CN100462006C - Use of ginkgoic acid in preparation of biological pesticide for killing Oncomelania snail and preventing schistosomiasis - Google Patents
Use of ginkgoic acid in preparation of biological pesticide for killing Oncomelania snail and preventing schistosomiasis Download PDFInfo
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Abstract
The invention relates to the use of ginkgoic acid in preparation of biological pesticide for killing Oncomelania snail and preventing schistosomiasis, wherein experiment result shows that, the ginkgolic acid can effectively inhibit key enzyme and de-coupling reaction to mitochondria oxidation phosphorylation in mollusk energy supply biochemical metabolism, effectively inhibit tyrosinase during insect exuviation procedure and prostaglandin synthetase during breeding process, thus can eradicate unwanted mollusk harming agronomic crops.
Description
Technical field
The invention belongs to field of biological pesticide, relate to ginkgoic acid (ginkgolic acids), be specifically related to that ginkgoic acid is used to the purposes killing oncomelania, prevent schistosomiasis and as the application of novel biopesticide in the new purposes of preparation in the biopesticide.
Background technology
(Oncomelania hupensis O.hupensis) belongs to mollusk to described oncomelania, is the unique intermediate host who propagates the widely popular schistosomiasis of China.
Bilharzial miracidium can develop into mother sporocyst after invading breechblock, thereby thousands of times of ground propagation in breechblock, constantly produce cercaria in water or moist soil table, so that infected person and other animals are are so gone round and begun again and facilitated spreading and spreading of schistosomiasis.Killing oncomelania can cut off bilharzial route of transmission.So killing oncomelania is the internationally recognized important measures that prevent schistosomiasis.
Schistosomiasis endemic in the temperate zone, subtropics, torrid areas.Calendar year 2001 WHO estimates that the population that schistosomiasis is infected in the whole world is about 200,000,000 people, has 600,000,000 populations to be on the hazard, and has 76 countries influenced, and wherein the overwhelming majority is a developing country, this disease serious harm these national people's health and economic development.According to statistics in 2003, China had 12 provinces, 418 counties that schistosomiasis endemic is arranged, and schistosomiasis patient is about 1,000,000 people.Popular district total population is about 100,000,000 people, and the long-pending 3787km that reaches of spiral surface is arranged
2Though China is the mass control to schistosomiasis through 50～sixties, the popular of this disease is effectively controlled, in recent years, because factors such as floods, reservoir, dam construction, movements of population, schistosomiasis begins bounce-back.For example, almost all there is oncomelania Dongting Lake lake region, Hunan, and no swimming, and Yueyang along the lake, Changde, Yiyang Prefecture all are severely afflicated areas.Therefore, the control to schistosomiasis becomes one of strategic objective of national sustainable development.
The chemistry spiral shell that goes out remains and eliminates or reduce the most quick of schistosomiasis infection and propagation and one of effective method.
So far, using the chemosynthesis molluscicide is the main means of control oncomelania.Penta sodium pentachlorophenate is once used by many countries, and sn ail control effect is good, but to non-target biology, raising fish and shrimp class tool broad spectrum toxicity especially, and cause serious environmental to pollute, classified as the forbidding product by international community already.Niclosamidum (Niclosamidum) sn ail control effect is fine, be universally acknowledged molluscicide, but cost an arm and a leg, use in developing country's large tracts of land to be restricted, also can produce harm when high concentration is used, and also can cause the pesticide resistance of target biology to increase aquatiles such as fish.
In view of the drawback of chemosynthesis molluscicide, international community is natural to researching and developing, the molluscicide of plant origin has given very big concern.Therefore, the research and development sn ail control effect is good, cost is lower, biodegradable novel molluscacide has its necessity and urgency.
Ginkgo (Ginkgo biloba L.) is the rare tree of China, and China's gingko resource accounts for more than 70% of world's total amount.Ginkgo leaf, fruit all can be used as medicine.GINKGO BILOBA EXTRACT class in the ginkgo leaf and bilobalide-like have significant curative effect to the heart, cerebrovascular system disease, with it is that (Ginkgo biloba extract GBE) has become the autonomic drug of sales volume maximum on the international market to the ginkgo biloba p.e that is developed to of main pharmacodynamics composition.At present, China to the utilization of ginkgo biloba p.e except the demand that satisfies the domestic market, also with the medicinal extract material outlet abroad to satisfy the demand of international market.
Ginkgoic acid is the another kind of active component except that flavones, lactone in the ginkgo, is present in ginkgo leaf and the exosper.Ginkgoic acid is a class alkyl or alkenyl phenolic acid compound.
Ginkgoic acid is an important component of can not ignore in ginkgo leaf and fruit thereof, the exosper.Ginkgo is one of species the most ancient on the earth, and it is to leave over " living fossil " Jurassic Period.The ginkgo population study existing to China shows that its life-span reaches 2～3,000 years, is natural world rare " long-lived family ".One of major reason of ginkgo " longevity " is that the common damage by disease and insect of plant is had significant resistance ginkgoic acid is exactly important member in himself defense system.Ginkgo is a deciduous species, has every year a large amount of ginkgo leaves and exosper to enter natural world degraded, circulation, can not destroy ecotope.Thereby ginkgoic acid is an environmental compatible, biodegradable.
The extraction of ginkgoic acid and evaluation, referring to following paper:
van?Beek?TA，Wintermans?MS.Preparation?isolation?and?dual?columnhigh-performance?liquid?chromatography?of?ginkgolic?acids?from?Ginkgo?biloba.J?Chromatogr?A.2001；930：109～117
van?Beek，TA.Chemical?analysis?of?Ginkgo?biloba?leaves?and?extracts.J?Chromatogr?A.2002；967(1)：21-55
Fuzzati，N，Pace，R，Villa，F.A?simple?HPLC-UV?method?for?the?assay?of?ginkgolicacids?inGinkgo?biloba?extracts.Fitoterapia?2003；74(3)：247-56
Yang?Liu-qing，Wu?Xiang-yang，Chen?Jun.Determination?of?ginkgolic?acids?by?highperformance?liquid?chromatography.Yaoxue?Xuebao?2002；37(7)：555-558
Known, ginkgo biloba p.e has significant protective effect to the heart, cerebrovascular system, maincenter and peripheral neverous system, has been widely used in the control of clinical relevant disease.Referring to following monograph:
Francis?V.DeFeudis.Ginkgo?biloba?extract(EGb761)：from?chemistry?to?the?clinic.UllsteinMedical.Wiesbaden，1998；ISBN：3-86126-173-1；
[AHP]2003.American?Herbal?Pharmacopoeia.Ginkgo?Leaf，Ginkgo?Leaf?Dry?ExtractGinkgo?biloba?L.Standards?of?Analysis，Quality?Control，and?Therapeutics.ISBN：1-929425-17-1.PO?Box?66809，Scotts?Valley，CA?95067?USA)
Ginkgoic acid is the main matter that causes bad reaction in the GBE preparation.Because of its sensitization and cytotoxic effect, in standard preparation, be strictly controlled, need in extraction, preparation process, to be removed.Referring to following patent and paper: CN1145230-A; CN1073848-C.High content ginko leaves flavone lactone extractpreparation STATE CHINESE MEDICINE PHARM ENG RES; SHANGHAI XINGLINSCI ﹠amp; TECHNOLOGY PHARM.XIE D, WANG N, GAO Q, et al.
CN 99803683.8 compositions extracted from gingko biloba leaves and preparation method and application; Shed is grand etc., Xingling Science ﹠ Technology and Pharmaceutical Industry Co., Ltd., Shanghai;
CN1470487-A.Method?for?removing?ginkgolic?acid?from?ginkgo?leaf?extract?by?extractionmethod?YANGZIJIANG?PHARM?IND?GROUP?JIANGSU.HUANG?S，WANG?D，LIUC。
There are some researches show that ginkgoic acid has wide biological activity, as to the inhibitory action of G (+) bacterium, anti-plant pathogenic fungi effect, antitumor action etc.
Ginkgoic acid accounts for 1～2% of ginkgo leaf dry weight, accounts for 3～5% of gingko episperm (doing).China obtains ginkgo leaf medicinal extract amount year and reaches 200 tons, and ginkgoic acid content is about 2.2% in ginkgo leaf medicinal extract amount, and the ginkgoic acid total amount of promptly removing from ginkgo leaf medicinal extract is about 4.4 tons.And the annual ginkgo fruit exosper (doing) that abandons reaches 1.2 ten thousand tons, wherein contains about 420 tons of ginkgoic acid.Hence one can see that, has every year the ginkgoic acid of a great deal of (ton surplus in the of 400) to be dropped.To the ginkgoic acid research and development, can fully utilize, turn waste into wealth ginkgo leaf and exosper, produce huge social and economic benefit.
Summary of the invention
The purpose of this invention is to provide the new purposes of ginkgoic acid in the preparation biopesticide, be specifically related to the purposes of ginkgoic acid in preparation kill harmful molluscan biopesticide, relate in particular to ginkgoic acid and kill oncomelania in preparation, prevent the purposes in the biopesticide of schistosomiasis.
The present invention is through harmful mollusk experiment, and the result shows, ginkgoic acid can effectively suppress some key enzyme in the mollusk energy supply biochemical metabolism and the mitochondria oxidative phosphorylation produced to separate coupled effect, causes its energy supply to interrupt and produces killing action; Ginkgoic acid can effectively suppress the key enzyme-tyrosinase in the insect molting process and the important enzyme-prostaglandin synthetase and the prostaglandin endoperoxides synzyme of insect reproductive process, thereby suppress breeding and the growth of insect, above-mentioned experimental result confirms that ginkgoic acid can be used for killing the harmful mollusk that spreads disease or harm the crops.
The present invention is through experiment confirm, and ginkgoic acid can be killed unique intermediate host---the oncomelania in the widely popular Japanese schistosomiasis of China effectively, and ginkgoic acid can be used for the control of schistosomiasis to the killing action of the oncomelania of propagation schistosomiasis.The present invention has opened up the new purposes that ginkgoic acid is killed oncomelania, prevented schistosomiasis, not only can turn waste into wealth, ginkgo leaf is fully utilized, and also can further prepare the new bio molluscicide and prevent schistosomiasis.
Ginkgoic acid involved in the present invention is the alkyl or alkenyl phenolic acid compound, has the structure of general formula (I), and its R is the fat chain, it is the main active of killing oncomelania, mainly be present in the leaf and exosper of deciduous species ginkgo, can enter natural world degraded, circulation, can not cause environmental pollution.The ginkgoic acid structure is different with the plant molluscicide structure (as: hemiterpene class, triterpenes, osajin etc.) of known chemosynthesis molluscicide or natural origin, its structure is comparatively simple, be easy to synthesize or make structure of modification, carbon (C) atomicity on its R base can be 8～20 (C8～20); Double key number on its R base can be 0～4, as: C13: 0, C13: 1, etc.; Carbon (C) atom on its R base can be replaced as O, S, N or halogen equipotential by other atoms.
Described ginkgoic acid is by chemically separated method, obtain from the crude extract or pure the carrying the thing of ginkgo leaf and gingko episperm and fruit thereof, or ginkgo biloba p.e reclaimed in the remaining waste material in extracting process again, fully utilize, or obtain from the each several part extract that other plant contains this compounds; Or obtain by the method for chemosynthesis, be that raw material is synthesized with the salicylic acid; Or further by high performance liquid chroma-tography technology (HPLC) with other correlation analysis technology are analyzed, evaluation and Quality Control.
The medicine that comprises The compounds of this invention can be the crude extract or the pure thing of carrying of the leaf, exosper and the fruit thereof that derive from ginkgo, also can be to derive from the each several part extract that other plant contains this compounds.
The medicine that comprises The compounds of this invention can be separately uses as molluscicide, also can share with other molluscicides, to strengthen molluscicidal effect or to reduce its toxic and side effect, reduce cost.
The pharmaceutical composition that comprises The compounds of this invention can be a solid form, as pulvis, granule, liposome etc.Its solid carrier can be nontoxic, biodegradable, hydrophily or lyophobic dust.The pharmaceutical composition that comprises The compounds of this invention can be a liquid form, as solution, emulsion, suspension etc.Its liquid-carrier can be water or hydrophily or hydrophobic organic solvent and with the mixture of various ratios in water.
The pharmaceutical composition that comprises The compounds of this invention can be sprayed on the waters or soil of containing oncomelania.Can spray once in 3～4 days at interval, to strengthen killing effect oncomelania.
Mentioned all publications, patent application, patent and other lists of references of this paper all is incorporated herein by reference.
Embodiment
(C13: 0) component is killed the effect of oncomelania to embodiment 1 ginkgoic acid
1. the extraction of ginkgoic acid:
With the gingko episperm of 100g drying, pulverizing, add benzinum (60～90 ℃ of boiling ranges) ultrasonic extraction 1h in 1: 5 ratio, filter, filter residue repeats to extract twice, merges to extract, and flings to benzinum and gets exosper crude extract medicinal extract.
With the medicinal extract gradation with a small amount of petroleum ether dissolution application of sample (silica gel 120g on silicagel column, silicagel column is the glass column of Φ 32 * 400mm, wet method dress post), use benzinum: ether: formic acid=89: 11: 1 (V/V/V) wash-out, fluorescence occurring at the 254nm place with thin-layer chromatography is detection, collect the ginkgoic acid component, be concentrated into dried; Repeated post once, the ginkgoic acid component that obtains is washed to neutrality, is concentrated into driedly, or uses anhydrous Na
2After the SO4 drying, get the ginkgoic acid mixture.
2. the purifying of ginkgoic acid and analysis (rp-hplc method):
15cm C18RP-HPLC post carries the logotype mutually of silver-colored cation HPLC exchange column with 20cm, use methyl alcohol: water=93: 7 (with 0.1% formic acid acidifying) is flowing phase, and the ultraviolet detection wavelength is 311nm, each component of ginkgoic acid can be carried out separation and purification, and and standard control, do quantitative assay.Zhi Bei C13 according to said method: 0 purity can reach more than 95%.
With ginkgoic acid (C13: 0) component is soaked the extremely effect of oncomelania:
(C13: 0) component is through above-mentioned purifying for ginkgoic acid, purity is 95% white crystal, use a small amount of anhydrous alcohol solution, be diluted to variable concentrations with the running water that boils the back cooling again: 50mg/L, 25mg/L, 12.5mg/L, 6.25mg/L, 3.13mg/L (the ethanol final concentration all＜0.5%).With the positive contrast medicine of niclosamidum (producing), the niclosamide ethanolamine wetting powder is diluted to desired concn with the running water that boils the back cooling by Huainan pharmaceutical factory.Simultaneously, establishing with the running water and 0.5% ethanolic solution that boil the back cooling is contrast.
Oncomelania: pick up from the popular district of yueyang, hunan blood fluke.Remove positive oncomelania with colony's ease larva of a tapeworm or the cercaria of a schistosome method, at laboratory cultures 72h.Selecting spiral shell age is that 8～9 one-tenth spiral shells that revolve are standby.
Add in the 25ml beaker in each beaker of different soups (promptly filling it up with to rim of a cup) of the above-mentioned preparation of 30ml and put 10 oncomelanias, rim of a cup covers nylon wire, in case oncomelania climbs out of.
To being soaked in the corresponding liquid, being contained in the oncomelania in the beaker on the horizontal table top, under room temperature (22 ± 1 ℃), observe the death condition of oncomelania during respectively at 24h and 48h.Observe its active situation with needle point method in conjunction with hammering method, judge whether it is dead.
Experimental result sees Table 1.The hypoergia of oncomelania is not obvious in water and the ethanol control group, and (C13: 0) the oncomelania energy of component processing descends essentially no climbing phenomenon through ginkgoic acid.
Table 1: (C13: 0) component is soaked the effect (48h) of oncomelania extremely to ginkgoic acid
| Water | Ethanol (0.5%) | Niclosamidum (mg/L) | Ginkgoic acid-C13 (50mg/L) | Ginkgoic acid-C13 (25mg/L) | Ginkgoic acid-C13 (12.5mg/L) | Ginkgoic acid-C13 (6.25mg/L) | Ginkgoic acid-C13 (3.13mg/L) | |
| Death toll | 0 | 0 | 10 | 10 | 9 | 4 | 1 | 0 |
| (inhibiting rate %) | 0 | 0 | 100 | 100 | 90 | 40 | 10 | 0 |
Above-mentioned experiment is through repeated test, as a result basically identical.Ginkgoic acid (C13: 0) kill oncomelania as calculated
LD50＝14.51mg/L(R＝0.9549)
(C15: 1) component is killed the effect of oncomelania to embodiment 2 ginkgoic acids
Ginkgoic acid (C15: extraction 1):
With above-mentioned exosper crude extract medicinal extract with a small amount of petroleum ether dissolution application of sample (silica gel 120g on silicagel column, silicagel column is the glass column of Φ 32 * 400mm, wet method dress post), use benzinum: ether: formic acid=89: 11: 1 (V/V/V) wash-out, fluorescence occurring at the 254nm place with thin-layer chromatography is detection, collect the wherein component of spot maximum, be concentrated into dried, ginkgoic acid (C15: 1) component.
2. analyze and purity testing (rp-hplc method):
15cm C18RP-HPLC post carries the logotype mutually of silver-colored cation HPLC exchange column with 20cm, use methyl alcohol: water=93: 7 (with 0.1% formic acid acidifying) is flowing phase, and the ultraviolet detection wavelength is 311nm, and (C15: 1) component is identified to above-mentioned ginkgoic acid, and and standard control, make purity testing.(C15: 1) component purity is 60.63% to the gained ginkgoic acid after measured.
With ginkgoic acid (C15: 1) component soaks the extremely effect of oncomelania:
With the above-mentioned ginkgoic acid that obtains (C15: 1) component, use a small amount of anhydrous alcohol solution, be diluted to variable concentrations with the running water that boils back cooling again: 50mg/L, 25mg/L, 12.5mg/L, 6.25mg/L, 3.13mg/L (the ethanol final concentration all＜0.5%).With the positive contrast medicine of niclosamidum (producing), the niclosamide ethanolamine wetting powder is diluted to desired concn with the running water that boils the back cooling by Huainan pharmaceutical factory.Simultaneously, establishing with the running water and 0.5% ethanolic solution that boil the back cooling is contrast.
Oncomelania: pick up from the popular district of yueyang, hunan blood fluke.Remove positive oncomelania with colony's ease larva of a tapeworm or the cercaria of a schistosome method, at laboratory cultures 72h.Selecting spiral shell age is that 8～9 one-tenth spiral shells that revolve are standby.
Add in the 25ml beaker in each beaker of different soups (promptly filling it up with to rim of a cup) of the above-mentioned preparation of 30ml and put 10 oncomelanias, rim of a cup covers nylon wire, in case oncomelania climbs out of.
To being soaked in the corresponding liquid, being contained in the oncomelania in the beaker on the horizontal table top, under room temperature (22 ± 1 ℃), observe the death condition of oncomelania during respectively at 24h and 48h.Observe its active situation with needle point method in conjunction with hammering method, judge whether it is dead.
Experimental result sees Table 2.The hypoergia of oncomelania is not obvious in water and the ethanol control group.(C15: 1) the oncomelania energy of component processing descends essentially no climbing phenomenon through ginkgoic acid.
Table 2: (C15: 1) component soaks the effect (48h) of oncomelania extremely to ginkgoic acid
| Water | Ethanol (0.5%) | Niclosamidum (mg/L) | Ginkgoic acid-C13 (50mg/L) | Ginkgoic acid-C13 (25mg/L) | Ginkgoic acid-C13 (12.5mg/L) | Ginkgoic acid-C13 (6.25mg/L) | Ginkgoic acid-C13 (3.13mg/L) | |
| Death toll | 0 | 0 | 10 | 10 | 5 | 2 | 0 | 0 |
| (inhibiting rate %) | 0 | 0 | 100 | 100 | 50 | 20 | 0 | 0 |
Above-mentioned experiment is through repeated test, as a result basically identical.(C15: 1) (60.63%) component is killed the LD50=20.46mg/L (R=0.9568) of oncomelania to ginkgoic acid as calculated
Embodiment 3 gingko episperm crude extract medicinal extract are to the killing action of oncomelania
1. the preparation of gingko episperm crude extract:
With the gingko episperm of 100g drying, pulverizing, add benzinum (60～90 ℃ of boiling ranges) ultrasonic extraction 1h in 1: 5 ratio, filter, filter residue repeats to extract twice, merges to extract, and flings to benzinum and gets exosper crude extract medicinal extract.
2. the content analysis (rp-hplc method) of ginkgoic acid in the gingko episperm crude extract:
Measure with above-mentioned RP-HPLC method, the content of total ginkgoic acid is 5.46% in the gingko episperm crude extract.
3. gingko episperm crude extract medicinal extract soaks the effect of oncomelania extremely:
With above-mentioned gingko episperm crude extract, use a small amount of anhydrous alcohol solution, being diluted to crude extract content with the running water that boils the back cooling again is 62.5mg/L (ethanol final concentration＜0.5%).With the positive contrast medicine of niclosamidum (producing), the niclosamide ethanolamine wetting powder is diluted to desired concn with the running water that boils the back cooling by Huainan pharmaceutical factory.Simultaneously, establishing with the running water and 0.5% ethanolic solution that boil the back cooling is contrast.
Oncomelania: pick up from the popular district of yueyang, hunan blood fluke.Remove positive oncomelania with colony's ease larva of a tapeworm or the cercaria of a schistosome method, at laboratory cultures 72h.Selecting spiral shell age is that 8～9 one-tenth spiral shells that revolve are standby.
Add in the 25ml beaker in each beaker of different soups (promptly filling it up with to rim of a cup) of the above-mentioned preparation of 30ml and put 10 oncomelanias, rim of a cup covers nylon wire, in case oncomelania climbs out of.
To being soaked in the corresponding liquid, being contained in the oncomelania in the beaker on the horizontal table top, under room temperature (22 ± 1 ℃), observe the death condition of oncomelania during respectively at 24h and 48h.Observe its active situation with needle point method in conjunction with hammering method, judge whether it is dead.
Experimental result is: gingko episperm crude extract 62.5mg/L 24h mortality of snails is 60%, and the 48h lethality is 100%.The hypoergia of oncomelania is not obvious in water and the ethanol control group, and the oncomelania energy of handling through medicinal extract descends essentially no climbing phenomenon.
(C15: 1) component and niclosamidum are to the collaborative killing action of oncomelania for embodiment 4 ginkgoic acids
1. ginkgoic acid (C15: the 1) preparation of component and niclosamidum mixed liquor
(C15: 1) component, measure its purity through HPLC is 60.63% to the ginkgoic acid of above-mentioned separation preparation.With ginkgoic acid (C15: 1) a small amount of anhydrous alcohol solution of component, again with the running water dilution of boiling the back cooling, mix with the niclosamide ethanolamine wetting powder of preparing with the running water that boils the back cooling, (C15: 1) final concentration of component and niclosamide ethanolamine wetting powder is respectively 10mg/L and 0.5mg/L to be diluted to ginkgoic acid with the running water that boils the back cooling again.
2. (C15: 1) component and niclosamidum mixed liquor soak the effect of oncomelania extremely to ginkgoic acid
With the positive contrast medicine of niclosamidum (producing), the niclosamide ethanolamine wetting powder is diluted to desired concn with the running water that boils the back cooling by Huainan pharmaceutical factory.Simultaneously, establishing with the running water and 0.5% ethanolic solution that boil the back cooling is contrast.
Oncomelania: pick up from the popular district of yueyang, hunan blood fluke.Remove positive oncomelania with colony's ease larva of a tapeworm or the cercaria of a schistosome method, at laboratory cultures 72h.Selecting spiral shell age is that 8～9 one-tenth spiral shells that revolve are standby.
Add in the 25ml beaker in each beaker of different soups (promptly filling it up with to rim of a cup) of the above-mentioned preparation of 30ml and put 10 oncomelanias, rim of a cup covers nylon wire, in case oncomelania climbs out of.
To being soaked in the corresponding liquid, being contained in the oncomelania in the beaker on the horizontal table top, under room temperature (22 ± 1 ℃), observe the death condition of oncomelania during respectively at 24h and 48h.Observe its active situation with needle point method in conjunction with hammering method, judge whether it is dead.
Experimental result is: (C15: 1) in component and the niclosamidum mixed liquor, the 24h mortality of snails is 80%, and the 48h lethality is 100% at ginkgoic acid.The hypoergia of oncomelania is not obvious in water and the ethanol control group, and the oncomelania energy of handling through medicinal extract descends essentially no climbing phenomenon.
Embodiment 5 ginkgoic acids are to the coupled effect of separating of oncomelania mitochondria oxidative phosphorylation
1. ginkgoic acid (C15: the 1) preparation of component
(C15: 1) component, measure its purity through HPLC is 60.63% to the ginkgoic acid of above-mentioned separation preparation.(C15: 1) a small amount of anhydrous alcohol solution of component, the running water with cooling after boiling dilutes again, and is standby with ginkgoic acid.
2. the coupled effect of separating of mitochondrial preparation of oncomelania and oxidative phosphorylation is measured
Get a collection of energetic adult oncomelania, remove positive oncomelania with colony's ease larva of a tapeworm or the cercaria of a schistosome method, at laboratory cultures 72h.Selecting spiral shell age is that 8～9 one-tenth spiral shells that revolve are standby.With broken spiral shell method broken shell, the cephalopodium portion of getting oncomelania is by centrifugal its mitochondria of purchasing of literature method.Adopt oxygen electrode method to measure mitochondrial respiratory control ratio (RCR), and (C15: 1) component is to the uncoupling of mitochondria oxidative phosphorylation reaction to adopt the variation of peacock green method mensuration phosphorus and the method research ginkgoic acids such as activity of ATP enzyme.
Result of study shows, (C15: 1) component is at used dosage range (10mg/L for ginkgoic acid, 20mg/L, 40mg/L) be dose dependent and improve the content of Phos in the reactant liquor and the activity of ATP enzyme, reduce its RCR, show that (C15: 1) oxidative phosphorylation has and separates coupled effect preferably component ginkgoic acid to the oncomelania mitochondria.
Claims (4)
1. ginkgoic acid is killed oncomelania in preparation, prevents the purposes in the biopesticide of schistosomiasis, and described ginkgoic acid is the ginkgoic acid of following general formula (I),
Wherein, R is the fat chain, and the carbon number on its R base is 13 or 15; And when carbon number is 13,
Double key number 0 on the R base; When carbon number is 15, the double key number 1 on the R base.
2. according to the purposes of claim 1, it is characterized in that described biopesticide is a molluscicide.
3. according to the purposes of claim 2, it is characterized in that described molluscicide is a liquid preparation.
4. according to the purposes of claim 2, it is characterized in that described molluscicide uses separately or share with niclosamidum.
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| CNB2006100240405A CN100462006C (en) | 2006-02-21 | 2006-02-21 | Use of ginkgoic acid in preparation of biological pesticide for killing Oncomelania snail and preventing schistosomiasis |
| PCT/CN2007/000461 WO2007095842A1 (en) | 2006-02-21 | 2007-02-15 | Use of ginkgolic acids in preparing biotic pesticide for killing snails and preventing schistomiasis |
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| CNB2006100240405A CN100462006C (en) | 2006-02-21 | 2006-02-21 | Use of ginkgoic acid in preparation of biological pesticide for killing Oncomelania snail and preventing schistosomiasis |
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| CN102010328B (en) * | 2009-09-04 | 2013-07-10 | 复旦大学 | Long-chain alkyl salicylic compound as well as preparation method application thereof |
| CN101911938B (en) * | 2010-07-29 | 2013-04-17 | 江苏大学 | Application of ginkgoic acid and ginkgol compounds in killing Ampullaria gigas |
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| CN103520187B (en) * | 2013-11-04 | 2016-03-09 | 中国疾病预防控制中心寄生虫病预防控制所 | The application of ginkgoic acid in anti-Cryptosporidium |
| CN107087608B (en) * | 2017-05-16 | 2019-11-05 | 江苏省血吸虫病防治研究所 | Chlorophenoxyacetic acid is preparing the application in snail-killing medicine |
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| US20050136136A1 (en) * | 2001-07-11 | 2005-06-23 | The Trustees Of Columbia University | Method for isolating terpene trilactones (ginkgolides, bilobadide) from leaves and pharmaceutical powders of ginkgo biloba |
| WO2005105122A1 (en) * | 2004-04-29 | 2005-11-10 | Goetect Gmbh | Ginkgo biloba extract and method for producing the same |
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