596 results about "Isocratic elution" patented technology

The invention provides an industrial method for extracting natural L-citrulline from plant tissues of watermelons or citrullus lamatus, and the like, which belongs to the technical field of extracting natural products in the food industry and the medicine industry. The watermelons, the citrullus lamtus or the stems and leaves thereof are taken as raw materials, and the extracting process comprises the following steps: firstly, the raw materials are washed clean and broken to separate the watermelon peel from the watermelon pulp for respectively extracting juice; the two juices are mixed, and micro-filtration and ultra-filtration are carried out to remove insoluble substances and macromolecular substances; and then reverse osmosis and nano-filtration are carried out to obtain the filtrate; the pH value of the filtrate is regulated and then a macroporous strong acid cation exchange resin column is added; and then ammonia water is taken as an eluant for isocratic elution; the eluent obtained is decompressed, distilled, deaminized, discolored, condensed and crystallized to obtain a white crystalline product of the natural L-citrulline extracted from the watermelons. The prepared product is a pure natural product without harmful substances and has high purity and good quality, thus the natural L-citrulline can be directly taken as a food additive and raw material of health products or medicines.

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 discloses a mixed fatty acid high performance liquid phase chromatography method and application thereof. In order to solve the problems that the prior art has single method in separating linolenic acid, palmitic acid, linoleic acid and other three fatty acid mixtures with very similar chemical and physical properties with HPLC, and defects in elution method, mobile phase composition selection and the like, the invention provides a method for simultaneously separating six fatty acid mixtures, i.e. linolenic acid, palmitic acid, linoleic acid, oleic acid, palmitic acid and stearic acid with HPLC. The method esterifies fatty acid solution to be tested with the acetone solution of omega-bromoacetophenone and the acetone solution of triethanolamine, and separates the solution with HPLC. The HPLC adopts an isocratic elution method, and the mixture of acetonitrile and CTAB solution or acetonitrile-water solution is selected to serve as the mobile phase. Compared with the priorart, the invention has the advantages of simple and feasible operation, good separation effect, high detection sensitivity, accurate and reliable results, and simultaneous separation of 6 fatty acids.

The invention provides bacillus subtilis natto ST188 with CGMCC No. 8400 and a high-yield method for purifying vitamin menadione-7 by using the bacterial strain. The high-yield method comprises the following steps: (1) carrying out spray drying on natto fermentation liquor, and extracting or leaching bacillus natto fermentation liquor spray drying powder by using a solvent so as to obtain a leaching solution; (2) concentrating the leaching solution obtained in the step (1) so as to obtain extract; (3) carrying out column chromatography on the extract obtained in the step (2), carrying out gradient or isocratic elution, and concentrating collected liquid, thus obtaining a menadione-7 crude product; and (4) crystallizing and purifying the menadione-7 crude product obtained in the step (3), thus obtaining the pure vitamin menadione-7 product.

The invention discloses a method for measuring organic acid in hawthorn tincture used in cigarettes, which is characterized by comprising the steps of: adopting methanol, water, glacial acetic acid and triethylamine as a quaternary mobile phase, isocratic-eluting, and detecting by adopting a high-performance liquid chromatography-diode array detection method. In the method for detecting the content of oleanolic acid and ursolic acid in hawthorn tincture, when the sample is pretreated, a hawthorn tincture sample only needs to be diluted with the methanol, and then filtered with a 0.45-Mum filter membrane for liquid chromatographic separation and analysis without interference of other impurities; in addition, the method has the advantages of being simple, convenient and fast, having wide linear range, good separation effect and accurate and reliable data, and the like.

The invention provides a high performance liquid chromatography for simultaneously detecting the content of ten organic acids in a fruit, comprising the following steps of: carrying out high performance liquid chromatography detection on a prepared standard sample solution and a solution to be detected under the following chromatography conditions: a hydrogen ion exchange column is adopted as an analysis column, flowing phases include a 0.001-0.005mol/L sulfuric acid solution as a water phase A and acetonitrile as an organic phase B, wherein the volume percentage of A is 100-96%, the volume percentage of B is 4-0%, the flow rate is 0.4-0.5mL/min, isocratic elution is used as an elution form, the detection wavelength is 210-280nm, the detection temperature is 25-50 DEG C, the feeding volume is 10-20muL, and the detection time is 30min; and finally, quantitatively analyzing and calculating the content of the ten organic acids in the fruit by using an external standard method. The method has the following characteristics of easiness and fastness in operation.

The invention relates to a liquid phase separation detection method of afatinib and enantiomer of the afatinib, and belongs to the technical field of separation detection. The method comprises the step of carrying out isocratic elution by adopting 250mm*4.6mm and 5 micrometers CHIRALPAK OZ-H chiral chromatographic column as a separation chromatographic column and adopting a mixed solution of n-hexane, ethanol, methanol and diethylamine as a flow phase. By adopting the method, the afatinib and an enantiomer impurity of the afatinib can be effectively separated and detected, and the peak pattern of a chromatographic peak is good.

The invention discloses a method for detecting Cr (VI) in cigarette paper by using an inhibition electrical conductivity detector characterized by ion chromatographic separation and large volume sample injection. The method comprises the specific steps as follows: shearing a cigarette paper sample into pieces, and accurately weighing cigarette paper pieces into a container; adding an extracting solution for vibrating extraction, and filtering the extracting solution by an aquo-system filter film, so as to obtain a to-be-detected solution; preparing a Cr (VI) standard solution, and testing and establishing a standard curve; and moreover, passing a filter liquor through an ion exchange column, and after isocratic elution, adopting the inhibition electrical conductivity detector and the ion chromatograph for analysis. According to the invention, as the large volume sample-injection electrical conductivity detector is adopted for detection, the derivation step is avoided, the preliminary treatment process is simplified, and the time, the labor and the materials for detection are saved; and further, the method can efficiently detect Cr (VI) in the cigarette paper, is rapid and reliable, is simple and convenient and easy to carry out, and fills the technical void in detecting Cr (VI) in the cigarette paper directly by the ion chromatography.

The invention discloses a method for efficiently extracting carotenoids in yellow peach fruits and determining the carotenoids in the yellow peach fruits by a liquid phase. Extracting conditions are as follows: the material-liquid ratio of pulp to an extracting agent acetone is 1 to 6 (g/mL), and ultrasonic light-shielding assisted extraction is carried out for 1 hour; chromatographic conditions are as follows: a YMC-C30 type chromatographic column is adopted; a mobile phase comprises methyl tert-butyl ether methanol with a ratio being 30 to 70; an isocratic elution manner is adopted; a flow speed is equal to 1.0mL/min; the wavelength is 450nm; the column temperature is 25 DEG C; a sample feeding amount is 20 microliters; and the time lasts for 18 minutes. According to the method for efficiently extracting the carotenoids in the yellow peach fruits and determining the carotenoids in the yellow peach fruits by the liquid phase, the extraction material-liquid ratio is 1 to 6 (g/mL), the ultrasonic light-shielding assisted extraction is carried out for 1 hour and the methyl tert-butyl ether and the methanol with the ratio of 30 to 70 are used as the mobile phase for carrying out the isocratic elution; the three conditions are combined so that a plurality of types of carotenoids in the yellow peach fruits can be effectively and completely extracted and rapidly and accurately determined; and the method is easy to operate, free of saponification and gradient elution, good in repeatability of a determined result, high in accuracy and short in liquid phase time.

The invention relates to the technical field of analytical chemistry and discloses a method for detecting afatinib dimaleate isomer and main degradation impurities through a high performance liquid chromatography.According to the method, afatinib dimaleate raw materials or preparations are compounded into a detection solution with diluent, bonding amylose-trichlorobenzene carbamic acid ester serves as a stationary phase, a normal hexane-ethanol-organic base solution serves as a mobile phase for isocratic elution, and an HPLC spectrogram is recorded under afatinib absorption wavelength.According to the method, a chromatographic column with trichlorobenzene carbamic acid ester serving as a filling material, the normal hexane-ethanol-organic base solution serves as the mobile phase, the content of isomers existing in afatinib dimaleate and preparations thereof can be measured quantitatively, afatinib, the isomers and the main impurities can be separated completely, the peak pattern is good, the method meets the specifications in Chinese Pharmacopoeia, quality of afatinib dimaleate and afatinib dimaleate preparation products is effectively controlled, and the method is great in specificity, high in sensitivity and good in accuracy.

The invention relates to the technical field of detection, in particular to a qualitative and quantitative detection method of one or more of cannabidiol, cannabidiolic acid and tetrahydrocannabinol.The qualitative and quantitative detection method of one or more of the cannabidiol, the cannabidiolic acid and the tetrahydrocannabinol comprises the following steps that: ultrasound extraction is conducted on smashed hemp flower leaves through methyl alcohol, and a test solution is obtained; the test solution is separated and detected through high performance liquid chromatography, a moving phase of the high performance liquid chromatography is a mixture of acetic acid water solution and acetonitrile, and isocratic elution is conducted; and the content of one or more of the cannabidiol, thecannabidiolic acid and the tetrahydrocannabinol are detected through an external standard method. According to the qualitative and quantitative detection method of one or more of the cannabidiol, thecannabidiolic acid and the tetrahydrocannabinol, specific wave length and a moving phase are selected, the peak pattern is good, sensitivity is high, research is conveniently conducted, and qualitative and quantitative treatment of the cannabidiol, the cannabidiolic acid and the tetrahydrocannabinol can be detected quickly and accurately under treatment of the same sample.

The invention relates to a method for extracting compound amino acid from watermelons or seeding watermelons, which belongs to the technical field of natural product extraction in the food and medicine industry. The invention uses watermelons or seeding watermelons as raw materials and comprises the steps: firstly, watermelon peel and watermelon pulp are separated and are respectively extracted for juice; the obtained juice is mixed, micro filtered and ultrafilter to remove insoluble substances and large molecular substances,; reverse osmosis and nanofiltration are carried out to the juice; after the ph value of the permeate liquid is adjusted, macroporous and strong acidic cation exchange resin columns are added in the permeate liquid; isocratic elution is carried out by using ammonia as an eluant; the eluant is distilled by pressure reduction for deaminase and decoloration; a concentrated solution is dried by a high-speed centrifugal spraying drier to obtain compound amino acid which is rich in amino acid up to over 20 types, such as citrulline, gamma-aminobutyric acid, and the like. The obtained product is a pure natural product, does not contain harmful substances, has good purity quality and can be directly used as food additives and raw materials of health food or medicine.

The invention discloses an obeticholic acid and a detection method for related substances in a preparation of the obeticholic acid. The detection method includes the specific steps that the high performance liquid chromatography method is adopted, and a C18 column is adopted; a diode array detector is adopted, and the column temperature is 20 DEG C to 30 DEG C; a phosphate buffering liquid-acetonitrile-methyl alcohol solution serves as a moving phase for isocratic elution. By means of the detection method, the conditions of impurities and degradation products of the obeticholic acid can be rapidly and accurately detected, effective separation of the obeticholic acid and all known impurities can be achieved, the interference of all kinds of impurities generated in the synthesis and preparation processes of the obeticholic acid in the product purity is avoided, and the quality of raw material medicine and the quality of the preparation can be fully and reliably controlled. The detection method is easy and convenient to operate, high in sensitivity and good in separation degree, and an effectively analysis method is provided for controlling the quality of the product.

The invention relates to a method for simultaneously detecting multiple sugars and sugar alcohols in food. An HPLC-differential refraction detection method is adopted as the method; the method is characterized by comprising the HPLC chromatographic conditions that an amide-bonded column is adopted as a chromatographic column, single mobile phases of acetone, water and triethylamine are adopted as mobile phases, isocratic elution is performed, the column temperature is 70 DEG C, the flowing velocity is 0.8 ml/min, and the sample injection amount is 10 milliliters. The detection method can be suitable for the food such as honey, wine and baijiu; detection can be performed only by simply processing a sample, the method is rapid, high in recovery rate, wide in linear range, good in correlation coefficient and low in detection limit, and the blank of simultaneous detection on the sugars and sugar alcohols in the food in China is filled up.

The invention relates to a quantitative determination method of chlorogenic acid and three kinds of isochlorogenic acid in lonicera flower medicinal material and preparation thereof, which is characterized in that: ordinary isocratic elution is adopted at the first time, acetonitrile-methanol-0.1% phosphoric acid with the volume ratio of (15 to 14): (6.5 to 6): (78.5 to 80) is adopted as a flow phase, and the content of the chlorogenic acid and three kinds of isochlorogenic acid in the lonicera flower medicinal material and preparation thereof is simultaneously determined at 326 minus or plus2nm. The method is simple, convenient and rapid to operate and is easy to popularize and master; separation of each peak of a quantitative chromatogram map of each component on three different chromatographic columns is good, the basic line is stable, and the peak appearance is completed within 20 to 30 minutes. Compared with the gradient elution, the cost of instruments is greatly reduced, the popularization rate of the method is improved, and the determination time is reduced. Due to the adoption of the quantitative determination method, a determination method and reference data can be supplied for evaluating the quality of the lonicera flower medicinal material and the preparation thereof through multiple indexes.

The invention provides a method for detecting phosphatidylcholine, which is used for determining the chemical composition and quantifying the phosphatidylcholine in yolk lecithin by using a method of using both high performance liquid chromatography and an ultraviolet detector. The method is characterized in that a silicagel column ZORBAX RX-SIL (4.6 mm X 250 mm, 5 microns) manufactured by Agilent is adopted as an analytical column and methanol/acetic acid is used as a mobile phase to carry out isocratic elution, and other elution conditions are an ultraviolet detection wavelength of 200 nm, a column temperature of 30 DEG C, a sample feeding quantity of 15 microliters and a flowing speed of 0.5 ml/min. The mobile phase of the method has the characteristics of binary system, simpler composition, lower flowing speed of 0.5 ml/min, faster peak time within 14 minutes and linear range of a phosphatidylcholine standard curve of 10-160 microgram/ml; the added 3.75% acetic acid solution improves a peak shape of the phosphatidylcholine and effectively inhibits serious tailing phenomenon; and the use cost of a solvent is saved. It is proved by a methodological replication experiment that the method has the advantages of high precision, good stability and higher accuracy.

The invention discloses an analytic method for related substances in raw material and a preparation of trelagliptin succinate. A high performance liquid chromatography is adopted, and chromatographic conditions include that a chromatographic column is octadecyl silane bonded silica gel chromatographic column, the mixed solution with the pH of 3.3-3.7 and the volume ratio of phosphate buffer aqueous solution to acetonitrile of (88-92):(8-12) is taken as a mobile phase A, the mixed solution with the pH of 3.3-3.7 and the volume ratio of phosphate buffer aqueous solution to acetonitrile of (18-22):(78-82) is taken as a mobile phase B, detection wavelength is 218-222nm and isocratic elution is carried out. The analytic method disclosed by the invention can detect many impurities and can rapidly, effectively and accurately monitor the related substances in trelagliptin succinate.

The invention provides a method for extracting a cordycepin component from Cordyceps militaris. The method comprises the following steps: ultrafinely pulverizing Cordyceps militaris fruiting body to obtain ultrafine powder; extracting the ultrafine powder with ultrapure water, collecting supernatant, drying, precipitating with ethanol, centrifuging, collecting supernatant, performing rotary evaporation, and drying to obtain crude extract; dissolving the crude extract with 15% methanol-water solution to 100 mg/mL, separating with preparative liquid phase DAC reversed phase chromatographic column with reversed phase separation filler of C18, performing gradient elution with 5-15% methanol-water solution, performing isocratic elution with 15% methanol-water solution, setting detection wavelength of 260 nm, performing in vitro antitumor activity screening of components, and selecting the component with the highest activity, i.e. target cordycepin component. The method provided by the invention is simple and controllable, and has good repeatability; the content of cordycepin in the obtained component is high, the batch quality is stability, and the cordycepin component can be used for preparing antitumor drugs.

The invention relates to a fractionation detection method for tenofovir alafenamide and isomer thereof, and belongs to the technical field of fractionation detection. The detection method comprises the following steps: taking a chromatographic column filled with octadecylsilane chemically bonded silica; and carrying out isocratic elution by taking methanol and an acidic aqueous solution as mobilephases under certain conditions. By the method, the tenofovir alafenamide and the isomer thereof can be effectively detected in a fractionation manner, the peak pattern of chromatographic peaks is good, the appearance time is short, and the degree of fractionation is good.

The invention relates to a method for separating and purifying corn anti-hypertensive peptides, which belongs to a method for separating corn peptides from corn protein powder. The method for separating and purifying the corn anti-hypertensive peptides comprises the following steps of: hydrolyzing the corn protein powder the corn yellow pigment of which is removed by using alkali protease, filtering by adopting a plate frame filter to remove impurities, then filtering by a film, afterwards determining the molecular weight of a target product by the analysis of high-performance liquid chromatography, selecting a hydrolysate with the activity of lowering the blood pressure, and separating and purifying the hydrolysate with the activity of lowering the blood pressure by adopting a chromatographic separation system of a simulated moving bed so as to obtain the corn anti-hypertensive peptides. The separation and the purification are carried out by adopting an isocratic elution mode, wherein the feeding flow rate is 5 to 100ml/min; the flow rate of a mobile phase is 6.5 to 130ml/min, and the concentration of the mobile phase is 40 percent of ethanol; the flow rate of an outlet of a product is 4.5 to 90ml/min; and the flow rate of a circulating pump is 10 to 200ml/min, and the switching time is 6 to 120min. On the basis of achieving an industrialized level, the invention can separate the corn anti-hypertensive peptides with relatively higher purity.

The invention discloses an extraction process of ginseng saponin Rd. The extraction process comprises the following steps: S1, grinding ginseng saponin Rd-containing herbal materials into herbal powder, performing reflux extraction by using 70-95% ethanol, recycling the ethanol, and concentrating to obtain a total extract; S2, taking the total extract, enabling the total extract to pass through D101 macroporous adsorption resin, eluting by using 40-80% ethanol, recycling the ethanol, and performing deconperssion concentration to obtain total saponin; S3, taking the total saponin, enabling the total saponin to pass through a silica gel chromatography column, performing isocratic elution by using a mixture of n-butanol, ethyl acetate and water, performing thin layer chromatography, combining fractions to obtain crude ginseng saponin Rd; S4, taking the crude ginseng saponin Rd, enabling the crude ginseng saponin Rd to pass through an Rp-C18 silica gel column, performing isocratic elution by using a mixture of methanol and water, performing thin layer chromatography, combining required components, separating out precipitate to obtain the ginseng saponin Rd. By the extraction process, the ginseng saponin Rd separating and purifying effects can be improved, use of a toxic organic reagent is avoided, the operation is simple and convenient and a production process is easy to control; therefore, the extraction process is suitable for industrial production.

The invention relates to a separation and detection method for astaxanthin in a haematococcus pluvialis extract. The separation and detection method comprises the following steps of 1 extraction of carotenoid in the haematococcus pluvialis extract; 2 enzymolysis of the carotenoid; 3 astaxanthin separation and detection through a normal-phase high performance liquid analysis method, wherein the liquid-phase on-device conditions comprise the detection wavelength is 474 nm, a chromatographic column is Luna 3micro Silica(2), the chromatographic column temperature ranges from 20 DEG C to 25 DEG C, the flow speed is 1-1.2 ml/min, a flow phase is prepared from n-hexane and acetone according to the volume ratio of 75%:25%-90%:10%, and isocratic elution is conducted. According to the separation and detection method, the pretreatment efficiency is high, the intersolubility between the extracting reagent acetone and the astaxanthin is good, the enzymolysis time is shorter than the saponification time, and the efficiency is high; the high performance liquid chromatography on-device conditions are good; isocratic elution is achieved, a base line is easier to stabilize, the peak shape is good, the separation degree is high, and more isomers can be separated out; the peak flowing time is short, and the method is not prone to be influenced by outside light and heat and more suitable for large-scale detection.

The invention provides a method for extracting N6-(2-ethoxyl) adenosine from cordyceps militaris. The method comprises the following steps: carrying out superfine comminution on cordyceps militaris fruiting body to obtain superfine powder; adding ultrapure water into the superfine powder, heating and extracting at the temperature of 100 DEG C, centrifuging, taking and drying the supernatant; adding a little ultrapure water to dissolve, adding 95% ethyl alcohol till the final concentration is 80%, carrying out ethanol precipitation, collecting and drying the supernatant overnight; dissolving by using a 15% methanol-water liquid till the concentration is 200mg/ml, separating through a preparative liquid chromatography, carrying out isocratic elution for 30-80 minutes by using the 15% methanol-water liquid, collecting the target chromatographic peak, concentrating and drying to obtain the N6-(2-ethoxyl) adenosine. The method is simple, controllable and good in repeatability; the purity of the extracted N6-(2-ethoxyl) adenosine can be up to 98%; and the N6-(2-ethoxyl) adenosine is proved to have the effect of resisting insomnia and can be used for preparing insomnia-resisting drugs.

The invention discloses a high performance liquid chromatography (HPLC) method for detecting contents of flavonoid components in herba houttuyniae. The method comprises the following steps: (1) preparing a test solution, taking herba houttuyniae powder, extracting the herba houttuyniae powder with lower alcohol, and filtering to obtain the test solution; (2) preparing a reference solution, takingreference substances, i.e., hyperoside and quercitrin, mixing, adding the lower alcohol to prepare the mixed reference solution; (3) respectively injecting the test solution and the reference solutioninto a high performance liquid chromatographic instrument for detecting, wherein the chromatographic conditions are as follows: chromatographic column: C18 chromatographic column; mobile phases: a mobile phase A is acetonitrile, and a mobile phase B is 0.1% phosphoric acid aqueous solution; elution procedure: isocratic elution is carried out for 30 minutes, and the volume ratio of A to B is equalto (18-22) to (82-78); detection wavelength: 205nm; (4) calculating according to the detection result so as to obtain the contents of the hyperoside and the quercitrin in the herba houttuyniae. The detection method provided by the invention is accurate and reliable as well as simple and quick, and lays a foundation for improving the quality control level of the herba houttuyniae.

The invention provides a continuous gradient elution system based on a stimulated moving bed (SMB) and a treatment method of the continuous gradient elution system. The continuous gradient elution system comprises the SMB and a matched device connected with a single chromatographic column in an elution area of the SMB, and the matched device comprises five liquid storage tanks, thirteen valves and five pumps. Gradient elution liquid can be continuously and periodically supplied to each chromatographic column in the elution area of the SMB. Each time when a turnplate of the SMB rotates by a cell, and both the gradient elution process and a mode of gradient elution liquid concentration increase with time of each chromatographic column are completely coincident with that of the previous chromatographic column, so that gradient elution of each chromatographic column in the elution area is realized. The continuous gradient elution system has good elution and separation effect; with equal elution flow, single-stage separation factor of a to-be-separated target component is greater than isocratic elution, band migration distance needed for separating to obtain a target product is short, separation period is short, process operation is simple, and programmed operation control can be realized easily.