CN1264822C - Process for preparing high purity huperzine A - Google Patents
Process for preparing high purity huperzine A Download PDFInfo
- Publication number
- CN1264822C CN1264822C CN 200410052688 CN200410052688A CN1264822C CN 1264822 C CN1264822 C CN 1264822C CN 200410052688 CN200410052688 CN 200410052688 CN 200410052688 A CN200410052688 A CN 200410052688A CN 1264822 C CN1264822 C CN 1264822C
- Authority
- CN
- China
- Prior art keywords
- huperzine
- preparation
- solvent system
- water
- phase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- ZRJBHWIHUMBLCN-UHFFFAOYSA-N Shuangyiping Natural products N1C(=O)C=CC2=C1CC1C(=CC)C2(N)CC(C)=C1 ZRJBHWIHUMBLCN-UHFFFAOYSA-N 0.000 title claims abstract description 40
- ZRJBHWIHUMBLCN-YQEJDHNASA-N huperzine A Chemical compound N1C(=O)C=CC2=C1C[C@H]1\C(=C/C)[C@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-YQEJDHNASA-N 0.000 title claims abstract description 40
- ZRJBHWIHUMBLCN-BMIGLBTASA-N rac-huperzine A Natural products N1C(=O)C=CC2=C1C[C@@H]1C(=CC)[C@@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-BMIGLBTASA-N 0.000 title claims abstract description 40
- ZRJBHWIHUMBLCN-SEQYCRGISA-N Huperzine A Natural products N1C(=O)C=CC2=C1C[C@H]1/C(=C/C)[C@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-SEQYCRGISA-N 0.000 title claims abstract description 16
- 238000004519 manufacturing process Methods 0.000 title 1
- 239000012071 phase Substances 0.000 claims abstract description 33
- 238000002360 preparation method Methods 0.000 claims abstract description 28
- 239000002904 solvent Substances 0.000 claims abstract description 21
- 238000005070 sampling Methods 0.000 claims abstract description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 9
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 6
- 239000007790 solid phase Substances 0.000 claims abstract description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 5
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 5
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims abstract description 4
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 claims abstract description 3
- OFBQJSOFQDEBGM-UHFFFAOYSA-N Pentane Chemical compound CCCCC OFBQJSOFQDEBGM-UHFFFAOYSA-N 0.000 claims abstract description 3
- 230000005526 G1 to G0 transition Effects 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 14
- VZUHPKQBNDMOCH-UHFFFAOYSA-N butan-1-ol;hexane;hydrate Chemical compound O.CCCCO.CCCCCC VZUHPKQBNDMOCH-UHFFFAOYSA-N 0.000 claims description 3
- NDEJLRZXIVXVFQ-UHFFFAOYSA-N heptane;propan-2-one;hydrate Chemical compound O.CC(C)=O.CCCCCCC NDEJLRZXIVXVFQ-UHFFFAOYSA-N 0.000 claims description 2
- ZLSZUXNHKLRERC-UHFFFAOYSA-N hexane;methanol;hydrate Chemical compound O.OC.CCCCCC ZLSZUXNHKLRERC-UHFFFAOYSA-N 0.000 claims description 2
- 238000000034 method Methods 0.000 abstract description 12
- 238000002425 crystallisation Methods 0.000 abstract description 7
- 230000008025 crystallization Effects 0.000 abstract description 7
- 238000011084 recovery Methods 0.000 abstract description 6
- 239000000178 monomer Substances 0.000 abstract description 5
- ZQPQGKQTIZYFEF-WCVJEAGWSA-N Huperzine Natural products C1([C@H]2[C@H](O)C(=O)N[C@H]2[C@@H](O)C=2C=CC=CC=2)=CC=CC=C1 ZQPQGKQTIZYFEF-WCVJEAGWSA-N 0.000 abstract description 3
- 150000002191 fatty alcohols Chemical class 0.000 abstract description 3
- 150000001335 aliphatic alkanes Chemical class 0.000 abstract 2
- 102000011842 Serrate-Jagged Proteins Human genes 0.000 abstract 1
- 108010036039 Serrate-Jagged Proteins Proteins 0.000 abstract 1
- 230000008020 evaporation Effects 0.000 abstract 1
- 238000001704 evaporation Methods 0.000 abstract 1
- 239000004615 ingredient Substances 0.000 abstract 1
- APLHEOBEIBHCHW-YQEJDHNASA-N Selagine Natural products O=C1NC2=C([C@]3(N)/C(=C/C)/[C@@H](CC(C)=C3)C2)C=C1 APLHEOBEIBHCHW-YQEJDHNASA-N 0.000 description 24
- 238000000926 separation method Methods 0.000 description 10
- 239000003814 drug Substances 0.000 description 9
- 208000024827 Alzheimer disease Diseases 0.000 description 7
- 238000010262 high-speed countercurrent chromatography Methods 0.000 description 7
- 206010039966 Senile dementia Diseases 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 238000004128 high performance liquid chromatography Methods 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- BFKJFAAPBSQJPD-UHFFFAOYSA-N tetrafluoroethene Chemical group FC(F)=C(F)F BFKJFAAPBSQJPD-UHFFFAOYSA-N 0.000 description 5
- 238000005303 weighing Methods 0.000 description 5
- 230000001713 cholinergic effect Effects 0.000 description 4
- 238000002955 isolation Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 102100033639 Acetylcholinesterase Human genes 0.000 description 3
- 108010022752 Acetylcholinesterase Proteins 0.000 description 3
- 206010012289 Dementia Diseases 0.000 description 3
- 230000002411 adverse Effects 0.000 description 3
- 210000004556 brain Anatomy 0.000 description 3
- 230000019771 cognition Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 229960001685 tacrine Drugs 0.000 description 3
- YLJREFDVOIBQDA-UHFFFAOYSA-N tacrine Chemical compound C1=CC=C2C(N)=C(CCCC3)C3=NC2=C1 YLJREFDVOIBQDA-UHFFFAOYSA-N 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 241001504070 Huperzia Species 0.000 description 2
- 229960004373 acetylcholine Drugs 0.000 description 2
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 2
- 229930013930 alkaloid Natural products 0.000 description 2
- 150000003797 alkaloid derivatives Chemical class 0.000 description 2
- -1 biology Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000000544 cholinesterase inhibitor Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000004064 dysfunction Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- ASUTZQLVASHGKV-JDFRZJQESA-N galanthamine Chemical compound O1C(=C23)C(OC)=CC=C2CN(C)CC[C@]23[C@@H]1C[C@@H](O)C=C2 ASUTZQLVASHGKV-JDFRZJQESA-N 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 238000000622 liquid--liquid extraction Methods 0.000 description 2
- 230000015654 memory Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- GGRLKHMFMUXIOG-UHFFFAOYSA-M 2-acetyloxyethyl(trimethyl)azanium;hydroxide Chemical compound [OH-].CC(=O)OCC[N+](C)(C)C GGRLKHMFMUXIOG-UHFFFAOYSA-M 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 208000000044 Amnesia Diseases 0.000 description 1
- 108010053652 Butyrylcholinesterase Proteins 0.000 description 1
- 102000017927 CHRM1 Human genes 0.000 description 1
- 108010009685 Cholinergic Receptors Proteins 0.000 description 1
- 102100032404 Cholinesterase Human genes 0.000 description 1
- 101150073075 Chrm1 gene Proteins 0.000 description 1
- 241001090156 Huperzia serrata Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 208000026139 Memory disease Diseases 0.000 description 1
- UQOFGTXDASPNLL-XHNCKOQMSA-N Muscarine Chemical compound C[C@@H]1O[C@H](C[N+](C)(C)C)C[C@H]1O UQOFGTXDASPNLL-XHNCKOQMSA-N 0.000 description 1
- 206010029350 Neurotoxicity Diseases 0.000 description 1
- PIJVFDBKTWXHHD-UHFFFAOYSA-N Physostigmine Natural products C12=CC(OC(=O)NC)=CC=C2N(C)C2C1(C)CCN2C PIJVFDBKTWXHHD-UHFFFAOYSA-N 0.000 description 1
- 206010044221 Toxic encephalopathy Diseases 0.000 description 1
- 102000034337 acetylcholine receptors Human genes 0.000 description 1
- 229940022698 acetylcholinesterase Drugs 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000006933 amyloid-beta aggregation Effects 0.000 description 1
- 239000002260 anti-inflammatory agent Substances 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- WJJMNDUMQPNECX-UHFFFAOYSA-N dipicolinic acid Chemical compound OC(=O)C1=CC=CC(C(O)=O)=N1 WJJMNDUMQPNECX-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 230000003181 encephalopathic effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000001076 estrogenic effect Effects 0.000 description 1
- STVYPZZQOXPZBX-UHFFFAOYSA-N ethanol;hexane;hydrate Chemical compound O.CCO.CCCCCC STVYPZZQOXPZBX-UHFFFAOYSA-N 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 229960003980 galantamine Drugs 0.000 description 1
- 210000004884 grey matter Anatomy 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- 230000007087 memory ability Effects 0.000 description 1
- 230000006984 memory degeneration Effects 0.000 description 1
- 230000006386 memory function Effects 0.000 description 1
- 208000023060 memory loss Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 206010028417 myasthenia gravis Diseases 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 230000007135 neurotoxicity Effects 0.000 description 1
- 231100000228 neurotoxicity Toxicity 0.000 description 1
- 239000002858 neurotransmitter agent Substances 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 229960001697 physostigmine Drugs 0.000 description 1
- PIJVFDBKTWXHHD-HIFRSBDPSA-N physostigmine Chemical compound C12=CC(OC(=O)NC)=CC=C2N(C)[C@@H]2[C@@]1(C)CCN2C PIJVFDBKTWXHHD-HIFRSBDPSA-N 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 239000000018 receptor agonist Substances 0.000 description 1
- 229940044601 receptor agonist Drugs 0.000 description 1
- 230000000452 restraining effect Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 210000000225 synapse Anatomy 0.000 description 1
- 231100001274 therapeutic index Toxicity 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Nitrogen Condensed Heterocyclic Rings (AREA)
Abstract
The present invention relates to a preparing method for separating huperzine A with high purity from huperzine containing plant huperzine serrate by using high-speed countercurrent chromatograph (HSCCC), which comprises a solvent system for preparing a top phase of solid phase and a bottom phase of mobile phase. The solvent system comprises normal alkanes of normal heptane, hexane or normal pentane, fatty alcohols of methanol, etanol, normal butyl alcohol or acetone, and water, and the three parts have the following proportions by volume: 1 to 5 portions of normal alkane, 0.1 to 2 portions of fatty alcohol, and 1 to 5 portions of water. The conditional temperature is 20 DEG C to 30 DEG C, a column is filled with solid phase, the rotary speed of a host machine is 600 to 1000 rpm, and the mobile phase is pumped into the column at a flow speed of 0.5 to 4 mL/min. After the system shows a state of dynamic balance, sample introduction is carried out from a sampling valve, and target ingredients are received according to a monitor. Crystallization is carried out after fraction, concentration and evaporation to dry, and monomers of huperzine A are finally obtained. The method is suitable for high-speed countercurrent chromatographs with various models and preparation of huperzine A with various contents, and has the characteristics of large fractional dose, high recovery rate and easy and convenient operation.
Description
Technical field
The present invention relates to adopt high-speed countercurrent chromatography (HSCCC) from the plants of Huperzia Herba Lycopodii serrati, to prepare the preparation method of the method, particularly a kind of high-purity huperzine A of natural alkaloid.
Background technology
Along with the aging of social population, the senile dementia sickness rate rises relatively, and the dementia morbidity of over-65s is 5%～10% at present, and still among increasing.According to statistics, the world today has more than 5,000 ten thousand the elderlys to suffer from various degree senile dementia.China more than 60 years old population expect 2010 and will account for 15.7% of total population, the dementia problem is the inevitable problem that we faced equally.Therefore seek the treatment senile dementia disease active drug oneself be subjected to the great attention of various countries, become the task of top priority.In the senile dementia 50%～60% be Alzheimer (Alzheimer ' s Disease, AD), it be a kind of be the primary grey matter encephalopathic of principal feature with memory loss and cognition dysfunction.Clinical manifestation is a progressive dementia, and its cause of disease is still indeterminate, and hypothesis has multiple, as neurotransmitter defective, inflammation, radical damage, amyloid, neurotoxicity, hormonoprivia, apoptosis etc.The purpose of treatment AD is to improve cognition dysfunction, improves patient's viability.The seventies, research mainly concentrates on the relation of vagusstoff and AD; Mainly be the influence of research anticholinesterase to AD the eighties; The nineties, research not only has acetyl choline receptor agonists, has also comprised oestrogenic hormon, antiphlogistic drug, has influenced the medicine of Radical Metabolism and the medicine of inhibition amyloid beta deposition.Up to the present, the clinical treatment of AD remains a global problem that remains to be broken through but for various reasons.In recent years, Chinese scholars is being done a large amount of fundamental researchs and clinical application research aspect the natural drug effective constituent treatment senile dementia using, because cholinergic system and people's study, memory function is closely-related in the brain.The early stage brain cholinergic system of discovering patient AD has been subjected to infringement, causes the decline of cynapse position acetyl choline content, thereby patient's study, memory capability exerted an influence.Therefore, at the acetyl choline content that improves in patient's AD brain, promote the medicine of cholinergic nerve function to arise at the historic moment.The research of this class medicine now concentrates on anticholinesterase and muscarine M1 receptor stimulant.Selagine (huperzine A) is a kind of new alkaloid that is separated to from plants of Huperzia Herba Lycopodii serrati Huperzia serrata (Thunb.) Tre v. in recent years for the potent acetylcholinesterase of reversibility (AChE) inhibitor of China's initiative.The clinical study of carrying out at present shows that it is better than tacrine (tacrine) in cholinergic synapses to the restraining effect of AChE.Animal ethology studies show that selagine can improve the learning and memory ability of animal, and therapeutic index is higher than tacrine, Physostigmine and lycoremine.In the double blind trial in clinical 8 weeks by a definite date, oral 0.2mg/d, has 58% patient significantly to improve all tested memories, cognition and capacity parameter P＜0.05 at every day 2 times, and serious reaction do not occur, so selagine is the medicine of a kind of rising treatment AD.To senile dementia, simple dysmnesia and myasthenia gravis treatment have significant curative effect, and duration of efficacy is long, and no serious adverse reaction is the most effective medicine of domestic and international above-mentioned illness, is paid close attention to deeply, domestic approved listing.Be listed in one of inhibitor of s-generation Pseudocholinesterase in the world.
The structural formula of selagine is as follows:
Chemical name be (5R, 9R, 11E)-5-amino-11-ethidine-5,6,9,10-tetrahydrochysene-7-methyl-5,9-methylene ring suffering is (b) pyridine-2 (1H) ketone also
(Hup A) is extremely low at the occurring in nature amount for selagine, and contains a large amount of impurity, has a large amount of researchists exploring so far both at home and abroad and seeks ideal separation way.Once the someone utilized methods such as solvent extraction, silica gel column chromatography to separate, though can obtain few products, eventually because of time-consuming, yield is low and be difficult to large-scale production.And the synthetic product are difficult to obtain the ideal curative effect.The adverse current chromatogram method for separating and preparing can reach the purity height, and yield is high and be better than traditional separation methods such as column chromatography.High-speed countercurrent chromatography (High-SpeedCountercurrent Chromatography, HSCCC) be that a kind of successive that grew up in nearly 30 years need not the efficient of any solid support, liquid liquid distribution chromatography isolation technique fast, its principle is the centrifugal force that utilizes borded pile to produce when planetary motion, immiscible two-phase is constantly mixed, keep a phase (solid phase) wherein simultaneously, utilize constant flow pump to import another phase (moving phase) continuously, the solute that enters borded pile with moving phase distributes between two-phase repeatedly, press the order of partition ratio, quilt is wash-out successively.The elder generation that allocation proportion is big in moving phase is by wash-out, otherwise, solid mutually in big back of allocation proportion by wash-out.The variety of issue that it has avoided solid state adhesion body or carrier to bring---sample is adsorbed easily, loss and sex change, when other liquid chromatography technology amount of being prepared is separated, its allocative efficiency can obviously reduce, solvent-oil ratio is big, HSCCC guarantees higher peak type resolution, fractional dose is big, sample is lossless, the rate of recovery is high, isolating environment relaxes, and saves solvent.High-speed counter-current chromatograph can directly advance a large amount of crude extract samples or synthetic mixture, separating resulting can reach quite high purity, even can directly connect instruments such as mass spectrograph, be widely used in the preparation separation and the purifying of field chemical substances such as biology, medicine, environmental protection.Therefore, HSCCC has easy and simple to handle, and the theoretical rate of recovery is 100%, favorable reproducibility and separation efficiency height, the characteristics that fractional dose is bigger.
Summary of the invention
The technical problem to be solved in the present invention is the defective that overcomes traditional isolation technique, adopts high-speed countercurrent chromatography (HSCCC), and a kind of preparation method of high-purity huperzine A is provided.This method has rate of recovery height, the big and advantages of simple operation of fractional dose.
Technical scheme of the present invention comprises the steps:
Adopt the high speed adverse current chromatogram preparation method, it comprises that preparation constitutes the solid phase, the solvent system of moving phase: be filled with the counter current chromatograph pillar with stationary phase earlier, regulate engine speed, moving phase is pumped in the post; After treating that whole system is set up running balance, by the sampling valve sample introduction; According to monitor uv-spectrogram receiving target composition.At first select the two combined of a solvent system, solvent system is made of three components, the A component can be selected from normal paraffins such as normal heptane, normal hexane, Skellysolve A, the B component can be selected from Fatty Alcohol(C12-C14 and C12-C18) and aliphatic ketones such as methyl alcohol, ethanol, propyl carbinol, acetone, the C component is a water, preferentially selects normal hexane-n-butanol-water system.
According to solubility constant, do not destroying under the system equilibrated situation, regulate A, B, C, the volume ratio of three components is followed successively by: 1-5: 0.1-2: 1-5.For example 2: 1: 2; 5: 1: 4; 20: 8: 20 etc.; Being stationary phase mutually on the above-mentioned system, is moving phase down mutually.And obtain the pure product of huperzine through flash liberation, purity is more than 99%.
Experiment condition is fit to temperature 20-30 ℃, and in the said temperature scope, when temperature was higher, appearance time slightly shifted to an earlier date, and separation efficiency changes little, and peak shape is not had much influences.
Engine speed and flow velocity need be controlled within the specific limits, by volume solvent system are placed separating funnel, shake up standing demix.Behind the ready to balance certain hour, upper and lower phase is separated, adopt the TBE-300A high-speed counter-current chromatograph, this type column volume is 300ml, and the sample introduction circle is 20ml, is furnished with AKTA Prime pump, monitor and registering instrument.Before the sample introduction, earlier be filled with whole pillar with stationary phase, the adjustment engine speed is 600-1000rpm, with the flow velocity of 0.5-4mL/min moving phase is pumped in the post.
Since the unstable of selagine, the target component that separates and collect, and the concentrated evaporate to dryness of horse back cut carries out crystallization and obtains monomer.
The inventive method has adopted the high speed adverse current chromatogram isolation technique, be a kind of successive need not any solid support efficiently, liquid liquid distribution chromatography isolation technique fast, it has overcome solid state adhesion body or carrier irreversible adsorption, loss and sex change, make separated object rate of recovery height, theory reaches 100%, and the present invention is actual to be reached more than 95%.Because adopt preferred solvent system, the processing condition of control experiment condition temperature, adjustment engine speed and flow velocity can high efficiencyly be separated, and obtain high-purity huperzine A monomer (reaching more than 99%) again.The inventive method is fit to separate the highly purified selagine monomer of preparation from the different content selagine crude product of various technology approach preparations.The counter current chromatograph that is used in various models separates preparation selagine monomer, can directly advance slightly get sample in a large number product or synthetic mixture, separating resulting can reach quite high purity, guarantee higher peak type resolution, fractional dose is big, the rate of recovery is high, isolating environment relaxes, save solvent, easy and simple to handle.
Embodiment
Embodiment 1
Choose normal hexane-n-butanol-water system separation and purification selagine on half preparation type counter current chromatograph.By 5: 1: 5 volume ratios above-mentioned solvent composition is disposed in the separating funnel earlier, shakes up the back standing demix.Ready to balance after for some time separates upper and lower phase.Adopt half preparation type counter current chromatograph, be furnished with the tetrafluoroethylene post, 20ml sampling valve, column volume are 200ml, AKTA Prime pump, detector and registering instrument.Taking by weighing 200mg selagine study is dissolved in the 20ml stationary phase stand-by.Before the sample introduction, earlier be filled with whole pillar with stationary phase, the adjustment engine speed is 950rpm, with the flow velocity of 3.0ml/min moving phase is pumped in the post; After treating that whole system is set up running balance, by the sampling valve sample introduction; Then according to the detector uv-spectrogram, the receiving target composition.Obtain the selagine fraction, carry out crystallization behind the concentrated evaporate to dryness, its HPLC purity reaches about 99%.
Embodiment 2
Choose normal hexane-methanol-water system separation and purification selagine on half preparation type counter current chromatograph.Experiment condition is fit to 25 ℃ of temperature, by 2: 1: 4 volume ratios above-mentioned solvent composition is disposed in the separating funnel earlier, shakes up the back standing demix.Ready to balance after for some time separates upper and lower phase.Adopt half preparation type counter current chromatograph, be furnished with the tetrafluoroethylene post, 20ml sampling valve, column volume are 200ml, AKTA Prime pump, detector and registering instrument.Taking by weighing 150mg selagine study is dissolved in the 20ml stationary phase stand-by.Before the sample introduction, earlier be filled with whole pillar with stationary phase, the adjustment engine speed is 750rpm, with the flow velocity of 4.0ml/min moving phase is pumped in the post; After treating that whole system is set up running balance, by the sampling valve sample introduction; Then according to the detector uv-spectrogram, the receiving target composition.Obtain the selagine fraction, carry out crystallization behind the concentrated evaporate to dryness, its HPLC purity reaches about 99%.
Embodiment 3
Choose normal hexane-ethanol-water system separation and purification selagine on half preparation type counter current chromatograph.Experiment condition is fit to 30 ℃ of temperature, by 2: 1: 4 volume ratios above-mentioned solvent composition is disposed in the separating funnel earlier, shakes up the back standing demix.Ready to balance after for some time separates upper and lower phase.Adopt half preparation type counter current chromatograph, be furnished with the tetrafluoroethylene post, 20ml sampling valve, column volume are 200ml, AKTA Prime pump, detector and registering instrument.Taking by weighing 150mg selagine study is dissolved in the 20ml stationary phase stand-by.Before the sample introduction, earlier be filled with whole pillar with stationary phase, the adjustment engine speed is 750rpm, with the flow velocity of 4.0ml/min moving phase is pumped in the post; After treating that whole system is set up running balance, by the sampling valve sample introduction; Then according to the detector uv-spectrogram, the receiving target composition.Obtain the selagine fraction, carry out crystallization behind the concentrated evaporate to dryness, its HPLC purity reaches about 99%.
Embodiment 4
Choose normal heptane-acetone-water system separation and purification selagine on half preparation type counter current chromatograph.Experiment condition is fit to 22 ℃ of temperature, by 4: 1: 3 volume ratios above-mentioned solvent composition is disposed in the separating funnel earlier, shakes up the back standing demix.Ready to balance after for some time separates upper and lower phase.Adopt half preparation type counter current chromatograph, be furnished with the tetrafluoroethylene post, 20ml sampling valve, column volume are 200ml, AKTA Prime pump, detector and registering instrument.Taking by weighing 150mg selagine study is dissolved in the 20ml stationary phase stand-by.Before the sample introduction, earlier be filled with whole pillar with stationary phase, the adjustment engine speed is 650rpm, with the flow velocity of 3.0ml/min moving phase is pumped in the post; After treating that whole system is set up running balance, by the sampling valve sample introduction; Then according to the detector uv-spectrogram, the receiving target composition.Obtain the selagine fraction, carry out crystallization behind the concentrated evaporate to dryness, its HPLC purity reaches about 99%.
Embodiment 5
Choose Skellysolve A-methanol-water system separation and purification selagine on half preparation type counter current chromatograph.Experiment condition is fit to 20 ℃ of temperature, by 2: 1: 1 volume ratios above-mentioned solvent composition is disposed in the separating funnel earlier, shakes up the back standing demix.Ready to balance after for some time separates upper and lower phase.Adopt half preparation type counter current chromatograph, be furnished with the tetrafluoroethylene post, 20ml sampling valve, column volume are 200ml, AKTA Prime pump, detector and registering instrument.Taking by weighing 180mg selagine study is dissolved in the 20ml stationary phase stand-by.Before the sample introduction, earlier be filled with whole pillar with stationary phase, the adjustment engine speed is 1000rpm, with the flow velocity of 3.0ml/min moving phase is pumped in the post; After treating that whole system is set up running balance, by the sampling valve sample introduction; Then according to the detector uv-spectrogram, the receiving target composition.Obtain the selagine fraction, carry out crystallization behind the concentrated evaporate to dryness, its HPLC purity reaches about 99%.
Claims (7)
1. the preparation method of a high-purity huperzine A, it comprises that preparation constitutes the solid phase, the solvent system of moving phase: be filled with the counter current chromatograph pillar with stationary phase earlier, regulate engine speed, moving phase is pumped in the post; After treating that whole system is set up running balance, by the sampling valve sample introduction; According to uv-spectrogram monitor receiving target composition, it is characterized in that: described solvent system is by normal heptane, normal hexane, Skellysolve A and methyl alcohol, ethanol, propyl carbinol or acetone, form with water, the volume ratio of three components is followed successively by: 1-5: 0.1-2: 1-5, on be the solid phase mutually, is moving phase mutually down.
2. the preparation method of a kind of high-purity huperzine A according to claim 1, it is characterized in that: described engine speed is 600-1000rpm, the flow velocity that described moving phase pumps in the post is 0.5-4mL/min.
3. the preparation method of a kind of high-purity huperzine A according to claim 1 and 2 is characterized in that; Described solvent system is normal hexane-n-butanol-water, and the volume ratio of three components was followed successively by 5: 1: 5, engine speed 950rpm, and the flow velocity that moving phase pumps in the post is 3ml/min.
4. the preparation method of a kind of high-purity huperzine A according to claim 1 and 2, it is characterized in that: described solvent system is normal heptane-acetone-water, the consumption volume ratio is followed successively by: 4: 1: 3, described engine speed 650rpm, the flow velocity that described moving phase pumps in the post is 3ml/min.
5. the preparation method of a kind of high-purity huperzine A according to claim 1 and 2, it is characterized in that: described solvent system is Skellysolve A-methanol-water, and the consumption volume ratio is followed successively by: 2: 1: 1.
6. the preparation method of a kind of high-purity huperzine A according to claim 1 and 2, it is characterized in that: described solvent system is normal hexane-alcohol-water, the consumption volume ratio is followed successively by: 2: 1: 4, described engine speed 750rpm, the flow velocity that described moving phase pumps in the post is 4ml/min.
7. according to the preparation method of the described a kind of high-purity huperzine A of arbitrary claim in the claim 1 to 2, it is characterized in that: described solvent system is normal hexane-methanol-water, and the consumption volume ratio was followed successively by 2: 1: 4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410052688 CN1264822C (en) | 2004-07-09 | 2004-07-09 | Process for preparing high purity huperzine A |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410052688 CN1264822C (en) | 2004-07-09 | 2004-07-09 | Process for preparing high purity huperzine A |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1587260A CN1587260A (en) | 2005-03-02 |
| CN1264822C true CN1264822C (en) | 2006-07-19 |
Family
ID=34602559
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200410052688 Expired - Lifetime CN1264822C (en) | 2004-07-09 | 2004-07-09 | Process for preparing high purity huperzine A |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1264822C (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102627605B (en) * | 2012-03-22 | 2014-09-10 | 中国科学院上海药物研究所 | Huperzine A polymorph, its preparation method, medicinal composition containing huperzine A polymorph and its application |
| CN105085523B (en) * | 2015-09-02 | 2017-03-08 | 中南大学 | A kind of high speed adverse current chromatogram separates the method preparing high-purity huperzine third |
-
2004
- 2004-07-09 CN CN 200410052688 patent/CN1264822C/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| CN1587260A (en) | 2005-03-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100497344C (en) | Preparation of high-purity ginkgolide | |
| CN101864191B (en) | Preparation method of high-purity monascus pigment component | |
| CN105669631B (en) | A kind of potentilla plants extract and the method for therefrom separating four kinds of tanninses compounds | |
| CN102030733B (en) | The preparation method of a kind of high purity costuslactone and dehydro-α-curcumene | |
| CN101607956B (en) | Method for splitting chiral medicine of hydrochloric acid lomefloxacin | |
| CN102234305A (en) | Method for preparing high-purity anemoside B4 | |
| CN110194758A (en) | A method of the fast separating and purifying Aristolochic Acid compound from caulis aristologhiae manshuriensis | |
| CN1394870A (en) | Method for separating and purifying tanshinone | |
| CN101565437B (en) | Separation and preparation method of patuletin-3-O-glucoside and astragalin | |
| CN1264822C (en) | Process for preparing high purity huperzine A | |
| CN100500689C (en) | Method of separating and preparing wild jujube seed saponin | |
| CN107629140A (en) | A kind of method of ionic liquid double-aqueous phase system extraction Goods-Flow Plan | |
| CN101805352B (en) | Method for preparing eriocalyxin B | |
| CN1261434C (en) | Method for separating high-purity galanthamine from short-tube lycoris crude extract | |
| CN1781907A (en) | Method for producing high purity huperzine A | |
| CN102532147B (en) | Preparation method of high purity dictamnine monomer | |
| CN100526329C (en) | Production of high-purity peiminine and fritimine | |
| CN1176887C (en) | Separating prepn process of carotenol | |
| CN109369382B (en) | A method for preparing ginkgolic acids | |
| CN1193001C (en) | Separation preparing method for aloe-emodin and parietic acid | |
| CN102167693B (en) | Method for separating and preparing alpha-terthienyl, thianthrene and p-terphenyl | |
| CN101759640B (en) | Method for preparing high purity benzoylmesaconine | |
| CN101070335A (en) | Process for preparing high-purity gentiopicrin | |
| CN1594261A (en) | Preparation method of high purity physcion and chrysophanol | |
| CN112552150B (en) | Method for preparing asarone monomer based on coordination effect |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: SHANGHAI TONGTIAN BIOTECHNOLOGY CO., LTD. Free format text: FORMER OWNER: TONGTIAN BIOCHEMICAL TECHNOLOGY CO., LTD., SHANGHAI Effective date: 20061201 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20061201 Address after: 201203 room 326, 301 Edison Road, Zhangjiang hi tech park, Shanghai Patentee after: SHANGHAI TAUTO BIOTECH Co.,Ltd. Address before: 200122, room 1311, Huarong Building, No. 1279 Pudong South Road, Shanghai, Pudong New Area Patentee before: SHANGHAI TAUTO BIOTECH Co.,Ltd. |
|
| C56 | Change in the name or address of the patentee |
Owner name: SHANGHAI TAUTO BIOTECH. CO., LTD. Free format text: FORMER NAME: SHANGHAI TONGTIAN BIOTECHNOLOGY CO., LTD. |
|
| CP01 | Change in the name or title of a patent holder |
Address after: 201203 room 326, 301 Edison Road, Zhangjiang hi tech park, Shanghai Patentee after: SHANGHAI TAUTO BIOTECH Co.,Ltd. Address before: 201203 room 326, 301 Edison Road, Zhangjiang hi tech park, Shanghai Patentee before: SHANGHAI TAUTO BIOTECH Co.,Ltd. |
|
| CP03 | Change of name, title or address |
Address after: Room 101-2, 102-1, 301, 301-1, 326 Edison Road, Pudong New Area, Shanghai, 201203 Patentee after: SHANGHAI TAUTO BIOTECH Co.,Ltd. Address before: 201203 room 326, 301 Edison Road, Zhangjiang hi tech park, Shanghai Patentee before: SHANGHAI TAUTO BIOTECH Co.,Ltd. |
|
| CP03 | Change of name, title or address | ||
| CX01 | Expiry of patent term |
Granted publication date: 20060719 |
|
| CX01 | Expiry of patent term |