554 results about "CMC chromatography" patented technology

The invention discloses a method for detecting the residual quantity of dextran sulfate sodium in an antibody. Specifically, the invention provides a method for detecting the residual quantity of dextran sulfate sodium in an antibody, which comprises the following steps: removing protein in the antibody, and reacting with a 2-aminopyridine solution; a high performance liquid chromatography method is adopted to detect a test sample and sodium dextran sulfate standard substances with different concentrations. The detection method disclosed by the invention is low in detection limit, high in accuracy and suitable for analyzing the residual quantity of dextran sulfate in the antibody.

The invention discloses hawthorn leaf pectic polysaccharide with lipid-lowering activity and a preparation method and application thereof, and belongs to a preparation method of plant polysaccharide, the preparation method comprises the following steps: refluxing hawthorn leaves with ethanol, discarding monosaccharide and small molecule compounds, refluxing with hot water to extract medicinal materials, removing protein through an enzymolysis method and a Sevage reagent method, decolorizing with polyamide resin, and drying to obtain the hawthorn leaf pectic polysaccharide with lipid-lowering activity. The crude polysaccharide is separated through DEAE-52 column chromatography, and finally, the crude polysaccharide is further purified through Sephadex G-100, so that the uniform purified polysaccharide is obtained. Tests on pancreatic lipase inhibition, taurocholate binding capacity and glycocholate binding rate show that the compound has remarkable lipid-lowering activity and is expected to become a potential natural lipid-lowering preparation.

The present disclosure provides systems and methods for separating and characterizing oligonucleotides. Fixed dose combinations of small interfering RNAs can be subjected to denaturing ion pairing reverse phase liquid chromatography using optimal conditions to separate single RNA strands for purity analysis.

The present application provides a method for preparing a liquid composition containing Compound I, which comprises a step of purifying a crude product containing Compound I by high performance liquid chromatography, and its use in myocardial perfusion PET imaging, wherein the mobile phase used in the purification step by high performance liquid chromatography contains ethanol and water. By optimizing the process parameters and process flow, it can be applied to large-scale batch activity production and meet the needs of automation. The resulting myocardial perfusion PET contrast agent has high radiochemical purity, high activity concentration, good product stability, and high and stable yield or output.

The invention relates to a method for detecting the encapsulation efficiency of an injection of a cytomegalovirus inhibitor. Comprising the following steps: mixing an injection of the cytomegalovirus inhibitor with diethyl ether according to a certain volume ratio, carrying out extraction separation, and carrying out quantitative analysis by using high performance liquid chromatography after separation. The detection method is simple and rapid, and can ensure the accuracy of the detection result of the encapsulation efficiency without using a complex instrument and almost without damaging the structure of the inclusion compound. Meanwhile, the method meets the requirements of accuracy and repeatability.

The invention provides a construction method and application of a fingerprint spectrum of a liver-warming decoction, and belongs to the technical field of medicine detection.The construction method includes the steps that a test solution is prepared through the liver-warming decoction, acetonitrile (A)-0.08-0.12 wt% formic acid aqueous solution (B) serves as a mobile phase, and the fingerprint spectrum of the liver-warming decoction is obtained through high performance liquid chromatography detection; and a standard fingerprint spectrum is established and is used for quality evaluation or control in the whole process of research / development / production / clinical application of the liver-warming decoction. According to the construction method of the fingerprint spectrum of the liver-warming decoction, main medicinal material components in the liver-warming decoction can be detected through high performance liquid chromatography, so that the quality condition of the liver-warming decoction can be reflected more comprehensively.

The invention provides a specific chromatogram construction method and application of polygonum-folium isatidis formula granules. The specific chromatogram construction method comprises the following steps: acquiring a specific chromatogram of a to-be-detected substance by adopting a high performance liquid chromatography; the specific chromatographic conditions of the high performance liquid chromatography are provided, octadecylsilane chemically bonded silica is used as a chromatographic column of a filler, the detection wavelength is 305-315 nm, acetonitrile is used as a mobile phase A, a 0.04%-0.06% phosphoric acid aqueous solution is used as a mobile phase B, and elution is carried out according to a specific gradient program. The specificity of the material information corresponding to the characteristic peak in the characteristic spectrum obtained by the construction method is stronger, the related information in the characteristic spectrum represented by the method is more comprehensive, and the overall quality control of the polygonum-folium isatidis can be better realized.

The invention belongs to the technical field of pharmaceutical analysis, and particularly relates to a thrombopoietin peptidomimetic peptide diagram detection and analysis method which comprises the following steps: (1) performing buffer system replacement on thrombopoietin peptidomimetic through ultrafiltration centrifugation; (2) carrying out enzymolysis on the thrombopoietin peptidomimetic by adopting trypsin; and (3) detecting and analyzing the enzymolysis peptide fragment by using a reversed-phase high-performance liquid chromatography. The method disclosed by the invention has the beneficial effects that sample concentration and buffer solution replacement are performed through the ultrafiltration tube, and the sample subjected to enzymolysis is high in purity and can be directly detected by using reversed-phase high-performance liquid chromatography, so that the method for detecting and analyzing the peptide diagram of the thrombopoietin peptidomimetic is simple and convenient to operate, and the detection efficiency of thrombopoietin peptidomimetic drugs is improved; by controlling the chromatographic conditions, the accuracy and precision of high performance liquid chromatography detection are improved, and the quality analysis and control of thrombopoietin peptidomimetic drugs can be realized.

The invention relates to a method for detecting six second-line antituberculous drugs by high performance liquid chromatography. The method comprises the following steps: providing an internal standard stock solution and a standard stock solution of six second-line antituberculosis drugs; preparing an internal standard working solution and a standard curve working solution by utilizing the diluent; preparing a solution of a product to be detected by using the Tianlongitudinal sample extraction liquid TZ-002; a liquid chromatograph is used for detection, and the conditions are as follows: a mobile phase A is a Tianlongitudinal sample releasing agent TZ-D004; a mobile phase B is acetonitrile; a mobile phase C is water; and gradient elution. According to the method, the plasma concentration of the second-line antituberculous drug taken by a tuberculosis patient is measured by applying the reversed-phase high-performance liquid chromatography, the cost is relatively low, the analysis time is short, the sensitivity is high, and the accuracy is high. The kit can be used for simultaneously detecting the blood concentration of six second-line antituberculosis drugs such as isopropylthioamide, levofloxacin, linezolid, moxifloxacin, deramanib and bedaquiline in human serum, and is expected to provide a more reliable basis for clinicians to adjust the dosage of the antituberculosis drugs.

A method for purifying a protein comprising an antibody, antibody fragment, or immunoglobulin single variable domain, from a solution containing at least one contaminant by superantigen chromatography comprising: a) adsorbing the protein to the superantigen immobilized on a solid support; b) removing the at least one contaminant by contacting the immobilized superantigen containing the adsorbed protein with a first wash buffer comprising an aliphatic carboxylate; and c) eluting the protein from the superantigen immobilized on the solid support.

The present invention belongs to the field of pharmaceutical chemical technology, and specifically relates to a method for analyzing chiral impurities of L-carnitine key intermediate L-carnitine. The method first uses (S)-2-(6-methoxy-2-naphthyl) propionyl chloride as a derivatization reagent to derive L-carnitine, and then uses high performance liquid chromatography to separate and detect the derivatized L-carnitine. The present invention proposes for the first time to use HPLC to determine the chiral purity of L-carnitine, which has simple steps and mild conditions, and is conducive to industrial promotion and application; the method of the present invention has strong specificity, high sensitivity, and good durability, and can be used for the accurate determination of the chiral purity of L-carnitine.

The invention provides an antibody screening method for flow cytometry, and belongs to the technical field of flow cytometry. Antibody purity analysis and concentration determination are carried out through high performance liquid chromatography; the binding characteristic of the antibody and the target cell is researched by adopting an equal-ratio dilution method; analyzing the combined signal by using an improved Hilbert-Huang decomposition algorithm; evaluating the combination stability through covariance matrix decomposition; collecting cell event data by using a flow cytometer; constructing a fluorescence intensity contribution matrix and performing signal optimization; calculating a signal stability contribution value to determine an effective detection area; a 0-1 knapsack optimization method is adopted to carry out multiple detection combination optimization; finally, repeatability verification is carried out through a bootstrap resampling method, the stable-performance antibody for flow detection is screened out, and the technical problems that in the prior art, the signal stability of antibody screening for flow detection is poor, and specificity is insufficient are solved.

Provided is a method for large-scale preparation of a purified preparation of a recombinant lentiviral vector at the GMP grade. The method comprises: (a) providing raw material feed liquid to be purified that comprises recombinant viral vectors; (b) carrying out a microfiltration treatment on the feed liquid to obtain a microfiltered filtrate comprising the recombinant viral vectors; (c) optionally concentrating the filtrate to obtain a concentrated filtrate; (d) purifying the filtrate obtained in the previous step by means of chromatography to obtain a crude pure product comprising the recombinant viral vectors; and (e) subjecting the crude pure product obtained in the previous step to liquid exchange and elaborate purification to obtain the purified recombinant viral vectors.

The invention provides a method for detecting the contents of collagen and regenerated silk fibroin in a composite material, which comprises the following steps: (1) hydrolyzing the composite material for a certain time, and neutralizing to obtain a solution to be detected; (2) preparing a tyrosine reference solution; and (3) detecting the peak areas of the solution to be detected and the tyrosine reference substance solution by adopting high performance liquid chromatography, and calculating to obtain the content of regenerated silk fibroin in the composite material, and (4) detecting the total nitrogen content of the composite material by using a Kjeldahl method, and calculating to obtain the collagen content in the composite material. According to the method, the high performance liquid chromatography is combined with the Kjeldahl determination method, accurate determination of two components can be realized without derivatization treatment, the operation is simple and convenient, and the detection cost is low; the limit of quantitation of the method reaches 10.35 mu g / mL (S / N is greater than or equal to 16.59), the precision RSD is less than 1%, and the method has high sensitivity and accuracy.

Provided herein are methods for purifying polypeptides (e.g., antibodies) comprising an Fc region by Protein A chromatography; methods for using wash solutions comprising benzoate and / or benzyl alcohol during Protein A chromatography; and methods for conditioning the harvest using sodium benzoate prior to Protein A chromatography. [Selected Figure] Figure 1

The present invention generally pertains to methods of characterizing crosslinking sites of a protein of interest. In particular, the present invention pertains to the use of size exclusion chromatography, peptide mapping and subunit analysis to identify and quantify crosslinking sites of a protein of interest and determine a contribution of crosslinking to the formation of high molecular weight species.

The invention discloses a method for determining related substances in a fluralamoxidectin drop, the related substances comprise an impurity A, an impurity B, an impurity C, an impurity D, an impurity G and an impurity J. The method is carried out by adopting a high performance liquid chromatography method, and the chromatographic conditions are as follows: an ammonium dihydrogen phosphate buffer solution with the pH value of 4.5-5.2 is taken as a mobile phase A, the mobile phase B is taken as a mobile phase C, and the mobile phase C is taken as a mobile phase D; acetonitrile is used as a mobile phase B for gradient elution, and the concentration of the ammonium dihydrogen phosphate buffer solution is 0.02 mol / L-0. 1 mol / L; the flow velocity is 0.7 mL / min to 1.3 mL / min; the column temperature is 40-50 DEG C; and the detection wavelength is 242 nm. According to the method, related substances in the fluralamoxidectin drops can be simultaneously determined under the same chromatographic condition, simultaneous detection and separation of various impurities are realized, the detection time is shortened, the method can be used for quality research and quality control of fluralamoxidectin drop products, and the drug safety risk is reduced.

The invention relates to the field of medicine detection, and particularly provides a method for detecting the residual content of triethanolamine by ion exchange-high performance liquid chromatography, and the chromatographic conditions in the method are as follows: a chromatographic column adopts a bonded cation exchange medium sulfonic group or carboxyl as a filler; a mobile phase is a phosphate buffer solution, and isocratic elution is carried out. According to the method disclosed by the invention, the triethanolamine does not need to be subjected to pre-column derivatization, and macromolecular protein and triethanolamine separation development (direct flow-through of macromolecular protein) is also not needed, so that the problem that the macromolecular protein is precipitated and removed to detect the triethanolamine when the gas chromatography is used is solved; and the problem that other detector modules such as a mass spectrum detector, an evaporative light detector and an electric conductivity detector need to be equipped and maintained is also solved. The method has the advantages of simple operation, accurate result, small interference and high sensitivity.

The invention discloses a method for detecting the content of nicotinamide mononucleotide, ergothioneine and nicotinamide. According to the method, the content of nicotinamide mononucleotide, ergothioneine and nicotinamide in a sample is analyzed by adopting a high performance liquid chromatography; according to the method, a CAPCELL PAK ADME chromatographic column is used, and the molecular weight of the CAPCELL PAK ADME chromatographic column is 100 And an acetonitrile-buffer solution is used as a mobile phase. Chromatographic peaks obtained by the method are sharp and symmetrical in shape, good in area reproducibility, good in separation degree, simple, rapid, high in specificity, short in detection period and high in sensitivity; the contents of nicotinamide mononucleotide, ergothioneine and nicotinamide in a sample can be rapidly, efficiently, accurately and simultaneously detected.

The invention discloses a method for determining chemical components in a medicine for treating vitiligo and a method for establishing a fingerprint spectrum. Comprising the following steps: S1, preparing a to-be-detected solution and a reference substance solution: grinding Bailing tablets, dissolving the Bailing tablets in a solvent, and extracting to prepare the to-be-detected solution; dissolving a paeoniflorin reference substance in a solvent to prepare a reference substance solution; wherein the extraction solvent is a methanol-water mixed solution; s2, respectively obtaining chromatograms of the to-be-detected solution and the reference substance solution under the same detection conditions by adopting ultra-high performance liquid chromatography, and carrying out qualitative analysis and / or quantitative analysis on chemical components in the chromatograms of the to-be-detected solution; wherein the detection conditions are as follows: the wavelength of a detector is 201 + / -2nm; the elution conditions are as follows: acetonitrile is used as a phase A, water is used as a phase B, and elution is performed through a gradient elution procedure. According to the method, the chemical components in the medicine for treating the leucoderma can be accurately determined, so that the establishment of the fingerprint spectrum database of the medicine for treating the leucoderma can be better realized.

The invention provides a method for detecting related substances of dihydroxypropyl arginine hydrochloride, which is characterized in that a high performance liquid chromatography method is adopted, a chromatographic column is a reversed-phase, cation exchange and hydrophilic interaction mixed type chromatographic column, and a mobile phase is used for gradient elution. The detection method provided by the invention can effectively separate and detect main components, starting materials and process impurities in dihydroxypropyl arginine hydrochloride, has the advantages of high sensitivity, good degree of separation, simple operation, fast appearance time, short detection time, good method reproducibility and stability, and strong specificity, meets the requirements of product quality control, and is suitable for industrial production. The method has the advantages of high sensitivity and high precision, is favorable for ensuring the safety and effectiveness of the product, reduces the production cost, and has remarkable technical advantages and application value.

The current invention reports a method for purifying an antibody by reducing the content of a host cell protein. The method employs a wash step with a low conductivity aqueous solution in an affinity chromatography.

The invention provides a method for simultaneously detecting the purity of an antibody conjugate and the coupling ratio of a drug antibody based on a size exclusion chromatography method and application of the method. The method comprises the following steps: detecting and analyzing the purity of an ADC (Analog to Digital Converter) sample to be detected and the coupling ratio of the drug antibody by adopting an SEC-UPLC (Single Emission Computation-Ultra Performance Liquid Chromatography) serially connected with an ultraviolet detector; collecting a signal of the sample under the antibody characteristic ultraviolet absorption wavelength and the small molecule characteristic ultraviolet absorption wavelength at the same time; calculating the DAR value and the monomer purity of the ADC molecule according to the chromatographic absorption peak area; a mobile phase adopted by the SEC-UPLC is a sodium chloride-phosphate buffer solution containing 0-10vol% of an organic solvent; the ADC sample to be detected is an ADC molecule coupled with Dxd type small molecules. The invention provides an analysis method which is widely applicable to determination of the DAR value and the monomer purity of the ADC molecule coupled with the Dxd small molecules. The method can simultaneously solve the problems that the DAR value detection and analysis flux of the Dxd small molecules is low and the applicability is limited.

The invention belongs to the technical field of medicine detection, and particularly relates to a method for determining the content of impurities in OAB-14 by using high performance liquid chromatography. The method comprises the following steps: preparing a test sample labeling solution, a test sample solution and a contrast solution of OAB-14, respectively injecting the test sample labeling solution, the blank solution, the contrast solution and the test sample solution into a liquid chromatograph, recording chromatograms, and calculating according to a main component self-contrast method to obtain the content of impurities in the test sample solution; wherein the impurities are one or more of an impurity A, an impurity B, an impurity C, an impurity D, an impurity E or an impurity F. According to the method, the impurities in the OAB-14 can be qualitatively and quantitatively measured at the same time, the stability and the reproducibility are good, the separation degree between a main peak and each impurity peak is good, the precision is high, and an analysis means is provided for quality control of the OAB-14.

The invention discloses a quality detection method of a gout clearing mixture for treating hyperuricemia, which adopts a separation analysis technology of ultra-high performance liquid chromatography to detect eight chemical components in the gout clearing mixture. Comprising the following components: 3-O-feruloylquinic acid, 5-O-caffeoylshikimic acid, paeoniflorin, polydatin, neoastilbin, astilbin, berberine hydrochloride and emodin-8-beta-D-glucopyranoside. According to the method, eight chemical components of six active components in the gout clearing mixture are quantitatively analyzed at the same time, and the method has the advantages of being easy and convenient to operate, good in stability and easy to popularize. The invention can provide a comprehensive and reliable quality control basis for the gout clearing mixture, and has important significance for ensuring the clinical curative effect of the gout clearing mixture.

The invention discloses a method for detecting the contents of main components of proteins and composite adjuvants in a recombinant varicella-herpes zoster vaccine and application of the recombinant varicella-herpes zoster vaccine. Comprising the following steps: preparing a protein calibration standard solution sample; preparing a QS-21 calibration standard solution sample; preparing a dioleoyl phosphatidylcholine and cholesterol calibration standard solution sample; preparing a detection sample; loading the sample into a liquid chromatograph, and operating an analysis method; and calculating the contents of main components of proteins and adjuvants in the vaccine. The method is a high performance liquid chromatography method capable of simultaneously detecting the contents of protein, dioleoyl phosphatidylcholine, cholesterol and saponin QS-21 in the vaccine. According to the method, the contents of protein, dioleoyl phosphatidylcholine, cholesterol and saponin QS-21 in the vaccine can be accurately detected only through one-time detection, and the vaccine verification efficiency is greatly improved.

The invention belongs to the technical field of medical analysis, and particularly relates to a method for separating chemical components in hemsley rockvine root. According to the present invention, various chromatography methods are adopted to carry out separation and purification, such that sitroside (I), vanillyl glycolII (II), 5-hydroxymaltol (III), vanillic acid (IV), afzelin (V), 2, 3-dihydroxypyrazole (9E, 11E)-13-hydroxypyrazole (9, 11-dienoate), and 2, 3-dihydroxypyrazole (10E, 12E)-9-hydroxypyrazole (10, 12-dienoate) are obtained from hemsley rockvine root. Wherein the compounds II, III, VI and VII are separated from the hemsley rockvine root for the first time, and development and quality control of the hemsley rockvine root medicinal material are facilitated.

The disclosed methods are directed to preparing and detecting polypeptides using neutrophil elastase, such as human neutrophil elastase. The polypeptides are optionally denatured, reduced, and / or alkylated before being subjected to a first digestion. A second digestion comprises the use of neutrophil elastase, such as human neutrophil elastase. The prepared fragments are then analyzed chromatographically, electrophoretically, or spectrometrically, or a combination of these methods. The methods are especially useful for the preparation of therapeutic polypeptides for analysis, especially those that are not easily cleaved, such as some bi-specific T-cell engager (BiTE®) molecules.

The invention belongs to the technical field of pharmaceutical analysis, and discloses a method for determining two potential genotoxic impurities in carbon [13C]-urea by gas chromatography. Comprising the following steps: (1) preparing a sample solution; (2) preparing a reference solution; (3) taking nitrogen as carrier gas, and detecting by adopting a medium-polarity or weak-polarity chromatographic column separation system; and (4) calculating by a peak area through an external standard method to obtain the accurate contents of methyl carbamate and ethyl carbamate in the carbon [13C]-urea. The method disclosed by the invention is simple to operate, good in specificity, linearity, precision and accuracy and high in sensitivity, and can realize accurate detection of residual potential genotoxic impurities, namely methyl carbamate and ethyl carbamate, in the carbon [13C]-urea.

Provided herein are fusion proteins and methods for purifying nucleic acids from a composition. The disclosed compositions and methods allow for faster, more efficient purification of nucleic acids compared to traditional affinity chromatography.