735 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 a rhizoma menispermi medicinal material and formula granule UPLC (Ultra Performance Liquid Chromatography) identification method, which is characterized in that specified values of relative retention time of the rhizoma menispermi medicinal material and formula granule are determined through a high performance liquid chromatography, and the relative retention time of a corresponding test solution is respectively within + / -10% of the corresponding specified values of the rhizoma menispermi medicinal material and formula granule. The method can effectively identify the rhizoma menispermi medicinal material and the formula granules, can be used for identifying rhizoma menispermi and counterfeit berba aristolochiae mollissimae and subprostrate sophora medicinal materials and formula granules, and can also be used for identifying rhizoma coptidis, cortex phellodendri, caulis sinomenii and other medicinal materials containing alkaloid components and formula granules. The method overcomes the defects of the existing method, and has the characteristics of simplicity in operation, good stability, rich chromatographic peak information amount, good repeatability and high stability.

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.

Methods of detecting and analyzing N-nitrosocytisine in a cytisine sample using chromatography is provided herein. N-nitrosocytisine potentially present in the cytisine sample may be detected and analyzed using mass spectrometry.

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 invention provides a SmbHLH130 gene and an application of the SmbHLH130 gene in increasing the content of tanshinone and salvianolic acid in salvia miltiorrhiza. According to the invention, a transcription factor SmbHLH130 is obtained through multi-omics data analysis in the early stage. A CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-associated 9) technology is utilized, a SmbHLH130 gene editing carrier is constructed, mutant hairy roots of gene knockout SmbHLH130 are obtained through agrobacterium tumefaciens transformation mediated by agrobacterium rhizogenes ATCC15834, and whether knockout of the mutant hairy roots is successful or not is identified through sequencing. And determining the content of tanshinone and salvianolic acid in the hairy roots of the transgenic salvia miltiorrhiza by using a high performance liquid chromatography (HPLC). The tanshinone content and the salvianolic acid content of the obtained gene knockout hairy roots are remarkably improved. Real-time fluorescent quantitative PCR (RT-qPCR) analysis shows that the expression quantity of a key enzyme gene participating in the biosynthesis of the active component of the salvia miltiorrhiza is also remarkably increased. The invention provides a method for increasing tanshinone and salvianolic acid contents of hairy roots of salvia miltiorrhiza, which can provide a new thought and target gene for production of active components of salvia miltiorrhiza and molecular design and breeding of salvia miltiorrhiza, and has higher application value and research value.

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 provides a separation and detection method of a cis-isomer in olotinib. According to the method, chiral chromatographic column high performance liquid chromatography is adopted, cellulose derivative bonded silica gel is taken as a stationary phase, and n-hexane and ethanol are taken as a mobile phase. The method provided by the invention can effectively separate and measure the cis-isomer in the olotinib, and has the advantages of high efficiency, simplicity, convenience, high sensitivity, good separation effect and separation degree of 5.42. The invention provides a detection method for developing and marketing the olatinib and accurately detecting the cis-isomer in the olatinib, and ensures the product quality.

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 present subject matter is related to identification / extraction and quantification of mass separated chromatographic peaks obtained from chromatography coupled mass spectrometry instruments This includes both liquid and gas chromatography as well as high resolution and triple quadrupole type mass analyzers. This also includes the case when these are operated in conjunction with ion mobility separation devices. The raw data generated by the instrument is first converted into an open source format such as the mzML format, obvious noise is removed by applying ad hoc filters, mass scans are centroided, regions of interest (ROI) are built in Retention Time dimension, ROIs are binned into 3 or more buckets, longer ROIs are broken into special smaller ROIs using overlapping and sliding window approach, ROIs are classified as peak or noise, peak boundaries identified if classified as peak, longer ROIs formed from special smaller ROIs, and chromatography coupled mass spectrometry data Features are quantified and reported in Retention Time dimension. The ROI classification and peak boundary detection steps make use of specially trained deep learning methods.

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 method for detecting related substances in a tandospirone citrate product, which adopts high performance liquid chromatography and can effectively detect the related substances in the tandospirone citrate product. The detection method provided by the invention has the advantages of no baseline interference, high separation degree, low detection limit, strong specificity, high sensitivity, and good accuracy, precision and durability. According to the invention, new compounds, namely a compound IV, Tand-3, Tand-4, Tand-5, Tand-base-4, Tand-base-6 and Tand-base-8, are also found in the invention. The compound can be used as a reference substance for content analysis and / or quality control of a product containing tandospirone citrate or tandospirone.

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 field of pharmaceutical analysis, and particularly relates to a method for establishing a traditional Chinese medicine fingerprint spectrum of a gastrodia elata and uncaria decoction. The method is realized by the following steps: firstly, preparing a reference substance solution and a test solution, then detecting by adopting a high performance liquid chromatography, and establishing a fingerprint spectrum. According to the method provided by the invention, by reasonably selecting the components of the reference substance and adopting a multi-wavelength method according to the maximum absorption wavelength of the components, the change of the component amount between the samples can be effectively represented, so that the quality of the samples is more accurately judged; the invention further provides a screening and optimizing method for the preparation method of the gastrodia elata and uncaria decoction, the prepared method is good in repeatability and stability, the quality of the gastrodia elata and uncaria decoction can be monitored more comprehensively, and the method provides valuable reference for quality evaluation and standardization of traditional Chinese medicine decoction.

The invention discloses a method for detecting rafenasin related substances, which comprises the following steps: detecting by adopting a high performance liquid chromatography under the chromatographic conditions that a stationary phase is an octadecyl silane bonded silica gel chromatographic column, a mobile phase A is a phosphate buffer aqueous solution, a mobile phase B is methanol, and gradient elution is carried out; the gradient elution procedure is as follows: within 0-10 minutes, the volume fraction of the mobile phase A is gradually changed from 88-92% to 73-77%; within 10-43 min, the volume fraction of the mobile phase A is gradually changed from 73-77% to 38-42%; within 43-45 min, the volume fraction of the mobile phase A is gradually changed from 38-42% to 88-92%; within 45-50 min, the volume fraction of the mobile phase A is kept at 88-92%. According to the chromatographic conditions disclosed by the invention, effective separation between the main peak of the rafenasin and each impurity can be realized, the specificity is good, and the sensitivity is high.

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.

The invention discloses a chiral analysis method of a series of semeglutide intermediates, the analysis method is used for rapidly determining the chiral purity of a series of semeglutide intermediates, a high performance liquid chromatography (HPLC) is combined with a chiral column, and chromatographic conditions and elution procedures are optimized, so that the analysis method is simple and convenient to operate, rapid, high in universality, high in sensitivity, high in sensitivity and high in accuracy. According to the method, the chromatographic conditions and the elution procedure are optimized, the chiral separation effect and resolution are improved, efficient and accurate chiral separation and analysis of the semeglutide series intermediates shown in the formula 1 are achieved, and the method is suitable for chiral quality control of the semeglutide series intermediates shown in the formula 1.

The present invention provides a detection kit for detecting 25-hydroxyvitamin D and a detection method thereof. Formulations of a dissociating agent and an antibody dry powder are known through screening and optimization, such that self-mixing and dissociation of a sample are realized without being affected by human factors and depending on any mixing equipment, ensuring a 25-hydroxyvitamin D antibody is fully reacted with a substance to be tested. In addition, an operating process of an immunochromatography assay is improved, such that dissociation of the sample and bonding of the 25-hydroxyvitamin D antibody and the substance to be tested are completed in one reaction tube and completely separated from a test strip; and the test strip is employed for sample injection and detection after completion of reaction, and a specific control line is arranged. This significantly improves the accuracy and sensitivity of portable detection of 25-hydroxyvitamin D with the immunochromatography assay, and the detection sensitivity of the 25-hydroxyvitamin D reaches 1 ng / mL.

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 relates to the field of analysis, in particular to a separation method for mixing different oligonucleotide sequences. The method for separating oligonucleotides mixed with different sequences comprises the step of separating and detecting samples mixed with different oligonucleotides by adopting a high performance liquid chromatography, and the chromatographic conditions are as follows: octadecylsilane chemically bonded silica is used as a filler, and an HPLC A liquid is used as a mobile phase A; a mixed solution of an HPLC (High Performance Liquid Chromatography) A solution and acetonitrile is used as a mobile phase B, and the volume ratio of the HPLC A solution to the acetonitrile is (60-80): (20-40); the HPLC A liquid is 0.1 M TEAA (Tetraethyl Acrylic Acid) Performing gradient elution; according to the method, oligonucleotides with different sequences are separated and mixed through the selected chromatographic conditions, N-X impurities and N + X impurities can be effectively removed, especially N-X impurities and N + X impurities with X smaller than or equal to 2, the finally obtained oligonucleotide medicine is high in purity, and meanwhile the high yield is kept.

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.