158 results about "Tetrodotoxin" patented technology

This invention relates to a stable pharmaceutical composition of freeze-dried tetrodotoxin powder which contains trace amount of tetrodotoxin, substances which can stabilizes tetrodotoxin, including disaccharide(s) or polyglucose(s) or analogues thereof and solvent(s), and solvents which can help tetrodotoxin dissolve.

The present invention relates to matter extraction from animal material and is the extraction process of tetrodotoxin. The present invention provides the technological method of raising yield of tetrodotoxin. The technological process includes the steps of: cutting internal organs of globefish, grinding into slurry, press filtering, heating to eliminate protein and oil, filtering to obtain clear liquid, concentration, elution of crystal inchromatographic column to obtain pure tetrodotoxin product.

The invention relates to a method for separating Aeromonas molluscorum producing tetrodotoxin from Takifugu fasciatus tissues and a fermentation culture method of the Aeromonas molluscorum as well as a detection method of the produced tetrodotoxin. The method for separating the Aeromonas molluscorum producing the tetrodotoxin from the Takifugu fasciatus tissues comprises a bacteria separation method, a bacteria identification method, a bacteria fermentation culture method, a competitiveness ELISA (Enzyme Linked Immunosorbent Assay) detection method and an LC-MS (Liquid Chromatograph-mass Spectrometry) detection method of the tetrodotoxin produced by the Aeromonas molluscorum, and the like. In the invention, by separating bacteria from Takifugu fasciatus ovaries and adopting a TCBS (Thiosulfate Citrate Bile (salt) Sucrose (agar)) culture medium for screening, the Aeromonas molluscorum capable of producing the tetrodotoxin is separated from blowfish bodies for the first time; the separated bacteria is identified by adopting a 16 SrDNA method; the competitiveness ELISA detection method and the LC-MS detection method are both firstly adopted for the tetrodotoxin produced by the separated Aeromonas molluscorum; and the produced tetrodotoxin and tetrodotoxin extracted from the blowfish bodies are determined to be the same substance; in addition, the separated Aeromonas molluscorum can massively separate the tetrodotoxin from the bacteria after being cultured.

A process for preparing the liquid extract of tetrodotoxin includes such steps as quickly freezing the viscera of globefish, cutting to become small blocks, immersing in deionized water, fermenting, quick heating, quick cooling, primary filtering and fine filtering. It has high output rate.

The externally applied compound tetrodotoxin ointment is prepared with globe fish liver oil, vaseline, frankincense, myrrh, dragon's blood, notoginseng, safflower, catechu, borneol, vitamin B1, vitamin B2, vitamin C, moroxydine, dexamethasone and lidocaine in certain weight proportion. It has very high curative rate on trauma and traumatic infection, including intractable ulcer and fistula, high effective rate and curative rate on skin diseases caused by bacteria, fungi, viruses, etc, and certain treating effect on serious furuncle, luetic balanitis, etc.

The invention discloses a process for preparing leftover fertilizer processed through globefish by adopting an enzymolysis and fermentation method. A composite enzymatic hydrolysis and multi-strain fermenting mode is adopted for degrading macromolecule substances difficult to utilize in leftovers processed through the globefish into micromolecule polysaccharide, polypeptide and nitrogen-containing compounds, a large number of soil beneficial bacteria and various special-effect metabolites are generated, the biological activity of the leftover fertilizer processed through the globefish is improved, and pesticide and the fertilizer are combined into one, which is beneficial for pushing the development of the ecological agriculture and increasing the resource comprehensive utilization rate and the economic benefits of the globefish processing industry. The number of the effective active bacteria in the fertilizer is larger than or equal to 20 billion/mL. The fertilizer integrates natural nutrient ingredients, antibiotic substances and zoohormone, and has a composite effect of organic fertilizer, microbial fertilizer and microelement fertilizer; the fertilizer contains a certain amount of tetrodotoxin, and can effectively kill pests in soil, the pesticide and fertilizer combination is achieved, the duration time for the pest killing effect is long, and the application prospect is wide.

The invention discloses a method for determining tetrodotoxin in marine organisms by utilizing immunoaffinity column purification-liquid phase chromatography-tandem mass spectrometry. The method is characterized in that the tetrodotoxin in the marine organisms can be rapidly, simply and high efficiently determined by utilizing the unique selection identifiability of the immunoaffinity column on the basis of the high sensitivity and accuracy of the liquid phase chromatograph tandem mass spectrometry.

The invention provides a set of single stranded DNA (ssDNA) aptamers DA-01 and TTX-27 which can simultaneously recognize domoic acid (DA) and tetrodotoxin (TTX), including one ssDNA aptamer DA-06 which specifically recognizes DA, one ssDNA aptamer TTX-07 which can specifically recognize TTX, and one ssDNA aptamer STX-41 which can specifically recognize saxitoxin (STX). According to the invention,magnetic reduced graphene oxide (MRGO) is adopted to assist in separated systematic evolution of ligands by exponential enrichment (SELEX), properties of MRGO of being capable of adsorbing free ssDNA,but not adsorbing ssDNA combined with target molecules are utilized, and magnetic separation enrichment characteristics of MRGO are further used to separate ssDNA with affinity and without affinity to target toxins; primary structure homology and secondary structure similarity on obtained ssDNA are analyzed after 16 repeated rounds of incubation, separation, amplification and in-vitro screening for single-strand preparation, then the affinity and the specificity are measured, and finally one set of the aptamers with high affinity and specificity is obtained. The set of the aptamers has broadapplication prospects in the aspects of analysis, detection, separation, enrichment, removal and purification for DA, TTX and STX in aquatic products.

The invention discloses a rapid cultivation feed for takifugu rubripes. The rapid cultivation feed comprises the following components in parts by weight: 20-40 parts of domestic fish meal, 10-20 parts of shrimp powder, 10-20 parts of meat and bone meal, 1-10 parts of chicken oil, 10-20 parts of corn starch, 1-5 parts of soybean lecithin, 0.1-5 parts of an oyster shell extract, 0.1-8 parts of cylindrotheca fusiformis powder, 2-6 parts of sodium alginate, 1-15 parts of functional additive components, 0.1-3 parts of multivitamins and 0.1-5 parts of multiminerals. A preparation method of the rapid cultivation feed disclosed by the invention comprises the following steps: (1) manufacturing raw materials; (2) synthesizing a biological material; (3) converting to a cooked material; (4) puffing and granulating; (5) drying; (6) spraying and coating; and (7) performing vacuum drying. According to the feed disclosed by the invention, by scientific matching, the body color of adult takifugu rubripes can be improved, the growth can be promoted, and the enrichment of tetrodotoxin can be reduced.

The domesticating and culturing process of dark-grained Eastern globefish with controlled tetrodotoxin includes the steps of artificial propagation, fry culture, globefish culture, etc. The present invention makes it possible to control the tetrodotoxin content under fresh water culture condition simply without pollution.

The invention relates to a method for preparing a tetrodotoxin-degrading substance from a tetrodotoxin-producing bacteria fermentation broth, and a preparation and detection method of an extract liquid. The method comprises a method for artificially causing plasmid loss, a tetrodotoxin-degrading substance extraction method, and a tetrodotoxin degradation detection method. According to the invention, for a first time, the existence of a tetrodotoxin-degrading substance in aeromonas molluscorum is found, and an extraction method of the substance is established. The method has certain potential to be used in large-scale productions of the tetrodotoxin-degrading substance. The extract of the bacteria fermentation broth has a tetrodotoxin-degrading effect, and can be used for preparing medicines used for treating tetrodotoxin intoxication. Also, a raw material can be provided for related fields such as scientific research and pharmaceutical industries.

The invention relates to a tetrodotoxin preparation for treating drug addiction and relieving pain. The preparation comprises small-dose tetrodotoxin serving as a medicinal active ingredient and a pharmaceutical permissible cosolvent or support agent, wherein the cosolvent is amino acid. The amino acid is used as the cosolvent, so the amino acid has the function of the cosolvent, increases the water solubility of the tetrodotoxin, and can also form highly water-soluble salt solution with the tetrodotoxin. The tetrodotoxin used in the invention is prepared by a specific method; and the purity of the prepared tetrodotoxin reaches 98 percent and the yield is high. The preparation consisting of the tetrodotoxin prepared by the method and the amino acid has the advantages of high water solubility, high stability, safety, reliability, stable quality, no need of preservation in a refrigerator at a specific temperature, storage life of more than two years at room temperature, capability of greatly saving the storage and transportation cost and facilitating medical use, obvious treatment effect, and cure rate of 100 percent when particularly used for physiological detoxification.

The invention discloses a preparing method for tetrodotoxin standard biological samples. The preparing method includes the steps that nassarius samples are extracted with an acetic-acid methanol solution, a crude extracting solution is purified with a carbon-nano-tube solid-phase extraction column, the purified venom is subjected to high-performance-liquid-chromatography purification, and then isfrozen, dried and concentrated, crystals are separated out, and the tetrodotoxin standard biological samples are obtained. According to the preparing method for the tetrodotoxin standard biological samples, in the TTX extraction aspect, the purifying aspect and the pure-product preparing aspect, the nassarius serves as a raw material, extraction is carried out with the acetic-acid methanol solution, the crude extracting solution is purified through the large-volume-carbon-nano-tube solid-phase extraction column, then the purified venom is concentrated, and the samples are purified and preparedthrough high-performance-liquid sample feeding. The sample preparing method and process are easy and convenient to operate, efficient and suitable for batch production and technology preparation of the tetrodotoxin standard biological samples.

The invention discloses a making method of a semi-quantitative tetrodotoxin detection card. The method comprises the following steps: 1, preparing a colloidal gold sample combination pad; 2, chromatographic membrane processing: uniformly drawing a tetrodotoxin artificial antigen forming a detection line and a quality control antibody forming a quality control line in the middle of a chromatographic membrane by using a membrane drawing instrument; and 3, assembling: splicing a carrier pad, the chromatographic membrane, the colloidal gold sample combination pad and a water absorption pad to form a layered structure, wherein the carrier pad is positioned at the bottom, the chromatographic membrane is arranged on the carrier pad, and the colloidal gold sample combination pad and the water absorption pad are respectively placed at two ends of the chromatographic membrane. The above obtained product allows tetrodotoxin to be rapidly semi-quantitatively analyzed through ocular estimation, and has the characteristics of small size, light weight, easy preservation and transportation, and realization of detection of a detected sample without separation or purification.

The method for extracting tetrodotoxin from liver of globefish includes the following steps: cutting liver of globefish, soaking at low temp., stirring, filtering, freezing, repeatedly soaking for several times to obtain crude venom, freezing and standing still, precipitating, heating and boiling, discharging out venom, freezing, and separating out supernatant venom, regulating pH value, adding active carbon, stirring, filtering, mixing cation 110 resin and said clear liquor, stirring, filtering, making said clear liquor pass through cation 110 resin, saturating and using acid to make elution, collecting eluant, regulating pH value, cooling, standing still, crystallizing, sucking out mother liquor, filtering, more regulating pH, freezing, standing still, crystallizing and repeatedly regulating pH for several times to obtain white powder crystal.

The invention relates to a method for massively and extremely efficiently extracting high-purity tetrodotoxin from viscera of globefishes, and aims at overcoming defects, in existing various methods for extracting and purifying tetrodotoxin, that the efficient liquid phase method with high product purity is high in cost of equipment (a high-pressure liquid phase instrument), and the dipping fractionation method is low in efficiency and low in product purity. The tetrodotoxin is applicable to drug rehabilitation, analgesia and pressure reduction drug preparations. The method is performed for massively extracting and preparing tetrodotoxin. The method is simple in processes, high in efficiency, high in yield, and high in quality; the product purity is up to 97-99% and directly meets the requirements of biological study and clinical application. Additionally, the invention aims at providing various preparations containing the tetrodotoxin prepared by the method.

The present invention relates methods for the biosynthesis of tetrodotoxin (TTX) involving the steps of obtaining a culture possessing one or more of a Vibrio species, such as through a seed culture/ inoculating the culture and tissue extract from a textrodotoxin-bearing organism in a fermenter medium, and isolating and purifying tetrodotoxin from said fermenter, resulting in a yield of TTX of about 0.5 g TTX/L in about 3 to 5 days, such TTX being at least 90% pure.

The invention discloses a method for rapidly detecting the content of tetrodotoxin in fish flesh through liquid chromatography-mass spectrometry. The method comprises (1) sample preprocessing and (2) chromatography-mass spectrometry detection. Diatomite is added in the preprocessing process against the effect problem of a matrix in a fish flesh sample and combining the chemical characteristics of tetrodotoxin, and the tetrodotoxin is separated from muscle tissues in an ultralow temperature freezing manner. A sample undergoes extraction by 1% acetic acid-methanol in a cell crasher at 60 DEG C, obtained extract is purified by a Carbon-GCB SPE small column, and the purified extract is directly introduced and analyzed, so the concentration time is saved. Compared with 0.2% acetic acid-water solution extraction, other SPE purification and ultrafiltration methods in the prior art, the method disclosed in the invention, adopting direct introduction and analysis of the sample, has the advantages of simplicity and low consumption in the processing process, good accuracy, convenience in operation, and good reappearance.

The invention provides a tetrodotoxin enteric-coated and sustained-release pellet and a preparation method thereof, and relates to the field of a medicine preparation technology and application. The tetrodotoxin enteric-coated and sustained-release pellet sequentially comprises a blank pellet core, an upper medicine layer, a sustained-release layer, an isolation layer and an enteric layer from inside to outside, wherein the upper medicine layer is tetrodotoxin containing a cosolvent and a bonding agent; according to a weight ratio, the weight of the sustained-release layer accounts for 20 percent to 40 percent of the weight of the medicine containing pellet; the weight of the isolation layer accounts for 2 percent to 8 percent of the weight of the sustained-release pellet; the weight of the enteric layer accounts for 10 percent to 30 percent of the weight of the isolation pellet. The blank pellet core is used as a medicine carrier; a fluidized bed medication method is used for preparing the tetrodotoxin medicine-carried pellet; the sustained-release layer, the isolation layer and the enteric layer are sequentially sprayed to obtain the tetrodotoxin enteric-coated and sustained-release pellet. The advantages of medicine release controllability, a long-period pain relieving effect, good clinic use complaisance, high safety and the like are realized. In addition, the blank pellet core fluidized bed medication and coating method is applicable to the preparation of the ultra-low-specification tetrodotoxin medicine oral sustained-release preparation; the medication rate is high; the content uniformity is good.