CN102127089A - Method for extracting camptothecin - Google Patents
Method for extracting camptothecin Download PDFInfo
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- CN102127089A CN102127089A CN2010105294412A CN201010529441A CN102127089A CN 102127089 A CN102127089 A CN 102127089A CN 2010105294412 A CN2010105294412 A CN 2010105294412A CN 201010529441 A CN201010529441 A CN 201010529441A CN 102127089 A CN102127089 A CN 102127089A
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Abstract
The invention relates to the field of medicament preparation, in particular to a method for extracting camptothecin, aiming at solving the problems of low conversion rate, low yield and poor product quality in the preparation of the camptothecin in the prior art. The method comprises the steps of: step 1, extracting by an alcohol extraction method; step 2, further extracting the extracted substances obtained in the step 1 by an alkali extraction method; and step 3, carrying out over-column purification. in the method, the production efficiency and conversion rate are greatly improved, and more active substances in raw material are promoted to be converted into the camptothecin, so that the product quality is further improved, the product purity is high, and the method is suitable for industrialized production.
Description
Technical field
The present invention relates to the Chinese medicine extraction of effective components, be specifically related to the extracting method of extraction camptothecine from camplotheca acuminata.
Background technology
Camptothecine English name: Camptothecin, chemical name: 1H-Pyrano[34:6,7] indolizino[1,2-6] puinoline-3,14 (4H, 12H-)-dione, 4-ethyl-4-hydroxy-(s)-, molecular formula: C20H16N2O4 molecular weight: 348.34
Source: fruit, root, the leaf of Nyssaceae plant camptotheca acuminata Camptotheca acuminata Decne.
(camptothecin CPT), is a kind of alkaloid contained in China endemic tree camplotheca acuminata to camptothecine.Research is recently pointed out that camptothecine and compound thereof have and is suppressed cell proliferation, apoptosis-induced and promote the effect of differentiation, camptothecine can suppress the propagation of vascular endothelial cell, apoptosis-induced, thereby stop the formation of new vessel, having notable antitumor activity, is that find up to now unique a kind of brings into play Cytotoxic cancer therapy drug by suppressing topoisomerase I specially.Camptothecine has considerable production and researching value as a kind of bulk drug.
The chemical structure of camptothecine is very unique, chemical property is also different with general alkaloid, it is to common alkaloid detection reagent such as phenol (iron trichloride) and Dragendorff reagent etc., all be negative reaction, do not form crystalline salt, but it can open lactonic ring running under the situation of alkali, runs into the reducible camptothecine that obtains in acid back with various acid, the organic solvent that camptothecine is water insoluble and common is to light, thermo-responsive easy decomposition.
In the prior art following several mode is taked in the extraction of camptothecine substantially, as the Soxhlet reflux extraction, ultrasonic wave reflux extraction, homogenate method, alkaline process; These several extracting method all have difference separately, the operation of Soxhlet reflux extraction is more loaded down with trivial details, extraction time is long, extraction efficiency is low, the strong concussion that the ultrasonic wave reflux extraction produces owing to ultrasonic wave in leaching process, not only can destroy the structure of camptothecine, and can strengthen the probability that camptothecine and other material contact and react, the homogenate method also has similar defective; Alkaline process extraction operation is more loaded down with trivial details, and extraction time is long, and extraction yield is low.
Summary of the invention
An object of the present invention is to solving above-mentioned prior art extraction time long, transformation efficiency is low, problems such as technological operation complexity provide reliability, the product recovery rate height of a kind of simple operation, conversion, can reach the extracting method of the camptothecine that suitability for industrialized production requires.
Present method comprises the steps:
Step 1. is followed the example of with alcohol extracting and is extracted;
The material that step 2. will go up the step extraction further extracts with alkaline extraction;
Step 3. is crossed column purification.
Further, the extracting method of a kind of camptothecine of the present invention carries out according to following steps:
1. alcohol extracting is followed the example of: will clean Camptotheca acuminata root or fruit after drying, and through pulverizing, by mass, the ethanol that adds the mass percentage concentration 40-70% that 3-8 doubly measures, soak at room temperature 0.5-2 hour.Then with mixture heating up to 60-80 ℃, be incubated 1.5-2 hour, extract twice, extracting liquid filtering, concentrating under reduced pressure becomes medicinal extract;
2. alkaline extraction: in medicinal extract, add the dilute sodium hydroxide of the mass percentage concentration 0.3%-5% of 10 times of amounts, be heated to 70 ℃ and kept temperature 0.2-2 hour, filter then.Filtrate is acidified to pH2-5 with acid, leaves standstill, and filtration drying gets the camptothecine finished product.
3. purifying: cross column chromatography, use the 0.5-10% ethanol elution, must the pure product of camptothecine.
Follow the example of with alcohol extracting in step 1. and to carry out after extracting solution is condensed into medicinal extract in the leaching process ethanol of medicinal extract with 40-70% being left standstill 6-14 hour, concentrating under reduced pressure becomes medicinal extract again, reclaims ethanol.
Further, the extracting method of a kind of camptothecine of the present invention carries out according to following steps:
1. alcohol extracting is followed the example of: will clean Camptotheca acuminata root or fruit after drying, and through pulverizing, by mass, soak 0.5-2 hour at the alcohol at normal temperature that adds the mass percentage concentration 50-55% that 5-7 doubly measures.With mixture heating up to 70 ℃, be incubated 1.5-2 hour then, extract twice, extracting liquid filtering, concentrating under reduced pressure becomes medicinal extract.The ethanol of medicinal extract with 40-70% was left standstill 12 hours, and concentrating under reduced pressure becomes medicinal extract again, reclaims ethanol.
2. alkaline extraction: in medicinal extract, add the dilute sodium hydroxide of the mass percentage concentration 0.3%-5% of 10 times of amounts, be heated to 70 ℃ 0.5 hour, filter then.Filtrate, is left standstill to pH3-4 with hcl acidifying, and filtration drying gets the camptothecine finished product.
3. purifying: alumina column chromatography, use 30% ethanol elution, the pure product of camptothecine.
The phytic acid or the xitix that can add the 0-1% of raw materials quality in the process that pure method is carried are made antioxidant, avoid the oxidation of active reaction thing, improve yield.
The preferred 0.01%-0.2 of the amount of phytic acid or xitix, more preferably 0.05-0.1%, most preferably 0.08%.
Phytic acid, chemical name: phytinic acid is a kind of organic phosphoric acid compounds that extracts from plant seed.Molecular formula: C6H18O24P6 molecular weight: 660.04.
Xitix is the common name of the bioactive compound of xitix, is a kind of water-soluble vitamins, has another name called vitamins C, and content is abundant in the fruits and vegetables.In the redox metabolic reaction, play regulating effect, lack it and can cause vitamin C deficiency.
Percentage composition among the present invention, part or multiple are the quality percentage composition, mass parts or quality multiple.
Beneficial effect of the present invention is:
Because the water insoluble and common organic solvent of camptothecine is to light, thermo-responsive easy decomposition.The present invention through research improvement repeatedly, under the situation of temperature control, adding antioxidant, adopts alcohol extracting method to combine with alkaline process on the basis of the work of summing up forefathers and oneself exploration, effectively raises extraction efficiency and purity.
In long alkali lye leaching process, because camptothecine is very unstable in sodium salt, be easy to combine and generate more stable material with other materials, so the time that shortens the alkali lye lixiviate can make the gain in yield of final product undoubtedly, and how to shorten extraction time, it is an also insurmountable difficult problem in the prior art, the present invention is by carrying out the method for alcohol extracting earlier, under the prerequisite of not destroying effective constituent in the raw material, tentatively extract, make the subtraction of back extract and just can finish by the very of short duration time, owing in preparation process, having added phytic acid or xitix is made antioxidant, avoid the oxidation of active reaction thing, improved yield.The unstable of camptothecine is well-known, but how to strengthen its stability is a problem demanding prompt solution, the contriver is through a large amount of experiments, trial by different approaches, finally find to have added phytic acid or the such method of xitix and can avoid the oxidation of active reaction thing, improve yield, though avoiding the oxidation of active reaction thing is the characteristic property of phytic acid or xitix, but be used in the Chinese medicine preparation report is not also arranged, when initial contriver proposes this thinking and prepares to test, also there are the association area personnel to propose to add phytic acid or xitix may react with the material in the raw material, produce precipitation or dregs, thereby influence the extraction of camptothecine; But the contriver is by having experimental results demonstrate that this is a kind of thinking mistaken ideas.
Technology of the present invention is simple, production cost is low, and product purity is high more than 98%, the product yield height, and can reach suitability for industrialized production.Greatly improve production efficiency and transformation efficiency by method of the present invention, impelled the active substance in the raw material more to be converted into trans camptothecine, made quality product further improve, and the product purity height, be fit to suitability for industrialized production.
Embodiment
Further specify the present invention below in conjunction with embodiment, but not as a limitation of the invention.
Embodiment 1
Carry out according to following technology
1. alcohol extracting is followed the example of: will clean the Camptotheca acuminata root 10kg after drying, and through pulverizing, to 80 orders by mass, soak 1 hour at the alcohol at normal temperature that adds 60kg mass percentage concentration 50%.With mixture heating up to 70 ℃, be incubated 1.5 hours then, extract twice, extracting liquid filtering, concentrating under reduced pressure becomes medicinal extract.Medicinal extract was left standstill 12 hours with 40% ethanol, and concentrating under reduced pressure becomes medicinal extract again, reclaims ethanol.
2. alkaline extraction: in medicinal extract, add the dilute sodium hydroxide of the mass percentage concentration 0.3% of 10 times of amounts, be heated to 70 ℃ 0.5 hour, filter then.Filtrate, is left standstill to pH3 with hcl acidifying, and filtration drying gets the camptothecine finished product.
3. purifying: alumina column chromatography, use 30% ethanol elution, the pure product of camptothecine.
Embodiment 2
1. alcohol extracting is followed the example of: will clean Camptotheca acuminata root or fruit after drying, and through pulverizing, by mass, soak 2 hours at the alcohol at normal temperature of the mass percentage concentration 55% that adds 7 times of amounts.0.1% the phytic acid that adds raw materials quality is made antioxidant, avoids the oxidation of active reaction thing, improves yield.With mixture heating up to 70 ℃, be incubated 2 hours then, extract twice, extracting liquid filtering, concentrating under reduced pressure becomes medicinal extract.Medicinal extract was left standstill 12 hours with 70% ethanol, and concentrating under reduced pressure becomes medicinal extract again, reclaims ethanol.
2. alkaline extraction: in medicinal extract, add the dilute sodium hydroxide of the mass percentage concentration 5% of 10 times of amounts, be heated to 70 ℃ 0.5 hour, filter then.Filtrate, is left standstill to pH4 with hcl acidifying, and filtration drying gets the camptothecine finished product.
3. purifying: alumina column chromatography, use 30% ethanol elution, the pure product of camptothecine.
Embodiment 3-10
The amount of phytic acid or xitix is as shown in the table, and all the other are with embodiment 2
Comparative Examples 1
Roll up " the camptothecine Study on Extraction Method " that the second phase (2008.6) Sun Yanxia etc. carries out with reference to Chengdu college journal (natural science edition) the 27th, accurately take by weighing sample 1g and adopt 95% ethanol Soxhlet refluxing extraction, in apparatus,Soxhlet's, extract 6h, cooling is settled to 100ml, qualitative filter paper filters, again through the filtering with microporous membrane in 0.45 μ m aperture,, go the analysis of filtrate sample presentation to try to achieve camptothecine content in the sample after the collection filtrate by 1.3 chromatographic conditions.
The prior art and the embodiment of the invention are extracted camptothecine content, and each scheme is carried out three times, and averaging compares:
By comparative result as can be seen, the inventive method is carried out the extraction of camptothecine, obviously is better than prior art.
Claims (9)
1. the extracting method of a camptothecine, it is characterized in that: described technology is as follows:
Step 1. is followed the example of with alcohol extracting and is extracted;
The material that step 2. will go up the step extraction further extracts with alkaline extraction;
Step 3. is crossed column purification.
2. the extracting method of a kind of camptothecine as claimed in claim 1, it is characterized in that: described technology is as follows:
Step 1. alcohol extracting is followed the example of: will clean Camptotheca acuminata root or fruit after drying, through pulverizing, by mass, at the ethanol that adds the mass percentage concentration 40-70% that 3-8 doubly measures, soak at room temperature 0.5-2 hour, then with mixture heating up to 60-80 ℃, be incubated 1.5-2 hour, extract twice, extracting liquid filtering, concentrating under reduced pressure becomes medicinal extract;
Step 2. alkaline extraction: add the diluted sodium hydroxide solution of the mass percentage concentration 0.3%-5% of 10 times of amounts in medicinal extract, be heated to 70 ℃ and kept temperature 0.2-2 hour, filter then, filtrate is acidified to pH2-5 with acid, leaves standstill, and filtration drying gets the camptothecine finished product;
Step 3. purifying: cross column chromatography, use the 0.5-10% ethanol elution, get the pure product of camptothecine.
3. the extracting method of a kind of camptothecine as claimed in claim 2, it is characterized in that: described technology is as follows: during step 1. alcohol extracting is followed the example of, can after making medicinal extract the ethanol of medicinal extract with 40-70% be left standstill 6-14 hour, concentrating under reduced pressure becomes medicinal extract once more, reclaims ethanol.
4. the extracting method of a kind of camptothecine as claimed in claim 3, it is characterized in that: described technology is as follows: step 1. alcohol extracting is followed the example of: will clean Camptotheca acuminata root or fruit after drying, through pulverizing, by mass, soaked 0.5-2 hour at the alcohol at normal temperature that adds the mass percentage concentration 50-55% that 5-7 doubly measures, then with mixture heating up to 70 ℃, be incubated 1.5-2 hour, extract twice, extracting liquid filtering, concentrating under reduced pressure becomes medicinal extract, and the ethanol of medicinal extract with 40-70% was left standstill 12 hours, concentrating under reduced pressure becomes medicinal extract once more, reclaims ethanol;
Step 2. alkaline extraction: add the dilute sodium hydroxide of the mass percentage concentration 0.3%-5% of 10 times of amounts in medicinal extract, be heated to 70 ℃ of insulations 0.5 hour, filter then, filtrate, is left standstill to pH3-4 with hcl acidifying, and filtration drying gets the camptothecine finished product;
Step 3. purifying: alumina column chromatography, use 30% ethanol elution, get the pure product of camptothecine.
5. as the extracting method of the arbitrary described a kind of camptothecine of claim 1-4, it is characterized in that: the phytic acid or the xitix that in the process that pure method is carried, add the 0-1% of raw materials quality.
6. the extracting method of a kind of camptothecine as claimed in claim 5 is characterized in that: add phytic acid or xitix before extracting when soak at room temperature or through the soak at room temperature post-heating.
7. the extracting method of a kind of camptothecine as claimed in claim 6, it is characterized in that: the amount of phytic acid or xitix is the 0.01%-0.2% of raw materials quality.
8. the extracting method of a kind of camptothecine as claimed in claim 7, it is characterized in that: the amount of phytic acid or xitix is the 0.05-0.1% of raw materials quality.
9. the extracting method of a kind of camptothecine as claimed in claim 8, it is characterized in that: the amount of phytic acid or xitix is 0.08% of a raw materials quality.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201010529441.2A CN102127089B (en) | 2010-11-03 | 2010-11-03 | Method for extracting camptothecin |
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| CN201010529441.2A CN102127089B (en) | 2010-11-03 | 2010-11-03 | Method for extracting camptothecin |
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| CN102127089A true CN102127089A (en) | 2011-07-20 |
| CN102127089B CN102127089B (en) | 2014-04-16 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102603758A (en) * | 2012-02-12 | 2012-07-25 | 张苑金 | Extraction method of camptothecin and 9-hydroxymethyl camptothecin from tender leaves and twigs of Nothapodytes nimmoniana |
| CN107873742A (en) * | 2017-12-06 | 2018-04-06 | 昭平县科学技术实验场 | Composite insecticide containing wilfordine and camptothecine and preparation method thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1121920A (en) * | 1994-10-26 | 1996-05-08 | 四川省广汉市松林生化厂 | Method for extracting camptothecin |
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- 2010-11-03 CN CN201010529441.2A patent/CN102127089B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1121920A (en) * | 1994-10-26 | 1996-05-08 | 四川省广汉市松林生化厂 | Method for extracting camptothecin |
Non-Patent Citations (6)
| Title |
|---|
| 《中国优秀硕士论文集（基础科学辑）》 20071215 李连强 "喜树叶中CPT和HCPT含量的动态变化及CPT分离与纯化" 第10页 1-9 , 第6期 * |
| 《中草药》 20050612 王瑞芳等 "大孔吸附树脂对喜树果中喜树碱和喜果苷的分离纯化研究" 849-851 3-9 第36卷, 第6期 * |
| 《山地农业生物学报》 20041228 赵东亮等 "从喜树果中分离提取喜树碱的工艺条件" 第494页 1-9 第23卷, 第6期 * |
| 李连强: ""喜树叶中CPT和HCPT含量的动态变化及CPT分离与纯化"", 《中国优秀硕士论文集（基础科学辑）》 * |
| 王瑞芳等: ""大孔吸附树脂对喜树果中喜树碱和喜果苷的分离纯化研究"", 《中草药》 * |
| 赵东亮等: ""从喜树果中分离提取喜树碱的工艺条件"", 《山地农业生物学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102603758A (en) * | 2012-02-12 | 2012-07-25 | 张苑金 | Extraction method of camptothecin and 9-hydroxymethyl camptothecin from tender leaves and twigs of Nothapodytes nimmoniana |
| CN107873742A (en) * | 2017-12-06 | 2018-04-06 | 昭平县科学技术实验场 | Composite insecticide containing wilfordine and camptothecine and preparation method thereof |
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| CN102127089B (en) | 2014-04-16 |
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