CN1306960A - Method for extracting huperzine A as acetylcholinesterase depressant - Google Patents
Method for extracting huperzine A as acetylcholinesterase depressant Download PDFInfo
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- CN1306960A CN1306960A CN 00101469 CN00101469A CN1306960A CN 1306960 A CN1306960 A CN 1306960A CN 00101469 CN00101469 CN 00101469 CN 00101469 A CN00101469 A CN 00101469A CN 1306960 A CN1306960 A CN 1306960A
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- chloroform
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Abstract
The extraction process includes the steps of soaking medicine material in heated dilute acid; treating with dilute acid and alkali and extraction with chloroform; concentration. extraction and eliminating impurity with dilute alkali; back extraction with dilute acid; decoloring with active carbon; alkali regulation, chlorolform extraction and concentration to dry; recrystallization in acetone. It has the advantages of less organic solvent used, short production period, high yield and easy industry application.
Description
The present invention relates to a kind of new method for extracting of acetylcholinesterase depressant selagine.
Selagine (is called for short Hup A, structural formula is as follows) have another name called Fuding alkali, be from Huperziaceae stone araucaria feet added to a snake by an ignorant artist China firs (Herba Lycopodii serrati), to separate a kind of natural alkaloid that obtains, it is potent reversibility acetylcholinesterase (Ach E) inhibitor, degenerative brain disorder, simple dysmnesia and myasthenia gravis treatment are had significant curative effect, and duration of efficacy is long, no serious adverse reaction, be the present the most effective medicine of domestic and international above-mentioned illness, paid close attention to deeply.Because its therapeutic index height, and long action time have been listed in one of inhibitor of s-generation acetylcholinesterase in the world.But Hup A is extremely low at the occurring in nature amount, coexistence has a large amount of impurity with it, though someone once attempted to utilize the way of synthetic to obtain, but abandon because of the obvious no above-mentioned biologic activity of sintetics, have a large amount of researchists constantly inquiring into so far both at home and abroad for seeking comparatively ideal extraction and separation method.
Molecular formula: C
15H
18N
2O molecular weight: 242.32
Up to now, the production technique that can be used for selagine reluctantly is:
Acid solution fully soak Herba Lycopodii serrati xeraphium → alkali acid treatment chloroform extract → be condensed into fluid extract → silica gel column chromatography → heterogeneous solvent elution repeatedly and collect cut → be concentrated into dried → recrystallization ends until obtaining qualified product repeatedly.
Process characteristic:
1. a large amount of poisonous organic solvent methyl alcohol and chloroforms of using;
2. time-consuming (one-period needs 1 month approximately);
3. yield low (less than 5/1000000ths);
4. can not batch production production.
The new method for extracting that the purpose of this invention is to provide a kind of acetylcholinesterase depressant selagine.
In order to achieve the above object, the present invention takes following measures:
The extraction step of acetylcholinesterase depressant selagine is as follows:
1. the medicinal material diluted acid adds heat soaking: 0.5～1.5% tartrate fully soaked Herba Lycopodii serrati dry powder 12～18 hours under 50～60 ℃, and centrifugation gets supernatant liquor;
2. diluted acid alkaline purification chloroform extraction: with diluted alkaline ammoniacal liquor adjust pH, chloroform extracts three times repeatedly, concentrates and reclaims chloroform, concentrated solution should be 1/10～1/15 of original volume, strip with 0.5～1.5%NaOH, transfer alkali with ammoniacal liquor behind the combining extraction liquid, chloroform extracts three times again;
3. concentrate: 40 ℃～50 ℃ left and right sides vacuum concentration reclaim chloroform, become medicinal extract;
4. diluted alkaline abstraction impurity removal:, abandon water layer with the dilute ammonia solution equal proportion extraction concentrated solution secondary of pH8.0～8.2;
5. diluted acid is stripped: with the above-mentioned chloroform layer 4. of HCl aqueous solution equal proportion extraction of 0.2～0.5N, remove the pigment impurity that coexists with selagine, abandon chloroform layer;
6. activated carbon decolorizing: get the sour water extraction liquid, 10～20 times of dilute with waters, HCl transfers to pH1～2, activated carbon dosage stirs decolouring 5～10 minutes for 10～20% of the dilution front volume, filter colourless transparent liquid;
7. alkali accent, chloroform extraction and be concentrated into dried: destainer is transferred pH9～10 with strong aqua, and with the equal-volume chloroform extraction once, vacuum concentration reclaims chloroform to doing, the blush solid;
8. acetone recrystallization: add proper amount of acetone in above-mentioned red solid, reflux makes dissolving, after being cooled to room temperature,, get the off-white color solid in 0～10 ℃ of following crystallization, purity＞90%, after adding the minimum of chloroform dissolving again,, get white solid with the method acetone recrystallization, purity＞97% is with product purity＞99% after three crystallizations of method.
Advantage of the present invention:
1. significantly reduced the use of poisonous organic solvent;
2. the cycle is lacked (being generally 3～4 days);
3. yield height (greater than 5/100000ths);
4. be easy to batch production production.
Elaborate below in conjunction with embodiment:
The technological process of production of selagine:
The diluted acid heating is fully soaked Herba Lycopodii serrati xeraphium → diluted alkaline acid treatment chloroform extraction → concentrated → diluted alkaline abstraction impurity removal → diluted acid reextraction → activated carbon decolorizing → alkali accent, chloroform extraction and is concentrated into dried → acetone recrystallization → pure product.
Embodiment 1
The extraction step of selagine is as follows:
1. the medicinal material diluted acid adds heat soaking: 0.5% tartrate fully soaked Herba Lycopodii serrati dry powder 12 hours under 50 ℃, and centrifugation gets supernatant liquor.The common room temperature lixiviate of traditional technology is more than 36 hours, and is time-consuming.
2. diluted acid alkaline purification chloroform extraction: with diluted alkaline ammoniacal liquor adjust pH, chloroform extracts three times repeatedly, concentrates to reclaim chloroform, and concentrated solution should be 1/10 of original volume, strips with 0.5%NaOH again, transfers alkali with ammoniacal liquor behind the combining extraction liquid, and chloroform extracts three times again.
3. concentrate: 40 ℃ of left and right sides vacuum concentration reclaim chloroform, become medicinal extract.
4. diluted alkaline abstraction impurity removal:, abandon water layer with the dilute ammonia solution equal proportion extraction concentrated solution secondary of pH8.0.
5. diluted acid is stripped: with the above-mentioned chloroform layer 4. of HCl aqueous solution equal proportion extraction of 0.2N, remove the pigment impurity that coexists with selagine, abandon chloroform layer.One of gordian technique that this step is a this patent.
6. activated carbon decolorizing: get the sour water extraction liquid, 10 times of dilute with waters, HCl transfers to pH1.0, and activated carbon dosage is 10% of the dilution front volume.Stir decolouring 5 minutes, filter colourless transparent liquid.
7. alkali accent, chloroform extraction and be concentrated into dried: destainer is transferred pH9.0 with strong aqua, and with the equal-volume chloroform extraction once, vacuum concentration reclaims chloroform to doing, the blush solid.
8. acetone recrystallization: add proper amount of acetone in above-mentioned red solid, reflux makes dissolving, be cooled to room temperature after, in 0 ℃ of following crystallization, the off-white color solid, purity＞90%.After adding the minimum of chloroform dissolving again,, get white solid, purity＞97% with the method acetone recrystallization.With product purity＞99% after three crystallizations of method.Finishing above-mentioned whole process generally needs 3～4 days, and calculating recovery rate by medicinal material is about 5/100000ths.
Embodiment 2
The selagine extraction step is as follows:
1. the medicinal material diluted acid adds heat soaking: 1% tartrate fully soaked Herba Lycopodii serrati dry powder 15 hours under 55 ℃, and centrifugation gets supernatant liquor;
2. diluted acid alkaline purification chloroform extraction: with diluted alkaline ammoniacal liquor adjust pH, chloroform extracts three times repeatedly, concentrates to reclaim chloroform, and concentrated solution should be 1/12 of original volume, strips with 1%NaOH again, transfers alkali with ammoniacal liquor behind the combining extraction liquid, and chloroform extracts three times again;
3. concentrate: 45 ℃ of left and right sides vacuum concentration reclaim chloroform, become medicinal extract;
4. diluted alkaline abstraction impurity removal:, abandon water layer with the dilute ammonia solution equal proportion extraction concentrated solution secondary of pH8.0;
5. diluted acid is stripped: with the above-mentioned chloroform layer 4. of HCl aqueous solution equal proportion extraction of 0.2N, remove the pigment impurity that coexists with selagine, abandon chloroform layer;
6. activated carbon decolorizing: get the sour water extraction liquid, 15 times of dilute with waters, HCl transfers to pH1.5, and activated carbon dosage is 15% of the dilution front volume.Stir decolouring 8 minutes, filter colourless transparent liquid;
7. alkali accent, chloroform extraction and be concentrated into dried: destainer is transferred pH9.5 with strong aqua, and with the equal-volume chloroform extraction once, vacuum concentration reclaims chloroform to doing, the blush solid;
8. acetone recrystallization: add proper amount of acetone in above-mentioned blush solid, reflux makes dissolving, after being cooled to room temperature,, get the off-white color solid in 5 ℃ of following crystallizations, purity＞90%, after adding the minimum of chloroform dissolving again,, get white solid with the method acetone recrystallization, purity＞97% is with product purity＞99% after three crystallizations of method;
Embodiment 3
The extraction step of selagine is as follows:
1. the medicinal material diluted acid adds heat soaking: 1.5% tartrate fully soaked Herba Lycopodii serrati dry powder 18 hours under 60 ℃, and centrifugation gets supernatant liquor.The common room temperature lixiviate of traditional technology is more than 36 hours, and is time-consuming.
2. diluted acid alkaline purification chloroform extraction: with diluted alkaline ammoniacal liquor adjust pH, chloroform extracts three times repeatedly, concentrates to reclaim chloroform, and concentrated solution should be 1/15 of original volume, strips with 1.5%NaOH again, transfers alkali with ammoniacal liquor behind the combining extraction liquid, and chloroform extracts three times again.
3. concentrate: 50 ℃ of left and right sides vacuum concentration reclaim chloroform, become medicinal extract.
4. diluted alkaline abstraction impurity removal:, abandon water layer with the dilute ammonia solution equal proportion extraction concentrated solution secondary of pH8.2.
5. diluted acid is stripped: with the above-mentioned chloroform layer 4. of HCl aqueous solution equal proportion extraction of 0.5N, remove the pigment impurity that coexists with selagine, abandon chloroform layer.One of gordian technique that this step is a this patent.
6. activated carbon decolorizing: get the sour water extraction liquid, 20 times of dilute with waters, HCl transfers to pH2, and activated carbon dosage is 20% of the dilution front volume.Stir decolouring 10 minutes, filter colourless transparent liquid.
7. alkali accent, chloroform extraction and be concentrated into dried: destainer is transferred pH10 with strong aqua, and with the equal-volume chloroform extraction once, vacuum concentration reclaims chloroform to doing, the blush solid.
8. acetone recrystallization: add proper amount of acetone in above-mentioned blush solid, reflux makes dissolving, be cooled to room temperature after, in 10 ℃ of following crystallizations, the off-white color solid, purity＞90%.After adding the minimum of chloroform dissolving again,, get white solid, purity＞97% with the method acetone recrystallization.With product purity＞99% after three crystallizations of method.Finishing above-mentioned whole process generally needs 3～4 days, and calculating recovery rate by medicinal material is about 5/100000ths.
Product quality analysis:
(1) high performance liquid chromatography (HPLC) is analyzed:
Day island proper Tianjin LC-10A type HPLC instrument, moving phase: methyl alcohol: distilled water: trolamine=500: 500: 1, equal proportion wash-out, flow are 0.8ml/min; 35 ℃ of column temperatures; Chromatographic column is ODS-C
18(150mm * 6mmid.); Sensitivity is 0.01aufs; The ultraviolet detection wavelength is 310nm; Qualitative with reference to the reference substance retention time, external standard method is quantitative.The HPLC analytical results, products obtained therefrom purity＞99%.
(2) UV scanning: adopt day visible ultraviolet scanner of island proper Tianjin UV-2110 type, 200～400 scope interscans, two absorption peaks are respectively 231nm and 313nm.
(3) fusing point test:
Adopt the pharmacopeia prescriptive procedure, getting the product fusing point is 229 ℃.
Claims (2)
1. the new method for extracting of an acetylcholinesterase depressant selagine is characterized in that its step is as follows:
1. the medicinal material diluted acid adds heat soaking: 0.5～1.5% tartrate fully soaked Herba Lycopodii serrati dry powder 12～18 hours under 50～60 ℃, and centrifugation gets supernatant liquor;
2. diluted acid alkaline purification chloroform extraction: with diluted alkaline ammoniacal liquor adjust pH, chloroform extracts three times repeatedly, concentrates and reclaims chloroform, concentrated solution should be 1/10～1/15 of original volume, strip with 0.5～1.5%NaOH, transfer alkali with ammoniacal liquor behind the combining extraction liquid, chloroform extracts three times again;
3. concentrate: 40 ℃～50 ℃ left and right sides vacuum concentration reclaim chloroform, become medicinal extract;
4. diluted alkaline abstraction impurity removal:, abandon water layer with the dilute ammonia solution equal proportion extraction concentrated solution secondary of pHg.0～8.2;
5. diluted acid is stripped: with the above-mentioned chloroform layer 4. of HCl aqueous solution equal proportion extraction of 0.2～0.5N, remove the pigment impurity that coexists with selagine, abandon chloroform layer;
6. activated carbon decolorizing: get the sour water extraction liquid, 10～20 times of dilute with waters, HCl transfers to pH1～2, activated carbon dosage stirs decolouring 5～10 minutes for 10～20% of the dilution front volume, filter colourless transparent liquid;
7. alkali accent, chloroform extraction and be concentrated into dried: destainer is transferred pH9～10 with strong aqua, and with the equal-volume chloroform extraction once, vacuum concentration reclaims chloroform to doing, the blush solid;
8. acetone recrystallization: add proper amount of acetone in above-mentioned blush solid, reflux makes dissolving, after being cooled to room temperature,, get the off-white color solid in 0～10 ℃ of following crystallization, purity＞90%, after adding the minimum of chloroform dissolving again,, get white solid with the method acetone recrystallization, purity＞97% is with product purity＞99% after three crystallizations of method.
2. the new method for extracting of a kind of acetylcholinesterase depressant selagine according to claim 1 is characterized in that its step is as follows:
1. the medicinal material diluted acid adds heat soaking: 1% tartrate fully soaked Herba Lycopodii serrati dry powder 15 hours under 55 ℃, and centrifugation gets supernatant liquor;
2. diluted acid alkaline purification chloroform extraction: with diluted alkaline ammoniacal liquor adjust pH, chloroform extracts three times repeatedly, concentrates to reclaim chloroform, and concentrated solution should be 1/12 of original volume, strips with 1%NaOH again, transfers alkali with ammoniacal liquor behind the combining extraction liquid, and chloroform extracts three times again;
3. concentrate: 45 ℃ of left and right sides vacuum concentration reclaim chloroform, become medicinal extract;
4. diluted alkaline abstraction impurity removal:, abandon water layer with the dilute ammonia solution equal proportion extraction concentrated solution secondary of pH8.0;
5. diluted acid is stripped: with the above-mentioned chloroform layer 4. of HCl aqueous solution equal proportion extraction of 0.2N, remove
5. diluted acid is stripped: with the above-mentioned chloroform layer 4. of HCl aqueous solution equal proportion extraction of 0.2N, remove the pigment impurity that coexists with selagine, abandon chloroform layer;
6. activated carbon decolorizing: get the sour water extraction liquid, 15 times of dilute with waters, HCl transfers to pH1.5, and activated carbon dosage is 15% of the dilution front volume.Stir decolouring 8 minutes, filter colourless transparent liquid;
7. alkali accent, chloroform extraction and be concentrated into dried: destainer is transferred pH9.5 with strong aqua, and with the equal-volume chloroform extraction once, vacuum concentration reclaims chloroform to doing, the blush solid;
8. acetone recrystallization: add proper amount of acetone in above-mentioned blush solid, reflux makes dissolving, after being cooled to room temperature,, get the off-white color solid in 5 ℃ of following crystallizations, purity＞90%, after adding the minimum of chloroform dissolving again,, get white solid with the method acetone recrystallization, purity＞97% is with product purity＞99% after three crystallizations of method;
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN00101469A CN1101381C (en) | 2000-01-21 | 2000-01-21 | Method for extracting huperzine A as acetylcholinesterase depressant |
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| CN00101469A CN1101381C (en) | 2000-01-21 | 2000-01-21 | Method for extracting huperzine A as acetylcholinesterase depressant |
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| CN1306960A true CN1306960A (en) | 2001-08-08 |
| CN1101381C CN1101381C (en) | 2003-02-12 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100383122C (en) * | 2006-06-15 | 2008-04-23 | 河南太龙药业股份有限公司 | Process of extracting lycopdine A from plant |
| CN100441200C (en) * | 2004-11-26 | 2008-12-10 | 谢德隆 | China fir extracts with lycopodine A and B composition and their preparing method |
| CN103951618A (en) * | 2014-05-09 | 2014-07-30 | 自贡天健生物科技有限公司 | Huperzine A crystal, and preparation method and application thereof |
| CN110078667A (en) * | 2019-04-28 | 2019-08-02 | 云南汉德生物技术有限公司 | A method of extracting huperzine |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1035503A (en) * | 1989-03-11 | 1989-09-13 | 军事医学科学院毒物药物研究所 | Productive technology for real cholinesterase inhibitor-fuding alkali |
-
2000
- 2000-01-21 CN CN00101469A patent/CN1101381C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100441200C (en) * | 2004-11-26 | 2008-12-10 | 谢德隆 | China fir extracts with lycopodine A and B composition and their preparing method |
| CN100383122C (en) * | 2006-06-15 | 2008-04-23 | 河南太龙药业股份有限公司 | Process of extracting lycopdine A from plant |
| CN103951618A (en) * | 2014-05-09 | 2014-07-30 | 自贡天健生物科技有限公司 | Huperzine A crystal, and preparation method and application thereof |
| CN110078667A (en) * | 2019-04-28 | 2019-08-02 | 云南汉德生物技术有限公司 | A method of extracting huperzine |
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| CN1101381C (en) | 2003-02-12 |
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