20 results about "Hydrophilic interaction chromatography" patented technology

The invention discloses a method for rapidly determining the content of breast milk oligosaccharide in dairy products, which comprises the following steps: sampling and pretreating different types of dairy products according to sample states, carrying out ultrasonic extraction, refrigerated centrifugal degreasing and centrifugal ultrafiltration protein purification on the obtained sample, and reducing the purified sample solution by using sodium borohydride to obtain a sample solution to be detected; reducing the breast milk oligosaccharide standard substance by sodium borohydride to obtain a standard solution to be detected; determining the standard substance solution and the to-be-detected sample solution by adopting ultra-high performance liquid chromatography-tandem mass spectrometry, establishing a standard curve, and carrying out quantitative analysis on the to-be-detected sample solution; the chromatographic conditions are as follows: an ACQUITY UPLC BEH Amide hydrophilic interaction chromatographic column is adopted; the mobile phase A is 20 mmol / L ammonium formate; the mobile phase B is acetonitrile; and gradient elution.

The invention belongs to the technical field of detection and analysis, and provides a method for detecting the average coupling rate of a high-hydrophilicity small-molecule high-glycosylation antibody coupling drug and application of the method for detecting the average coupling rate of the high-hydrophilicity small-molecule high-glycosylation antibody coupling drug. The method comprises the following steps: firstly, carrying out desugaring treatment on a high-glycosylation coupling medicine, then coupling desugarized high-hydrophilicity toxin molecules with an antibody medicine, analyzing by using hydrophilic interaction chromatography conditions, and calculating to obtain the accurate average coupling rate of the antibody coupling medicine. According to the method, through desugaring treatment and hydrophilic interaction chromatography detection in the analysis process, the defect that the separation degree of high-hydrophilicity micromolecules with different coupling ratios is low due to hydrophobicity difference in a traditional method is overcome, so that the measured average coupling rate result is more accurate and real. The method is suitable for coupling a high-hydrophilicity payload drug with a high-glycosylation antibody drug, and is especially suitable for an antibody coupling drug which is high in micromolecule hydrophilicity and has obvious influence on the separation degree of a traditional DAR liquid phase analysis method.

The invention discloses a stationary phase of a novel zwitterionic monomer containing a multi-charge structure as well as a preparation method and application of the stationary phase, and belongs to the technical field of hydrophilic interaction chromatography (HILIC) stationary phases. According to the method, multi-charge zwitterions are used as hydrophilic monomers, the multi-charge structure of the multi-charge zwitterions endows the monomers with higher hydrophilic performance, and the multi-charge zwitterions are immobilized on the surface of a matrix by utilizing an active free radical polymerization method, so that a series of novel super-hydrophilic HILIC stationary phases are finally prepared. The multi-charge zwitterionic monomer adopted by the invention not only has better hydrophilic performance and higher charge density, but also can efficiently, simply and conveniently realize matrix surface grafting and modification. The developed stationary phase is suitable for HILIC column chromatography separation analysis, and the HILIC chromatographic performance can be greatly improved; the method can also be applied to the fields of biological sample pretreatment, material science, marker clinical diagnosis and the like, and has high economic and social values.

The invention discloses a method for detecting natural plant hormones and exogenous growth regulators of vegetables, efficient separation and accurate detection of the natural plant hormones and the exogenous growth regulators are realized through combination of hydrophilic interaction chromatography and mass spectrometry, a hydrophilic interaction chromatographic column is adopted, and conditions such as mobile phase composition and flow velocity are optimized; the selectivity of polar substances is improved by utilizing hydrophilic interaction, so that compounds with similar structures achieve baseline separation, and chromatographic peak overlapping interference is avoided; the mass spectrometric detection only responds to a specific ion signal by screening characteristic parent ion / daughter ion pairs of a target object, so that matrix impurities and non-target ion interference are effectively eliminated, the false positive risk is remarkably reduced, and the detection sensitivity and specificity are improved; the combined technology is combined with the efficient separation capacity of the hydrophilic interaction chromatographic column and a multi-reaction monitoring mode, synchronous separation of multiple compounds with similar polarities in single analysis is achieved, parallel detection of dozens of target objects is supported, and the analysis time is greatly shortened.

The present application relates to a kind of monodisperse core-shell type hybrid silica microspheres containing vinyl shell and its preparation and application. Specifically, using "two-phase method" strategy, monodisperse solid silica microspheres are used as matrix, vinyl triethoxysilane and tetraethyl orthosilicate are used as raw materials, lauryl amide propyl betaine and sodium subtiliporin are used as double-template pore-forming agent, under the catalysis of urea, sol-gel reaction occurs, a layer of porous hybrid shell containing vinyl functional group is constructed on the surface of solid silica microspheres, and core-shell type silica hybrid microspheres are formed. Because the shell contains a large number of vinyl functional groups, after hydrophilic modification with captopril, it is used as the adsorbent of hydrophilic interaction chromatography, and is applied to the selective enrichment of biological samples. The core-shell type silica hybrid microspheres prepared in the present application have uniform morphology, good monodispersity, and rich vinyl functional groups on the surface, which are easy to modify and apply subsequently.

The invention relates to a method for quantitatively detecting the content of sucrose in an mRNA (messenger ribonucleic acid) composition, in particular to a UPLC-CAD (Ultra Performance Liquid Chromatography-Computer Aided Design) sucrose residue detection method operated in an HILIC (Hydrophilic Interaction Chromatography) mode. According to the method, liquid chromatography (HILIC) suitable for hydrophilic interaction is adopted, a polyvinyl alcohol silica gel column and an electrospray detector (CAD) are selected, and acetonitrile-water is used as a mobile phase. The method is accurate, reliable, good in chromatographic separation degree and high in sensitivity, and the content of sucrose in the mRNA solution can be accurately measured in the mRNA composition.

The invention relates to the technical field of tobacco chemical analysis, in particular to a multistage mass spectrometry identification method for chemical structures of sucrose esters in tobacco and tobacco products, which comprises the following steps: crushing and extracting a tobacco product sample; carrying out solid-liquid separation and filtration to obtain a liquid to be detected; separating the liquid to be detected by adopting hydrophilic interaction chromatography; performing triple quadrupole-ion trap tandem mass spectrometry analysis on the components separated in the step S2 by adopting an electrospray ion source in a positive ion mode to sequentially obtain mass spectrometry information of parent ions, daughter ions and grandchild ions; and determining the chemical structure of the sucrose ester based on the mass spectrum information of the parent ion, the daughter ion and the grandchild ion measured in the step S3. According to the method, complex pretreatment steps are not needed, sucrose esters with different configurations can be identified more efficiently, interference can be effectively eliminated, and substituent group structure information of a low-content target object can be scientifically confirmed.

The invention relates to a method for quantitatively detecting ion residues in an mRNA (messenger Ribonucleic Acid) composition, in particular to a method for detecting metal ions by UPLC-CAD (Ultra Performance Liquid Chromatography-Computer Aided Design) operated in a HILIC (Hydrophilic Interaction Chromatography) mode. According to the method, liquid chromatography (HILIC) suitable for hydrophilic interaction is adopted, an unbonded silica gel column and an electrospray detector (CAD) are selected, and acetonitrile-water is used as a mobile phase. The method is accurate, reliable, good in chromatographic separation degree and high in sensitivity, and can simultaneously realize limit separation of four targets of sodium, potassium, magnesium and calcium in the mRNA solution.

The invention belongs to the technical field of food-borne functional factors, and particularly relates to a bioactive glycopeptide as well as a preparation method and application thereof. According to the application, casein glycomacropeptide is taken as a substrate, specific protease is adopted for carrying out directional enzymolysis reaction on the casein glycomacropeptide, and after primary separation and purification by hydrophilic interaction chromatography and secondary purification by cation exchange chromatography are carried out, the bioactive glycopeptide with the molecular weight of less than 1,500 Da and more than 80% of components and the random coil secondary structure content of more than 35% is obtained. A system performance test shows that in an in-vitro cell experiment, the bioactive glycopeptide prepared by the embodiment of the invention can remarkably improve the proliferation rate of preosteoblasts and lay a sufficient cell quantity foundation for bone cell generation; meanwhile, the activity of an osteogenic early differentiation key marker ALP is effectively up-regulated, and the differentiation process of preosteoblasts towards the osteoblast direction is accelerated; in addition, the expression quantity of osteogenesis-related regulatory protein calcium-sensitive receptors is remarkably increased, and the function improvement of cells to mature bone cells is further promoted.

This invention discloses a method for detecting γ-aminobutyric acid (GABA) and γ-hydroxybutyric acid (GHB) using liquid chromatography-tandem mass spectrometry (LC-MS / MS). Ammonium bicarbonate is added to both mobile phase A and mobile phase B, and ammonia is also added to mobile phase A. The chromatographic column is a zwitterionic column, and hydrophilic interaction chromatography mode is used. The detection method provided by this invention has high sensitivity, raising the limits of quantification for GABA and GHB to 20 nM. The retention and peak shape of GABA and GHB in the chromatogram are significantly enhanced, and the analysis speed is significantly improved. Furthermore, the use of PBS buffer as an alternative matrix expands the range of matrix applications for in vitro drug screening.

The application discloses a preparation method of spermine-bonded silica gel hydrophilic chromatographic stationary phase and application thereof, and belongs to the technical field of hydrophilic chromatography. The preparation steps are as follows: first, epoxy propyl silane modification is carried out on spherical silica gel to prepare epoxy propyl functionalized silica gel; then, spermine is uniformly mixed with the epoxy propyl functionalized silica gel in an organic solvent to carry out reaction, so that spermine-bonded silica gel chromatographic stationary phase Sil-SPM is prepared. It is proved by infrared spectrum and element analysis characterization that spermine has been successfully bonded to the surface of the silica gel. Chromatographic separation experiments show that the hydrophilic chromatographic stationary phase Sil-SPM prepared by the application has good hydrophilic chromatographic selectivity, and has excellent separation selectivity for base nucleosides. Meanwhile, the stationary phase can also be applied to the determination of the contents of uric acid and creatine in urine, and has potential application value in the separation of hydrophilic biological samples.

The present invention relates to a kind of amino-modified reduced graphene oxide and its preparation method and application, by first modifying the amino linker on the surface of reduced graphene oxide (rGO) to obtain a first product, then removing the Fmoc protecting group from the first product, obtaining the amino-modified reduced graphene oxide, having an ultra-large specific surface area and a flexible structure, which can increase the solid loading of N-sugar chains and reduce the contact steric hindrance between it and the enriched N-sugar chains, therefore, the amino-modified reduced graphene oxide of the present invention is combined with MALDI-TOF MS to realize the specific enrichment and identification of protein N-glycopeptides or N-sugar chains in complex biological systems. The amino-modified reduced graphene oxide material of the present invention is rich in N groups and has excellent hydrophilic properties. And the material can be directly combined with a MALDI matrix and a MALDI laser source to realize the effective release and identification of the enriched N-sugar chains. Compared with other protein N-glycan enrichment methods in the prior art, the material enrichment method based on hydrophilic interaction chromatography (HILIC) has obvious advantages, such as high biocompatibility, simple operation, short time consumption, high recovery rate, and can achieve efficient capture and purification of N-glycans in biological samples.

Described herein are methods of characterizing free thiol species in a protein or peptide. For example, described herein are methods of using hydrophilic interaction chromatography (HILIC) in combination with one or more mass spectrometers to characterize free thiol species in a protein or peptide. Also described herein are methods for characterizing free thiol species in an antibody, such as a therapeutic antibody.

Provided herein are methods of determining the size and purify of nucleic acids (e.g., mRNAs) by using hydrophilic interaction chromatography (HILIC)-based methods to separate the nucleic acids from a mixture, followed by mass spectrometry to determine the size of the nucleic acids.

The invention relates to the technical field of traditional Chinese medicine quality analysis and detection, in particular to a construction method and application of a trichosanthes kirilowii maxim polysaccharide fingerprint spectrum. According to the method, trichosanthes kirilowii maxim polysaccharide is obtained in an ultrasonic extraction and alcohol precipitation mode, then the trichosanthes kirilowii maxim polysaccharide is depolymerized through an ultrasonic-assisted acidolysis method, trichosanthes kirilowii maxim polysaccharide depolymerized matter is subjected to hydrophilic interaction chromatography-electrospray detector-electrospray-quadrupole-time-of-flight mass spectrometry (HILIC-CAD / ESI-Q-TOF / MS) analysis, and 16 chemical components are obtained in total. 16 chemical components are used as common peaks to establish the trichosanthes kirilowii maxim polysaccharide fingerprint spectrum, and a method and technical support are provided for trichosanthes kirilowii maxim polysaccharide quality evaluation and control.

The invention relates to poly-amide bonded hydrophilic interaction chromatography (HILIC) stationary phases and novel HILIC methods for use in the characterization of large biological molecules modified with polar groups, known to those skilled in the art as glycans. The invention particularly provides novel, poly-amide bonded materials designed for efficient separation of large biomolecules, e.g. materials having a large percentage of larger pores (i.e. wide pores). Furthermore, the invention advantageously provides novel HILIC methods that can be used in combination with the stationary phase materials described herein to effectively separate protein and peptide glycoforms by eliminating previously unsolved problems, such as on-column aggregation of protein samples, low sensitivity of chromatographic detection of the glycan moieties, and low resolution of peaks due to restricted pore diffusion and long intra / inter-particle diffusion distances.

The invention relates to the technical field of pharmaceutical analysis, in particular to a method for detecting purity of amoxicillin sodium and clavulanate potassium for injection. The method mainly solves the technical problems of serious system pollution, poor method stability and incapability of effectively separating and identifying unknown degradation impurities due to the fact that ion pair reagents are required to be used in the detection process of the conventional reversed-phase chromatography method. A hydrophilic interaction chromatography-high resolution mass spectrometry coupling technology is adopted, and an amide chromatographic column separation condition and mass spectrometry detection parameters are optimized, so that a detection method without an ion pair reagent is established. The method can simultaneously realize accurate quantification of main components of amoxicillin sodium and clavulanate potassium, and qualitative identification and quantitative analysis of related unknown impurities. The method has the advantages of high sensitivity, strong specificity and good reproducibility, effectively avoids the use of an ion pair reagent, and provides a reliable technical means for purity control and product quality evaluation of amoxicillin sodium and clavulanate potassium for injection.

Systems and methods for characterizing low molecular weight (LMW) protein drug product impurities are provided. One embodiment uses hydrophilic interaction chromatography (HILIC) coupled to mass spectrometry analysis. After removal of the N-linked glycans from the protein drug product, for example an antibody drug product, the elution of LMW impurities from the HILIC column was determined by the size of the molecular weight species. In some embodiments, the HILIC separation is performed under denaturing conditions, making the detection of LMW forms using this method highly comparable to both SDS-PAGE and CE-SDS methods. LMW drug product impurities include, but are not limited to light chain, half antibody, H2L, H2, HL, HC, peptide backbone-truncated species, and combinations thereof.

The present invention discloses a captopril-modified hollow nanosilica sphere and its preparation method and application, belonging to the technical field of nanomaterials. Among them, the preparation method of the captopril-modified hollow nanosilica sphere is as follows: using water as a solvent, F127 as a template agent, mesitylene as a micelle swelling agent, potassium sulfate as an auxiliary agent, and tetramethoxysilane and vinyltrimethoxysilane as silicon-based precursors, preparing vinyl-modified hollow nanosilica spheres through a sol-gel reaction; then, further modifying captopril on the hollow nanosilica spheres through a thiol-ene photoinitiated reaction. The beneficial effects of the present invention are as follows: The captopril-modified hollow nanosilica sphere is the first hydrophilic interaction chromatography stationary phase material prepared with captopril as a modifier monomer, having uniform particle size, good dispersibility, not easy to agglomerate, large specific surface area, and having good enrichment specificity and selectivity for N-glycopeptides, and can be used for the selective enrichment of N-glycopeptides in complex biological samples.

The present invention discloses a method for enriching O-GlcNAc glycopeptides based on reversible chemical enzymatic labeling-hydrophilic interaction chromatography enrichment. Specifically, it relates to: chemical enzymatic labeling of O-GlcNAc glycopeptides based on the transglycosylation activity of mutants of endoglycosidases such as Endo-M, transferring the N-sugar chain with an oxazoline structure as a whole to the O-GlcNAc glycosyl, and then using hydrophilic interaction chromatography to enrich the labeled O-GlcNAc glycopeptides in one step. For the enriched O-GlcNAc glycopeptides with introduced N-sugar chain structure, the labeled sugar chain is removed by wild-type endoglycosidase to achieve traceless enrichment of O-GlcNAc glycopeptides. Compared with other O-GlcNAc glycopeptide enrichment technologies, the present invention has the advantages of high O-GlcNAc glycopeptide enrichment efficiency and simple operation, and provides a new solution and enrichment method for the study of O-GlcNAc glycoproteomics.