CN101323618B - Method for separating and preparation of Gelsemium elegans alkaloid monomer by high speed counter current chromatography - Google Patents
Method for separating and preparation of Gelsemium elegans alkaloid monomer by high speed counter current chromatography Download PDFInfo
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Abstract
The invention discloses a method for separating and preparing a gelsemium alkaloid monomer from gelsemium by adopting high-speed counter current chromatography, which pertains to the separation method of medicinal plant monomers. The invention takes total alkaloids of gelsemium as a raw material and a high-speed counter current chromatograph as a separating device and the method comprises that a solvent system which is composed of stationary phase and mobile phase is prepared; the high-speed counter current chromatograph is filled with the stationary phase and then the mobile phase is pumped in and balanced; and then sampling is conducted by a sample valve; target components are collected according to the map of a detector or in combination of the detection methods of high efficiency liquid chromatography and thin layer chromatography, and distilled under reduced pressure and then recrystallized to obtain the gelsemium alkaloid monomer with high purity. By adjusting the specific proportional parameters of the solvent system, single or multiple target gelsemium alkaloid monomers can be separated by one step or step by step. The method is convenient and efficient, large in preparation amount, less in sample loss, good in separation effect and high in controllability and applicable to automatic production and the obtained gelsemium alkaloid monomer from separation has diverse pharmacological activities and application prospect to be prepared into medicines.
Description
Technical field
The present invention relates to the monomeric separation method of medicinal plant, use high-speed countercurrent chromatography specifically and from elegant jessamine, separate the method for preparing the elegant jessamine alkaloid monomer.
Background technology
(Gelsemium elegans Benth. is the loganiaceae plant herb of climing rattan recklessly GEB) to elegant jessamine, is divided into two kinds of Chinese elegant jessamine and North America elegant jessamines.Abound with in Zhejiang in China, ground such as Fujian, Guangdong, Guangxi, Hunan, Guizhou, Yunnan, have another name called Graceful Jessamine Herb, elegant jessamine, malicious root, big tea medicine etc.Abroad, H Gerrad in 1887 etc. are made into Galenicals, by some national Martindale-The Extra Pharmacopoeias are recorded; The record of elegant jessamine is all arranged in ADI and the Japan " medicine among the people in the world ".China is more long to the research history of elegant jessamine, and " legendary god of farming's book on Chinese herbal medicine ", Compendium of Material Medica are all on the books, and China is among the people to use the former plant of elegant jessamine always and treat all kinds of pain, especially chronic neuropathic pain and cancer pain.The main effective constituent of elegant jessamine is indoles alkaloid.Gelsemium alkaloids or crude extract have treatment Immunological diseases, antitumor, relieving inflammation and relaxing pain, anti-psoriatic, expand pupil, promote pharmacological action (Zhang Lanlan such as growth of animals or poultry, Lin Jingming, the Wuzhong. elegant jessamine chemical ingredients and pharmacological research progress, Chinese medicinal materials, 2003,26 (6): 451-453).Elegant jessamine toxicity is big, and clinical application is subjected to certain limitation, but under its significant clinical efficacy temptation, active day by day to its research and development in recent years, begin to aim at its alkaloid monomer.
The elegant jessamine alkaloid monomer has separated obtaining koumine, first, the third of the twelve Earthly Branches, the fourth of the twelve Earthly Branches, occasion, third, fourth, penta, oneself, more than 40 kind of monomers such as heptan and humantenmine, second, third, fourth at present, and its toxicity varies in size, and having many uses too of elegant jessamine alkaloid monomer is general.Has antitumor action (Wu Darong as koumine, Qin Rui, Cai Jing, Chi Debiao. the research of koumine antitumor action, Pharmacology and Clinics of Chinese Materia Medica, 2006,22 (5): 6-8), regulate immunity function (Sun Lisha, Lei Linsheng, Fang Fangzhi, Yang Shuqin, Wang Jian. koumine is to the restraining effect of mouse boosting cell proliferative response and humoral immune reaction, Pharmacology and Clinics of Chinese Materia Medica, 1999,15 (6): 10-12), treatment psoriatic (Zhang Lanlan, Huang Changquan, Zhang Zhongyi, Deng. the observation of curative effect of koumine treatment psoriasiform animal model. No.1 Military Medical Univ.'s journal, 2005,25 (5): 547-549) and antiplatelet aggregative activity (Fang Fangzhi, Dan Chunwen, Chen Pingyan. koumine influences the rabbit platelet accumulative, Pharmacology and Clinics of Chinese Materia Medica, 1998,14 (1): 21-24) etc.
Present bibliographical information, being used for from the method for gelsemium alkaloids separation elegant jessamine alkaloid monomer is column chromatography, as silica gel column chromatography, alumina column chromatography method.The weak point of column chromatography is: separation cycle is long, and the rate of recovery is low, and separating effect is very undesirable.High speed adverse current chromatogram (High-speed Counter-currentChromatography, HSCCC) be the new separation technology that since the eighties in 20th century, on the liquid luquid partition chromatography basis, has grown up in the world, it need not any solid-state supporter or carrier thereby overcome the irreversible adsorption effect of traditional isolation technique to sample component, material and solvent consumption have been saved, the sample recovery rate height, also have easy and simple to handle simultaneously, fractional dose is big, separation cycle is short, separation efficiency height and high repeatability and other advantages, easily form automatic production, be widely used in biology, medicine, the preparation separation and the purifying of the various materials in field such as environmental protection.
By literature search, the existing lot of documents report of gelsemium alkaloids extracting method (Xu Jian, Liang Chongzhen. the comparison of gelsemium alkaloids extracting method, the pharmacy circular, 1988,23 (6): 341-342), apply for a patent: the 1. extracting method of gelsemium alkaloids comprising 2 pieces, publication number: CN101011466A, open day: on August 8th, 2007; 2. a kind of Chinese medicine gelsemium alkaloids of anticancer analgesic and contain its pharmaceutical composition and preparation method thereof, publication number: CN1528767A, open day: on September 15th, 2004.But apply for a patent and do not relate to monomeric separation method for above-mentioned 2 pieces.In the existing document, do not see that high-speed countercurrent chromatography is used for the isolating domestic and international report of elegant jessamine alkaloid monomer.Therefore, the invention discloses high-speed countercurrent chromatography separates the elegant jessamine alkaloid monomer from elegant jessamine method.
Summary of the invention
In order to overcome existing deficiency of separating elegant jessamine alkaloid monomer technology, the object of the invention is to provide a kind of novel method of efficient, quick-acting separation elegant jessamine alkaloid monomer, promptly uses high-speed countercurrent chromatography to separate the method for preparing the elegant jessamine alkaloid monomer from elegant jessamine.
The technical solution adopted for the present invention to solve the technical problems is: a kind of high-speed countercurrent chromatography separates the method for preparing the elegant jessamine alkaloid monomer from elegant jessamine, it is characterized in that:
Raw material: gelsemium alkaloids.
Described raw material gelsemium alkaloids, be with reference to classical elegant jessamine alkaloid extracting method obtain (Chen faithful and upright person. the elegant jessamine alkaloid extraction with separate bulletin of Chinese materia medica, 1987,12 (5): 41; Xu Jian, Liang Chongzhen. the comparison of gelsemium alkaloids extracting method, pharmacy circular, 1988,23 (6): 341-342; Chen Jingfeng, Yuan Hui. the extraction of gelsemium total alkaloids, separation, evaluation and general toxicity, Agricultural University Of Hunan's journal (natural science edition), 2003,29 (5): 422-425).
Separating device: high-speed counter-current chromatograph
Described separating device high-speed counter-current chromatograph according to the difference of required target fractional dose, can be selected analysis mode, half preparation type or countercurrent chromatography instrument.
High speed adverse current chromatogram separates the method for preparing the elegant jessamine alkaloid monomer, comprises that preparation constitutes the solvent system of stationary phase, moving phase; High-speed counter-current chromatograph is filled full stationary phase; Pump into moving phase and balance; By the sampling valve sample introduction; According to the detector collection of illustrative plates or in conjunction with high performance liquid chromatography, thin-layered chromatography detection method, collect the purpose component, recrystallization obtains high purity elegant jessamine alkaloid monomer after the evaporated under reduced pressure.
Described solvent system can be: 1) alkane: fatty acid ester: Fatty Alcohol(C12-C14 and C12-C18): the two phase solvent system that water constitutes; 2) chloroform: Fatty Alcohol(C12-C14 and C12-C18): the two phase solvent system that water (containing finite concentration acid) constitutes; 3) and the polynary two phase solvent system of on above-mentioned two kinds of system bases, setting up.
Described separation method by adjusting the concrete proportioning parameter of solvent system, can be used for step separation single target elegant jessamine alkaloid monomer; Also can a step separate a plurality of target elegant jessamine alkaloid monomers; Also can separate the elegant jessamine alkaloid monomer step by step, promptly at first separate one or more more segregative elegant jessamine alkaloid monomers, the component that will fail to separate pumps from chromatographic instrument then, concentrates the back and adopts high-speed countercurrent chromatography to carry out secondary separation.
Isolated elegant jessamine alkaloid monomer can be measured fusing point by the fusing point instrument, high performance liquid chromatograph is measured purity, ultraviolet (Uultraviolet, UV) all-wave spectrum scanning spectra, infrared (Infrared, IR) scanning spectra, nuclear magnetic resonance spectrum (Nuclear Magnetic Resonance, NMR), (Mass Spectrum MS) waits its structure of method validation to mass spectrum.
Advantage of the present invention is:
1) convenient and swift, easy and simple to handle, separation cycle is short.Tradition method for separating and preparing complex operation, separation cycle are long, generally take 1～2 month, even for more time; Adopt this law high-speed countercurrent chromatography to separate the elegant jessamine alkaloid monomer, only needed get final product in 1～2 day, shorten separation cycle greatly.
2) preparation amount is big.According to required target sample amount, can select the high-speed counter-current chromatograph of different model to separate.Wherein, countercurrent chromatography instrument sample size can reach several grams even tens grams, and the amount of the elegant jessamine alkaloid monomer that flash liberation goes out is the reducible pilot scale level that reaches.
3) the sample loss is few, yield height, good separating effect.The used stationary phase of traditional column chromatography is a solid, is prone to the not high phenomenon of absorption, hangover, separation purity of sample; And the used stationary phase of high speed adverse current chromatogram partition method is a liquid, does not have adsorption phenomena, thereby separates the yield height, is difficult for producing hangover, and good separating effect can reach more than 98.5% through purity behind the recrystallization.
4) the repetition controllability is good.Traditional column chromatography often adopts gradient elution, and elutriant concentration, elution speed are difficult for accurately control, poor repeatability; And this law controllable parameters, good reproducibility is fit to automatic production, can be applicable.
5) separate the elegant jessamine alkaloid monomer obtain, have multiple pharmacologically active, as regulate immunity, effect such as antitumor, have the potential application that is prepared into medicine.This separation method then provides the industrialization preparation basis of realizing that the elegant jessamine alkaloid monomer is medicinal.
In a word,, can adopt high-speed countercurrent chromatography by one step to separate certain or plurality of target elegant jessamine alkaloid monomer, also can separate multiple elegant jessamine alkaloid monomer step by step by adjusting solvent system.Easy and simple to handle than traditional separation method, separation cycle is short, controllability is good, preparation amount is big, the sample loss is few, has to be used for automated preparation elegant jessamine alkaloid monomer Application Prospect, and the elegant jessamine alkaloid monomer has multiple pharmacological effect, be expected to be applied to prepare the medicine for the treatment of multiple disease, and this law can become the medicinal industrialization preparation basis of preparation elegant jessamine alkaloid monomer.
Description of drawings
Below in conjunction with drawings and Examples the present invention is further described.
Fig. 1 is the high speed adverse current chromatogram figure that separates koumine from gelsemium alkaloids of embodiment 1;
X-coordinate is represented the time among the figure; 1 is koumine, and 2 is gelsemine, and 3 is hexa-gelsemicine.
Fig. 2 is the high speed adverse current chromatogram figure that separates gelsemine and hexa-gelsemicine from gelsemium alkaloids of embodiment 2;
X-coordinate is represented the time among the figure; 1 is hexa-gelsemicine, and 2 is gelsemine.
Fig. 3 be embodiment 3 from gelsemium alkaloids, separate 4 kinds of monomeric high speed adverse current chromatogram figure such as koumine, gelsemine and hexa-gelsemicine;
X-coordinate is represented the time among the figure; 1 is that monomer 1,2 is a monomer 2: hexa-gelsemicine, 3 is monomer 3: koumine, 4 is monomer 4: gelsemine.
Embodiment
Below the invention will be further described by specific embodiment, but practical range of the present invention is not constituted any restriction.
High-speed countercurrent chromatography separates elegant jessamine alkaloid monomer koumine (see figure 1) from gelsemium alkaloids.
1. sample: gelsemium alkaloids, be with reference to classical elegant jessamine alkaloid extracting method (Chen faithful and upright person. the elegant jessamine alkaloid extraction with separate bulletin of Chinese materia medica, 1987,12 (5): 41; Xu Jian, Liang Chongzhen. the comparison of gelsemium alkaloids extracting method, pharmacy circular, 1988,23 (6): 341-342; Chen Jingfeng, Yuan Hui. the extraction of gelsemium total alkaloids, separation, evaluation and general toxicity, Agricultural University Of Hunan's journal (natural science edition), 2003,29 (5): 422-425) extract acquisition from banyan crotchet kiss rhizome.
2. instrument: TBE-300A high-speed counter-current chromatograph, Shanghai Tongtian Biotechnology Co., Ltd..
3. method:
3.1 the preparation solvent system, normal hexane: ethyl acetate: dehydrated alcohol: water (3: 3: 2: 3V/V) be total to 2200ml, fully standing stay-over demixion behind the mixing.Second natural gift are from stratified two phase liquid, and are last as stationary phase, following to moving phase, ultrasonic half an hour.
3.2 preparation sample solution: the about 300mg of hook taking kiss total alkaloids, with the moving phase dissolving that is slightly less than 20ml, ultrasonic 10min, standby.
3.3 balance high-speed counter-current chromatograph stationary phase, moving phase system: it is full to pump that stationary phase pumps into high-speed counter-current chromatograph with the 25ml/min flow velocity, and setting the constant temperature circulator temperature is 25 ℃; Go into moving phase to balance with the 1.5ml/min flow pump, pump moving phase is opened main frame simultaneously rapidly, transfers the main frame counter-rotating, and rotating speed is 900rpm.The retention rate ρ of this solvent system is 71.6% (ρ=(V during calculated equilibrium
Always-V
Go out)/V
Always* 100%, V wherein
Always: pipeline cumulative volume (ml); V
Go out: the stationary phase volume (ml) that moving phase is released in the equilibrium process).Balance and stability 10min.
3.4 last sample separates: open UV-detector and N2000 chromatographic working station software, sample is gone into instrument by the sampling valve suck-back, clicks software when sample introduction finishes rapidly and begins to gather the uv-absorbing data.40min begins to collect effusive sample with automatic Fraction Collector behind the sample introduction, and every 4min collects 1 pipe, collects 60 pipes altogether.After collection finishes, engine speed is transferred to 0, Shutdown Hosts and constant temperature circulator pump into stationary phase with the 25ml/min flow velocity, the equilibrium system solution in the main frame is pumped recovery concentrate standby.
4. the target monomer is collected purifying: the component of respectively managing that will collect detects with high performance liquid chromatography.Testing conditions is as follows: chromatographic column is Yi Lite Hypersil ODS2 performance liquid chromatographic column (5 μ m, 4.6mm * 250mm), 25 ℃ of a column temperature; Moving phase is methyl alcohol: water: n-Butyl Amine 99 (50: 50: 0.1V/V); Flow velocity is 1.0ml/min; UV-detector, the detection wavelength is 256nm.After testing, will contain the component merging of the higher identical component of purity, evaporated under reduced pressure, ethyl alcohol recrystallization, obtain a monomer component, be defined as koumine, purity＞98.5% through methods analysts such as the scanning of ultraviolet all-wave spectrum, infared spectrum scanning, nuclear magnetic resonance spectrum, mass spectrums.
High-speed countercurrent chromatography separates elegant jessamine alkaloid monomer gelsemine and hexa-gelsemicine.(see figure 2)
Present embodiment is to adopt high-speed countercurrent chromatography to separate the alkaloidal monomeric example of elegant jessamine step by step.The first step adopts high-speed countercurrent chromatography to separate koumine, and concrete grammar sees embodiment 1 for details; Second step, with the first step high-speed countercurrent chromatography fail isolating component collect concentrate after, as new sample separation, separate through high-speed countercurrent chromatography once more, can get gelsemine and hexa-gelsemicine.Present embodiment only stated for second step.
After 2.1 two components of gelsemine of failing to separate among sample: the embodiment 1 and hexa-gelsemicine (being 2,3 components among Fig. 1) were collected the liquid merging, 45 ℃ of evaporated under reduced pressure were as new sample separation.
2.2 instrument: TBE-300A high-speed counter-current chromatograph, Shanghai Tongtian Biotechnology Co., Ltd..
2.33. method:
3.1 the preparation solvent system, chloroform: methyl alcohol: 0.2mol/l hydrochloric acid (4: 3: 1.5V/V) be total to 1700ml, fully standing stay-over demixion behind the mixing.Second natural gift are from stratified two phase liquid, and are last as stationary phase, following to moving phase, ultrasonic half an hour.
3.2 preparation sample solution: get the about 300mg of above-mentioned sample, with the moving phase dissolving that is slightly less than 20ml, ultrasonic 10min, standby.
3.3 balance high-speed counter-current chromatograph stationary phase, moving phase system: it is full to pump that stationary phase pumps into high-speed counter-current chromatograph with the 25ml/min flow velocity, and setting the constant temperature circulator temperature is 25 ℃; Go into moving phase to balance with the 2ml/min flow pump, pump moving phase is opened main frame simultaneously rapidly, transfers the main frame counter-rotating, and rotating speed is 800rpm.The retention rate ρ of this solvent system is 66% during calculated equilibrium.Balance and stability 10min.
3.4 last sample separates: open UV-detector and N2000 chromatographic working station software, sample is gone into instrument by the sampling valve suck-back, clicks software when sample introduction finishes rapidly and begins to gather uv-absorbing data (the detection wavelength is 254nm).30min begins to collect effusive sample with automatic Fraction Collector behind the sample introduction, and every 4min collects 1 pipe, collects 100 pipes altogether.After collection finishes, engine speed is transferred to 0, Shutdown Hosts and constant temperature circulator pump recovery with nitrogen with the equilibrium system solution in the main frame and concentrate standby.
4. the target monomer is collected purifying: the component of respectively managing that will collect detects with high performance liquid chromatography.Testing conditions is as follows: chromatographic column is Yi Lite Hypersil ODS2 performance liquid chromatographic column (5 μ m, 4.6mm * 250mm), 25 ℃ of a column temperature; Moving phase is methyl alcohol: water: n-Butyl Amine 99 (50: 50: 0.1V/V); Flow velocity is 1.0ml/min; UV-detector, the detection wavelength is 256nm.After testing, will contain the component merging of the higher identical component of purity, evaporated under reduced pressure, ethyl alcohol recrystallization, obtain two monomer components, be defined as gelsemine and hexa-gelsemicine, purity＞98.5% through methods analysts such as the scanning of ultraviolet all-wave spectrum, infared spectrum scanning, nuclear magnetic resonance spectrum, mass spectrums.
High-speed countercurrent chromatography separates 4 kinds of monomers such as koumine, gelsemine and hexa-gelsemicine.(see figure 3)
1. sample: gelsemium alkaloids, be with reference to classical elegant jessamine alkaloid extracting method (Chen faithful and upright person. the elegant jessamine alkaloid extraction with separate bulletin of Chinese materia medica, 1987,12 (5): 41; Xu Jian, Liang Chongzhen. the comparison of gelsemium alkaloids extracting method, pharmacy circular, 1988,23 (6): 341-342; Chen Jingfeng, Yuan Hui. the extraction of gelsemium total alkaloids, separation, evaluation and general toxicity, Agricultural University Of Hunan's journal (natural science edition), 2003,29 (5): 422-425) extract acquisition from banyan crotchet kiss rhizome.
2. instrument: TBE-300A high-speed counter-current chromatograph, Shanghai Tongtian Biotechnology Co., Ltd..
3. method:
3.1 the preparation solvent system, chloroform: methyl alcohol: 0.2mol/l hydrochloric acid (2: 2: 1V/V) be total to 2000ml, fully standing stay-over demixion behind the mixing.Second natural gift are from stratified two phase liquid, and are last as stationary phase, following to moving phase, ultrasonic half an hour.
3.2 preparation sample solution: hook taking kiss total alkaloids 300mg, with the moving phase dissolving that is slightly less than 20ml, ultrasonic 10min, standby.
3.3 balance high-speed counter-current chromatograph stationary phase, moving phase system: it is full to pump that stationary phase pumps into high-speed counter-current chromatograph with the 25ml/min flow velocity, and setting the constant temperature circulator temperature is 25 ℃; Go into moving phase to balance with the 1.5ml/min flow pump, pump moving phase is opened main frame simultaneously rapidly, transfers the main frame counter-rotating, and rotating speed is 850rpm.The retention rate ρ of this solvent system is 73% during calculated equilibrium.Balance and stability 10min.
3.4 last sample separates: open UV-detector and N2000 chromatographic working station software, sample is gone into instrument by the sampling valve suck-back, clicks software when sample introduction finishes rapidly and begins to gather uv-absorbing data (the detection wavelength is 254nm).40min begins to collect effusive sample with automatic Fraction Collector behind the sample introduction, and every 4min collects 1 pipe, collects 110 pipes altogether.After collection finishes, engine speed is transferred to 0, Shutdown Hosts and constant temperature circulator pump recovery with nitrogen with the equilibrium system solution in the main frame and concentrate standby.
4. the target monomer is collected purifying: the component of respectively managing that will collect detects with high performance liquid chromatography.Testing conditions is as follows: chromatographic column is Yi Lite Hypersil ODS2 performance liquid chromatographic column (5 μ m, 4.6mm * 250mm), 25 ℃ of a column temperature; Moving phase is methyl alcohol: water: n-Butyl Amine 99 (50: 50: 0.1V/V); Flow velocity is 1.0ml/min; UV-detector, the detection wavelength is 256nm.After testing, the component that will contain the higher identical component of purity merges, evaporated under reduced pressure, ethyl alcohol recrystallization, obtain four monomer components, through methods analysts such as the scanning of ultraviolet all-wave spectrum, infared spectrum scanning, nuclear magnetic resonance spectrum, mass spectrum definite wherein three kinds be koumine, gelsemine and hexa-gelsemicine, purity＞98.5%.
Claims (5)
1. a high-speed countercurrent chromatography separates the method for preparing the elegant jessamine alkaloid monomer from elegant jessamine, it is characterized in that: be raw material with the gelsemium alkaloids, with the high-speed counter-current chromatograph is separating device, and this separation method comprises: preparation constitutes the solvent system of stationary phase, moving phase; High-speed counter-current chromatograph is filled full stationary phase; Pump into moving phase and balance; By the sampling valve sample introduction; According to the detector collection of illustrative plates or in conjunction with any detection method in high performance liquid chromatography and the tlc, collect the purpose component, recrystallization obtains the elegant jessamine alkaloid monomer after the evaporated under reduced pressure.
2. a kind of high-speed countercurrent chromatography according to claim 1 separates the method for preparing the elegant jessamine alkaloid monomer from elegant jessamine, it is characterized in that: described high-speed counter-current chromatograph is analysis mode, half preparation type or preparation type.
3. a kind of high-speed countercurrent chromatography according to claim 1 separates the method for preparing the elegant jessamine alkaloid monomer from elegant jessamine, it is characterized in that: described solvent system is one of following solvents system: 1) alkane: fatty acid ester: Fatty Alcohol(C12-C14 and C12-C18): the two phase solvent system that water constitutes; 2) chloroform: Fatty Alcohol(C12-C14 and C12-C18): the two phase solvent system that contains the water formation of finite concentration acid; 3) the polynary two phase solvent system of on above-mentioned two kinds of system bases, setting up.
4. a kind of high-speed countercurrent chromatography according to claim 1 separates the method for preparing the elegant jessamine alkaloid monomer from elegant jessamine, it is characterized in that: described separation method, by adjusting the concrete proportioning parameter of solvent system, be used for step separation single target elegant jessamine alkaloid monomer; Or a plurality of target elegant jessamine alkaloid monomers of step separation; Or separate the elegant jessamine alkaloid monomer step by step, and promptly at first separate one or more more segregative elegant jessamine alkaloid monomers, the component that will fail to separate pumps from chromatographic instrument then, concentrates the back and adopts high-speed countercurrent chromatography to carry out secondary separation.
5. a kind of high-speed countercurrent chromatography according to claim 1 separates the method for preparing the elegant jessamine alkaloid monomer from elegant jessamine, it is characterized in that: isolated elegant jessamine alkaloid monomer is verified its structure by fusing point instrument mensuration fusing point, high performance liquid chromatograph mensuration purity, ultraviolet all-wave spectrum scanning spectra, infrared scan collection of illustrative plates, nuclear magnetic resonance spectrum, mass spectrometry method.
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| US9078890B2 (en) | 2011-06-25 | 2015-07-14 | Fujian Medical University | Use of koumine and its homologues in preparation of medicament for treatment of autoimmune diseases of involved bones and joints |
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| CN102731515B (en) * | 2011-03-31 | 2016-01-20 | 上海中医药大学 | Preparation of koumine by silica gel chromatography |
| CN102321094B (en) * | 2011-07-24 | 2013-10-02 | 福建医科大学 | New Gelsmium elegans Benth. alkaloid compound as well as preparation method and application thereof |
| CN104833752B (en) * | 2015-05-12 | 2016-08-17 | 广西壮族自治区梧州食品药品检验所 | Measure the kounidine in elegant jessamine and the method for koumine |
| CN104991019B (en) * | 2015-06-29 | 2016-08-24 | 公安部物证鉴定中心 | Gelsemine and the liquid chromatography-tandem mass of koumine in biological material |
| CN107987087A (en) * | 2017-10-30 | 2018-05-04 | 福建医科大学 | The method that middle pressure preparative liquid chromatography isolates and purifies koumine |
| CN107936033A (en) * | 2017-10-30 | 2018-04-20 | 福建医科大学 | The method that industrially prepared liquid chromatogram isolates and purifies koumine |
| CN107894475B (en) * | 2017-11-20 | 2020-07-31 | 湖南农业大学 | Liquid chromatography-tandem mass spectrometry quantitative method for simultaneously detecting multiple effective components in gelsmium elegans |
| CN111560024B (en) * | 2020-05-13 | 2021-06-08 | 湖南农业大学 | Method for separating gelsemium elegans alkaloid monomer from gelsemium elegans alkaloid by combining high-speed counter-current chromatography with preparative liquid chromatography |
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