34 results about "Stepwise elution" patented technology

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 provides a preparation method of high-purity mixed sodium deoxyribonucleotide. The method comprises the following steps of: (1) loading an enzymolysis liquid onto a No.1 column and a No.2 column which are filled with regenerated anion exchange resin and are connected in series; (2) introducing water into the No.1 column, washing till the pH of an effluent liquid of the No.2 column is 7.0-9.0, introducing an acid into the No.1 column, and washing till the pH of an effluent liquid of the No.2 column is 3.0-3.5; (3) introducing a mixed solution of sodium chloride and an acid into the No.1 column, and introducing the effluent liquid of the No.2 column into a No.3 column filled with active carbon; and (4) washing the No.3 column with water, eluting with a cholamine solution, collecting a part of deoxyribonucleotide of which the concentration is higher than 20 g / L and the HPLC (High Performance Liquid Chromatography) is over 98 percent from an eluent, concentrating, and performing ultrafiltration and freeze drying to obtain a finished product. By adopting measures such as serial combination, stepwise elution and the like, a high-purity product which only contains four types of sodium deoxyribonucleotides and has a stable ratio can be obtained.

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 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 relates to a method for separating lactoperoxidase from bovine whey and belongs to the technical field of biochemical engineering. According to the method, the bovine whey serves as a raw material liquid, cation exchange chimeric composite crystal gel with multistage pores serves as a separating medium, and conventional treatment, such as crystal gel chromatography, bed column balancing, sampling and adsorbing, buffer solution flushing, saliferous eluent stepwise elution, subsequent desalting and freeze drying, is carried out, so that high-purity lactoperoxidase with the purity of 98% is obtained through separating, and the yield can reach 92%. The cation exchange chimeric composite crystal gel medium has very good selectivity to lactoferrin and lactoperoxidase, which are difficultly separated through conventional chromatography, in the whey. The method has the advantages that steps are few, the operation is simple, the separation is rapid, the cost is low, and the obtained lactoperoxidase is high in purity and yield and has broad application prospects in the respects of high-value utilization of whey and separation of active protein.

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 relates to a preparation method of water-soluble tawny natural pigments in camellia cleifera seed meal. By taking residues after squeezing tea oil from camellia cleifera seed as a raw material, the method for preparing the high purity tawny pigments by the following processes: preparing raw materials of camellia cleifera seed meal; crushing by a crusher; extracting by edible alcohol; filtering; concentrating an extracting solution; carrying out polyamide column chromatography; carrying out stepwise elution on a mixture containing 0.1% acetate solution and an edible alcohol solution; and obtaining a tawny pigment product. The product comprises various flavone glycoside compounds, the main component comprises two compounds, and the purity of flavonoid pigment components in the final product detected by HPLC can reach over 70-95%. The obtained tawny pigments can be added into foods or cosmetics, particularly transparent liquid shampoo or shower gels which are clear and natural in color and have sense of beauty and natural antioxidant activity.