CN102727599A - Trollius chinensis dreg fermented product with antifungal effect and process for preparing trollius chinensis dreg fermented product - Google Patents
Trollius chinensis dreg fermented product with antifungal effect and process for preparing trollius chinensis dreg fermented product Download PDFInfo
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- CN102727599A CN102727599A CN2012102361861A CN201210236186A CN102727599A CN 102727599 A CN102727599 A CN 102727599A CN 2012102361861 A CN2012102361861 A CN 2012102361861A CN 201210236186 A CN201210236186 A CN 201210236186A CN 102727599 A CN102727599 A CN 102727599A
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Abstract
The invention relates to a trollius chinensis dreg fermented product with an antifungal effect and a process for preparing the trollius chinensis dreg fermented product, and belongs to the technical field of treating fungal infection diseases. The technical scheme is as follows: the dried trollius chinensis dreg is ground into powder to be used as a raw material, the Anqi yeast is taken as a fermenting strain, the nutrition ingredients of peptone and glucose which are necessary to the yeast growth are added at the same time, and the trollius chinensis dreg fermented product is extracted; and the trollius chinensis dreg fermented product is an ethanol extract of a fermented product, and each gram of the dried trollius chinensis dreg fermented product contains 12.5-25 gram of dregs equivalently. The experimental result shows that the trollius chinensis dreg fermented product has a very good curative effect on an experimental animal infected with fungus, and no toxic and side effects are found out; and the trollius chinensis dreg fermented product has a good inhibition or sterilization effect on pathogenic or drug-resistant fungi strains which are clinically separated, and has the antifungal concentration which is consistent with that the clinically common antifungal drugs-fluconazole and jie'eryin. The trollius chinensis dreg fermented product provides a new choice for clinical or livestock antifungal drugs, and has the characteristics of low cost, convenience in application, high benefit and the like.
Description
Technical field
The present invention relates to a kind of Flos Trollii medicinal residues fermented product and preparation technology thereof of antifungic action, belong to treatment fungal infection disease technical field.
Background technology
China's Chinese medicine and pharmacy is with a long history, and Chinese medicinal material industry is one of China's conventional industries.Chinese medicine enterprise can produce a considerable amount of medicinal residues when producing a large amount of Chinese patent medicines, medicinal residues are generally wet stock, and very easily corrupt, taste is smelly different.Early stage Chinese medicine slag forms of treatment mainly comprises landfill, burning, FX stacking etc.These processing methods not only exhaust substantial contribution, have more caused the wasting of resources and serious pollution, and how handling Chinese medicine slag has become a problem demanding prompt solution.So, utilize biotechnology that the Chinese medicine medicinal residues are handled, turn waste into wealth in the hope of reaching, the utilization rate that improves natural resources of Chinese medicinal materials enjoys people's attention.Not only contain nascent materials such as rich in protein, cellulose in the medicinal residues; And because present extraction process more single (decocting boils); These still remain in the medicinal residues through the part active component in the Chinese medicine after extracting, decocting, and these residual nascent and secondary substances make medicinal residues have very high recycle value, therefore; Medicinal residues begin to receive people's attention as a kind of resource, have become one of important content of modernization of cmm.
Along with the application of the carrying out of application, bone marrow and the organ transplantation of a large amount of broad ectrum antibiotic, glucocorticoid and immunosuppressant, conduit interventional therapy, particularly AIDS popular, fungal infection diseases such as candidemia and skin, mucosa increase gradually.The fungal infection incidence rate constantly rises; Except the quantity of encountered pathogenic bacteria infection is on the increase; Some rare fungal infection have report also the time; To the situation that mycosis is on the rise, also increasing to the research of antifungal drug, but curative effect antifungal drug high, that side effect is low is few at present.Therefore, research and development colleges and universities, nontoxic, cheap, convenient application antifungal drug have profound significance.
Flos Trollii is the dry flower and alabastrum of ranunculaceae plant Flos Trollii (Trollius chinensis Bunge), is distributed widely in areas such as southwest, northwest, North China, northeast.Flos Trollii has long medicinal history, and among the people infusing with boiled water worked as tea, has the effect of heat-clearing and toxic substances removing.Order; Domestic have drugmaker of tens of family to produce Flos Trollii Chinese medicine; Preparations such as globeflower capsule, Tabellae Trollii, tropaeolum oral liquid, Chinese globe flower injection are arranged, the clinical inflammation such as treatment flu, fever, chronic tonsillitis, acute myringitis, urinary tract infection that are mainly used in.According to incompletely statistics, the whole nation utilizes the medicinal residues of Flos Trollii pharmacy generation that millions of tons are arranged every year, and the overwhelming majority goes out of use, and not only contaminated environment, and pharmaceutical factory also will consume a large amount of expenses for handling medicinal residues.Therefore how the Flos Trollii medicinal residues are made full use of, turn waste into wealth, of far-reaching significance.
Summary of the invention
The purpose of this invention is to provide a kind of Flos Trollii medicinal residues fermented product and preparation technology thereof of antifungic action, technological process is simple, with low cost, convenient in application, and is effective, solves the problems referred to above that exist in the background technology.
The objective of the invention is to realize like this: a kind of Flos Trollii medicinal residues fermented product preparation technology of antifungic action comprises following processing step:
1. dried Flos Trollii medicinal residues are ground into powder;
2. the yeast activation and proliferation is cultivated, and the enrichment liquid body of getting behind the autoclaving is cultivated based in the sterile chamber, adds Angel dry yeast 5-10 gram/100ml under the gnotobasis, puts 35-37 ℃ of constant incubator cultivation 5-7 h;
3. the value-added yeast liquid of activation being added the mass and size mark is 40-60% Flos Trollii medicinal residues powder, 4% peptone, 4% glucose, the fermentation of mix homogeneously continued;
4. fermentation condition is 30-37 ℃ of cultivation temperature, incubation time 72-96 hours;
5. after the fermentation ends, fermented product is added dehydrated alcohol, makes concentration of alcohol reach 60-80%, in extractor 70 ℃ the extraction 2-4h, with extracting solution 1500-2500 rev/min centrifugal 15-25 minute; Get supernatant and on Rotary Evaporators, reclaim ethanol, the 70 ℃ of water-baths of raffinate behind the recovery ethanol become paste, and 80 ℃ of baking boxs are dried, and promptly obtain Flos Trollii medicinal residues fermented product.
Described Flos Trollii medicinal residues fermented product extraction ratio is 4-8%, and promptly 100 gram Flos Trollii medicinal residues fermentation backs obtain 4-8 gram Flos Trollii medicinal residues fermented product with ethanol extraction, and every gram Flos Trollii medicinal residues fermented product quite contains medicinal residues 12.5-25g.
Described enrichment liquid body culture medium ratio is: peptone 40 g, yeast soak powder 20 g, glucose 40 g, place 1000 ml distilled water.
A kind of Flos Trollii medicinal residues fermented product of antifungic action; Dried Flos Trollii medicinal residues are ground into powder as raw material; With the Angel Yeast bacterium is fermented bacterium, adds necessary peptone, the glucose nutritional labeling of yeast growth simultaneously, extracts Flos Trollii medicinal residues fermented product; Said Flos Trollii medicinal residues fermented product is the ethanol extraction of fermentation article, and the every gram Flos Trollii medicinal residues fermented product in dry back quite contains medicinal residues 12.5-25g.
Flos Trollii medicinal residues fermented product is processed different dosage form as required, as antifungal drug, is used for the inside and outside fungal infection treatment of body and the disease treatment of poultry fungal infection of human body.Behind said Flos Trollii medicinal residues fermented product abrasive flour, can be made into dosage forms such as tablet, capsule or granule, be used for oral medication whole body fungal infection disease; Also can process cream, ointment, lotion or spray, fungal infection such as treatment skin, mucosa.
With different approaches fungal infection model in the Flos Trollii medicinal residues fermented product treatment animal body, observe the therapeutic effect of the inside and outside fungal infection of its animal body, observed its toxicity simultaneously.The result shows that Flos Trollii medicinal residues fermented product has good therapeutic effect to the laboratory animal of fungal infection, does not find any toxic and side effects; External direct antifungal experiment, the result shows has good inhibition or killing effect to the clinical isolating pathogenic drug resistance fungal bacterial strain that reaches, and its antifungal concentration is starkly lower than conventional Folium Artemisiae Argyi water decoction and ethanol extraction.The present invention holds antifungal drug for clinical or domestic animal provides a kind of new selection, has characteristics such as with low cost, convenient in application, benefit height.
Good effect of the present invention is: (1) Flos Trollii medicinal residues aboundresources; The medicinal residues that extract Flos Trollii behind the medicinal ingredient make full use of effective combination with garbage improvement and resource thereof; The fundamental state policy that meets sustainable development and recycling economy; All useful to country, enterprise, consumer, have popularizing application prospect, can produce economic benefit and social benefit; (2) Flos Trollii medicinal residues fermentations medical value is high, and it is that decocting boils and carries the thing effective ingredient that pharmacy corporation is produced the Flos Trollii preparation overwhelming majority, and the part effective ingredient remains in the medicinal residues.Using microbe of the present invention fermentation can make some compositions further transform through the effect of microbial enzyme, and some effective ingredient can further extract with ethanol on the other hand, for the utilization again and the new purposes of Flos Trollii resource provides scientific basis; (3) preparation technology is simple: this method technological process is simple, does not need specific apparatus and equipment, and cost is low, and is with short production cycle, is suitable for suitability for industrialized production; (4) convenient in application, effect is obvious: the said preparation consumption is little, potency ratio is high, it is all convenient to store and use.And can obviously increase the therapeutic effect of fungal infection laboratory animal; (5) reduce pollution, increase economic benefit: Flos Trollii medicinal residues water content is higher, and is very easily corrupt; Can cause serious pollution to environment; And still contain protein, polysaccharide, various trace element and vitamin and a certain amount of composition capable of using in the garbage of medicinal residues, in time it is fully utilized, not only can the contaminated solution problem; Also can bring certain economic benefits, have broad prospects.
The specific embodiment
Below, the present invention is described further through embodiment.
A kind of Flos Trollii medicinal residues fermented product preparation technology of antifungic action comprises following processing step:
1. dried Flos Trollii medicinal residues are ground into powder;
2. the yeast activation and proliferation is cultivated, and the enrichment liquid body of getting behind the autoclaving is cultivated based in the sterile chamber, adds Angel dry yeast 5-10 gram/100ml under the gnotobasis, puts 35-37 ℃ of constant incubator cultivation 5-7 h;
3. the value-added yeast liquid of activation being added the mass and size mark is 40-60% Flos Trollii medicinal residues powder, 4% peptone, 4% glucose, the fermentation of mix homogeneously continued;
4. fermentation condition is 30-37 ℃ of cultivation temperature, incubation time 72-96 hours;
5. after the fermentation ends, fermented product is added dehydrated alcohol, makes concentration of alcohol reach 60-80%, in extractor 70 ℃ the extraction 2-4h, with extracting solution 1500-2500 rev/min centrifugal 15-25 minute; Get supernatant and on Rotary Evaporators, reclaim ethanol, the 70 ℃ of water-baths of raffinate behind the recovery ethanol become paste, and 80 ℃ of baking boxs are dried, and promptly obtain Flos Trollii medicinal residues fermented product.
In an embodiment, the present invention is fermented the Flos Trollii medicinal residues by above-mentioned preparation technology, in the scope of regulation, has carried out the preparation of several schemes, and its tunning extract has been carried out external and interior animal experiment, all obtains consistent useful result.
Embodiment one:
1. the yeast activation and proliferation is cultivated: enrichment liquid body culture medium: yeast=100ml:5g, 35 ℃ of constant incubators are cultivated 7 h.
2. activation yeast and Flos Trollii medicinal residues ratio: activation yeast liquid: Flos Trollii medicinal residues=100ml:40g.
3. fermentation condition is: 30 ℃ of temperature, 96 hours time.
4. fermented product method for distilling: fermented product adds dehydrated alcohol, makes concentration of alcohol reach 60%, 70 ℃ of extraction 2h, 1500 rev/mins of centrifugal 25min of extracting solution.Supernatant reclaims ethanol, and the 70 ℃ of water-baths of raffinate behind the recovery ethanol become paste, and 80 ℃ of baking boxs are dried, and promptly obtain Flos Trollii medicinal residues fermented product extract.
Embodiment two:
1. the yeast activation and proliferation is cultivated: enrichment liquid body culture medium: yeast=100ml:8g, 36 ℃ of constant incubators are cultivated 6 h.
2. activation yeast and Flos Trollii medicinal residues ratio: activation yeast liquid: Flos Trollii medicinal residues=100ml:50g.
3. fermentation condition is: 34 ℃ of temperature, 84 hours time.
4. fermented product method for distilling: fermented product adds dehydrated alcohol, makes concentration of alcohol reach 70%, 70 ℃ of extraction 4h, 2000 rev/mins of centrifugal 20min of extracting solution.Supernatant reclaims ethanol, and the 70 ℃ of water-baths of raffinate behind the recovery ethanol become paste, and 80 ℃ of baking boxs are dried, and promptly obtain Flos Trollii medicinal residues fermented product extract.
Embodiment three:
1. the yeast activation and proliferation is cultivated: YPD fluid medium: yeast=100ml:10g, 37 ℃ of constant incubators are cultivated 5 h.
2. activation yeast and Flos Trollii medicinal residues ratio: activation yeast liquid: Flos Trollii medicinal residues=100ml:60g.
3. fermentation condition is: 37 ℃ of temperature, 72 hours time.
4. fermented product method for distilling: fermented product adds dehydrated alcohol, makes concentration of alcohol reach 80%, 70 ℃ of extraction 3h, 2500 rev/mins of centrifugal 15min of extracting solution.Supernatant reclaims ethanol, and the 70 ℃ of water-baths of raffinate behind the recovery ethanol become paste, and 80 ℃ of baking boxs are dried, and promptly obtain Flos Trollii medicinal residues fermented product extract.
With embodiment one, two, three grind into powders, process capsule, tablet, particulate oral respectively and be used for experiment in the animal body, process cream, ointment, lotion external and carry out mucocutaneous antifungal experiment.
External antifungal experiment
Adopt test tube method and Plating to observe Flos Trollii medicinal residues fermented product extract (sample) to Candida albicans and the external antifungic action of microsporum canis; And, measure minimal inhibitory concentration (MIC) and MBC (MBC) with the positive contrast medicine of fluconazol; The result sees table 1.
Table 1 result shows that sample is almost close with fluconazol to oidiomycetic inhibitory action of white and killing action, only differs a gradient; Sample is consistent with fluconazol with killing action to the inhibitory action of microsporum canis.Its minimum antifungal concentration MIC and antifungal concentration are all lower.
Table 1. sample is to the MIC and the MBC of Candida albicans and microsporum canis
2. oral antifungal experiment in the body
The healthy kunming mice of SPF level, male and female half and half are selected in experiment for use.At first cause immune function of mice low, use Candida albicans 10 afterwards with cyclophosphamide
7/ ml bacterium liquid tail vein injection (0.2ml/ is only), the injection fungus is each dosage form of mouse stomach sample simultaneously, by 100,50, three dose groups of 25mg/kg, 20 every group, irritate stomach every day once, continuous 7 days, observe the dead mouse situation of respectively organizing.The result sees table 2.
Table 2 sample is to the influence (n=20) of candida albicans infection mice dying rate (%)
Table 2 result shows that blank group mice does not have death, and the animal dead rate of model control group is 95.0% (P < 0.01); Each sample group mouse death rate is along with sample dosage increases, and its mortality rate reduces, and is similar with fluconazol, explains that sample has obvious antifungic action, has protective effect to the fungal infection dead mouse.
The experiment of topical application antifungal
Inject Candida albicans suspension (5 * 10 in the Kunming mouse intravaginal
6Individual/as ml) to build the vaginitis model, with sample emulsifiable paste, cream, lotion and the treatment of 10% JIEERYIN XIYE, successive administration 7 days was done smear for microscopic examination to the vaginal douche of mice on the 8th day respectively, and measured the fungus carrying capacity.The result sees table 3.
Table 3 sample is to mouse vagina irrigating solution fungus carrying capacity (* 10
3Individual/ml) comparison
(`c ± s, n=20)
Compare * with model group
P＜0.05, * *
P＜0.01
Table 3 result shows; Each sample topical application all has inhibitory action, also can obviously improve the symptoms such as vagina redness of mice Candida albicans; The curative effect of dialogue monilial vaginitis is similar with JIEERYIN, shows that sample dialogue monilial vaginitis has the obvious treatment effect.
The present invention has obtained good effect through external antifungal and animal experimental observation, draws to draw a conclusion:
1. the present invention develops the Flos Trollii drug resource fully effectively, and the environmental pollution that not only can avoid discarded medicinal residues to bring has also solved a large amount of expenses of handling medicinal residues simultaneously, makes Flos Trollii Chinese medicine make full use of, and medicinal residues are turned waste into wealth, and are significant.
2. Flos Trollii medicinal residues fermented product extract of the present invention has tangible antifungic action, and whole body fungal infection and local fungal infection are all had the good curing effect, can be used for the treatment of fungal infections medicine.
Claims (6)
1. the Flos Trollii medicinal residues fermented product preparation technology of an antifungic action is characterized in that comprising following processing step:
1. dried Flos Trollii medicinal residues are ground into powder;
2. the yeast activation and proliferation is cultivated, and the enrichment liquid body of getting behind the autoclaving is cultivated based in the sterile chamber, adds Angel dry yeast 5-10 gram/100ml under the gnotobasis, puts 35-37 ℃ of constant incubator cultivation 5-7 h;
3. the value-added yeast liquid of activation being added the mass and size mark is 40-60% Flos Trollii medicinal residues powder, 4% peptone, 4% glucose, the fermentation of mix homogeneously continued;
4. fermentation condition is 30-37 ℃ of cultivation temperature, incubation time 72-96 hours;
5. after the fermentation ends, fermented product is added dehydrated alcohol, makes concentration of alcohol reach 60-80%, in extractor 70 ℃ the extraction 2-4h, with extracting solution 1500-2500 rev/min centrifugal 15-25 minute; Get supernatant and on Rotary Evaporators, reclaim ethanol, the 70 ℃ of water-baths of raffinate behind the recovery ethanol become paste, and 80 ℃ of baking boxs are dried, and promptly obtain Flos Trollii medicinal residues fermented product.
2. according to the Flos Trollii medicinal residues fermented product preparation technology of the said a kind of antifungic action of claim 1; It is characterized in that described Flos Trollii medicinal residues fermented product extraction ratio is 4-8%; Promptly 100 gram Flos Trollii medicinal residues fermentation backs obtain 4-8 gram Flos Trollii medicinal residues fermented product with ethanol extraction, and every gram Flos Trollii medicinal residues fermented product quite contains medicinal residues 12.5-25g.
3. according to the Flos Trollii medicinal residues fermented product preparation technology of claim 1 or 2 said a kind of antifungic actions; It is characterized in that described enrichment liquid body culture medium ratio is: peptone 40 g, yeast soak powder 20 g, glucose 40 g, place 1000 ml distilled water.
4. the Flos Trollii medicinal residues fermented product of an antifungic action; It is characterized in that dried Flos Trollii medicinal residues are ground into powder as raw material; With the Angel Yeast bacterium is fermented bacterium, adds necessary peptone, the glucose nutritional labeling of yeast growth simultaneously, extracts Flos Trollii medicinal residues fermented product; Said Flos Trollii medicinal residues fermented product is the ethanol extraction of fermentation article, and the every gram Flos Trollii medicinal residues fermented product in dry back quite contains medicinal residues 12.5-25g.
5. according to the Flos Trollii medicinal residues fermented product of the said a kind of antifungic action of claim 4; It is characterized in that said Flos Trollii medicinal residues fermented product as antifungal drug, be used for whole body or local fungal treatment of infection and the poultry whole body of human body or the treatment that local fungal catches.
6. according to the Flos Trollii medicinal residues fermented product of the said a kind of antifungic action of claim 5, it is characterized in that said Flos Trollii medicinal residues fermented product abrasive flour after, process tablet, capsule or granule dosage form, be used for oral medication whole body fungal infection disease; Process cream, ointment, lotion or spray, fungal infection such as treatment skin, mucosa.
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| CN2012102361861A CN102727599B (en) | 2012-07-10 | 2012-07-10 | Trollius chinensis dreg fermented product with antifungal effect and process for preparing trollius chinensis dreg fermented product |
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Non-Patent Citations (4)
| Title |
|---|
| 周勇等: "《复合中药渣发酵功能饲料的研究》", 《中国酿造》 * |
| 唐津忠等: "《金莲花中黄酮类化合物的提取及其抗氧化性研究》", 《食品科学》 * |
| 孙静等: "《中药发酵技术研究进展》", 《药物评价研究》 * |
| 葛喜珍等: "《酵母菌发酵川乌、附子的初步研究》", 《时珍国医国药》 * |
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