47 results about "Shellfish toxins" patented technology

The invention discloses a paralytic shellfish poisoning (PSP) standard sample and a preparation method and an application thereof. Positive shellfish raw materials of PSP are initially frozen and dried after being acidized and homogenized to obtain a crude sample, and then the crude sample is frozen and dried once again after being ground and sieved to obtain the product in the invention with good uniformity and stability. The standard sample can be used for capability assurance activities of PSP test items in labs, for PSP qualitative and quantitative detection and for verification of a detection method, correction of a tester and quality control and examination of test results. The invention has low raw material cost and simple preparation method, and the obtained standard sample is a material standard sample, is even and stable and easy for storage, and is beneficial for effective monitoring on shellfish poisoning.

The invention discloses a method for detecting diarrhea shellfish toxin, which comprises the following steps: A) establishing a standard curve of depolymerization of okadaic acid (OA) induced HL-7702 hepatocyte F-actin; B) carrying out the calculation on the concentration of the OA according to the standard curve of the F-actin depolymerization of the HL-7702 hepatocyte, establishing a detection method of the hepatocyte F-actin of the diarrhea shellfish toxin to determine the possible detection limit range; C) selecting one or more of components (STX, DA and YTX) which are possibly coexist with the diarrhea shellfish to act on the HL-7702 hepatocyte, determining the specificity of the method for detecting the OA content by detecting the degree of damaging the polymerizing power of the HL-7702 hepatocyte F-actin by the above components. The detection method of the invention has the advantages: (1) taking cells as experimental subjects to avoid the using of experimental animals and conforming to the rule of 3R; (2) having high sensitivity; (3) having good specificity; (4) having good repeatability; and (5) having simple and convenient sample extraction and low matrix effects.

The invention discloses a method for preparing a composite shellfish toxin adsorbent by coating porous shell particles with chitosan. The method comprises the steps of conducting diluted acid washing and water washing on shells (fan shells, oyster shells, clam shell and the like) with shellfish meat removed to remove dirt on the surfaces of the shells, conducting stoving, electric furnace firing, high-temperature calcination, smashing and screening to obtain shell particles, soaking the shell particles in chitosan / acetic acid solution with certain concentration, and conducting draining and vacuum drying to obtain the composite adsorbent with chitosan coatings formed on the inner bore walls of the shell particles and on the surfaces of the particles. The invention further discloses a method for adsorbing shellfish toxins in aquatic products by means of the composite adsorbent and a method for regeneration of the composite adsorbent. By the adoption of the method for adsorbing shellfish toxins in aquatic products by means of the composite adsorbent, shellfish toxins can be removed efficiently, so that polluted aquatic products can be utilized and environment pollution can be reduced.

Provided is a method for massive culture of Dinophysis acuminata which is a marine dinoflagellate causing diarrhetic shellfish poisoning, and methods for extracting, isolating and purifying the shellfish toxin pectenotoxin-2 from the cultured Dinophysis acuminata. Particularly, Dinophysis acuminata is cultured massively in massive culture apparatus comprising polycarbonate water bath having the bottom sinking down toward the center of the bottom; acryl tube (E) containing fluorescent lamp laid long in the center of the water bath; air supplying device (B) supplying the air to the sinking center of the bottom of the water bath; and air purifying device containing one or more devices selected from the group consisting of UV lamp (C) and carbon cartridge filter (D) purifying the air supplied by the said air supplying device.

The invention discloses a diarrhetic shellfish toxic high throughput detection device and method based on image analysis. The device comprises a power supply adapter, a wide-angle lens, an intelligent mobile facility, a first darkroom, a second darkroom, a commercial reagent detection plate, a detection plate loading stage, and an electroluminescent panel. The method comprises the following steps: preparing a sample solution of diarrhetic shellfish toxin, then preparing a standard solution, using a commercial kit to prepare a detection plate to be detected; collecting and analyzing detection images to calculate the color ratio (CR) of R component of RGB space in a detection plate hole image; drawing a calibration curve of the commercial kit to detect the diarrhetic shellfish toxin through a least square method; obtaining the hole image CR value of the sample solution, and substituting the CR value of the sample solution into the calibration curve to calculate the concentration of the diarrhetic shellfish toxic of the sample solution. The provided device achieves the high throughput quantitative detection of diarrhetic shellfish toxic, and has the advantages of simple operation steps, low cost, and suitability for on-site detection.

The invention discloses a standard sample of diarrhetic shellfish toxin and a preparation method of the standard sample. Marine mussels are taken as materials, and research on a key preparation technology of a quality control sample of the diarrhetic shellfish toxin is emphasized to conduct from the aspects of sample impurity-removal and homogenization, freeze-drying, sample sub-packaging, determination and the like; on the basis, homogenization and stability testing is carried out; and the content reference value of the quality control sample of the diarrhetic shellfish toxin is determined by determination by a value fixing method and statistical treatment on fixed value data, and the standard sample of the diarrhetic shellfish toxin for biological detection is developed. The preparation of the standard sample has important practical significance and application value in comprehensively and deeply researching ingredient detection of diarrhetic shellfish toxin and the preparation technology and a stability ensuring technology of the standard sample, actively developing detection on the standard sample of the shellfish toxin, and filling up the blank in the detection field.

The invention relates to a preparation method of a marine shellfish toxin monoclonal antibody, which is characterized in that: the antigen which is used for preparing small molecule shellfish toxin monoclonal antibody for animal immunization and hybridoma screening is the same conjugate, thereby reducing the trouble and the waste in the different preparation of immunogen and a detection source in the preparation method of a conventional small molecule monoclonal antibody; and a screening method combined with a positive screening method and a negative screening method is adopted for monoclonal antibody hybridoma screening, so that the consumption of a large amount of standard antigen can be reduced and the antibody secreted by the screened hybridoma has better antigen competition activity. The marine shellfish toxin monoclonal antibody preparation method has the benefits of simple antigen preparation, small used amount of standard antigen, less cost and good antibody competition activity.

The invention discloses a method for extracting paralytic shellfish toxins from shell tissues. The method comprises the following steps: 1, mixing shell raw materials with acetic acid until the pH value of the obtained mixture is 2-4, carrying out ultrasonic crushing, heating the crushed mixture in boiling water for 5-10min, cooling, filtering, and collecting the obtained filtrate; 2, carrying out reduced pressure concentration on the filtrate, freezing, centrifuging, taking the obtained supernatant, extracting the supernatant by using trichloromethane, uniformly mixing the obtained upper layer extract liquid with ethanol, stirring at room temperature for 20-24h, carrying out secondary centrifuge, collecting the obtained new supernatant, carrying out reduced pressure concentration, drying, dissolving the obtained dried material in acidic methanol, filtering, collecting the obtained new filtrate, drying, dissolving the dried new filtrate in an aqueous acetic acid solution with the pH value of 2-4, adding the obtained sample to a gel column, carrying out column chromatography, and collecting the fraction with the retention time of 160-250min. The method maximally improves the extraction rate of the paralytic shellfish toxins, can effectively remove fats, proteins, carbohydrates and other impurities in the tissues, and maximally reduces the bioactive loss of the toxins.

The invention relates to ionic liquid bonded silica gel used for enriching and purifying shellfish toxins. N-phenylbenzimidazole is added into a toluene solution of gamma-chloropropylmethyldiethoxysilane, and is evenly stirred; then, reflux reaction is performed under nitrogen atmosphere for 48 hours; activated silica gel is added after cooling, and the reflux reaction is continued for 24 hours. The N-phenylbenzimidazole ionic liquid bonded silica gel material is prepared by the invention for the first time; compared with a common ionic liquid supported silica gel material and a conventional ionic liquid bonded silica gel material, the N-phenylbenzimidazole ionic liquid bonded silica gel material is capable of achieving a goal of enriching and purifying various shellfish toxins in a sample by reacting with a target object through two modes of hydrophobic interaction and ion exchange, so that the defect that isolated objects are single is changed.

The invention discloses a nucleic acid apt (aptamer) biosensor for rapidly detecting STX (saxitoxin) and a preparation method of the nucleic acid apt biosensor. The apt sequence of STX is 5'SH-(CH2)6-CCGTGGAAACATGTTCATTGGGCGCACTCCGCTTTCTGT-3'. According to the biosensor, STX detecting sensitivity is improved, and STX at a minimum of 0.05 ng / mL can be detected. The advantages of fastness and ultra-sensitivity of colorimetric analysis are closely combined with the characteristics of high affinity and high selectivity of the nucleic acid apt, the biosensor has the characteristics of being simpleto operate and portable of sensors, and the requirements of ultra-sensitivity, high specificity, rapid detection and on-site detection of STX in a seawater shellfish complex matrix can be met. Research results can provide technical support for research and development of rapid detection technology and equipment for shellfish toxins in seawater shellfish, and have great scientific research significance and market value.

The invention belongs to the technical field of aptamer design, and particularly relates to an aptamer design method based on single-nucleotide molecular docking. Starting from an experimental structure of a target small molecule, positions of nucleotides combined around the target small molecule are obtained through hydration docking; then, the discrete nucleotides are assembled into a complete aptamer; and finally, the binding capacity of the designed aptamer and the target small molecule is detected by adopting an MST experiment. The aptamer design method in the invention has been used in the design of aptamers of toxin small molecule okadaic acid (OA); the toxin is a widely distributed marine biological toxin, and can cause diarrhea type shellfish poisoning, so that the detection of OAin marine products is of great significance to food safety; an aptamer OA-D1 targeting OA is obtained by design through a calculation method; the MST experiment verifies that the equilibrium dissociation constant of the aptamer and OA is 75.6 nM; and therefore, the calculation design method is reasonable and effective.

A simple solid phase extraction (SPE) method for continuous sequestering and concentration of waterborne cues from sea water conditioned with aquatic source organisms that induce toxin formation in dinoflagellates, and a method of increasing this toxin formation.

The invention discloses a fluorescence characteristic standard spectrum library for identifying paralytic shellfish poisoning-producing microalgae as well as a construction method and application of the fluorescence characteristic standard spectrum library. The method comprises the following steps of: extracting three-dimensional fluorescence spectrum information of PSP-producing algae and non-PSP-producing algae growing under different environmental conditions, extracting characteristic peaks of experimental algae by using a Db7 wavelet function, performing clustering analysis by using a system clustering method, removing abnormal spectrums, and screening standard spectrums to obtain a Db7 fluorescence characteristic standard spectrum library. When a discriminating function established according to the fluorescence characteristic standard spectrum library is used for discriminating the PSP-producing algae and the non-PSP-producing algae, the discriminating correct rates are 84.6% and95.6% respectively, the correct rates are very high, and the purpose of quickly and accurately identifying the poisoning-producing algae is basically achieved. The fluorescence characteristic standardspectrum library can be used for preparing a red tide algae identification sensor or a portable algae fluorescence identification instrument.

The invention discloses an application of a three-dimensional fluorescence spectrometry in identification of paralytic shellfish toxin-producing microalgae. The method comprises the following steps: extracting three-dimensional fluorescence spectrum information of PSP-producing algae and PSP-non-producing algae growing under different environmental conditions, extracting characteristic peaks of experimental algae by using a Coif2 wavelet function, performing clustering analysis by using a system clustering method, removing abnormal spectrums, and screening standard spectrums to obtain a Coif2fluorescence characteristic standard spectrum library; when the discriminating function established according to the fluorescence characteristic standard spectrum library is used for discriminating the PSP-producing algae and the PSP-non-producing algae, the discriminating correct rates are respectively 77.3% and 84.5%, the correct rates are very high, and the purpose of quickly and accurately identifying the toxin-producing algae is basically achieved. The invention provides a research result of the application of the three-dimensional fluorescence spectrometry in identification of paralyticshellfish toxin-producing microalgae for the first time, and the three-dimensional fluorescence spectrometry can be applied to the aspects of red tide algae identification sensors or portable algae fluorescence identification instruments and the like.

The invention discloses a saxitoxin nondestructive rapid detection method and system based on a hyperspectral image technology, relates to the technical field of saxitoxin detection, and is used for solving the problems that an existing saxitoxin detection method is long in shellfish detection time and complex in operation, and a sample needs to be pretreated. The method is characterized by comprising the following steps: acquiring hyperspectral images of a plurality of shellfish samples; performing spectral data extraction and preprocessing on the hyperspectral image; obtaining an optimal characteristic wave band subset based on a characteristic selection algorithm of fuzzy clustering and particle swarm optimization; inputting the optimal characteristic wave band subset into a saxitoxin nondestructive testing model based on a cost-sensitive probabilistic neural network for training to obtain a trained testing model; and inputting the spectral data of the preprocessed shellfish sample to be detected into the trained detection model to obtain a detection result. According to the method, the hyperspectral image technology is applied to nondestructive testing of saxitoxin, and a new way is provided for solving the food safety problem of shellfish.

The invention relates to the field of food safety, in particular to a preparation method of chitosan microspheres for adsorbing shellfish toxins. The invention adopts the following technical scheme: the preparation method of chitosan microspheres for adsorbing shellfish toxins is characterized by comprising the following steps: first, gradually adding 120-160 parts by weight of acetic acid solution to 3-4 parts by weight of chitosan to ensure that the chitosan is dissolved in the acetic acid solution; then carrying out low-temperature refrigeration for 18-20 hours; then sucking chitosan into asyringe; then dropwise adding the chitosan in the syringe into coagulation liquid; after the chitosan is coagulated into spheres, repeatedly washing the spheres to be neutral by deionized water; andthen carrying out drying. The chitosan microspheres provided by the invention have the advantages of high adsorption efficiency and good adsorption effect.

The invention relates to the technical field of marine toxin removing, and particularly relates to a method for removing shellfish toxin. The method comprises the following steps: soaking shellfish ina pretreatment solution; performing antioxidant solution treatment, and draining and packing the shellfish; freezing the shellfish at the temperature of negative 2 to negative 4 DEG C, and performingirradiation treatment; and performing freezing treatment at the temperature of negative 2 to negative 4 DEG C, and performing storage under the condition of 4-8 DEG C. According to the method, heavymetals and shellfish toxin in shellfish bodies can be effectively removed, and the shellfish can meet the safety requirement of foods; and furthermore, the taste and color of shellfish products are not influenced in the treatment process, antioxidant addition is not required in the preparing process, the shellfish product prepared by the method has the advantages of long preservation period and noother adverse smell, and is suitable for large-scale popularization and application.

The invention relates to a method for quickly screening and identifying various paralytic shellfish toxins in red tide algae. The method comprises the following steps: weighing a certain quantity of red tide algae samples, putting into a centrifugal tube, adding a certain quantity of extracting solvents, crushing by adopting a cell crusher or an algae grinding method, then extracting the various paralytic shellfish toxins and combining into an extracting solution; performing centrifugal separation to remove a precipitate, absorbing supernatant liquid, filtering through a micro-porous filtering membrane and injecting into a sample bottle for later use; performing separation analysis on the paralytic shellfish toxins in the red tide algae crude extracting liquid by adopting a high-performance liquid chromatography-high resolution mass spectrometry; through comparing information related to the accurate molecular weight of the common paralytic shellfish toxins in the red tide algae, the quick identification on the various common paralytic shellfish toxins in the red tide algae is realized. The method provided by the invention can be used in a department of marine environmental management to quickly determine whether the paralytic shellfish toxins are contained in the red tide algae or not and is further used for judging the influence of the marine shellfish toxins on aquatic products to ensure the consumption safety of the aquatic products.

The present invention relates to a process for extracting and purifying shellfish toxins. First, the purification, identification and expanded cultivation of toxin-producing algal species are carried out. Natural seawater samples are continuously separated, purified and cultivated, and the species is determined morphologically by scanning electron microscopy. The full-length 18SrDNA sequence was obtained from the PCR product and sequenced, and the sequence comparison and analysis was carried out at NCBI through Blast to confirm its species, and then optimize the culture conditions of the target algae species, such as the composition of the culture medium, pH value, temperature, Light, salinity, culture time, etc., and use optimized conditions to expand the culture, and then extract and purify to obtain shellfish toxins. The process provided by the invention not only increases the content of the shellfish toxin in the culture solution and is beneficial to later extraction, but also has simple extraction and purification steps, high yield and low cost, and is beneficial to large-scale industrial application.

The invention discloses a raw oyster health care wine, which is prepared from raw oyster 20-25 parts, kaoliang wine 120-150 part, the fruit of Chinese wolfberry 5-7 part, Poria cocos 12-16 part, aronia melanocarpa 5-10 parts, and natural spring water 60-100 parts. The fresh oyster is purified by removing microbial pathogenic bacteria pollution, heavy metal pollution and shellfish toxin pollution,the surface of the oyster meat can be compressed, the nutrition can be loc and the smell can be eliminated by soaking the oyster meat in low-temperature saline water. The supernatant separated from the broken oyster meat can be directly added into the wine, and the oyster cake made from the residue is soaked in wine slowly to release the nutrients contained in the oyster meat to the maximum extent. Meanwhile, the fruit of Chinese wolfberry, Poria cocos and aronia melanocarpa can strengthen the physique and cooperate with raw oyster to play the role of tonifying essence, strengthening kidney, consolidating the essence and tranquilizing mind, and have better health care effect on the deficiency of body and weakness of qi and male fertility.

The invention belongs to the technical field of food safety detection, and particularly relates to an amnesic shellfish toxin immunoaffinity column and a preparation method and application thereof. The preparation method includes: taking sepharose gel activated by adopting an epoxy chloropropane activation method as a carrier; coupling genetic recombinant protein G onto the carrier to obtain protein G-sepharose; connecting an amnesic shellfish toxin antibody onto the protein G-sepharose to obtain antibody-protein G-sepharose; using a crosslinking agent for crosslinking to obtain an antibody-protein G-spharose filler; packing the antibody-protein G-sepharose filler to form the amnesic shellfish toxin immunoaffinity column having high purity and affinity. The characteristic that the geneticrecombinant protein G is combined with the amnesic shellfish toxin antibody specifically is fully utilized, and Fab segment of the amnesic shellfish toxin antibody is exposed outside, so that antibodycombining capacity of the genetic recombinant protein G, capturing ability of amnesic shellfish toxin and purifying efficiency of the amnesic shellfish toxin are improved substantially.

The invention relates to a method for extracting and purifying a shellfish toxin from a toxic algae culture solution. The method comprises four steps of filter membrane filtration, adsorption and desalting, extraction and column chromatography. An extraction agent is prepared from n-hexane, chloroform, ethyl acetate and n-butyl alcohol. Normal phase silica gel eluents are prepared from ethyl acetate, n-hexane and chloroform. Reversed phase silica eluents are prepared from water, n-butanol, acetonitrile and methanol. High performance liquid chromatography eluents are prepared from acetonitrileand methanol. By optimizing technology processes, the extraction agent and eluents are reasonably chosen, all steps are interacted collaboratively, product yield and purity are comprehensively improved, yield reaches 2.69g, and purity reaches to 99%. The method is simple and efficient, reduces costs, improves the yield and purity, and is beneficial to large-scale industrial application.

The invention discloses a filtering device for extracting shellfish toxin, and aims to provide a filtering device capable of effectively solving the problem that the negative pressure in a closed cavity is not enough and filtering process of the sample extracting fluid is influenced for the sealing between a filtering cylinder and a sealing box body is poor, and outside gas enters a vacuumized closed cavity. The filtering device includes a sealing box body, an assistant jacking mechanism, a sleeve body placed on an outer top face of the sealing box body, a filtering cylinder plugged in the placing sleeve body, and a self-adaptive pre-tightening sealing mechanism arranged in the sealing box body; a first interface interconnected to the outside face of the sealing box body is arranged on the inside face of the inner cavity of the sealing box body.

The invention discloses a manufacturing method and application of quantitative paralytic shellfish toxin detection test paper. Firstly, a carrier pad is arranged at the bottommost layer; a nitrocellulose membrane is paved on the carrier pad; a sample combination pad and a water absorption pad are respectively arranged at two ends of the nitrocellulose membrane; one end of the sample combination pad and one end of the water absorption pad are partially overlapped with the nitrocellulose membrane, an adhesive tape layer covers one end of the sample combination pad, a detection line on the nitrocellulose membrane is close to one end of the sample combination pad, and a quality control line is close to one end of the water absorption pad, wherein the detection line comprises a T1 line and a T2line which are respectively formed by coating an amino terminal with an artificial antigen and coating a guanidyl terminal with an artificial antigen, and the quality control line is formed by a goat-anti-mouse secondary antibody; the sample combination pad is sprayed with a guanidyl-end artificial antigen preparation monoclonal antibody marked with colloidal gold and an amino-end artificial antigen preparation monoclonal antibody marked with colloidal gold. The test paper has the characteristics of good specificity, high sensitivity and simple operation technology, accords with the industrialization development trend, and has the potential of wide popularization.

The invention provides an early warning method for paralytic saxitoxin in bivalve mollusks. Whether the risk that the content of paralytic saxitoxin exceeds the standard exists or not is determined by detecting the content of paralytic saxitoxin in digestive glands of the bivalve mollusks. According to the method, the digestive gland is used as a monitoring sample, in toxin extraction operation, compared with integral soft tissue used in a conventional detection method, homogenate grinding is easier, operation is easy and convenient, and time and labor are saved. The volume of a detection sample is smaller, and toxin extraction is more economical. The accumulation of the digestive adenosine toxin is quicker and the accumulation amount is high, so that the toxin is more easily and sensitively detected. Meanwhile, compared with monitoring of the toxic algae level in a water body and a shellfish organism biomarker, the digestive gland is used as a toxin monitoring sample, interference of other factors is small, and the shellfish toxin level can be reflected more directly.