3737 results about "Gradient elution" patented technology

The invention discloses a method for separating high purity phycocyanin from spirulina, comprising the following steps: adopting 0.05-0.5M of phosphate buffer to elute a spirulina powder raw material by an elution method, enabling the phycocyanin to be effused from spirulina cells, adding soluble chitosan to phycocyanin crude extract, controlling the mass concentration of chitosan in the phycocyanin solution to be 0.1-1%, and collecting supernate by centrifuging; adding activated carbon to the supernate, centrifuging and collecting supernate, adopting ammonium sulfate precipitation with the mass concentration of 30-60% to precipitate the phycocyanin in the solution, adopting ion-exchange chromatography, and performing gradient elution by phosphate buffer containing sodium chloride to obtain the high purity phycocyanin solution. The invention has the advantages of low cost, short elapsed time, convenient and rapid operation, low energy consumption, and easy massive preparation, is an efficient method for massively preparing phycocyanin with high purity and high activity and is favor of scaled development and utilization of the phycocyanin in our country.

The invention discloses a preparation method of high pure crocin and geniposide. The preparation method comprises following main steps: selecting raw materials of gardenia ellis plants, extracting with water or an alcohol-water mixed solution, merging the extracting solutions, condensing, filtering, subjecting the obtained extracting solution to an adsorption treatment by macroporous resin, then performing gradient elution with a ethanol-water mixed solution, obtaining geniposide and gardenia yellow refined extracts; subjecting the geniposide refined extract to a recrystallization treatment with ethyl acetate so as to obtain the geniposide product; subjecting the gardenia yellow refined extract to go through column chromatography, which has been pressurized and stuffed with modified silica gel, to separate, and obtaining the high pure crocin product (color value larger than 600) and a chlorogenic acid component. The products have the advantages of high purity, simple technology, strong operability and convenience for automation, comprehensive utilization of the plant resources is achieved, the solvent is convenient to recycle and reuse, and the preparation method is easy to apply to the industrial amplification.

The invention relates to a screening method for 43 artificial synthetic pigments in an aquatic product. The method comprises the following steps: screening by virtue of quadrupole tandem time-of-flight mass spectrometry of liquid chromatogram; calling an established mass spectrometry screening database to automatically scan and retrieve in a molecular matching mode; through a C18 analytic column, carrying out gradient elution by taking an acetonitrile solution containing 0.1% of formic acid in a 5mmol/L ammonium acetate aqueous solution as a moving phase, wherein the flow rate is 0.3ml/minute; and detecting under a negative ion mode by using an electrospray ionization source, extracting a suspicious sample, purifying the sample by virtue of a matrix dispersion method, carrying out liquid chromatographic separation, quantitatively determining by virtue of an external standard method, and verifying. The screening method has the advantages that the detection method is simple and quick to operate and high in sensitivity. By adopting the matrix dispersion method to purify the sample, the interference of matrix components is effectively reduced. Misjudgment events such as false positive events are greatly reduced by virtue of the qualitative function of the quadrupole tandem time-of-flight mass spectrometry, so that the monitoring ability of the detection mechanism on the risk of the aquatic product is extremely enhanced.

The invention discloses an analysis method for simultaneously measuring residues of 12 sulfonamide medicaments, 19 quinolone medicaments and 8 benzimidazole medicaments and metabolites thereof in chicken liver by quick, easy, cheap, effective, rugged and safe (QuEChERS) extraction and purification-ultrahigh performance liquid chromatography-tandem mass spectrometry (QuEChERS-UPLC-MS/MS). The method comprises the following steps of: extracting a sample by using 1 percent acetic acid-acetonitrile solution, purifying the sample by using NH2 adsorbent, and degreasing the sample by using n-hexane;and then performing separation by using a Kromasil Eternity C18 chromatographic column (100mm*2.1mm, 2.5mum), performing gradient elution by using 0.1 percent formic acid and methanol as mobile phases, performing ionization in an electron spray positive ion (ESI<+>) mode, performing detection in a multi-reaction monitoring (MRM) mode, and performing quantification by an internal standard method. The 39 medicaments have good linearity (r is more than 0.98) in a blank adding concentration range of 5 to 100mug/kg, the average recovery rate of the medicaments in the adding level range of 10 to 50mug/kg, the relative standard deviation (RSD) is 1.5 to 23.4 percent, the limit of detection (LOD) of the 39 medicaments is 5mug/kg, and the low limit of quantification (LOQ) is 10mug/kg. The method is simple, convenient, quick, sensitive, accurate and rugged, and is suitable for confirmation and quantitative determination of the residues of the residues of the sulfonamide, quinolone and benzimidazole medicaments in the chicken liver.

During the extraction of resveratrol from giant knotwood, giant knotwood glycoside is hydrolyzed and enriched through organic synthesis. The extraction process includes mixing giant knotwood raw material and composite stuffing, high pressure chromatography in a high-pressure chromatographic column apparatus, gradient elution with chloroform and ethyl acetate and thin chromatographic tracking detection. The said process can reach a kilogram level yield and high product purity. The present invention may be used in extracting resveratrol and other similar product from giant knotwood material.

The invention discloses a method for extracting oligomeric proanthocyanidins, which comprises the following steps of: extracting a plant raw material containing the oligomeric proanthocyanidins with a solvent; enabling a crude extract to pass through a macroporous adsorbent resin column; eluting with water to remove protein and polysaccharide impurities; carrying out gradient elution with water and one or more selecting from alcohol, methyl alcohol and acetone as an eluant; collecting mixed solution with the volume ratio of n-butyl alcohol, acetic acid and water of 4:1:5; standing for demixing, wherein the solution at upper layer is a developing solvent; and carrying out silica gel thin-layer chromatography, wherein the elution part with the Rf value of 0.2-0.5 on a thin-layer chromatography plate is the oligomeric proanthocyanidins extract. Compared with the prior art, the invention has the advantages that the purity and the yield are both remarkably improved, the yield of the oligomeric proanthocyanidins can reach more than 80 percent, the product purity is no less than 50 percent and is basically more than 80 percent, and the method has simple and convenient process, low cost and easy industrialization.

The invention discloses a purification and preparation method of high-purity Daptomycin, which comprises the following steps of compounding a buffer solution and coarse Daptomycin into a sample solution, absorbing by using composite YT-01reverse phase silica gel columns and carrying out gradient elution or constant elution with the water solution of a strong polar solvent, which is used as a resolution agent. The chromatographic purity is higher than 98%.

Methods and apparatus for characterizing a polymer sample and in preferred embodiments, libraries of polymer samples, in a comprehensive, directly-coupled multi-dimensional liquid chromatography system are disclosed. The first and second dimensions are preferably high-performance liquid chromatography dimensions, such as for example, a first dimension adapted for determining composition (e.g. adapted for mobile-phase gradient elution chromatography, including reverse phase chromatography, adsorption chromatography and the like), and a second dimension adapted for determining molecular weight or particle size (e.g., adapted for size exclusion chromatography, including gel permeation chromatography).

The invention relates to a liquid chromatography for detecting anabolic hormone drug residue in animal-derived food, which is characterized by first sampling: animal musculature is taken off the fat and connective tissue, then minced and evenly ground, and the sample is weighed; extracting: anabolic hormone extract is extracted from the weighed sample by methanol ultrasonic extraction, the extract is evaporated to dryness in water bath through a rotary evaporator, and the residue is dissolved by methanol aqueous solution; purifying: C18 Solidoid extraction column on the extract is carried out solid phase extraction; and testing by devices: the filtrate is tested by opposite phase high efficiency liquid chromatography, quantified by external reference method-peak area, and then carried out with binary gradient elution. In the invention, the detected sample is complex biological sample; the established method can complete one detection in about one hour and is simple and fast, with reliable sensitivity and low cost; the method can analyze more veterinary hormone drug species than the synchronous usage of the existing GC or HPLC methods with easier operation, and has lower cost than the existing GC/MS and LC/MS or LC/MS/MS methods with low solvent toxicity.

A system for identifying Shenqi Fuzheng injection includes a mechanism for establishing a profile of a sample to be tested; a mechanism for establishing a characteristic fingerprint profile of Shenqi Fuzheng injection as a standard fingerprint profile; and a mechanism for comparing the profile of the sample to be tested with the standard fingerprint profile to distinguish between authentic Shenqi Fuzheng injection and counterfeit Shenqi Fuzheng injection.

A method for establishing a Shenqi Fuzheng injection fingerprint spectrum, including: employing an ultra-high voltage liquid chromatography mass spectrometer to test the Shenqi Fuzheng injection, the chromatography conditions including: chromatographic column: Agilent Zorbax Eclipse Plus C18, 2.1 mm×100 mm, 1.8 μm; mobile phase: mobile phase A is 0.1% formic acid aqueous solution, and mobile phase B is 0.1% formic acid acetonitrile solution; and employing gradient elution procedure.

The invention provides a method for establishing a fingerprint spectrum of a liver-enhancing medicine. The liver-enhancing medicine is composed of oriental wormwood, isatis root, angelica, white paeony root, red-rooted salvia root, Radix curcumae, Astragalus mongholicus, Codonopsis pilosula, rhizoma alismatis, sealwort, rehmannia, yam, hawthorn, large-leaved gentian, liquorice and medicated leaven. The establishing method comprises the step of detecting paeoniflorin in the liver-enhancing medicine by using a high efficiency liquid chromatography method, wherein conditions are as follows: a chromatographic column takes octadecylsilane chemically bonded silica as a filling material; mobile phases comprise a mobile phase A which is acetonitrile and a mobile phase B which is an acidic water solution, and the mobile phases are subjected to gradient elution; the flow velocity is 1.0mL/min; the column temperature is 30 DEG C; the detection wavelength is 210nm to 400nm; and the number of theoretical plates is calculated according to the paeoniflorin peak and should not be less than 6000. The obtained fingerprint spectrum is a chromatographic peak which mainly takes active ingredients of the large-leaved gentian, the white paeony root, the liquorice, the red-rooted salvia root, the oriental wormwood and the astragalus mongholicus in raw materials of the liver-enhancing medicine as main parts. According to the establishing method provided by the invention, the fingerprint spectrum capable of comprehensively representing the active ingredients of the liver-enhancing medicine can be effectively obtained and has the characteristics of high precision, stability and repeatability. The obtained fingerprint spectrum can be used for guaranteeing the stability and consistency of the product quality, so that the safety and effectiveness of a product are guaranteed.

The utility model discloses a high efficiency liquid phase chromatogram method which can detect the content of various polyphenolic compounds inside fruits at the same time by adopting liquid phase chromatogram containing ultraviolet detector and reversed-phase bonded silica chromatographic column; organic solvent mobile phase A and acidic aqueous mobile phase B with certain Ph value are adopted as gradient eluant. The utility model quantitatively detects various polyphenolic compounds inside fruits at the same time only on normal high efficiency liquid phase chromatogram. The utility model builds a method of detecting the content of various polyphenolic compounds inside fruits which combines microwave extraction and high efficiency liquid phase chromatogram. The utility model has the advantages of advanced and accurate method, simple and quick sample processing; meanwhile, Carrez reagent is used as scavenging agent of fruits sample and is firstly used as fruit polyphenols testing.

The invention relates to a method for detecting oligosaccharides in breast milk. The method includes a step of breast milk sample pretreatment, namely taking a breast milk sample, centrifugally removing upper fat, taking lower liquid, adding alcohol solvent, freezing for 5-15min at a temperature ranging from -40 DEG C to -100 DEG C, centrifuging and taking supernatant. The invention further provides a method for qualitative and quantitative detection of oligosaccharides in the pretreated breast milk according to a liquid chromatography-mass spectrometry method. The method for simultaneous detection of various oligosaccharides in non-reducing breast milk by liquid chromatography-mass spectrometry is established and has advantages of effectiveness, quickness and simplicity in pretreatment. AQ Exactive detector is adopted for mass spectrometry, liquid-phase gradient elution parameters and mass spectrometric detection parameters are determined, ion information of non-reducing breast milkoligosaccharides in mass spectrometric detection under a positive ion mode is given, and the method can be well applied to qualitative and quantitative detection of the oligosaccharides in the breastmilk.

The invention discloses a quick qualitative and quantitative method for oligosaccharide in breast milk. The quick qualitative and quantitative method mainly includes steps of 1, pretreating samples, to be more specific, removing fat and proteins from 150-250 micro-l of breast milk to obtain ultimate supernatant, adding ultra-pure water into the supernatant and diluting the supernatant to obtain the loaded samples; 2, establishing standard curves for standard substances by the aid of ultrahigh-performance liquid chromatography and mass spectrometry; 3, separating different components of the oligosaccharide in the breast milk in the loaded samples by the aid of ultrahigh-performance liquid mass spectrometry and carrying out quantitative analysis by the aid of mass spectrometry combined with the standard curves so as to obtain the content of the oligosaccharide in the breast milk. The ultrahigh-performance liquid chromatography is implemented by the aid of amino chromatographic columns with the sizes of 2.1*100 mm and 1.7 micrometers, 8-10 mmol/L of ammonium formate solution (A) and acetonitrile (B), and the ammonium formate solution (A) and the acetonitrile (B) are used as mobile phases; gradient elution programs are carried out by the aid of 95%-75% of B for 0-10 min or are carried out by the aid of 75% of B for 10-15 min or are carried out by the aid of 75%-65% of B for 15-20 min or are carried out by the aid of 65%-10% of B for 20-21 min or are carried out by the aid of 10% of B for 21-24 min or are carried out by the aid of 10%-95% of B for 24-25 min or are carried out by the aid of 95% of B for 25-35 min; the flow rates are 0.3 mL/min, and the column temperatures are 40-60 DEG C. The quick qualitative and quantitative method has the advantage that 12 types of oligosaccharide in the breast milk can be quickly detected by the aid of the quick qualitative and quantitative method and can be quantified by the aid of the quick qualitative and quantitative method.

The invention discloses a method for determining a radix notoginseng extract and contents of five types of ginsenosides in a preparation of the radix notoginseng extract by a Fourier transform near-infrared spectrograph. The method comprises the steps that a sample is detected by an ultra-high performance liquid chromatography, moving phase builds acetonitrile and water gradient elution parameters with separation degrees higher than 1.7 in chromatographic peaks of Rg1 and Re, and the spectrum preprocessing adopts the combination of 2 to 3 methods in Savitzky-Golay polynomial smoothing, second-order differential, Noriis derivative filtering and data normalization; and three preferred wave bands of the five types of ginsenosides are modeled, and a calibration model is built by any one of a partial least squares regression method, a principal component regression method and a multiple linear regression method. The method is used for determining the contents of the five types of ginsenosides in the sample to be detected, the determining result is consistent with a result determined by the ultra-high performance liquid chromatograph basically, the requirements of 'Chinese pharmacopoeia' are met, the accuracy is high, and the determination speed is increased substantially.

The invention relates to a preparation process of rosmarinic acid, which adopts the following steps for preparing the rosmarinic acid: combining aqueous extraction and ethanol extraction, carrying out ion exchange, placing materials on a chromatographic separation medium for gradient elution, and carrying out secondary decolourization, nanofiltration membrane concentration and recrystallization. The method solves the problems of large extraction solvent amount, much chemical residual, poor color intensity and low extraction rate. Products produced by adopting the method have the advantages of good quality, high purity, high extraction rate, simple operation and convenient scale production.

The invention relates to a method for quickly determining the content of additive of a plurality of types in food. An adopted technical scheme is as follows: a solid sample is accurately weighed into a centrifuge tube, petroleum ether or n-hexane is additionally arranged to become homogenate, and supernatant fluid is removed by centrifugation; after the petroleum ether or the n-hexane in the residue are volatilized, the mixed solvent of ethanol, ammonia and water is additionally arranged, ultrasonic extraction is carried out, and the supernatant fluid is obtained by centrifugation as sample liquid for testing; the sample liquid for testing is adopted, potassium ferrocyanide solution and zinc acetate solution are additionally arranged, the mixture is mixed uniformly and centrifuged; the supernatant fluid is concentrated in water bath at 60DEG C to 100DEG C; concentrated solution is transferred to a measuring flask with water; the pH value is regulated to 5 to 7, water is additionally arranged to fix the capacity to a scale; the mixture is mixed uniformly, is filtered by a 0.45mum microfiltration membrane and arranged in a sample bottle as sample liquid; and the sample liquid is tested in a liquid chromatograph by gradient elution under variable wavelengths. According to the invention, the method for quickly determining the content of additive of a plurality of types in food is simple and easy, has high precision, high accuracy, high sensitivity, has the recovery rate being above 80 percent, and is used for determining the content of additive of a plurality of types in food at the same time.

The invention discloses a method for extracting high-purity sulforaphane from broccoli seeds; and the method comprises the following steps of: (1) after crushing dry broccoli seeds, adding water to the dry broccoli seeds to hydrolyze the broccoli seeds, so as to obtain a solid-liquid mixture; (2) adding anhydrous ethanol to the mixture; ultrasonically extracting the mixture; and carrying out suction filtration or double-layer gauze filtering on the mixture; (3) concentrating and evaporating ethanol from a filter liquor in a vacuum state; (4) carrying out chromatography on a material liquor by using a macroporous resin column and eluting the material liquor by using an ethanol solution in a gradient way; (5) collecting an eluting solution containing sulforaphane; carrying out chromatography on the eluting solution by using an alumina column; and concentrating a sulforaphane chromatography solution to form an extract so as to obtain a crude sulforaphane product; (6) adding water to the obtained crude product so as to dissolve the crude product; and extracting the dissolved crude product by using a polar solvent; and (7) collecting an organic layer; and concentrating the organic layer in a vacuum state so as to obtain an oily extract, so that the high-purity sulforaphane is obtained. As the high-purity sulforaphane can be directly extracted from the broccoli seeds and broccoli related materials, the method for extracting the high-purity sulforaphane from the broccoli seeds, disclosed by the invention, has the advantages of simple process, stable product, simple equipment and safety in production and is suitable for enterprises for industrially extracting the sulforaphane.

The invention discloses a method for extracting and separating curcumin as an effective constituent of turmeric, which belongs to the extraction and separation of traditional Chinese medicine effective constituents and technical application. The structural formula of the compound is as shown in a figure. The method for extracting and separating curcumin comprises the steps of grinding turmeric, adding alkali liquid for cold extraction, filtering, performing column chromatography through macroporous resin, performing gradient elution by use of ethanol and water, collecting eluent, reducing pressure, evaporating, concentrating and vacuum-drying the eluent, and obtaining the effective constituent, namely curcumin. The method has the advantages that the macroporous resin used in the method can be repeatedly utilized, and the method is economical and simple by taking water and ethanol as solvents, can conveniently obtain the extract with the effective constituent content not lower than 90 percent, and is easy to industrialize.

The invention provides a technological method suitable for the industrialized purification of Carbetocin, a reversed-phase efficient liquid chromatography is used to purify the Carbetocin to result in high purity and good yield in order to meet industrialized demands. A crude peptide solution of the Carbetocin is filled with materials by a reversed-phase chromatographic column to be a stationary phase, a phosphate buffer solution is regarded as phase A and acetonitrile is regarded as phase B to implement gradient elution purification, wherein, the gradient: B%:20-40%, the pH of the phase A is 2.5-3.5; an anion exchange method is employed to convert the phosphate and trifluoroacetate to acetate. The invention employs one-step reversed-phase efficient liquid chromatography to purify followed by the acetate conversion by an anion exchange column in one step, thus obtaining high-purity acetate Carbetocin at high yield and offering an efficient purification technology for the massive purification and preparation of the peptide raw drugs.