124 results about "Preparative hplc" patented technology

The invention provides a preparation method of cyaniding-3-O-glucoside chloride. Specifically, the method comprises the steps of: subjecting purple crops as the raw materials to an extraction with an extract of ethanol water solution with a volume fraction of 30-80% and a pH value of 1-4, leaving the obtained anthocyanidin crude extract to an ultrafiltration treatment through an ultrafilter membrane with a molecular weight cut-off of 1000-5000 D, and freeze drying the filtrate so as to obtain filtrate dry powder with a molecular weight less than 1000-5000 D, preparing a column-loading solution of 0.05-5wt% with the filtrate dry powder and water, purifying the filtrate dry powder preliminarily on a gel chromatographic column, conducting elution and collecting the eluent located at a position of 520nm and with an absorption peak, and reserving fractions containing cyaniding-3-O-glucoside chloride; purifying the fractions containing cyaniding-3-O-glucoside chloride further with preparative HPLC (high performance liquid chromatography), and collecting cyaniding-3-O-glucoside chloride absorption peaks. According to the invention, by improving the conditions and operations of extraction separation and purification, sugar, protein, other types of anthocyanin and flavonoid pigments in extracts can be effectively removed. Meanwhile, with simple steps and safe reagents, the method of the invention is beneficial for obtaining high purity cyaniding-3-O-glucoside chloride products.

The invention discloses a preparation method of polypeptide synthesis carbetocin. The technical scheme of the method comprises the steps of taking Rink Amide MBHA (methylbenzene hydrogen amine) resin as an initial raw material; taking Fmoc-protected amino acid as a monomer; sequentially connecting amino acid, wherein Cys adopts Fmoc-Cys (CH2CH2CH2COOR)-OH, and the last amino acid uses Fmoc-Tyr (Me)-OH, and obtaining protected nonapeptide resin; removing the Fmoc azyl protected group of Tyr; cutting straight chain peptide from the resin by peptide cutting reagents such as FTA (fault tree analysis) and the like; precipitating a straight chain peptide rough product by adding diethyl ether; performing ring formation reaction by adding BOP (butyl octyl phthalate) /HOBt (oxhydryl benzotriazole)/DMF (dimethyl formamide); and purifying the preparation type HPLC (high performance liquid chromatography) in a separating way to obtain a target product. After the method is used, the each-step peptide connecting yield is more than 99%, the peptide cutting yield is 96.2%, and the gross yield is as high as more than 35%. The preparation method is convenient to industrial implementation, and has a higher industrial prospect.

The invention relates to a method for extracting and purifying 6-gingerol from ginger, which comprises the following steps: (1) extraction and concentration of gingerol: heating sliced ginger in ethanol under reflux, and concentrating to obtain a ginger extract; (2) leaching of gingerol: leaching the ginger extract to obtain a leaching solution, and recycling the extract under reduced pressure to obtain a gingerol crude extract; (3) silica gel column chromatography: dissolving the gingerol crude extract in ethyl acetate, adding silica gel, mixing, and after the solvent is volatilized, passing through a silica gel column by a dry process; after carrying out isocratic elution and thin-layer chromatography (TLC) detection, collecting the eluate part containing 6-gingerol, recycling the eluate under reduced pressure to obtain a 6-gingerol crude product; passing the 6-gingerol crude product through the silica gel column, and carrying out elution, detection, collection and solvent recycling to obtain the higher-purity 6-gingerol; and (4) purification by preparative HPLC (high performance liquid chromatography): dissolving the higher-purity 6-gingerol, purifying by preparative HPLC, carrying out isocratic elution, collecting the part with the maximum chromatogram peak, and drying by distillation to constant weight, thereby obtaining the high-purity 6-gingerol. The technique is simple and easy to operate; and the product has the advantages of high yield and good quality.

The invention provides a synthetic method for salvianolic acid A. The synthetic method is characterized by comprising a step of subjecting a salvianolic acid A synthetic precursor (10) to demethylation so as to prepare salvianolic acid A. A synthetic route is described in the specification. The prepared salvianolic acid A has a purity of 90 to 94%; and the purity of the prepared salvianolic acid A can be increased to 97% after preparative HPLC purification.

The invention discloses a preparation method of fucoxanthin and relates to the fucoxanthin. The preparation method of the fucoxanthin comprises the steps that a cleaned brown algae material is aired, and soaked by an organic solvent for extraction; an obtained extracting solution is condensed, and a fucoxanthin crude extract is obtained; the fucoxanthin crude extract is subjected to column chromatography separation; a fraction of the fucoxanthin is collected and condensed, and a fucoxanthin raw material is obtained; the obtained fucoxanthin raw material is dissolved by the organic solvent and transferred to a sample introduction bottle of a semipreparative/preparative HPLC (high performance liquid chromatograph); the semipreparative/preparative HPLC is started for separation and purification; fucoxanthin purification liquid is collected automatically through ultraviolet on-line detection or by a fraction collector triggered by a mass spectrum signal; and after the collected fucoxanthin purification liquid is decompressed, condensed, frozen and dried, the high-purity fucoxanthin is obtained. Through testing, the purity of the prepared fucoxanthin is greater than 99%.

The present invention discloses a method of purification of prostaglandins including fluorine atoms by using preparative HPLC. Tafluprost and Travoprost are prostaglandins including fluorine. The chemical structure of the impurities in crude Tafluprost and crude Travoprost also contain fluorine, therefore, the removal of the impurities is difficult. Purification by using preparative high performance liquid chromatography (HPLC) can achieve high-quality liquid bulk drugs.

The invention discloses a method for extracting a numb-taste substance from Zanthoxylum armatum DC. The method comprises the following steps: processing Zanthoxylum armatum DC. to obtain a Zanthoxylum armatum DC. powder, carrying out 95% ethanol reflux extraction to obtain an extract liquor, firstly carrying out silica-gel column chromatography, then carrying out ODS column chromatography and finally purifying by the use of Sephadex LH-20; suspending a numb-taste substance monomer obtained after preparative HPLC separation in water for 1-2 h, removing a volatile matter by high vacuum, and freeze-drying twice to obtain a powdery numb-taste substance monomer; and keeping the product in a dark place after vacuum packaging. According to the method, a numb-taste substance monomer with a large number of varieties and high purity can be obtained by an advanced separation purification technology, and a plurality of requirements can be met.

An improved analytical method for analysis of dianhydrogalactitol preparations provides a method for determining the purity of dianhydrogalactitol and detecting impurities in preparations of dianhydrogalactitol, as well as identifying any such impurities. The method employs high performance liquid chromatography (HPLC), in particular, HPLC with evaporative light scattering detection (ELSD); the HPLC can be followed by tandem mass spectroscopy. The method can further comprise the step of performing preparative HPLC collection of at least one specific substance peak present in a preparation of dianhydrogalactitol.

The invention discloses an 18F-labeled compound and a pod protease targeted PET imaging probe, which relates to the technical fields of radiopharmaceuticals and nuclear medicine. The invention provides a compound shown as a formula (I), asparagine site shearing and disulfide bond reduction are carried out in a tumor microenvironment with high expression of pod protease, and a radioactive dimer 18F-1-dimer is formed by virtue of a biocompatible CBT-Cys click condensation reaction, so that the tumor imaging effect is improved. The compound with a structure shown in the formula (I) is synthesizedby adopting a one-step ion exchange 18F labeling method, the method is simple to operate, and preparative HPLC for further purification is not needed. The invention further provides the pod proteasetargeted PET imaging probe which is a compound shown in (I). The PET imaging probe has the advantages of high stability, high sensitivity, strong specificity, good safety and the like.

The invention discloses a preparation method of solid phase peptide synthesis atosiban which includes the following steps: taking Rink Amide resins, Rink Amide MBHA resins or Rink Amide AM resins as starting materials and taking Fmoc amino acids as monomers, amino acids are grafted one by one and mercaptopropionic acids (Map(SX)) are protected by the last peptide chain; after protected nonapeptide resins are obtained, the acellular side-chain protective groups and cutting peptides are synchronized; then cutting peptides is carried out, and reduced crude atosiban is collected; the pH value is adjusted to 7.5 to 10.0, and oxidized crude atosiban is collected; target products are obtained by the separation and purification by preparative HPLC(C18 or C8 column). The preparation method is convenient in material source, simplifies technology and reduces cost; and the preparation method is low in the pollution of three wastes and is high in yield, and the preparation method is convenient for being industrialized and has good industrialization prospect.

An improved analytical method for analysis of dianhydrogalactitol preparations provides a method for determining the purity of dianhydrogalactitol and detecting impurities in preparations of dianhydrogalactitol, as well as identifying any such impurities. The method employs high performance liquid chromatography (HPLC), in particular, HPLC with evaporative light scattering detection (ELSD); the HPLC can be followed by tandem mass spectroscopy. The method can further comprise the step of performing preparative HPLC collection of at least one specific substance peak present in a preparation of dianhydrogalactitol.

The invention relates to a preparation method for extraction of catechin from tea, belonging to the field of separation and extraction technology. The method comprises the following steps: drying tea and grinding the dried tea to obtain a tea sample; weighing the tea sample, adding an ethanol solution, placing the obtained mixture in a microwave digestion instrument for extraction and carrying out filtering and washing so as to obtain extract; adding chloroform into the extract for extraction so as to obtain an aqueous solution, then extracting the aqueous solution with ethyl acetate, removing a water phase, subjecting an ester phase to pressure-reduced distillation so as to recover ethyl acetate and carrying out vacuum concentration and freeze drying so as to obtain crude catechin extract; and subjecting the crude catechin extract to chromatographic pre-separation at first and then to preparative HPLC purifying and refining so as to obtain high-purity catechin. The method provided by the invention is simple in operation process, low in the usage amount of solvents, high in extraction rate and applicable to both small-scale preparation in laboratories and large-scale industrial production.

The invention discloses a recycling preparative HPLC (high performance liquid chromatograph) provided with a multi-ported valve as well as a method used for preparation, separation and purification. The recycling preparative HPLC provided with the multi-ported valve comprises a liquid accumulator M, a high-pressure pump P, a sampler S, chromatographic columns C1 and C2, the multi-ported valve, detectors D1 and D2, three-way switching valves T1 and T2, liquid pipelines and the like. The recycling preparative HPLC has the outstanding effects as follows: 1, to-be-separated mixed components are circularly separated only in the two chromatographic columns and are not required for circulation through the high-pressure pump, chromatographic peak broadening is light, the separation efficiency is high, and the separation effect is good; 2, a system is provided with two detectors, peak cutting and circular separation can be accurately controlled, and the separation effect is improved; 3, eluent without the separated components is completely recycled, the consumption of eluent is greatly reduced, and the separation cost is very low.

The invention relates to a disposable auxiliary device and a method for preparing a radioactive medicament, and belongs to the technical field of radioactive medicament preparation devices. The disposable auxiliary device consists of the following parts: three radiation chemical reaction five-union three-way valves and a product preparation five-union three-way valve which are connected in sequence; the left end of a first three-way valve of the radiation chemical reaction five-union three-way valves is connected to an air supply port through a Luer taper and a silicone tube; the upper end ofthe first three-way valve is connected to a target material recycling port; a second three-way valve is connected with a conical container which is filled with a radionuclide F-18 through a small ionexchange column; a thirteenth three-way valve of the radiation chemical reaction five-union three-way valves is connected with a semi-preparation type HPLC sampling port through another Luer taper. The device provided by the invention is applicable to purifying semi-preparation type HPLC of the radioactive medicament, ensures that a reagent is completely added into or pumped out from a reaction bottle, can ensure the consistency of using amounts of the reagent, is beneficial to standardization of radioactive medicament production, and can reduce the using amount of the reagent.

The invention discloses a method for preparing algae haematochrome through polygenic combination expression, and in particular relates to a preparation method for producing the algae haematochrome, which can be applied in preparation of fluoroimmunoassay or functional food, by a polygenic combination expression technology, and application of the algae haematochrome. The preparation method comprises the specific steps of cloning key algae haematochrome synthesis enzyme genes (hoxl and pebS) from spiral seaweeds, and introducing the two key enzyme genes into a recipient cell by the polygenic combination expression technology to build a biological algae haematochrome synthesis way in the recipient cell; converting heme in escherichia coli to synthesize the algae haematochrome through two-step enzymatic catalysis reaction; finally performing extraction and preparative HPLC purification to obtain the high-purity algae haematochrome with the purity over 95 percent. The maximum absorption wavelength of the algae haematochrome is 550nm, and the maximum fluorescence-emission wavelength is 570nm. The algae haematochrome prepared by the method disclosed by the invention can be used as a fluorescence probe or a pigment antioxidant applied in preparation of effective components of the functional food.

The crude extract of Plectranthus Amboinicus (PA) has anti-inflammatory effects and can inhibit AP-1 binding in vitro. The incubation with PA crude extract resulted in significant inhibition of the LPS-induced expression of IL-6, IL-12, MCP- 1, and RANTES in HUVEC cells. After the crude extract was further fractionated using preparative HPLC, fraction 8, 9, 10 and 11 were identified to inhibit the AP-1 binding activity. The active component of fraction 8 is Mena 987; fraction 9 is Mena 998; fraction 10 is Mena 9102; and fraction 11 is rosmarinic acid and the synthestic rosmarinic acid analogues. Other compounds showed inhibitory activities as well. These compounds have inhibitory effects on AP-1 activity and are useful as preventive or therapeutic agent for diseases in which excessive expression or activities of AP-1 are involved.

The invention discloses a refining method for bleocin, comprising purifying the crude product of bleocin by preparative HPLC technology; wherein, the immobile phase of a preparation column is silica gel column of C8 or C18, the mobile phase is mixed liquid of sulphate aqueous solution with pH value between 3.5 and 5 and polar organic solvent. In the method, the preparative HPLC technology is adopted to increase the yield (85% generally) and the degree of automation of A2 and A3 which are the active ingredients of bleocin and greatly lower cost, thus can be applied to industrial production.

The invention discloses a ginkgo endophytic fungus and an application thereof. The ginkgo endophytic fungus fusarium proliferatum DZHQ1 with anti-tumor activity is separated from ginkgo bark, the species of a strain is judged with a colony morphology and 18sRNA sequencing combination method, the anti-cervical cancer activity of a crude extract of the strain is detected with MTT assay, and finally,secondary metabolites of the strain with inhibition rate being higher than 50% are separated with a semi-preparative HPLC method. Accordingly, compound monomers with anti-tumor activity can be further screened.

The present invention provides a method for producing a novel β-lactam antibiotic from a protoplast fusion strain. The method is to fermentatively culture the protoplast fusion strain of Penicillium chrysogenum and Cephalosporium acremonium. The ferment filtrate is isolated, lyophilized, and extracted by acetone or acetone/methanol. The extract is concentrated by decompression, and then analyzed by preparation type HPLC to isolate the active antibiotic compound.

The invention relates to a method for separating and enriching limonin compounds. The method is characterized by comprising the following steps: heating ground narrow-leaf cortex dictamni with an ethanol solution, performing reflux extraction for 3-4 times, filtering, merging the filtrate, respectively extracting with petroleum ether, dichloromethane and ethyl acetate after concentrating, selecting the extraction fraction of the dichloromethane, combining silica gel column chromatography, Sephadex LH-20, preparative HPLC (High Performance Liquid Chromatography), recrystallization and other separation and purification means, and separating the limonin compounds. The method is high in separation efficiency, slight in pollution and low in cost and has excellent popularization and applicationvalue.

The invention relates to an epothilone B separating and extracting method based on a membrane filtration technology. The method comprises the following steps: adsorbing a fermentation liquid by using a macroporous resin; then leaching the absorbed macroporous resin by using ethyl acetate and collecting a leaching liquid; filtering the leaching liquid by using an ultrafiltration membrane; washing and filtering the leaching liquid by using the ethyl acetate; combining filtrates; concentrating; performing detection through preparative HPLC (High Performance Liquid Chromatography); and crystallizing the epothilone B component obtained in a separating manner, thereby obtaining the epothilone B. The method is moderate in condition, simple and convenient to operate, few in separating steps and good in selectivity, and can be used for obviously increasing the yield of the epothilone B, thereby providing conditions more favorable for industrial production. The method with combination of membrane filtration and crystallization is adopted, so that a part of macromolecular impurities can be effectively removed; meanwhile, the application of an organic reagent is avoided, so that the industrial production pollution is reduced. Thus, the loss of the epothilone B in other complicated preparation processes is avoided. The epothilone B prepared in a crystallization manner has relatively high purity. Thus, the epothilone B is broad in application prospect and economic benefit.

The invention provides a preparation method and application of mucor-fermented soybean bitter peptide. The preparation method comprises the following steps of preparing powder A, powder B and powder C; dissolving the powder C into deionized water; by taking aqueous solution of A phase trifluoroacetic acid with the mass fraction of 0.05 percent and B phase trifluoroacetate acetonitrile with the mass fraction of 0.05 percent as eluent, performing gradient elution by adopting a semi-preparative HPLC (High Performance Liquid Chromatography) inverse phase C18 column, collecting a target peak, and performing freeze drying to obtain powder D; dissolving the powder D into the deionized water; by taking the aqueous solution of the A phase trifluoroacetic acid with the mass fraction of 0.05 percent and the B phase trifluoroacetate acetonitrile with the mass fraction of 0.05 percent as the eluent, performing gradient elution by adopting an analytical HPLC inverse phase C18 column, collecting a target peak, and performing freeze drying to obtain bitter peptide I. The prepared bitter peptide I is applied to food, and can be used for providing the bitterness of soybean in food, and increasing the delicate flavor of the food; the bitter peptide is easy for chemical synthesis; in addition, a synthesized product is high in purity and good in stability; the application range of the bitter peptide can be widened.

The invention discloses a polypeptide purification method and belongs to the technical field of protein and polypeptide purification. A preparative HPLC (high performance liquid chromatograph) is used for purifying polypeptides, 2,2,2-trifluoroethanol or tetrahydrofuran is added to an organic phase of a mobile phase of the preparative HPLC, the elution gradient, flow velocity and detection wavelength are set, and high-purity peptides are obtained. The polypeptide purification method has the advantages that the elution time is shortened, the use quantity of an organic solvent is reduced, the purity is high, the separation effect is good, and the cost is saved.

The invention belongs to the field of medicines, and relates to an application of a novel compound, which has effects of preventing and treating Alzheimer's disease and is extracted and separated fromdried shells of a plant xanthoceras sorbifolia, which belongs to the Xanthoceras Sapindaceae. In the invention, the novel compound is extracted and separated mainly through a traditional extraction and separation methods with combination of modern instrumental analytic methods, wherein the method mainly includes heating reflux extraction, macroporous adsorption resin column chromatography, silicagel column chromatography, ODS opened column chromatography, and preparative HPLC separation. A pharmacological experiment proves that the compound has excellent nervous protection effect on PC12 cell damage induced by A[beta]25-35; the compound can be used for further development and application of an anti-Alzheimer's disease medicine.

The invention discloses a colistin sulphate purification method. The method disclosed by the invention is implemented by using a semi-preparative HPLC (high performance liquid chromatography) separation and purification system and taking a reversed-phase C18 as packing and taking a salt solution and acetonitrile as moving phases. The preparation of colistin sulphate according to the method is simple in operation, less in experimental steps, low in cost and short in cycle, and the purity of prepared colistin sulphate can be more than or equal to 98.0%.