219 results about "Sephadex column" patented technology

The invention provides a triple-enzyme hydrolysis preparation method of anti-tumor polypeptides of spirulina. The method comprises the following steps: preparing a solution having a concentration of 5 percent from spirulina powder with ultrapure water; repeatedly freezing and thawing, homogenizing and performing ultrasonic treatment, centrifuging to obtain supernate, freezing and drying for later use; adding the prepared protein fluid into pepsin for enzyme hydrolysis, controlling the pH value, adding trypsin for enzyme hydrolysis, adding chymotrypsin for enzyme hydrolysis, controlling the pH value, deactivating enzyme, cooling, and centrifuging to obtain supernate; sequentially filtering with ultrafiltration membranes having molecular weight cut-off of 10KD, 5KD and 3KD respectively to obtain three kinds of spirulina protein enzymatic hydrolysates; carrying out sephadex G15 column chromatography on 0-3K of enzymatic hydrolysate, eluting with water, and collecting four polypeptide ingredients, namely anti-tumor polypeptides of spirulina. The obtained spirulina polypeptide and monopeptide ingredients can be used for establishing a theoretical basis for development and utilization of anti-tumor medicines and health foods.

The invention provides a preparation method of a small water turtle anti-tumor polypeptide. The method comprises the following steps: mixing 3 to 20g of meat paste of small water turtle and ultrapure water; agitating to obtain a mixed solution at concentration of 10 to 50% (w/v); adding trypsin to perform enzymolysis; deactivating enzyme after enzymolysis; cooling until reaching room temperature; centrifuging enzymatic hydrolysate; collecting supernate; sequentially filtering through ultrafiltration membranes which respectively have molecular cut off of 10KD, 5KD and 3KD, so as to obtain three enzymatic hydrolysates; performing sephadex G-15 column chromatography for 0-3K enzymatic hydrolysate; eluting with water; collecting four polypeptide components under a certain detection wavelength so as to obtain small water turtle anti-tumor polypeptide. The prepared small water turtle anti-tumor polypeptide is beneficial for the development and utilization of anti-tumor drugs and health foods.

The invention discloses a fluorescent probe, which is prepared by the following method. The method comprises the steps of: firstly, purifying fluorescence quantum dots synthesized in a carboxymethyl chitosan template, and connecting the fluorescence quantum dots with IgG (Immunoglobulin G) antibodies of a human by coupling effect of carbodiimide and N-hydroxysuccinimide eater; and then separating and purifying by using a sephadex column to obtain the fluorescent probe of the fluorescence quantum dots synthesized by immune-modified carboxymethyl chitosan. The invention further discloses a method for detecting staphylococcus aureus by using the fluorescent probe, comprising the detailed steps of: culturing the fluorescent probe and bacteria to be detected for 30 minutes, removing the fluorescent probe with nonspecific absorption by a plurality of times of centrifugal washing, finally, suspending bacteria marked by the fluorescent probe in a phosphate buffer solution, determining the fluorescence intensity and absorption intensity of bacterial suspension by adopting fluorescence spectrum and ultraviolet and visible absorption spectrum, and realizing quantitative determination of the staphylococcus aureus according to linear relation of the fluorescence intensity and the concentration of the bacterial suspension. The method disclosed by the invention is clean, simple and rapid and is suitable for being applied in rapid detection under emergency.

The invention provides a method for preparing multiple oligosaccharides by separating and purifying Chinese dates. The method comprises the following steps of extracting by a water extraction method, separating by a membrane separation technology, decoloring a strong-base anion exchange resin, and purifying by a DEAE-cellulose (Diethylaminoethyl Cellulose) column and a dextrangel column to obtain four oligosaccharides with edible Chinese dates or imperfect Chinese dates as raw materials. The invention provides a new direction for development of deep processing products of Chinese dates and provides new resources for development of the oligosaccharides. The method, provided by the invention, has the advantages of simple operation process and is suitable for industrial production. The prepared oligosaccharides have higher purity and can be used for functional research and development of function food and medicines.

The invention discloses ceramide-containing tea seed extract and a preparation method thereof. The preparation method comprises the following steps of: crushing raw materials, extracting by using water or low-carbon alcohol, and recycling an extraction solvent from the obtained extracting solution so as to obtain extract, wherein the raw materials are seeds of theaceae camellia plants, namely camellia oleifera abel, camellia or common camellia, or residues of the seeds of the plants subjected to oil component extraction; adding alkaline solution into the extract, and hydrolyzing at the temperature of between 10 and 80 DEG C for 0.5 to 20 hours; and separating the obtained hydrolyzate by using a macroporous resin column, performing chromatography through a silica gel column or a sephadex column, eluting by using 40 to 60 volume percent aqueous solution of methanol or ethanol, performing thin layer chromatography, collecting fractions, mixing the fractions, and drying to obtain the ceramide-containing tea seed extract. The raw materials are readily available and the method is environmentally-friendly; the preparation process is simple; the product quality is stable; the production cost is low; and the content of ceramide in the extract is high.

The invention discloses a method for extracting solanesol, cembrane diterpene, vitamin E and phytosterol from tobaccos simultaneously. The method comprises the steps that 1, extraction is carried out; 2, saponification is carried out, wherein saponified solanesol and other fat-soluble active ingredient extracts are obtained; 3, sephadex gel column separation is carried out, wherein solanesol, vitamin E and phytosterol (mixture) and cembrane diterpene crude extract are obtained respectively; 4, silica-gel column chromatography is carried out, wherein after silica-gel column chromatography, high-purity solanesol, vitamin E and phytosterol are obtained respectively; the obtained cembrane diterpene is subjected to silica-gel column chromatography, and alpha-4,8,13-cembratriene-1,3-diol and beta-4,8,13-cembratriene-1,3-diol are obtained respectively. Comprehensive extraction of tobacco fat-soluble active ingredients is achieved, tobacco residue obtained after fat-soluble active ingredients are extracted can be used for extraction of tobacco protein, chlorogenic acid, rutin, polysaccharide and nicotine, gradient utilization of tobacco active ingredients is achieved, and the utilization value of tobacco resources, particularly waste tobaccos is improved.

The invention relates to a method for simply preparing a cell factor from placenta mesenchymal stem cells. The invention discloses a method for simply preparing a placenta mesenchymal stem cell exosome. Specifically, the invention relates to a method for separating and extracting the exosome from the placenta mesenchymal stem cells, and the method comprises the following steps: culturing placentamesenchymal stem cells to obtain a culture supernatant; carrying out sephadex column chromatography on the culture supernatant to obtain a chromatographic solution; concentrating the chromatographic solution by using a tangential flow ultrafiltration system to obtain an exosome concentrate; and carrying out cryopreservation or freeze drying on the exosome concentrate. The method of the present invention exhibits excellent properties as described in the specification.

The invention provides a method for preparing spirulina polypeptide-chitosan nano particles. The method mainly comprises the following steps: extracting spirulina proteins by employing a method of combining multigelation and ultrasonication, hydrolyzing the spirulina proteins through trypsin, separating and purifying by a sephadex column, filtering through an ultrafiltration centrifugation tube, and preparing the spirulina polypeptide-chitosan nano particles by employing a sodium tripolyphosphate ion exchange method. The obtained spirulina polypeptide-chitosan nano particles Y2-CS have the following antitumor activities that the spirulina polypeptide-chitosan nano particles Y2-CS have a certain function of inhibiting in-vitro growth of human breast cancer cell MCF-7 and have a certain dose relationship; and when the concentration is 250mug/mL, the highest inhibition rate can be 26.05 percent. Therefore, the spirulina polypeptide-chitosan nano particles contribute to developing and utilizing antitumor health-care foods and medical products.

The invention discloses a preparation method for dressing polyamidoamine dendrimer by amino acid. The reaction is performed in organic solvent, hydroxyl benztriazole HOBt, 2-(1H-benzotriazole)-N, N N`, N`-tetramethyluroniumhexafluorophosphate HBTU, N and N-diisopropylethylamine DIPEA are taken as catalyzer, to ensure that 2, 2, 4, 6 and 7-pentamethyldihydrobenzofuran-5-sulfonyl Pbf and 9-fluorenylmethyloxycarbonyl Fmoc dual-protection amino acid and terminal hydroxyl group PAMAM dendrimer can generate the condensation reaction. The refinement is performed through adopting dextran gel chromatography, to ensure that trifuoroacetic acid and piperidine can respectively divest of the Pbf protecting group and the Fmoc protecting group of the amino acid and the depurant amino acid can be obtained to dress the PAMAM dendrimer of the terminal hydroxyl group. The operation of the preparation method is simple, the condition is moderate, the price is low, the purer end product can be obtained, the preparation process can not damage the activation of the guanidino in the amino acid, and the preparation method is suitable for a great quantity of preparation and industrialized production.

The invention discloses an extracting method of passion flower fruit pigment. The extracting method comprises the steps of selecting materials, cleaning, draining, extracting ethanol, extracting absolute ether, extracting ethyl acetate, absorbing and purifying macroporous resin, absorbing and purifying polyamide chromatographic column, absorbing and purifying sephadex columns, separating and purifying preparative liquid chromatography, cooling, drying and packaging. According to the method, the operation is simple, a used solvent and an absorbent can be used repeatedly, the energy loss is low, the real-time control is easy, and the continuous automatic cleaning production can be realized.

The invention discloses a preparation method of a selenium-containing protein in a selenium-enriched yeast, which mainly comprises the following steps: (1) performing high-pressure homogenizing and crushing on a selenium-enriched yeast, and then centrifuging; (2) adding 75-90% (percentage by mass) of ammonium sulfate into the centrifuged supernatant, stirring, then centrifuging to collect precipitate, dissolving the precipitate in a buffer solution, dialyzing a dialysis bag, performing dewatering concentration with polyethyleneglycol to a smaller volume, and performing vacuum freeze-drying to obtain a crude protein; and (3) dissolving a little dry crude protein powder in a Tris-HCl buffer solution, passing through a 0.22 mu m microporous filtering film, then loading onto a Sephadex column, eluting under the conditions that the flow rate is 0.5-3 ml/min, the detection wavelength is 280 nm and the eluate is a 50-150 mmol/L Tris-HCl buffer solution, and collecting a protein component. Compared with the prior art, the preparation method achieves the advantages of high crushing ratio, sufficient crude protein extraction, high selenium content in the purified protein and high purity of the targeted selenium-containing protein.

The invention provides a method for preparing sea cucumber fucoidan and sea cucumber glycoprotein. According to the method, acaudina molpadioides serves as the raw material, a mild stepped enzymolysis processing method is mainly adopted, an ultrafiltration membrane is utilized to separate out a mixture of crude polysaccharide and glycoprotein, then separation, purification and preparation of the sea cucumber fucoidan and the sea cucumber glycoprotein are achieved through a Q-Sepharose-F-F ion exchange column and an Sephadex G-150 sephadex column, a prepared sea cucumber fucoidan product is fawn, and the sea cucumber glycoprotein is faint yellow. According to the method, a co-production method is adopted, main ingredients such as polysaccharide and glycoprotein in the acaudina molpadioides can be prepared in a jointed mode, emission of waste is reduced in the extraction process, and production cost is lowered.

The invention discloses a marine fungi penicillium crustosum bacterial strain, quinolinone compounds derived from marine fungi penicillium crustosum, and a preparation method and applications of the quinolinone compounds. Specific structural formulas of the quinolinone compounds are represented by formula R1-formula R6. According to the preparation method, a culture medium is inoculated with the marine fungi penicillium crustosum bacterial strain AP2T1 CGMCC No.8516 for static fermentation; an obtained fermentation broth is extracted with ethyl acetate, and mycelium is extracted with an organic solvent; obtained crude extracts are mixed, an obtained mixture is subjected to sephadex column chromatography, reverse-phase silica-gel column chromatography, and thin layer chromatography detection using organic solvents; an obtained elution ingredient containing intermediates is subjected to methylation, and thin layer chromatography and reverse-phase silica-gel column chromatography separation and purification so as to obtain target compounds R1-R6. The target compounds R1-R6 possesses wide application prospect in preparation of antitumor drugs, antibacterial agents, and agricultural insecticides.

The invention relates to a method for simply preparing cytokines from placenta mesenchymal stem cells. On the one hand, the invention relates to a method for separating and extracting exosome and cytokines from placenta mesenchymal stem cells. The method comprises the following steps: culturing the placenta mesenchymal stem cells to obtain a culture supernatant; carrying out sephadex column chromatography on the cultured supernatant to obtain a chromatographic solution; concentrating the chromatographic solution by using a tangential flow ultrafiltration system to obtain an exosome concentrate, acquiring an ultrafiltration filtrate at the same time, concentrating the chromatographic solution again by using the tangential flow ultrafiltration system, and filtering and concentrating the ultrafiltration filtrate to obtain a cytokine concentrate; and carrying out cryopreservation or freeze drying on the exosome concentrate or the cytokine concentrate. The invention also relates to a methodfor preparing the cytokine concentrate. The method of the present invention exhibits excellent properties as described in the specification.

The invention discloses a Bacillus amyloliquefaciens WH3 strain, and a preparation method and application thereof. The preparation method comprises the following steps: 1, separation and identification of bacteria: separating bacteria resistant to rape sclerotinia rot from rape seedlings, and carrying out 16SrDNA and morphological identification to determine that the WH3 strain obtained by separation is Bacillus amyloliquefaciens; 2, separation, purification and identification of an antifungal active substance Safenour: fermenting the WH3 strain, extracting the antifungal active substance, separating and purifying through a sephadex column, and carrying out MALDI-TOF (matrix-assisted laser desorption/ionization-time of flight ) mass spectrometry on the active antifungal substance to inferthat the substance is a ring type polypeptide; and 3, application of the strain in the preparation of vaccines and immunological adjuvants. After the Safenour used as the adjuvant is mixed with a protein antigen and mice are respectively immunized through oral administration and injection of the mixture, effective body fluid and cellular immune response can be activated, and a high-titer specificantibody can be detected in the blood serum. The Safenour has low production cost and high stability, does not need to be emulsified when mixed with the antigen, and has a better immunoenhancement effect in comparison with Freund adjuvants and cholera toxin B subunits.

The invention discloses a method for synthesizing a swainsonine antigen, comprising the following steps of: firstly performing a reaction between halo alkyl aryl fatty acid and alcohol to generate halo alkyl aryl fatty acid ester, followed by a reaction between halo alkyl aryl fatty acid ester and swainsonine to generate a quaternary ammonium salt with a structure of aromatic nucleus, performing a reaction between the quaternary ammonium salt and sodium hydroxide for deesterification to generate a sodium salt, adjusting the pH value by the use of hydrochloric acid to generate SW-halo alkyl aryl fatty acid; condensing SW-halo alkyl aryl fatty acid with bovine serum albumin (BSA) to produce SW-BSA, sieving through a glucan gel column chromatography for desalination, freeze drying, determining the proportion of BSA and SW, and adding an immunoadjuvant for emulsification to obtain the SW immune antigen. The antigen is white and oily with the effective content being 5mg/mL and the effective immune period being 12 months. The effective protection rate reaches more than 90%.

The invention discloses a method for extracting quercetin from all-grass of grass jelly, which comprises the following steps: (1) getting a plurality of all-grass of grass jelly, adding 20 times of water to boil and extract, slowly adding limewater during agitation until the pH is 8 to 9, then adding 3 percent of a boric acid and boiling for 20 to 30 minutes under the pH condition; (2) filtering before getting cold, repeatedly depressurizing and filtering the residue 3 times and combining the filtrate, under the condition of 60 to 70 DEG C, adjusting a combined liquid to pH value of 5 by concentrated hydrochloric acid, pumping the acid liquid after placing for 24 hours; (3) washing the precipitate to neutrality by water, drying in 60 DEG C to derive a rutin crude product, recrystallizing by boiling water, and drying in 70 to 80 DEG C to derive the rutin pure product; (4) adding 2 percent of H2SO4 to the refined rutin by a proportion of 1:80, heating with fire for 2 hours, cooling and pumping; (5) washing the residual water to neutrality, and then implementing 95 percent ethanol recrystallization and sephadex chromatography separation (SphadexLH20), using carbinol as an eluant; (6) deriving quercetin yellowish green pure product, as the rutin can not be eluted. The invention has the advantages of simplicity, low cost and high yield, thus being suitable for industrial production.

The invention discloses a broccoli polypeptide extract as well as preparation method and application thereof. The preparation method comprises the following steps: preparing stems and leaves of broccoli, firstly enzymatically hydrolyzing the stems and the leaves by virtue of complex enzyme, then conducting alkaline extraction, and separating, precipitating, washing and drying so as to obtain broccoli protein powder; adding water, then adding alkaline protease and flavor protease for further enzymatic hydrolysis under the following conditions: temperature is 45-60 DEG C and pH value is 8.5-10, inactivating the enzyme, cooling, centrifuging, conducting rotary evaporation, concentrating and freeze-drying so as to obtain enzymatically hydrolyzed broccoli polypeptide; and ultra-filtering the enzymatically hydrolyzed broccoli polypeptide by virtue of a filter membrane and separating the enzymatically hydrolyzed broccoli polypeptide by virtue of a dextran gel column, eluting by taking a phosphate water solution as an eluent, collecting the eluent, filtering, evaporating to dryness, collecting and drying, so that the broccoli polypeptide extract is obtained. The broccoli polypeptide extract disclosed by the invention, which is prepared through a special preparation method, is significant in effect on enhancing immunity; therefore, the broccoli polypeptide extract can be used for preparing health care food and medicines capable of enhancing the immunity.

The invention discloses a stephanotis total alkaloid extract as well as a preparation method of the stephanotis total alkaloid extract. The preparation method of the stephanotis total alkaloid extract comprises the following steps of: grinding the dried stephanotis, heating and extracting, and decompressing and condensing the filtrate; diluting with heating water, and acidifying and filtrating to obtain a filtrate; alkalifying and filtrating to obtain the filtrate, and performing molecular weight cut off filtration on the obtained filtrate through a ceramic membrane filter; extracting with n-butyl alcohol, decompressing and condensing; spray drying the concentrated solution to obtain a dried powder extract; adding ethanol for ultrasonic extraction, mixing the extracting solution, absorbing the extracting solution by a sephadex column, eluting with 90% to 95% of ethanol, segmentally collecting, identifying through a high performance thin layer chromatography, mixing the parts containing alkaloids, freezing at a low temperature, and separating out an alkaloid crystal crude product; and adding ethanol for dissolving, standing and freezing to obtain a stephanotis total alkaloid pure product. A stephanotis total alkaloid medicinal preparation provided by the invention has the effect of adjuvant therapy on leucopenia, in particular to leucopenia of the tumor patients caused by radiotherapy and radiotherapy, and the leucopenia caused by other medicines.

The present invention discloses process of extracting and separating compounds I-VIII from ginseng-ophiopogon injection. These compounds have functions of inhibiting myocardial cell apoptosis, promoting the release of CO from vascular endothelial cell, suppressing oxidation caused by AAPH, etc; and this show that these compounds are main active components for the ginseng-ophiopogon injection to treat cardiac and cerebral vascular diseases. The extracting and separating process includes the steps of: chromatographic separation in macroporous resin column, MCI columm, silica gel column, glucosan gel column and reverse silica gel column; efficient liquid phase separation; sublimation; crystallization; etc. The compound structure identification is completed through infrared method, 1D and 2D nuclear magnetic resonance, mass spectrum and other spectroscopic method.

The invention discloses a refined part of total gardenia crocin with an anti-depression effect. The optimal process for preparing the refined part of the total gardenia crocin screened by a plurality of experiments adopts a method combining a macroporous resin column and a sephadex column to separate and prepare the refined part of the total gardenia crocin with relatively high purity. An experiment result shows that the refined part has a good effect of treating the depressive disorder; the acting time is obviously faster than that of positive medicine fluoxetine hydrochloride; the action effect is higher than that of fluoxetine; and the action time of the fluoxetine needs over three weeks in general. The clinical experiment result shows that the crocin is nontoxic, free of addiction and small in toxic and side effects, has a good application prospect, and is expected to be developed into novel purely natural anti-depression medicines or health products.

The invention relates to a preparation method of a yak milk casein antibacterial peptide and belongs to the technical field of biology. The preparation method includes steps of dissolving yak milk casein, adding trypsin for hydrolysis, inactivating the trypsin, precipitating casein, performing centrifugation to remove precipitation, subjecting yak milk casein to sephadex column chromatography seperation, determining molecular weight by mass spectra, performing ultrafiltration and performing spray drying, wherein ultrafiltration membranes with interception of 1000Da and 2500Da are used for collecting peptide fragments with the molecular weight range of 1000Da to 2500Da. According to the yak milk casein antibacterial peptide prepared by the method, peptide molecular weight distribution is concentrated, bacteriostatic activity is high, the minimum inhibition concentration of staphylococcus aureus is 270 mu g/mL, and the minimum inhibition concentration of Escherichia coli is 980 mu g/mL.

The invention relates to an ophiopogon japonicus extract, a preparation method and the hypoglycemic application thereof. The preparation method of the ophiopogon japonicus extract comprises: ophiopogon medicine is taken to be decocted for 1-3 times, added with water by 5-20 times every time, then decocted for 30-120min every time, and filtered and merged into water-decocted liquid; after being concentrated to the density of 1.05-1.35g/ml, the obtained water-decocted liquid is added with ethanol until the alcohol content has the volume percent of 50-80%, and is collected and precipitated after staying over night; the precipitation is separated and purified by sephadex column, and then processed by vacuum freeze drying, so that white loose flocculent crystal which is the ophiopogon japonicus extract can be obtained, and the extract has the sugar content of 90-98%( based on fructose) as well as the molecular weight within the range of 500-2000. Animal experiments prove that the ophiopogon japonicus extract has remarkable hyperglycemic effect, and the action mechanism of the ophiopogon japonicus extract can be further explained by vitro experiments; the result shows the ophiopogon japonicus extract can reduce blood sugar levels by inhibiting the activity of alpha-glucosaccharase in alimentary canal, reduce the absorption for glucose by intestinal villi and the protective action for islet cells. Therefore, the extract can be used for developing medicine for treating type II diabetes.

The invention provides a fritillaria flower extractive, a preparation method therefore and applications. The content of peimine and peiminine in the fritillaria flower extractive is 20%-60%(w/w) of the total extractive. The fritillaria flower extractive is used for preparation of medicines or health care products with cough relieving and phlegm resolving effects. The preparation method comprises ultrasonic extraction, filtrate condensation, alkalization, sephadex gel column separation and eluate condensation and drying, and the fritillaria flower extractive is obtained. The preparation method is simple to operate, effective components of fritillaria flowers can be collected effectively, and the fritillaria flower extractive has good development and application prospects.