49 results about "Reversed-phase chromatography" patented technology

The invention discloses a general secondary amine molecular cage-nanocellulose composite chiral stationary phase for positive and negative phases as well as a preparation method and application of the general secondary amine molecular cage-nanocellulose composite chiral stationary phase. A secondary amine molecular cage (RCC3-R) modified silica gel is used as a substrate, a nano cellulose derivative (NCDMPC) is loaded through a coating method to prepare two chiral stationary phases (CSP-1 and CSP-2), and aminopropyl silica gel is used as a substrate to prepare a control chiral stationary phase (CSP-3). By utilizing the three-dimensional cavity screening effect of RCC3-R and the synergistic effect of an NCDMPC chiral spiral skeleton, the common use of normal phase and reversed phase chromatography modes is realized, and the chiral recognition and separation performance is superior to that of CSP-3. 15 chiral compounds can be split in a normal-phase mode, 12 chiral compounds can be split in a reverse-phase mode, excellent splitting performance on alcohols, amines and chiral drugs is shown, a novel material can be provided for efficient separation of complex systems such as chiral drugs and pesticides, and the application prospect is wide.

The invention discloses a method for improving the detection recovery rate of ascorbyl palmitate in a compound product, which comprises the following steps: taking a sample, adding BHT (butylated hydroxytoluene), then adding an isopropanol-n-hexane mixed solution containing 0.8-1.2% by volume of glacial acetic acid as an extraction solvent, then fully mixing, extracting, freezing, centrifuging, taking supernate, and collecting the supernate to obtain the ascorbyl palmitate. The chromatographic conditions are as follows: a C18 reversed-phase chromatographic column is adopted, a mobile phase A and a mobile phase B are used for gradient elution, the flow velocity of the mobile phase is 0.8-1.2 mL / min, the column temperature is 28-32 DEG C, and the maximum absorption wavelength of ascorbyl palmitate is taken as the detection wavelength. According to the method, the extraction efficiency is improved by optimizing a sample pretreatment process, and the problem that the liquid phase detection error of the substance in the compound product is generally higher than 10% is effectively solved in combination with improved high performance liquid chromatography separation conditions.

The application discloses a lipoic acid high-performance liquid chromatography detection method, which comprises the following steps: S1, sample pretreatment: ultrasonic extraction is carried out on the sample by using a composite extraction liquid, then solid-liquid separation is carried out, and a test sample solution is obtained; the composite extraction liquid is an acetonitrile water mixed solution containing cysteine and EDTA salt; S2, HPLC detection: the chromatographic conditions are as follows: a C18 reversed-phase chromatographic column is used, a buffer solution-alcohol mixed system is used as a mobile phase, the pH of the buffer solution is 2.5-3.5, the detection wavelength is 210-220 nm in the first stage and is 275-285 nm in the second stage, the first stage is 0-7-9 min, and the second stage is 7-9 min-14-16 min. By optimizing the pretreatment method, optimizing the chromatographic conditions and adopting a dynamic detection wavelength, the lipoic acid content detection method with high sensitivity and strong anti-interference ability is obtained.

This invention discloses a purification method for Tirzepatide. The method includes: dissolving crude Tirzepatide peptide in ammonia or sodium bicarbonate solution, filtering, and purifying by reversed-phase chromatography to obtain a purified sample; eluting the purified sample using reversed-phase high-performance liquid chromatography (RP-HPLC) with a sodium or potassium buffer solution as mobile phase A3 and one or a mixture of acetonitrile, methanol, and isopropanol in any proportion as mobile phase B, preparing a sodium or potassium salt fraction; cooling the sodium or potassium salt fraction to -20 to 10°C and maintaining the temperature; then adding an organic solvent to the fraction, controlling the temperature at -20 to 10°C to precipitate the sample; after complete precipitation, solid-liquid separation is performed, the solid is collected, dissolved in water, filtered, and freeze-dried. This invention improves the concentration speed, ensures product purity, and strictly controls impurities.

The invention relates to the technical field of polypeptide medicine purification, in particular to a purification method of retaglutide. The method comprises the following steps: mixing a retaglutide crude product aqueous solution with salt to enable the final concentration of the salt to be 0.5-2.0 mol / L, and loading a sample to a chromatographic column filled with a hydrophobic interaction chromatographic filler; eluting by adopting a salt concentration decreasing gradient, and collecting to obtain a first purified solution; adjusting the pH and conductivity of the first purified liquid, and loading the first purified liquid to a chromatographic column filled with an ion exchange chromatographic filler; eluting by adopting a gradient of increasing salt concentration, and collecting to obtain a second purified solution; and loading the second purified solution to a chromatographic column filled with a reversed-phase chromatographic filler, taking an acidic modifier aqueous solution as a mobile phase A and an organic solvent as a mobile phase B, eluting by adopting a gradient of increasing concentration of the organic solvent, and collecting to obtain a final product. Through innovative process combination, the problems that in the prior art, impurities are difficult to deeply remove, and the consumption of an organic solvent is large are effectively solved.

The application provides a method for determining the content of biotin in a complex vitamin preparation. The method comprises detecting the content of biotin by high performance liquid chromatography, wherein a surface porous reversed-phase chromatographic column with modified amide groups is used for gradient elution, and a mobile phase A and a mobile phase B are used, the mobile phase A comprises potassium dihydrogen phosphate buffer and acetonitrile, and the mobile phase B comprises acetonitrile. The method can realize the non-interference separation and detection of the content of biotin in the complex vitamin preparation with a complex matrix, and has the advantages of simplicity, good specificity, rapid analysis, accurate result and sensitive response.

The invention discloses a high performance liquid chromatography (HPLC) detection method for enantiomers in moxifloxacin hydrochloride with high separation degree, and belongs to the technical field of medicine detection. The method aims at solving the problems that in the prior art, the leading edge of a main component peak is serious, and the detection accuracy and good peak pattern under the conditions of high separation degree and low content are difficult to meet at the same time, and the adoption of an expensive chiral chromatographic column is avoided. According to the detection method, chiral resolution is realized on a conventional high performance liquid chromatograph by adopting a reversed-phase chromatographic column (such as terminated octadecylsilane chemically bonded silica as a filler). According to the core technical scheme, the unique mobile phase composition is as follows: a water phase contains 0.54 g / L of zinc acetate and 1.17 g / L of L-valine, 1ml of diethylamine is creatively added, and the pH value is adjusted to 5.6 by using dilute sulfuric acid; and an organic phase adopts a methanol-ethanol mixed solution (15: 8) with a specific ratio. And finally, mixing the water phase and the organic phase according to the volume ratio of 77: 23. Under optimal chromatographic conditions (the column temperature is 40 DEG C and the detection wavelength is 295 nm), the method can realize efficient separation between moxifloxacin hydrochloride and enantiomers, the separation degree is greater than 4.5, and the detection accuracy of trace isomers can be realized. A methodological verification result shows that the method is high in sensitivity (the detection limit is 0.0096%, and the quantitation limit is 0.024%), and has a good linear relationship (a correlation coefficient rgt; and the accuracy is good (the recovery rate is between 80.0% and 120.0%). The method can be used for qualitative and quantitative analysis of the moxifloxacin hydrochloride bulk drug and enantiomers in the moxifloxacin hydrochloride preparation.

The present application belongs to the field of biopharmaceuticals. Disclosed is the use of a stem cell extract in the preparation of a drug for treating or preventing arthritis by regulating lysosomal and / or mitochondrial functions, wherein a method for preparing the stem cell extract comprises: S1) culturing mesenchymal stem cells and creating stress conditions to stimulate the mesenchymal stem cells; and S2) isolating and purifying a culture supernatant of the mesenchymal stem cells cultured in S1) to obtain a stem cell extract, wherein S2) comprises: S21) filtering the culture supernatant to obtain a filtrate; S22) concentrating the filtrate by means of 3KD ultrafiltration to obtain a crude sample; S23) purifying the crude sample by means of size-exclusion chromatography or reversed-phase chromatography to prepare a purified sample; and S24) preparing the purified sample into a preparation. In the present application, the culture supernatant containing secretions of the stressed stem cells is obtained by means of stress culturing of the stem cells, and in combination with a purification method, the supernatant is prepared into a preparation which can be used for treating arthritis.

The invention belongs to the technical field of analytical chemistry and medicinal chemistry, and particularly discloses a sucrose octasulfate separation and detection method based on high performance liquid chromatography. According to the method, a to-be-treated solution containing sucrose octasulfate is subjected to high performance liquid chromatography separation, and a detector is arranged for detection after high performance liquid chromatography separation to obtain the content of sucrose octasulfate; chromatographic conditions are as follows: a chromatographic column is a reversed-phase chromatographic column; the gradient elution procedure is carried out for 0-12 min, the ratio of a mobile phase A to a mobile phase B is 95: 5, 12-30 min, and the B is linearly increased to 20%; the linear recovery to the initial condition is carried out in 30-30.1 min, and the A: B = 95: 5 in 30.1-35 min; the column temperature is 35-37 DEG C; and direct amplification is carried out for preparative chromatography, so that separation and preparation of sucrose octasulfate can be realized. The separation or detection method disclosed by the invention has good separation degree, sensitivity and universality, and is high in analysis speed, good in repeatability and wide in application.

The invention relates to a method for purifying a non-steroidal GnRH agonist polypeptide, in particular to a method, a system and a product for preparing water-soluble high-purity delorelin acetate. According to the invention, a raw material medicine existing in a high acetate form is obtained through preparative-grade reversed-phase chromatography purification, ion conversion on a column and optimization of salt conversion and freeze-drying pretreatment. The HPLC purity of the product is greater than or equal to 98.5%, the acetic acid content is about 13.9%-14.7% (w / w), the solubility in pure water is greater than or equal to 120 mg / mL, and the product is kept clear for 24 hours; the retention time is consistent with that of a commercial reference substance. And compared with a contrast, the method has the advantages that the high-purity fraction yield is obviously improved, impurities and non-target pair ion residues are reduced, and the method is suitable for industrial amplification and preparation development.

The invention discloses an extraction method of oat polypeptide, and belongs to the technical field of polypeptide extraction. The method comprises the following steps: carrying out enzymolysis on an oat protein raw material to obtain a crude extract; carrying out oscillation adsorption on the crude extract by using a modified graphene material to enrich polypeptide; then desorbing to obtain a preliminarily enriched polypeptide solution; carrying out gradient elution by using a reversed-phase C18 chromatographic column, and carrying out fine separation; and finally, concentrating and drying to obtain a high-purity oat polypeptide product. The key preparation method of the modified graphene comprises the following steps: by taking an oat processing byproduct as a raw material, preparing sulfonic acid group carbon dots through a concentrated sulfuric acid hydrothermal method, and then grafting the sulfonic acid group carbon dots and graphene oxide through electrostatic self-assembly and hydrothermal reaction to obtain a three-dimensional network structure material with the surface rich in sulfonic acid groups. According to the method, the homologous modified material is used for enhancing specific adsorption, chromatographic refining is combined, the oat polypeptide is purified from the complex enzymatic hydrolysate in a high-efficiency, high-selectivity and low-cost mode, the process is green and environmentally friendly, and the obtained product is high in purity and good in biological activity.

The invention relates to the technical field of separation and purification of active ingredients of traditional Chinese medicines, and discloses a saponin compound derived from aralia elata and a preparation method thereof. The saponin compound has a structure as shown in a formula (I). The preparation method comprises the following steps: carrying out hydrous ethanol extraction on aralia mandshurica root bark, enriching through macroporous adsorption resin, carrying out directional enrichment on acidic saponin through anion exchange resin, carrying out coarse separation and preparative liquid phase refining by adopting reversed-phase C18 chromatography, and carrying out window collection in combination with online detection to obtain a target saponin compound. The method can improve the separation selectivity of the target component, reduce the co-elution phenomenon and improve the purity and batch stability of the final product, and is suitable for preparation of the aralia mandshurica sourced saponin compound.

The application relates to the technical field of medicines, and discloses application of a traditional Chinese medicine composition in serum neurotransmitters, which comprises the following steps: selecting 6-8-week-old mice to establish normal models, chronic fatigue syndrome models and functional dyspepsia models; blood sampling is conducted on the mice of the normal models, the chronic fatigue syndrome models and the functional dyspepsia models to obtain normal blood samples, CFS blood samples and FD blood samples respectively; protein precipitation operation is conducted to obtain normal deproteinized supernatants, CFS deproteinized supernatants and FD deproteinized supernatants; separation is conducted by using a reversed-phase chromatographic column, and then mass spectrometry is conducted to obtain normal serum neurotransmitter mass spectra, CFS serum neurotransmitter mass spectra and FD serum neurotransmitter mass spectra respectively; concentration value calculation and rhythm fluctuation analysis are conducted to obtain a neurotransmitter function index and rhythm stability quantitative score respectively. Therefore, the mechanism of the traditional Chinese medicine composition in serum neurotransmitters is clearly quantitatively characterized.

The invention provides a method for detecting the content of urea in a water sample by using high performance liquid chromatography. The method comprises the following steps: preparing a urea standard solution; performing filter membrane filtration pretreatment on the to-be-detected water sample; carrying out high performance liquid chromatography analysis according to optimized chromatographic conditions, and calculating the urea content by adopting a standard curve method; wherein the chromatographic conditions are as follows: a C18 type reversed-phase chromatographic column is adopted, a mixed solution of acetonitrile and ultrapure water is adopted as a mobile phase, isocratic elution is performed, and the detection wavelength is 190-200nm. According to the method, the mobile phase and the chromatographic column are improved, the urea content in the water sample is directly and accurately detected, operation is easy and convenient, the detection time is short, good specificity and stability are achieved, the detection limit is lower than 0.6 microgram / L, the recovery rate ranges from 95% to 105%, the relative standard deviation is smaller than 2%, and the method can be effectively applied to detection and analysis of the urea content in various environment water samples.

The application discloses a method for simultaneously detecting retinoids and lipid-soluble vitamin metabolic marker groups in biological samples and matching reagents. The application is based on magnetic bead solid-phase extraction technology, significantly improves sample purification effect, and reduces matrix interference; in combination with a high-aqueous-phase reversed-phase chromatography system and a positive-negative ion switching multi-reaction monitoring mode, simultaneous separation and detection of seven retinoid molecules and a full spectrum of lipid-soluble vitamins are realized. The method has high sensitivity, strong specificity and wide detection range, and for the first time realizes one-stop analysis across species and sample types, thereby providing reliable platform support for clinical and scientific research applications.

The invention discloses a detection and analysis method for an alpoxivan intermediate 3 and related impurities, and relates to the field of substance detection methods, the detection and analysis method comprises the following steps: S1, preparation of a test solution: taking a sample, adding a solvent into the sample, dissolving, and diluting for later use; s2, setting chromatographic conditions and a gradient elution program, wherein the chromatographic conditions of the chromatography comprise the use of a reversed-phase C18 chromatographic column, a mobile phase A and a mobile phase B; s3, taking the test solution, injecting the test solution into a liquid chromatograph, recording a chromatogram, and calculating the impurity content; wherein the mobile phase A is prepared from 0.15 to 0.25 weight percent of triethylamine solution, and the pH (Potential of Hydrogen) of the mobile phase A is 2 to 3; the mobile phase B comprises methanol. According to the method disclosed by the invention, the aim of effectively separating and quantifying various related impurities of the alpoxivan intermediate 3 is fulfilled through optimized chromatographic conditions.

The present application belongs to the field of fingerprint, and particularly relates to a method for detecting medicinal stems of Dendrobium candidum from Sichuan origin. The specific technical scheme is as follows: a method for constructing a Dendrobium fingerprint, comprising the following steps: mixing a Dendrobium powder to be tested with methanol, performing ultrasonic treatment, taking out, cooling, and filtering; evaporating the filtrate under reduced pressure, dissolving the residue with methanol, filtering, shaking, and obtaining a test solution; taking the test solution, performing high performance liquid chromatography detection, and recording a fingerprint; and the chromatography detection has the following chromatography conditions: a C 18 reverse-phase chromatography column; a column temperature of 30 DEG C; a detection wavelength of 270 nm; and a total flow rate of 0.8 mL / min. The method provided by the present application is simple to operate, has complete characteristic components, and has a stable test solution, high precision, and good reproducibility.

This invention belongs to the field of chromatographic detection and analysis, and discloses a reversed-phase chromatographic analysis method for the purity of enterokinase protein. High-performance liquid chromatography (HPLC) is used to detect enterokinase in the sample solution. In this HPLC method, the chromatographic column is selected from reversed-phase chromatographic packing material C18 or C8, with a particle size of 3.5~5 μm and a pore size of 120~1000 Å. The mobile phase is selected as follows: an aqueous solution of heptafluorobutyric acid (HFA) is used as mobile phase A, and an acetonitrile solution of HFA is used as mobile phase B. This invention improves the separation of the main peak and subsequent impurities in the enterokinase stock solution by screening suitable mobile phase systems and column types. It also optimizes chromatographic parameters such as the proportion of modifier in the mobile phase, column temperature, sample loading, mobile phase gradient, and wavelength, ultimately achieving effective separation of the main peak and subsequent impurities in the enterokinase stock solution. Furthermore, the analysis time is short and the repeatability is good, providing an effective analytical method for the quality control of enterokinase stock solution.

The invention discloses a kit for evaluating the effective use dosage of tripterygium glycosides for treating diabetic nephropathy, and belongs to the technical field of medicine, the kit comprises labeled demethylzeylasteral, an SPE 96 pore plate for serum sample treatment, and a reversed-phase chromatographic column for UPLC-MS (Ultra Performance Liquid Chromatography-Mass Spectrometry) analysis. According to the application, demethylzeylasteral is taken as a pharmacodynamic marker of tripterygium glycosides for treating diabetic nephropathy, so that the problem that the dosage of tripterygium glycosides is not matched with the curative effect and the safety in the treatment of diabetic nephropathy is solved, the treatment scheme can fit the individual condition of a patient, and accurate medication is realized; the effective dose of tripterygium glycosides for treating diabetic nephropathy can be judged, and poor curative effect caused by improper dose can be avoided; severe side effects possibly caused by the medicine are avoided, and the medication risk is reduced especially for patients who use the medicine for a long time.

The present application relates to a kind of quantitative determination electronic cigarette liquid trimellitic acid, p-phthalic acid, m-phthalic acid method, belong to electronic cigarette liquid component detection technical field, it is characterized in that: using to electronic cigarette liquid sample adds certain concentration of formic acid aqueous solution extraction and improves the acid-base environment of sample, further purify sample by vortex, filtration, using core-shell (surface porous) reversed-phase chromatographic column, realize the complete separation of three target objects, combined with liquid chromatography mass spectrometer technology, internal standard method quantitative, realize the accurate detection of trimellitic acid, p-phthalic acid, m-phthalic acid in electronic cigarette liquid.The present application has the advantages that simple operation, quantitative accurate, high sensitivity, target can be extracted from complex electronic cigarette liquid matrix and accurately quantified, can satisfy the accurate detection of trimellitic acid, p-phthalic acid, m-phthalic acid in electronic cigarette liquid.

The invention discloses a high performance liquid chromatography detection method of ethiprole, which comprises the following steps: preparing a standard solution, preparing a sample solution, and adopting a common solvent methanol for reversed-phase liquid chromatography as a diluent; a 0.5% acetic acid solution and acetonitrile are used as mobile phases for gradient elution; a C18 reversed-phase chromatographic column with porous particles on the surface and high-density coverage is adopted; respectively detecting the standard solution and the test solution according to the conditions, and comparing the spectrum and the main peak response value with the standard spectrum and the response value of the standard solution in the step S7 to calculate the mass fraction of ethiprole in the test solution. The ethiprole is efficiently and accurately detected through gradient elution optimization, acidic mobile phase combination, short column efficient separation, wavelength specificity and the like.

The invention discloses walnut undecapeptide as well as a preparation method and application thereof. The amino acid sequence of the walnut undecapeptide is as shown in SEQ ID NO. 1. The walnut undecapeptide has good immunoregulatory activity, and can be applied to preparation of immunoregulatory active drugs, food or health care products. The method comprises the following steps: carrying out protease enzymolysis on a walnut protein aqueous solution, carrying out solid-liquid separation after enzyme deactivation, carrying out membrane separation on the obtained supernatant, and drying to obtain a walnut protein hydrolysate; and purifying active peptides from the walnut protein hydrolysate by using a reversed-phase chromatographic column, collecting target active peptide components eluted by ethanol with a volume fraction of 7%, concentrating, purifying by using a sephadex chromatographic column, concentrating, and drying to obtain the walnut undecapeptide. The preparation method is of great significance in promoting deep processing and utilization of walnut protein, increasing the added value of products and promoting sustainable development of the industry.

The invention discloses walnut dodecapeptide as well as a preparation method and application thereof. The amino acid sequence of the walnut dodecapeptide is as shown in SEQ ID NO. 1. The walnut dodecapeptide has good immunoregulatory activity, and can be applied to preparation of immunoregulatory activity medicines, food or health care products. The method comprises the following steps: carrying out protease enzymolysis on a walnut protein aqueous solution, carrying out solid-liquid separation after enzyme deactivation, carrying out membrane separation on the obtained supernatant, and drying to obtain a walnut protein hydrolysate; and purifying active peptides from the walnut protein hydrolysate by using a reversed-phase chromatographic column, collecting a target active peptide component eluted by ethanol with a volume fraction of 10%, concentrating, purifying by using a sephadex chromatographic column, concentrating, and drying to obtain the walnut dodecapeptide. The preparation method is of great significance in promoting deep processing and utilization of walnut protein, increasing the added value of products and promoting sustainable development of the industry.

The invention belongs to the field of chemical detection and analysis, and particularly relates to a method for determining the content of sodium diisobutyl dithiophosphinate by using high performance liquid chromatography. According to the method, a C18 reversed-phase chromatographic column is adopted as a chromatographic column, an ultraviolet detector is adopted as a detector, a 20-30 mmol / L ammonium acetate aqueous solution is adopted as a mobile phase A, acetonitrile is adopted as a mobile phase B, a specific gradient elution procedure is adopted, and the content of sodium diisobutyl dithiophosphinate is separated and measured. According to the method, the sodium diisobutyl dithiophosphinate and the impurities thereof can be effectively separated, the content of the sodium diisobutyl dithiophosphinate can be rapidly and accurately measured by adopting the method, and a new solution thought is provided for the problems of purity detection and impurity control in the production of the sodium diisobutyl dithiophosphinate.

This invention discloses a method for detecting L-5-methyltetrahydrofolate content with magnesium ion interference resistance, comprising the following steps: S1. Sample pretreatment: S11. Using phosphate buffer containing antioxidants, disodium EDTA, and sodium citrate as the extraction reagent, the sample is ultrasonically extracted in an ice-water bath at 2-6°C under light-protected conditions; S12. Extraction is performed using an aqueous two-phase extraction system; S2. HPLC detection: A reversed-phase chromatographic column is used, with gradient elution using mobile phase A and mobile phase B; mobile phase A is an aqueous solution of potassium dihydrogen phosphate containing antioxidants, sodium citrate, and disodium EDTA, and mobile phase B is methanol. This invention solves the problems of peak tailing, signal suppression, and matrix interference in the detection of L-5-methyltetrahydrofolate in complex magnesium-containing systems by synergistically dissociating the magnesium-folate complex through pH control and a composite masking agent, combined with aqueous two-phase extraction to purify the matrix, and optimized HPLC separation conditions.

The invention relates to the technical field of test analysis, in particular to a cefditoren pivoxil related substance detection method, which adopts a high performance liquid chromatograph for testing, takes a sodium dihydrogen phosphate solution-acetonitrile-methanol system as a mobile phase, and adds an alkyl sodium sulfonate anion pair reagent into the mobile phase. A neutral ion pair with higher hydrophobicity is formed by the cefditoren acid and the-NH2 group in the cefditoren acid delta 3 isomer, and is in a molecular form, so that the retention time of the neutral ion pair on a chromatographic column is prolonged; meanwhile, the pH value of a sodium dihydrogen phosphate solution is controlled to enable a mobile phase to be acidic, so that-COOH ionization is inhibited, the interaction between ion pairs and a hydrophobic stationary phase in a reversed-phase chromatographic column is improved, proper retention of 14 related substances including the cefditoren acid and the cefditoren acid delta 3 isomer is realized, meanwhile, the 14 related substances are effectively separated, and the quality of the related substances is improved. The detection accuracy and the detection efficiency of the cefditoren pivoxil related substances are improved.

The invention discloses a method for efficiently preparing high-purity chrysolic acid based on torreya grandis oil, which comprises the following steps: dissolving urea in ethanol, adding free fatty acid, and carrying out inclusion reaction to obtain preliminarily purified torreya grandis oil fatty acid; dissolving the primarily purified torreya grandis oil fatty acid in ethanol, adding Lipozyme RM C lipase, and carrying out enzymatic reaction to obtain secondarily purified torreya grandis oil fatty acid; and dissolving the secondarily purified torreya grandis oil fatty acid in a mobile phase, injecting the obtained sample solution into a reversed-phase chromatographic column of octadecylsilane chemically bonded silica, collecting fractions, merging, and carrying out rotary evaporation until the fractions are dry, thereby obtaining the high-purity aureinic acid. According to the preparation method, urea inclusion preliminary enrichment, enzymatic selective esterification secondary enrichment and preparative chromatography fine purification are combined, the preparation method of the high-purity aureinic acid is reasonable in process route, remarkable in synergistic effect and extremely high in product purity, and the defects that in the prior art, the aureinic acid purification efficiency is low, and the purity is insufficient are overcome.

The present invention relates to a polysorbate separation method, comprising a polysorbate-containing solution to be measured flowing through a mixed-mode chromatograph and a reversed-phase chromatograph which are coupled in series, the use of the separation method in measuring the polysorbate content of said polysorbate-containing solution, a related apparatus, and a method for measuring the polysorbate content by using the separation method.

The invention discloses a detection and analysis method for an alpoxivan intermediate 6 and related impurities, and relates to the field of substance detection methods, and the detection and analysis method comprises the following steps: S1, preparation of a test solution: taking an alpoxivan intermediate sample, adding a solvent into the sample, dissolving, and diluting for later use; s2, setting chromatographic conditions and a gradient elution program, wherein the chromatographic conditions comprise the use of a reversed-phase C18 chromatographic column, a mobile phase A and a mobile phase B; s3, taking the test solution, injecting the test solution into a liquid chromatograph, recording a chromatogram, and calculating the impurity content; wherein the mobile phase A comprises acetonitrile and a 9-11 mmol / L potassium dihydrogen phosphate solution, the pH value of the potassium dihydrogen phosphate solution is 7.4-7.6, and the volume ratio of the potassium dihydrogen phosphate solution to the acetonitrile is (90-100): 5; the mobile phase B comprises acetonitrile. According to the method disclosed by the invention, the aim of effectively separating and quantifying various related impurities of the alpoxivan intermediates 6 is fulfilled through optimized chromatographic conditions.

The present disclosure provides chromatography compositions and methods for separation of biomolecules, in particular, stationary phases and chromatography columns for improved separation and resolution of DAR species in antibody drug conjugates (ADCs). In one example, a method comprises: (a) exposing the sample to a chromatographic stationary phase of a reverse phase chromatography column, and (b) applying an eluent to the chromatography column to elute the DAR species of antibody-drug conjugate in their native state, wherein the chromatographic stationary phase comprises: a solid support; and a polymer coating on a surface of the solid support; wherein the polymer coating comprises a copolymer comprising a first hydrophilic monomer and a second hydrophilic monomer, wherein the first hydrophilic monomer and second hydrophilic monomer have different hydrophilicity, wherein the copolymer has a tuned hydrophilicity determined by a molar ratio of the first hydrophilic monomer to the second hydrophilic monomer.