34 results about "Stepwise elution" patented technology

The invention discloses fuscoporia obliqua active ingredients capable of lowering blood sugar and a preparation method and application of the fuscoporia obliqua active ingredients. The preparation method takes fuscoporia obliqua fruit body as raw material and comprises the following steps: respectively extracting, filtering and concentrating the fuscoporia obliqua fruit body with normal temperature water and high temperature water; adding alcohol into concentrate and depositing to obtain crude polysaccharide; respectively pouring the polysaccharide extracted with normal temperature water and the crude polysaccharide extracted with high temperature water to flow through a (diethylaminoethanol) DEAE-52 cellulose column; carrying out subsection elution by using distilled water and NaCl solutions with different concentrations; and collecting stepwise elution peak sugar solution. Internal blood sugar reduction activity experiment shows that 0.2mol/L NaCl-section eluted sugar of the crude polysaccharide extracted with normal temperature water and 0.2mol/L NaCl-section eluted sugar of the crude polysaccharide extracted with high temperature water both have obvious blood sugar reduction activity, same blood sugar reduction activity with the blood sugar reduction medicine of metformin hydrochloride, and no obvious toxic or side effect.

The present invention provides a method and apparatus for analyzing compounds with amino group(s) contained in biological organisms. In this method, a compound with amino group(s) is derivatized and then the derivative of the compound with amino group(s) is eluted by liquid chromatography using a stepwise elution means on a concentration gradient. Subsequently, the derivative of the compound with amino group(s) eluted from the liquid chromatography is detected by mass spectrometry.

The invention relates to a preparation method of mangiferin. The preparation method adopts the process steps of saturated limewater solution soaking and extraction, macroporous resin adsorption, stepwise elution of water and ethanol solution of different concentration, concentration and crystallization under reduced pressure, and recrystallization by adjusting acidity through methanol. The obtained product has the advantages of high purity, high yield and low cost. The process is easy to popularize and industrialize.

The present invention relates to rapid detection of Cd<2 + > in rice, is a method for detection of Cd<2 + > in rice by combination of solid phase extraction, separation and fluorescent colorimetry, and the method is as follows: metal ions in a mixed solution obtained by digestion of rice can be enriched in filler on a sulfonated polymer filled column, Cu<2 + >, Ni<2 + >, Zn<2 + > and other heavy metal ions can be removed by stepwise elution, a mixed solution containing Cd<2 + >, Mn<2 + > and K<+ >, Na < + > ,Ca<2 + >, Mg<2 + > and other alkali metal and alkaline earth metal ions can be separated; the solution containing Cd<2 + > is added dropwise to test paper fixed with a cadmium ion fluorescence indicator; the cadmium ion fluorescence indicator is excited by light with the 365nm specific wavelength, electronic imaging equipment is used for capturing indicator fluorescent color changes before and after the addition of the Cd<2 + >. The color change values of red, green and blue (RGB) three channels in an image before and after the color change can be extracted for construction of a standard curve of Cd<2 + > and corresponding concentration gradient. In the testing process of a sample, the obtained sample change value is compared with values in the curve so as to perform quantitative analysis of Cd<2 + > in an unknown sample.

The invention relates to a method for separating and purifying (+)-isolariciresinol and (-)-lariciresinol from folium isatidis. The method comprises the following five steps of: (A) homogenate crushing enzymolysis: performing homogenate crushing of folium isatidis, and adding cellulase, pectinase or complex enzyme of the two for enzymolysis; (B) negative-pressure cavitation reinforced extraction: performing negative-pressure cavitation reinforced extraction of methanol, ethanol or alcohol-water mixed solution; (C) resin enrichment: loading a sample into a macroporous resin column, and performing stepwise elution and adsorption of macroporous resin column with target by use of 30-35% ethanol and 50-60% ethanol; (D) chromatographic separation: dissolving the sample with 90% methanol, performing gradient elution in a Toyopearl HW-40S gel resin column, and performing fractional collection of the fraction; and (E) crystallization: crystallizing a crude product of (+)-isolariciresinol and a crude product of (-)-lariciresinol with methanol to obtain the products of (+)-isolariciresinol and (-)-lariciresinol with purity over 95%. The method provided by the invention has the advantages of few steps, short time, little solvent consumption, high yield and relatively high economic value, and is more economical and environment-friendly and suitable for large-scale production.

The invention discloses a method for purifying chymotrypsin through affinity chromatography and stepwise elution. The method comprises the process steps as follows: (1) multiple crystallization; (2) activation; (3) salting out; (4) ultra-filtration; (5) pyrogen treatment; (6) dialysis; (7) affinity chromatography; and (8) freeze-drying and preparation of the chymotrypsin. According to the method, the production technology of the chymotrypsin is continuously improved, technological parameters are studied repeatedly through experiments and optimized continuously in particular, a more scientific large-scale production technology is established finally, the titer of the chymotrypsin purified through affinity chromatography is up to 1500-1800 iu/mg, and all indicators conform to regulations of Chinese Pharmacopoeia.

The invention discloses a method for efficiently separating immunocompetence polysaccharide of ganoderma atrum. Refined ganoderma atrum polysaccharide PSG serves as raw materials of the method, strong anion exchange resin Q-Sepharose Fast Flow is adopted for separating and purifying the PSG, sodium chloride solutions in different concentrations (0-2 M) are used for performing stepwise elution, different obtained components are sequentially dialyzed, frozen and dried, and ganoderma atrume acidic beta-(1->3, 1->6)-glucan polysaccharide components with immunocompetence are obtained. The method has the advantages that different chemical components in the PSG can be effectively separated by the strong anion exchange resin; the prepared acidic polysaccharide is high in sugar content, good in uniformity and superior to refined polysaccharide PSG in immunocompetence; the method is easy to operate, fast, efficient and good in reproducibility, elution liquid in use is free of toxin and pollution, and the method can be widely used for large-scale preparation of immunocompetence polysaccharide of ganoderma atrum.

The invention relates to a method for preparing total alkaloid from macleaya cordata and further separating the total alkaloid to obtain high content alkaloid, particularly a preparation method for high content sanguinarine and chelerythrine. The preparation method comprises: atomizing and extracting a macleaya cordata raw material by acid water, adsorbing and eluting an extraction liquid by weakly polar macroporous resin, and recovering a solvent from an eluant and drying the eluant to obtain the total alkaloid, the content of which is not less than 60%; dissolving the total alkaloid in low-degree ethanol, carrying out secondary adsorption by using polar macroporous resin, carrying out stepwise elution by using 60% and 80% ethanol, and recovering a solvent from an eluant and drying the eluant to separately obtain the chelerythrine and the sanguinarine, the contents of which are not less than 90%. The preparation method provided by the invention is simple in process step, short in production period, high in product yield and less in use level of solvent, the environment is slightly polluted, and the obtained product is high in content, stable in quality and low in cost.

The present invention relates to a urine proteomics sample pretreatment method and applications thereof, and belongs to the technical field of analysis. According to the present invention, the methodcan be used for enriching proteins in urine samples so as to achieve the deep analysis of proteomics in urine samples; the method is the novel urine sample pre-treatment method; the urine sample is subjected to primary enrichment through the material, and then a stepwise elution method is performed, such that the complexity of the urine sample can be effectively reduced; compared to the traditional urine sample pretreatment method, the method of the present invention has characteristics of convenience, easy performing, low cost and the like; and the method has good application prospect and practical value in proteomics.

The invention discloses a method for separating and preparing bisnorcholenaldehyde (BA) from crystallization mother liquor obtained after extraction of phytosterol fermentation liquor. The fermentation liquor obtained after fermentation of phytosterol through mycobacteria is further subjected to steps of extraction, crystallization and the like for extraction of androstenedione (4AD), a crystallization mother liquor condensed extract after the steps is completely dissolved by low-carbon alcohol, and then filtration is performed to obtain a filtrate containing BA. The filtrate containing BA is adsorbed by a small amount of resin, then is subjected to volatilization drying at a room temperature, and then is added into the top end of a chromatographic column filled with macroporous adsorption resin. Stepwise elution is performed by using mixed solutions of ethanol and water with different concentrations as eluents; and an eluent containing the BA is collected, and the eluent is subjected to evaporating concentration, then is completely dissolved by a mixed solution of methanol, acetonitrile and water, and then is crystallized at a low temperature, and the BA with the purity of 98.5% or more is obtained after filtration and crystal washing. The method is simple in process, the mother liquor can be purified and recycled, the cost is low, industrialization can be realized easily, the process is pollution-free and wastewater-free and is in line with environmental protection requirements, and an obtained product is in line with requirements of the pharmaceutical industry.

The invention discloses a method for extracting saponin from soapberry pericarp. The method comprises the following steps of extracting saponin from the soapberry pericarp to prepare a soapberry saponin extracting solution; concentrating the soapberry saponin extracting solution by adopting a membrane concentration method to obtain a concentrated solution; adsorbing the concentrated solution withX-5 macroporous resin to obtain macroporous resin adsorbed with soapberry saponin; performing stepwise elution on the macroporous resin adsorbed with the soapberry saponin, and collecting the eluent;and performing membrane concentration and drying on the eluent to obtain the soapberry saponin. Through the method disclosed by the invention, the energy consumption is greatly reduced through low temperature by adopting membrane concentration, and a large amount of foam is prevented from being generated, and a solvent recovery rate is high; the non-polar X-5 macroporous adsorption resin is adopted for adsorption, the elution is easy, and the stepwise elution is adopted, so that the purity of the obtained soapberry saponin is high, the waste can be changed into valuable, and the economic valueis high; and the whole production process is simple to operate, energy-saving and environment-friendly.

The invention discloses a method for simultaneously separating quercetin-3-O-sophoroside, isoquercetin and chlorogenic acid from Poacynum hendersonii leaves. The method comprises a macro-porous resincolumn adsorption process, an eluent stepwise elution process and a gel chromatographic column refining process. The Poacynum hendersonii leaves are used as a raw material, and are processed to obtainPoacynum hendersonii leaf extract, the extract is fully dissolved in distilled water, insoluble substances are filtered out, the obtained filtrate goes through a macro-porous resin column and undergoes stepwise elution with an eluent, and the obtained first-stage eluate is concentrated and freeze-dried to obtain chlorogenic acid having a purity of 52% or above; and the obtained second-stage eluate undergoes reduced pressure distillation, and the obtained product is dissolved in methanol, and is continuously purified by a gel chromatographic column to obtain the quercetin-3-O-sophoroside having a purity of 93% or above and isoquercetin having a purity of 97% or above. The above separation method allows highly pure quercetin-3-O-sophoroside (93% or above), isoquercetin (97% or above ) and chlorogenic acid (52% or above) to be simultaneously obtained in a simple process, so the method has the advantages of simple and feasible process, high production efficiency, high raw material utilization rate, suitableness for industrial production, and high production and practical values.

The invention discloses a method for purifying chymotrypsin through affinity chromatography stepwise elution and a pharmaceutical composition containing chymotrypsin. The method comprises the following process steps: (1) performing multiple-crystallization; (2) activating; (3) salting out; (4) performing ultrafiltration; (5) performing primitive heat treatment; (6) dialyzing; (7) performing affinity chromatography; and (8) draining, and preparing the chymotrypsin. The chymotrypsin production process is continuously improved, particularly various process parameters are subjected to repeated experiment research and are continuously optimized, and finally, a set of scientific large-scale production process is determined. The titer of the chymotrypsin obtained by affinity chromatography purification is 1500-1800iu/mg, and each index accords with the provision in the Chinese pharmacopoeia.

The invention discloses a method for conducting affinity chromatography, stepwise elution and purification on chymotrypsin and a medicine composition for improving the stability of chymotrypsin. The method includes the following process steps: 1, multiple crystallization; 2, activation; 3, salting out; 4, ultrafiltration; 5, pyrogen treatment; 6, dialysis; 7, affinity chromatography; 8, freeze-drying and preparation of the chymotrypsin. According to the method for conducting affinity chromatography, stepwise elution and purification on chymotrypsin and the medicine composition for improving the stability of chymotrypsin, the production process of the chymotrypsin is continuously improved, particularly repeated experimental study is conducted on all process parameters, continuous optimization is conducted, finally a scaled production process which is more scientific is established, the titer of the chymotrypsin obtained through affinity chromatography and purification reaches as high as 1500-1800 iu/mg, and all indexes meet the requirements of Chinese Pharmacopoeia.

The invention relates to a method of preparing taxol by separating 10-deacetyl taxol acylate, which adopts an efficient countercurrent chromatography for separation and is characterized in that the 10-deacetyl taxol (10-DAT) is used as a semi-synthetic precursor, and the acylate thereof is used as the raw material. The whole separation process adopts a stepwise elution mode, and both of the two sets of solvent systems are respectively hexane-ethyl acetate-methanol-water with the corresponding volume ratio of 0.9:1:0.9:1 and hexane-ethyl acetate-methanol-water with the corresponding volume ratio of 0.9:1:0.9:0.8. The invention has simple and controllable operation method, and efficient, cheap and clean process, and is suitable for preparing the chemical derivatives such as taxol or 10-deacetyl-10-propionyl taxol or 2'-acetyl taxol and the like of 10-deacetyl taxol.

The invention provides a gradient separation and comprehensive utilization method of an active ingredient of a puffball. The method comprises the following steps of: extracting the puffball with alcohol firstly to form a crude extract and extraction slag, performing evaporation and concentration on the crude extract to form alcohol and an extract, stepwise eluting the extract with solvents having different polarities to form different extraction parts, performing the evaporation and concentration on the different extraction parts to separate the solvents out to form a plurality of sub extracts, performing repeated stepwise elution on the sub extracts to form a single-component extract, and separating the solvents out of the single-component extract to form a pure monomeric compound. The crude extract, the extract, the various extraction parts, the sub extracts and the monomeric compound obtained by the method can serve as products of different levels for use according to different properties, and the extraction slag can also be used for manufacturing livestock feed, materials or organic fertilizer for edible mushroom cultivation and the like. The solvents can be evaporated and recycled. The method has a high utilization rate of the puffball, almost achieves zero discharge, and has no environmental pollution and high comprehensive benefits.

A composition which has high 4′-GL purity and can be used as a reference standard for various analyses can be obtained by a more convenient method than one conventionally used. A method for preparing a high-purity 4′-GL composition includes the steps of: (A) subjecting a 4′-GL-containing galacto-oligosaccharide to activated carbon column chromatography, and performing stepwise elution with plural organic solvent aqueous solutions, wherein the organic solvent aqueous solutions are used such that the concentration of the organic solvent in the organic solvent aqueous solution is higher than the concentration of the organic solvent in the immediately preceding organic solvent aqueous solution with respect to a series of elutions; and (B) adding an organic solvent to the final fraction eluted in step (A), and crystallizing the 4′-GL.

The invention discloses a preparation and purification method and applications of a Toona sinensis fruit polysaccharide, and relates to the technical field of extraction of Toona sinensis fruit polysaccharides. The preparation and purification method comprises: drying Toona sinensis fruits, crushing, leaching with hot water, sequentially carrying out rotary evaporation concentration, alcohol precipitation and freeze drying to obtain a crude polysaccharide, removing proteins with polyamide, decolorizing with active carbon, carrying out chromatography with a DEAE-Sepharose CL-6B anion exchange column, carrying out stepwise elution respectively with NaCl solutions with a concentration of 0, 0.1, 0.2 and 0.5 mol/L, collecting the eluent in the elution with the 0.2 mol/L NaCl, carrying out vacuum concentration, dialyzing with distilled water, and carrying out freeze drying to obtain the uniform Toona sinensis fruit polysaccharide STSP-3. According to the present invention, the prepared Toona sinensis fruit polysaccharide has high anticoagulant activity, the preparation method is simple, the potential safety hazards of the remaining chemical reagents are avoided, and the good developmentand application prospects are achieved.