis and Identification result shows, the Herba Ajugae volatile oil main chemical compositions is Terpane (14.89%), instead-nerolidol (7.10%), hydrogenation-1,1,4,7-tetramethyl--1H-cyclopropane [e] azulene-4a-alcohol (4.94%), gima ethylenic D (4.84%), palmitic acid (4.42%), 4-methyl isophthalic acid-different methyl-3-tetrahydrobenzene-1-alcohol, acetic ester (4.34%), 4-methyl isophthalic acid-sec.-propyl-3-hexanaphthene-1-alcohol (4.10%), 2-methyl-5-sec.-propyl two rings [3.1.0] oneself-2-alcohol (3.31%), 1-sec.-propyl-4,7-dimethyl-naphthane-4a-alcohol (2.83%), 2,6,6-trimethylammonium-two ring [3.1.1] oneself-2-alkene-4-alcohol, acetic ester (2.87%) etc., total content accounts for 89.83% of full oil.
Through evidence, the Tibetan medicine Herba Ajugae volatile oil, have broad-spectrum sterilization and bacteriostatic action, Herba Ajugae volatile oil is to bacteriums such as Staphylococcus, streptococcus, neisseria gonorrhoeae genus, enterobacter, escherichia coli genus, Salmonella genus, Shigella, Rhodopseudomonas, bacillus; Fungies such as Candida, genera cryptococcus, Aspergillus, Penicillium, Mucor, Epidermophyton, microsporum, trichophyton have certain effect of killing and/or suppressing to grow.Especially to common bacteria pathogenic bacteria streptococcus aureus, non-resistance streptococcus aureus, the resistance staphylococcus epidermidis, streptococcus pneumoniae, streptococcus acidi lactici, neisseria gonorrhoeae, aerogenesis intestines liver bacterium, intestinal bacteria, enterococcus faecalis, salmonella typhi, dysentery bacterium, pseudomonas aeruginosa, Bacillus subtilus, bacillus cereus, Candida albicans, cryptococcus neoformans aspergillus, withered mould, Mucor racemosus, acrothesium floccosum, Sabouraudites lanosus, trichophyton has the very strong effect of killing and suppressing to grow.Therefore, the Tibetan medicine Herba Ajugae volatile oil can be applicable to industries such as food, medicine, healthcare products, makeup, prepares the various forms of products with antibacterial and/or germicidal action.
Embodiment:
The effect of embodiment 1 Tibetan medicine Herba Ajugae volatile oil antibacterium
Bacterial classification and substratum
Experimental strain has Gansu Province's Center for Disease Control and Clinical Laboratory center, Gansu Province to provide.Be listed as follows:
Belong to Bacterial classification
Staphylococcus streptococcus neisseria enterobacter escherichia coli belongs to Salmonella belongs toShigella Rhodopseudomonas bacillus Streptococcus aureus ATCC 25923 (Stapylococcus aureus) non-resistance streptococcus aureus (Staphylococcus aureus) resistance staphylococcus epidermidis (Staphylococcus epidermidis), streptococcus pneumoniae (Streptococcus pneumoniae) streptococcus acidi lactici (Streptococcus lactis), neisseria gonorrhoeae ( Neisseria gonorrhoeae) Aerogenesis intestines liver bacterium( Enterbacter aerogenes) intestinal bacteria ATCC 25922 (Escherichia coh), Enterococcus faecalis( Enterococcus faecalis) salmonella typhi (Salmonella typhi) dysentery bacterium ( Shigellae dysenteriae) pseudomonas aeruginosa ATCC 25924 (Pseudomonas aeruginosa) Bacillus subtilus (Bacilhus subtilis) bacillus cereus (Bacillus cereus)
The experiment substratum: the streptococcus acidi lactici substratum, the neisseria gonorrhoeae substratum, bacterium MH substratum, the fungi sabouraud culture medium is all available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute.
2 experimental techniques:
2.1 fungistatic effect is measured the inhibition zone size with cylinder plate method
Activation bacterium liquid (fungi is collected spore, cultivates spore suspension) is diluted to 10 with stroke-physiological saline solution 5-10 6Cfu/ml is standby.Add 0.2ml bacterium liquid to exsiccant MH culture medium flat plate, smoothen.Place 5min, make substratum fully absorb bacterium liquid.Each dull and stereotyped 6 in sterilized Oxford cup of placing is placed 5min, makes Oxford cup adsorbed close in media surface.Add 0.1g/ml aqueous extract thing 100ul in the cup of Oxford.Positive control penbritin 0.1ug/ul, cephamycin 0.2ug/ul all adds 100ul.Negative control adds sterilized water.Then flat board is placed 35 ℃ of constant incubators to cultivate 22-24h, take out and measure antibacterial circle diameter with slide calliper rule, calculating mean value, D represents bacteriostatic activity with antibacterial circle diameter.The fungistatic effect judging criterion is: D≤8mm is insensitive, and 8mm＜D≤13mm is low responsive, and 13mm＜D≤19mm is a medium sensitivity, and D＞19mm is extremely sensitive.
2.2 the mensuration of minimal inhibitory concentration MIC and minimal bactericidal concentration MBC adopts Microdilution plate method
The test bacterial concentration is 10 6Cfu/ml.Good bacterium liquid is added in the 96 porocyte culture plates to draw dilution, every hole 100ul, every kind of medicine 100ul, contrast dilution 10 holes make each hole Chinese traditional medicine concentration be respectively 100mg/ml, 50mg/ml, 25mg/ml, 12.5mg/ml, 6.25mg/ml, 3.125mg/ml, 1.56mg/ml, 0.78mg/ml, 0.39mg/ml, 0.195mg/ml successively.Last hole does not add medicine (only adding substratum and bacterium) and adds dilution bacterium liquid 100ul, is the bacterial growth control wells.Stay a row hole not add bacterium (only adding substratum and medicine) and make the medicine control wells, positive control is a penbritin, and cephamycin is done doubling dilution by explanation.Place the 1min that vibrates on the vibrator, make in the hole behind the abundant mixing of solution, microwell plate is added a cover and is placed in the square enamel tray that is covered with wet gauze, hatches 18h in 37 ℃ of incubators, observes the contained lowest concentration of drug in no bacterial growth hole and is minimal inhibitory concentration.
The mensuration of minimal bactericidal concentration: get 〉=liquid culture (0.1ml) of MIC drug level is coated on the corresponding nutrient agar, hatched 18 hours in 37 ℃ of incubators, the survive counting of bacterium colony is judged MBC value (colony number is the MBC value less than the pairing lowest concentration of drug of 5 culture).
3 experimental results:
Table 2: Herba Ajugae volatile oil bacteriostatic experiment inhibition zone measuring result (mm)
Pathogenic bacteria Herba Ajugae volatile oil a Positive control b
The non-resistant Staphylococcus aureus Resistant Staphylococcus epidermidis of staphylococcus aureus streptococcus pneumonia streptococcus lactis neisseria gonorrhoeae aerogenesis intestines liver bacterium Escherichia coli enterococcus faecalis salmonella typhi shigella dysenteriae pseudomonas aeruginosa hay bacillus bacillus cereus 20.4 20.5 22.0 20.6 18.6 21.4 23.2 22.5 20.8 20.6 21.8 18.8 20.5 21.2 28.4 26.6 24.4 23.4 26.3 25.4 27.4 30.6 28.3 24.6 26.4 26.8 24.5 28.8
Annotate: a Herba Ajugae volatile oil add-on is the 1mg/ cup
The positive contrast of b levofloxacin.Add-on is 10 μ g/ cups.The negative contrast of 5%DMSO.
Table 3: Herba Ajugae volatile oil minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) (mg/ml of unit)
Pathogenic bacteria MIC MBC
The non-resistant Staphylococcus aureus Resistant Staphylococcus epidermidis of staphylococcus aureus streptococcus pneumonia streptococcus lactis neisseria gonorrhoeae aerogenesis intestines liver bacterium Escherichia coli enterococcus faecalis salmonella typhi shigella dysenteriae pseudomonas aeruginosa hay bacillus bacillus cereus 0.039 0.078 0.156 0.078 0.625 0.313 0.078 0.078 0.078 0.156 0.156 0.156 0.039 0.078 0.313 0.625 0.625 0.156 2.50 1.25 0.313 0.313 0.313 0.625 0.625 0.625 0.156 0.313
Embodiment 2: the external antifungic action of Herba Ajugae volatile oil
Bacterial classification and substratum
Experimental strain has Gansu Province's Center for Disease Control and Clinical Laboratory center, Gansu Province to provide.Be listed as follows:
Belong to Bacterial classification
Mycotoruloides Cryptococcus aspergillus Penicillium mucor, Epidermophyton Microsporon trichophyton White read coccus (Candida albicans) neogenesis cryptococcus (Cryptococcus neoformans) aspergillus (Aspertgillus niger); Withered mould (Penicillium citrinum) Mucor racemosus (Mucor ramosisiums); Acrothesium floccosum (Epidermophyton floccosum), microsporum canis (Micrisporum canis) Trichophyton rubrum (Trichophyton rubrum)
The experiment substratum: the streptococcus acidi lactici substratum, the neisseria gonorrhoeae substratum, bacterium MH substratum, the fungi sabouraud culture medium is all available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute.
2 experimental techniques:
2.1 fungistatic effect is measured the inhibition zone size with cylinder plate method
Experimental strain, substratum, experimental technique be with embodiment 1,
3 experimental results are as follows:
Table 4: Herba Ajugae volatile oil bacteriostatic experiment inhibition zone measuring result (mm)
Pathogenic bacteria Herba Ajugae volatile oil a Positive control b
White read coccus Xin give birth to Yin Qiu Jun the withered mould Mucor racemosus of aspergillus acrothesium floccosum microsporum canis Trichophyton rubrum Ureaplasma urealyticum 20.6 20.6 17.6 20.4 21.4 23.6 19.8 23.6 20.8 21.4 20.2 18.8 19.6 20.6 21.8 18.6 16.8 20.8
Annotate: a Herba Ajugae volatile oil add-on is the 1mg/ cup
The b fluconazole is the fungi positive control.Add-on is 10 μ g/ cups.The negative contrast of 5%DMSO.
Table 5: Herba Ajugae volatile oil minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) (mg/ml of unit)
Pathogenic bacteria MIC MBC
White read coccus Xin give birth to Yin Qiu Jun the withered mould Mucor racemosus of aspergillus acrothesium floccosum microsporum canis Trichophyton rubrum Ureaplasma urealyticum 0.625 1.25 1.25 0.625 1.25 1.25 0.625 0.625 0.156 1.25 2.50 5.00 1.25 2.50 2.50 2.50 1.25 0.313
The result shows: Herba Ajugae volatile oil has broad-spectrum antimicrobial and germicidal action.Bacterium, fungi, mycoplasma are extremely sensitive to Herba Ajugae volatile oil, to gram-positive microorganism: streptococcus aureus, resistance staphylococcus epidermidis, bacillus cereus, Bacillus subtilus, streptococcus acidi lactici, streptococcus pneumoniae; Gram negative bacterium: intestinal bacteria, enterococcus faecalis, Pseudomonas aeruginosa, neisseria gonorrhoeae, gas bacillus enterobacteria, salmonella typhi, dysentery bacterium; Deep infection fungi Candida albicans, cryptococcus neoformans, aspergillus, withered mould, Mucor racemosus; Superficial part fungal infection acrothesium floccosum, Sabouraudites lanosus, trichophyton; And the Ureaplasma urealyticum effect that killing all arranged and suppress to grow.Have important practice significance and using value.Minimal inhibitory concentration MIC scope is 0.039～1.25mg/ml, and minimal bactericidal concentration MBC scope is 0.313～5.00mg/ml.
By above concrete embodiment 1-2, broad-spectrum antimicrobial and germicidal action that Herba Ajugae volatile oil has are described, can in having food, healthcare products, makeup and the medicine of antibacterial and/or germicidal action, preparation use.Such as Herba Ajugae volatile oil the resistance staphylococcus epidermidis is had very strong inhibition and killing action, can be prepared as various forms of medicines, for clinical serious day by day antibiotics resistance plays an important role.Herba Ajugae volatile oil has very strong inhibition and killing action to Candida albicans, gonococcus, can be prepared as the healthcare products and the medicine of various forms of preventions and treatment venereal disease.Dracocephalum heterophyllum water volatile oil can be prepared as various forms of makeup and externally-applied ointment to powerful inhibition and the killing action of dermatophytes.Have important practice significance and application prospect.

Claims (7)

1. Tibetan medicine Herba Ajugae volatile oil extract is characterized in that this extract has sterilization and/or bacteriostatic action.
2. Tibetan medicine Herba Ajugae volatile oil extract according to claim 1 is characterized in that this extract is any part or the combination of several sections or the whole strain plant of dracocephalum heterophyllum plant.
3. according to claim 1 and 2 described Herba Ajugae volatile oil extracts, it is characterized in that it can being wet distillation, can also be the CO2 supercritical extraction.
4. the sterilization of Tibetan medicine Herba Ajugae volatile oil according to claim 1 and/or bacteriostatic action is characterized in that it being that bacterium and/or fungi are had inhibition and/or killing action.
5. bacterium according to claim 4, it is characterized in that bacterium is Staphylococcus, streptococcus, neisseria gonorrhoeae genus, enterobacter, escherichia coli genus, Salmonella genus, Shigella, Rhodopseudomonas, bacillus, preferred streptococcus aureus, non-resistance streptococcus aureus, resistance staphylococcus epidermidis, streptococcus pneumoniae, streptococcus acidi lactici, neisseria gonorrhoeae, aerogenesis intestines liver bacterium, intestinal bacteria, enterococcus faecalis, salmonella typhi, dysentery bacterium, pseudomonas aeruginosa, Bacillus subtilus, bacillus cereus.
6. fungi according to claim 4, it is characterized in that fungi is Candida, genera cryptococcus, Aspergillus, Penicillium, Mucor, Epidermophyton, microsporum, trichophyton, especially Candida albicans, cryptococcus neoformans, aspergillus niger, withered mould, Mucor racemosus, acrothesium floccosum, Sabouraudites lanosus, trichophyton.
7. according to the described Herba Ajugae volatile oil extract of claim 1 to 6, it is characterized in that this extract can use in preparation has food, healthcare products, makeup and the medicine of antibacterial and/or germicidal action.
CNA2005102007913A 2005-12-12 2005-12-12 Tibetan herb dracocephalum heterophyllum essential oil with bacteria inhibiting and/or killing functions Pending CN1789395A (en)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102618386A (en) * 2012-03-14 2012-08-01 东南大学 Method for preparing chenopodium vulvaria volatile oil and application of chenopodium vulvaria volatile oil
CN103436364A (en) * 2013-08-26 2013-12-11 兰州大学 Preparation method and application of dracocephalum heterophyllum benth volatile oil
CN104293481A (en) * 2014-09-29 2015-01-21 江苏奇力康皮肤药业有限公司 Method for preparing cow dung oil and application thereof
CN108567814A (en) * 2018-04-27 2018-09-25 江西中医药大学 A kind of Herba Moslae volatile oil is for sterilizing and/or antibacterial, antimicrobial agent application and preparation method thereof
DE102019130893A1 (en) * 2019-11-15 2021-05-20 Arne Trettin Mixture of substances for use on the skin

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102618386A (en) * 2012-03-14 2012-08-01 东南大学 Method for preparing chenopodium vulvaria volatile oil and application of chenopodium vulvaria volatile oil
CN103436364A (en) * 2013-08-26 2013-12-11 兰州大学 Preparation method and application of dracocephalum heterophyllum benth volatile oil
CN103436364B (en) * 2013-08-26 2016-05-25 兰州大学 A kind of preparation method of Herba Ajugae volatile oil and application thereof
CN104293481A (en) * 2014-09-29 2015-01-21 江苏奇力康皮肤药业有限公司 Method for preparing cow dung oil and application thereof
CN104293481B (en) * 2014-09-29 2016-08-10 江苏奇力康皮肤药业有限公司 A kind of preparation method and applications of cattle manure oil
CN108567814A (en) * 2018-04-27 2018-09-25 江西中医药大学 A kind of Herba Moslae volatile oil is for sterilizing and/or antibacterial, antimicrobial agent application and preparation method thereof
DE102019130893A1 (en) * 2019-11-15 2021-05-20 Arne Trettin Mixture of substances for use on the skin

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