110 results about "Hydrophilic interaction chromatography" patented technology

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 present invention provides an agglomerated multimodal chromatographic medium. the medium of the invention includes groups active in anion exchange, cation exchange and hydrophilic interaction chromatographic modalities. The invention provides methods of making these media and using them in separations of analytes. Also provided are separations devices incorporating the medium and systems incorporating these separations devices.

The invention discloses a hydrophilic interaction chromatography-tandem mass spectrometry detection method of phospholipids in Metapenaeus ensis. The method includes the steps of firstly, performing corresponding preprocessing on ground Metapenaeus ensis to obtain crude lipid extract; secondly, performing liquid phase separation on the crude lipid extract, wherein a chromatographic column is a YMC Triart diol HILIC column, a gradient elution method is used, a gradient system comprises a flowing phase A and a flowing phase B, the flowing phase A is an acetonitrile solution containing 53mmol/L formic acid, and the flowing phase B is an aqueous solution containing 60mM ammonium formate and 53mM formic acid; thirdly, performing mass spectrometric detection analysis on the elution liquid obtained in the second step. By the method, the total content of PE, PS and PC in the Metapenaeus ensis can be detected accurately, and mass spectrometry identification of various molecular species in the PC, PE and PS phospholipids can be achieved.

An exemplary multimodal chromatographic medium of the invention includes one or more strong anion exchange, weak anion exchange, strong cation exchange and/or weak cation exchange binding sites in combination with one or more reverse phase and/or hydrophilic interaction chromatography binding site. In an exemplary embodiment, the sites interact with one or more glycans in a mixture of glycans in a manner that allows separation of glycans in the mixture and analysis of the glycan mixture. The media are incorporated into devices and systems for chromatographic analysis. Also provided are methods of using the multimodal media of the invention to analyze glycans.

The invention discloses a honey adulteration detection method based on HPLC-ELSD and a partial least squares discriminant analysis method, and belongs to the technical field of food detection. The method comprises simple sample pretreatment, wherein the simple sample pretreatment comprises the steps of adopting a hydrophilic interaction chromatography separation-evaporation light scattering detector (HILIC-ELSD) method to analyze oligosaccharide and isomer components of oligosaccharide in authentic honey and a honey sample adulterated with fructose corn syrup or malt syrup, and utilizing difference of oligosaccharide component fingerprint spectrums of the authentic honey and the adulterated honey to conduct data processing through the partial least squares discriminant analysis method, so that adulteration identification is further achieved. The honey adulteration detection method based on the HPLC-ELSD and the partial least squares discriminant analysis method can effectively identify adulterated honey with C4 botanical syrup or C3 botanical syrup, the adopted analysis instrument is popular, and the method is convenient for popularization and application.

The present invention relates to an identification method for an O-glycosylation peptide fragment and a complete saccharide chain thereof. The technical process specifically comprises that a glycosylation protein sample is subjected to PNGase F digestion to remove N-glycosylation modification interference, enrichment is performed through hydrophilic interaction chromatography, and finally analysis is performed through time of flight mass spectrometer. According to the present invention, the method process has advantages of simple and rapid operation, high identification sensitivity, and the like; and the O-glycosylation peptide fragment and the complete saccharide chain thereof of the complex biological sample can be identified so as to achieve the research on the micro-uniformity of the O-glycosylation modification structure.

The invention provides a core-shell type hydrophilic chromatographic stationary phase with a metal organic framework material as the shell, a preparation method and application thereof. The stationary phase takes the metal organic framework prepared with an imidazole ionic liquid as the organic ligand to modify spherical silica gel particles, has wide application range, and can realize hydrophilic interaction chromatography separation analysis of most polar compounds. At the same time, the preparation method of the core-shell type hydrophilic interaction chromatography stationary phase provided by the invention has the advantages of simple preparation process, mild reaction conditions, good preparation reproducibility, and easy popularization and use.

The object of the present invention is to provide a packing material and a separation method that manifest excellent hydrophilic interactions.

The present invention provides a packing material for hydrophilic interaction chromatography consisting of a modified support treated with the surface modifier represented by the following formula (6) or (7).

(In this formula, m denotes 2-6 and n denotes 1-4. X₁, X₂, and X₃, independent of each other, denote a methoxy group, ethoxy group, or halogen. Up to two of X₁, X₂, and X₃ can be any of the following groups: a methyl group, ethyl group, propyl group, isopropyl group, butyl group, or isobutyl group.)

The invention discloses a method for purifying and separating oligosaccharides. A polar chromatographic column is prepared by taking a polar filler as a chromatographic stationary phase in the chromatographic mode of a hydrophilic interaction chromatography principle, wherein water, acetonitrile and a buffer salt serve as a mobile phase; water serves as a strong elution solvent; and an isocratic or gradient elution condition is used. In the method, parameters such as the type and concentration of the buffer salt, the gradient of the mobile phase, column temperature and the like are optimized, so that separation and preparation of acid, neutral and alkaline oligosaccharides are realized. The method has the characteristics of high stability, large loading amount, easiness, convenience and controllability in operating and the like, and is suitable for separating and purifying different types of oligosaccharides.

A method for extracting daptomycin from fermentation broth relates to fermentation broth. Supernatant liquor is obtained after the fermentation broth is centrifuged, the daptomycin enters an organic phase after the supernatant liquor is extracted by an organic solvent, sodium acetate-acetic acid buffer solution is used to perform re-extraction to reach a water phase so as to obtain strip-extraction liquid, the strip-extraction liquid is purified through hydrophobic interaction chromatography, isopropanol-sodium acetate-acetic acid buffer solution with constant intensity is used for elution to obtain hydrophobic interaction chromatography eluent, the eluent is separated and purified through ion-exchange column chromatography to obtain ion-exchange eluent, the sodium acetate-acetic acid buffer solution is used as basic liquid, and neutral salt is added for elution so as to complete extraction of the daptomycin from the fermentation broth. Organic solvent extraction and resin chromatography technology is applied comprehensively, and the daptomycin with purity more than 85% can be separated and purified from the fermentation broth by using the method. Compared with other separating and purifying technology, the method is moderate in operating condition, low in cost and capable of effectively reducing acidolysis of the daptomycin. The daptomycin serving as novel lipopeptide antibiotics has good medical market prospect.

The invention relates to a silica gel matrix high-performance liquid chromatography separation material. An electron deficient alkene monomer and sulfydryl silica gel are subjected to sulfydryl Michael addition to form a hydrophilic interaction chromatography stationary phase, a reversed phase chromatography stationary phase and an affinity chromatography stationary phase, and the structural formula is as shown in the description, wherein SiO2 is mesoporous silica gel or non-porous silica gel, R-X represents one of formulas as shown in the description, and X represents one of a polar functional group, a non-polar functional group or an affinity functional group. The chromatography separation material provided by the invention has a novel structure, contains a characteristic thioether functional group, also contains an amide, ester, sulfone or succinimide functional group, and can serve as a stationary phase of reversed phase chromatography, hydrophilic interaction chromatography or affinity chromatography to provide different action forces and improve the separation selectivity of the chromatography material. A preparation method provided by the invention is simple and efficient, and the obtained stationary phase has the advantages of large bonding amount, high stability and the like.

The invention relates to a rapid detection method for melamine and sulbactam in liquid milk, and belongs to the technical field of liquid milk analysis. In the rapid detection method, a sample is extracted through an acetic acid water solution, subjected to protein sedimentation through acetonitrile and diluted, Acquity UPLC BEH HILIC(100*2.1mm, i.d., particle diameter of 1.7 micrometers) separation is carried out, and electrospray ion source multi-reaction monitoring (MRM) mode tandem mass spectrum measurement is carried out. An extracting solvent, extracting time, purifying time, instrument measurement conditions and the like are optimized. Results prove that melamine and sulbactam have the good linear relation within the range of 0.02-20 ng/mL and 0.05-100 ng/mL respectively, and the correlation coefficient is larger than 0.99. In the high, middle and low addition levels, the recovery rate of practical samples is between 82.5% and 103.5%, the relative standard deviation (RSD n=6) is between 0.9% and 5.1%, and the method quantification limit of melamine and sulbactam is 10 micrograms per kilogram and 1.0 microgram per kilogram respectively. Pretreatment operation is simple and rapid, hydrophilic interaction chromatography-tandem mass spectrometry detection sensitivity is high, and qualitative and quantitative operation is accurate.

In one aspect, a method of performing electrostatic repulsion-hydrophilic interaction chromatography on a protein, peptide, or amino acid includes providing a column having an anion-exchange material at a pH of less than about 4, and eluting the compound using a mobile phase comprising an amount of organic solvent sufficient to substantially balance the electrostatic repulsion of the stationary phase with hydrophilic interaction. In another aspect, a method of performing electrostatic repulsion-hydrophilic interaction chromatography on a nucleic acid or nucleotide comprises providing a column having a cation-exchange material at a pH of less than about 3.4, and eluting the compound using a mobile phase comprising organic solvent sufficient to substantially balance the electrostatic repulsion of the stationary phase with hydrophilic interaction.

The invention discloses a method for detecting the content of L-hydroxyproline in a dairy product. The method comprises the following steps of: 1) pre-treating a sample, namely adding acid into the diary product and hydrolyzing to obtain test liquid; 2) analyzing the content of L-hydroxyproline by liquid chromatogram-tandem mass spectrometry, wherein hydrophilic interaction chromatography (HILIC) is adopted, a flowing phase is a mixed solution a flowing phase A and a flowing phase B, and the flowing phase A is a 10mM ammonium acetate aqueous solution of which the pH is 5.6; and the flowing phase B is acetonitrile, and the mass spectrometry condition is that: an electrospray ion source is adopted, a scanning mode is a positive ion scanning mode, a detection mode is a multi-reaction detection mode, electrospray voltage is 5,500V, the temperature of the ion source is 600 DEG C, the pressure force of a gas curtain is 20Psi, the atomization pressure force is 50Psi, and auxiliary pressure force is 30Psi. By the method, the detection accuracy and sensitivity can be improved obviously, and the content of the L-hydroxyproline in the dairy product can be determined easily, conveniently and quickly.

The invention relates to a high pressure liquid chromatography detection method for ergothioneine. According to the invention, an hydrophilic interaction chromatography (HILIC) analysis chromatographic column is employed for performing a high pressure liquid chromatography analysis on a sample containing ergothioneine, the chromatographic column employs a HILIC filler as a fixed phase, a solution of acetonitrile and water (v/v) with ratio of (80-95): (20-5) is taken as a mobile phase, the pH value of the solution is 5.6-8.0, and the column temperature is 25-45 DEG C. the invention also relates to a quantitative determination method of ergothioneine.

The invention discloses a hydrophilic interaction chromatographic stationary phase based on amide functional imidazole ionic liquid and a preparation method and application of the hydrophilic interaction chromatographic stationary phase, and belongs to the field of high performance liquid chromatography stationary phases. Ionic liquid is bonded to a 3-sulfydryl propyl modified silicon dioxide ball surface to prepare the hydrophilic interaction chromatographic stationary phase through a sulfydryl-double bond clicking chemical reaction. The preparation method comprises the steps that firstly, the amide functional imidazole ionic liquid of terminal alkenyl is prepared firstly, the silicon dioxide ball surface is grafted with 3-sulfydryl propyl groups through a silanization reaction, and the ionic liquid is bonded to the silicon ball surface modified by 3-sulfydryl propyl. The surface bonded amide imidazole ionic liquid serves as the hydrophilic interaction chromatography stationary phase to show good reservation and selectivity on typical hydrophilic compounds, and the column efficiency amounts to that of a commercial amide hydrophilic interaction chromatography stationary phase. In addition, due to the unique multiple interactions of the hydrophilic interaction chromatographic stationary phase, the stationary phase has especially good reservation and separation selectivity on flavonoid matter under the hydrophilic chromatography mode, and good application prospects are achieved.

The invention relates to the technical field of liquid chromatography, in particular to an online hydrophilic interaction chromatography/reversed phase chromatography serial interface device. The online hydrophilic interaction chromatography/reversed phase chromatography serial interface device comprises a high-pressure inert gas source and relevant pressure reduction and valve switching devices. Specifically, one end of a first-dimensional chromatographic column is connected with an inert gas (such as nitrogen) pressure reducing valve and a pump system through a switching valve and pipelines, and the other end of the first-dimensional chromatographic column is connected with a second-dimensional chromatographic column and waste liquid through a switching valve and pipelines; by switching the valves, the high-pressure inert gas replaces a solvent left in the first-dimensional chromatographic column, the problem of incompatibility of solvents in online series connection is solved, reservation of samples on the second-dimensional chromatographic column is guaranteed, and therefore hydrophilic interaction chromatography and reversed phase chromatography are connected in series. An interface is convenient to use, simple in operation, capable of saving time, efficient and high in universality; due to the fact that the hydrophilic interaction chromatography and the reversed phase chromatography are directly connected in series, the samples can be online, quickly and continuously operated, and losses of the samples are reduced.

The invention relates to high-performance liquid chromatography stationary phases, in particular to a hydrophilic interaction chromatography/ ion exchange chromatography mixed stationary phase and preparation and application of the hydrophilic interaction chromatography/ ion exchange chromatography mixed stationary phase. The structure of the hydrophilic interaction chromatography/ ion exchange chromatography mixed stationary phase is shown as the formula 1, and please see the formula 1 in the specifications, wherein X is -OCH3 or -OCH2CH3, Y1 is F or Cl or Br or I or BF4 or Tf2N or CF3SO3 or SCN or NO2 or NO3, Y2 is -NHCH3 or -N (CH3)2 or -N+ (CH3)3Y1 or -NHCH2CH3 or -N (CH2CH3)2 or -N+ (CH2CH3)3Y1, and n=2 or 3. According to the prepared mixed type stationary phase, raw materials are easy to obtain and the preparation procedure are simple; and the prepared mixed type stationary phase has good retaining performance and separation selectivity for multiple saccharides and has good application and popularization prospects.

The invention discloses a laurinol polyoxyethylene ether fully two-dimensional separation analysis method including the following steps: an ultrahigh pressure hydrophilic interaction chromatography-ion mobility mass spectrometry coupled method is adopted, a laurinol polyoxyethylene ether fully two-dimensional separation analysis system is constructed, and effective separation and accurate determination of a complex target compound system are realized; the method includes optimization of ultrahigh hydrophilic interaction chromatography conditions and ion mobility mass spectrometry conditions. According to the laurinol polyoxyethylene ether fully two-dimensional separation analysis method, through the ultrahigh pressure hydrophilic interaction chromatography-ion mobility mass spectrometry coupling technique, the laurinol polyoxyethylene ether fully two-dimensional separation analysis method is developed, and rapid separation detection of laurinol polyoxyethylene ether is realized.

The invention provides a detection method for sialic acid in a proteinic drug. Particularly, the invention provides a method for detecting the content of the sialic acid in protein. The method comprises the following steps: (1) providing a sample to be analyzed, wherein the sample contains the protein to be analyzed; (2) performing acid hydrolysis on the sample in the step (1), thus obtaining an acid hydrolysis product; (3) labeling the acid hydrolysis product obtained in the step (2) with a labeling reagent, thus obtaining a labeled mixture; (4) performing chromatographic pretreatment on the labeled mixture through an organic solvent, thus obtaining a pretreated mixture; (5) loading the pretreated mixture to a hydrophilic interaction chromatographic column for chromatographic separation, thus obtaining labeled sialic acid-containing eluent; (6) detecting the eluent, thus obtaining a measurement result of the content of the sialic acid in the protein.