CN112480040A - Preparation process of high-purity paclitaxel - Google Patents
Preparation process of high-purity paclitaxel Download PDFInfo
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- CN112480040A CN112480040A CN202011256740.3A CN202011256740A CN112480040A CN 112480040 A CN112480040 A CN 112480040A CN 202011256740 A CN202011256740 A CN 202011256740A CN 112480040 A CN112480040 A CN 112480040A
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Abstract
The invention relates to the technical field of medicine preparation, in particular to a preparation process of high-purity paclitaxel; in the invention, the polystyrene resin HPS-355 is chemically modified by the matching use of the ethylene glycol and the trihydroxy triethylamine; effectively expands the three-dimensional structure and volume thereof and obviously enhances the specific adsorption performance of the paclitaxel on the paclitaxel; so that the pigment and other impurities mixed in the paclitaxel can be effectively and fully separated from the paclitaxel; moreover, the invention can effectively remove impurity molecules by adopting the mutual matching use of the procedures of leaching, multiple times of extraction and concentration, macroporous resin elution and chromatographic separation, so that the purity of the prepared taxol is greatly improved; in addition, the alkaline impurities doped in the extraction liquid can be oxidized into derivatives with higher polarity by continuously introducing ozone into the extraction liquid, so that the final separation and purification operation of the paclitaxel is facilitated, and the paclitaxel with higher purity is prepared.
Description
Technical Field
The invention relates to the technical field of medicine preparation, in particular to a preparation process of high-purity paclitaxel.
Background
Paclitaxel, alias taxol, viologen, tertin, chemical name 5 beta, 20-epoxy-1, 2 alpha, 4, 7 beta, 10 beta, 13 alpha-hexahydroxy taxane-11-en-9-one-4, 10-diacetate-2-benzoate-13 [ (2'R, 3' S) -N-benzoyl-3-phenylisoserine ester ], molecular weight 853.92, molecular formula C47H51NO 14.
In 1963, the american chemists vanni and wal first isolated a crude extract of paclitaxel from a tree called pacific fir bark and wood grown in the western forest of the united states. In screening experiments, Wani and Wall found that crude paclitaxel extract had high activity against mouse tumor cells cultured ex vivo, and began to isolate this active ingredient. Since the content of the active ingredient in plants was very low, until 1971, they were not cooperative with McFall, a chemical professor of Duke university, and the chemical structure of the active ingredient, a tetracyclic diterpene compound, was determined by x-ray analysis and named paclitaxel.
Paclitaxel is required in increasing quantities and is scarce because of its outstanding therapeutic effect in treating cancer. However, the paclitaxel prepared in the prior art has low yield and relatively low purity, and cannot be directly used as an anticancer drug. Therefore, it is a technical problem to be solved by those skilled in the art to provide a process for preparing high-purity paclitaxel.
Disclosure of Invention
Aiming at the technical problems in the background technology, the invention provides a preparation process of high-purity paclitaxel, and the paclitaxel prepared by the invention has high content and high purity; the paclitaxel preparation process provided by the invention has wider market prospect and is more suitable for popularization.
Technical scheme
In order to achieve the purpose, the invention is realized by the following technical scheme:
a preparation process of high-purity paclitaxel comprises the following steps:
s1, cleaning the branches and leaves of the taxus chinensis with clear water, and then placing the branches and leaves of the taxus chinensis in a constant-temperature drying box for drying treatment; then crushing and grinding the dried taxus chinensis branches and leaves until the granularity is 60-90 meshes; storing the obtained fine powder of Taxus chinensis for use;
s2, accurately weighing 80-100 g of the fine taxus chinensis powder prepared in the step, soaking the fine taxus chinensis powder in a proper amount of extracting solution at the temperature of 40-50 ℃, performing ultrasonic extraction for 3-4 times, wherein the ultrasonic extraction time is 15-20 hours each time, and performing ultrasonic treatment on the extracting solution during extraction; then collecting and combining the leaching liquor obtained each time; storing and standby;
s3, pumping the leaching liquor obtained in the step S2 into supercritical extraction equipment equipped with an enhanced ultrasonic assembly, adding an organic extractant, setting the temperature of supercritical extraction to be 30-45 ℃, the pressure of supercritical extraction to be 3-5 MPa, and setting the time of supercritical extraction to be 2-6 h; after extraction is finished, collecting extract liquor; storing for later use;
s4, continuously inputting ozone into the extraction liquid obtained in the step S3; wherein, the amount of ozone introduced into each liter of reaction system per hour is 3-7 g, the temperature in the extraction liquid is set to be 10-40 ℃, and the continuous ozone introduction time is 2-3 h; concentrating the extractive solution under normal pressure until no solvent overflows, concentrating under reduced pressure under vacuum, and drying under reduced pressure to obtain paclitaxel extract;
s5, mixing the mixed components into macroporous resin with the same quantity as the paclitaxel extract, and loading the mixture onto a column by a sample dry method; then adding an eluent to elute the paclitaxel extract, and controlling each fraction to be 30-50 mL during elution; after obtaining the corresponding elution fraction, recovering the solvent from the elution fraction at normal pressure; then pumping into a concentration tank for reduced pressure concentration to obtain a concentrated solution;
s6, adding dichloromethane into the concentrated solution obtained in the step S5 to extract the concentrated solution, pumping the extract phase into a concentration tank to concentrate the extract to obtain an extract, namely a taxol extract crude product, dissolving the taxol crude extract obtained after chromatographic separation by using acetone, and putting the taxol crude extract into a preparative high performance liquid chromatograph to separate and purify the taxol crude extract; preparing paclitaxel fraction, concentrating under reduced pressure, and pumping to obtain paclitaxel product.
Further, the extracting solution used in step S2 is prepared by mixing ethyl acetate, acetone and the like by mass.
Furthermore, the ultrasonic leaching frequency in the step S2 is 25-30 kHz, and the ultrasonic power is 500-700W.
Furthermore, the organic extractant used in step S3 is any one of carbon dioxide, methane or ethylene; and the dosage standard of the extracting agent is V(organic extractant)：m(Leaching liquor)=25～35ml/g。
Further, the mixed components in the step S5 are prepared by mixing chloroform and methanol at a volume ratio of 95: 5.
Furthermore, the macroporous resin used in the step S5 is modified polystyrene resin HPS-355, and the preparation process comprises: placing the polystyrene resin HPS-355 in a reaction kettle, then respectively adding ethylene glycol and trihydroxy triethylamine into the reaction kettle, raising the temperature in the reaction kettle to 50-65 ℃, reacting at the constant temperature for 6-8 h, after the reaction is finished, naturally cooling the mixture in the kettle to room temperature, filtering, then sequentially washing the mixture with absolute ethyl alcohol and deionized water, and finally placing the obtained product in a vacuum drying oven to dry to constant weight to obtain a modified polystyrene resin HPS-355 finished product;
wherein the dosage of the ethylene glycol is 6-8 times of that of the polystyrene resin HPS-355, and the ratio of the trihydroxy triethylamine to the polystyrene resin HPS-355 is 4: 1.
Further, the flow rate of the eluent during the elution in the step S5 is 10-15 min per fraction.
Furthermore, in the S6, when separation and purification are carried out, the selected mobile phase group is prepared by mixing acetonitrile and water according to a volume ratio of 2: 3.
Furthermore, when the separation and purification are carried out in S6, the flow rate of the mobile phase is 1-3 mL/min, the column temperature is 30-35 ℃, and the detection wavelength is 234 nm.
Advantageous effects
Compared with the known public technology, the technical scheme provided by the invention has the following beneficial effects:
in the invention, the polystyrene resin HPS-355 is chemically modified by the matching use of the ethylene glycol and the trihydroxy triethylamine; wherein, the trihydroxy triethylamine and the polystyrene resin HPS-355 are subjected to grafting reaction, so that the trihydroxy triethylamine is successfully grafted on the surface of the polystyrene resin HPS-355 molecules, the three-dimensional space structure and the volume of the trihydroxy triethylamine are effectively expanded, and the specific adsorption performance of the trihydroxy triethylamine on paclitaxel is obviously enhanced. When the taxol molecules and the impurity molecules pass through the modified polystyrene resin HPS-355 molecules, because the speeds of the taxol molecules and the impurity molecules which are diffused into gel pores are different, the impurities can be fully separated from the taxol by the difference of the speeds of the chromatographic columns, and the pigment and other impurities (such as polysaccharide, tannin and the like which are mixed in the pigment and other impurities) which are mixed in the pigment and other impurities can be fully separated from the taxol. Furthermore, the invention can effectively remove impurity molecules by adopting the mutual matching use of the procedures of leaching, multiple times of extraction and concentration, macroporous resin elution and chromatographic separation, so that the purity of the prepared taxol is greatly improved. In addition, the alkaline impurities doped in the extraction liquid can be oxidized into derivatives with higher polarity by continuously introducing ozone into the extraction liquid, so that the final separation and purification operation of the paclitaxel is facilitated, and the paclitaxel with higher purity is prepared.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. It is to be understood that the embodiments described are only a few embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The present invention will be further described with reference to the following examples.
Example 1
A preparation process of high-purity paclitaxel comprises the following steps:
s1, cleaning the branches and leaves of the taxus chinensis with clear water, and then placing the branches and leaves of the taxus chinensis in a constant-temperature drying box for drying treatment; then crushing and grinding the dried taxus chinensis branches and leaves until the granularity is 60 meshes; storing the obtained fine powder of Taxus chinensis for use;
s2, accurately weighing 80g of the fine taxus chinensis powder prepared in the previous step, soaking the fine taxus chinensis powder in a proper amount of extracting solution at the temperature of 40 ℃, carrying out ultrasonic extraction for 3 times, wherein the ultrasonic extraction time is 15 hours each time, and carrying out ultrasonic treatment on the extracting solution during extraction; then collecting and combining the leaching liquor obtained each time; storing and standby;
s3, pumping the leaching liquor obtained in the step S2 into supercritical extraction equipment equipped with an enhanced ultrasonic assembly, adding an organic extractant, setting the temperature of supercritical extraction to be 30 ℃, the pressure of supercritical extraction to be 3MPa and the time of supercritical extraction to be 2 hours; after extraction is finished, collecting extract liquor; storing for later use;
s4, continuously inputting ozone into the extraction liquid obtained in the step S3; wherein, the ozone amount per liter of reaction system per hour is 3g, the temperature in the extraction liquid is set to be 10 ℃, and the ozone continuous introduction time is 2 h; concentrating the extractive solution under normal pressure until no solvent overflows, concentrating under reduced pressure under vacuum, and drying under reduced pressure to obtain paclitaxel extract;
s5, mixing the mixed components into macroporous resin with the same quantity as the paclitaxel extract, and loading the mixture onto a column by a sample dry method; then adding eluent to elute the paclitaxel extract, and controlling gradient elution to separate 30mL of each fraction during elution; after obtaining the corresponding elution fraction, recovering the solvent from the elution fraction at normal pressure; then pumping into a concentration tank for reduced pressure concentration to obtain a concentrated solution;
s6, adding dichloromethane into the concentrated solution obtained in the step S5 to extract the concentrated solution, pumping the extract phase into a concentration tank to concentrate the extract to obtain an extract, namely a taxol extract crude product, dissolving the taxol crude extract obtained after chromatographic separation by using acetone, and putting the taxol crude extract into a preparative high performance liquid chromatograph to separate and purify the taxol crude extract; preparing paclitaxel fraction, concentrating under reduced pressure, and pumping to obtain paclitaxel product.
The extract used in step S2 is prepared by mixing ethyl acetate, acetone and the like in mass.
The ultrasonic leaching frequency in the step S2 is 25kHz, and the ultrasonic power is 500W.
The organic extractant used in the step S3 is carbon dioxide; and the dosage standard of the extracting agent is V(organic extractant)：m(Leaching liquor)=25ml/g。
The mixed components in the step S5 are prepared by mixing chloroform and methanol according to the volume ratio of 95: 5.
The macroporous resin used in the step S5 is modified polystyrene resin HPS-355, and the preparation process comprises the following steps: placing the polystyrene resin HPS-355 in a reaction kettle, then respectively adding ethylene glycol and trihydroxy triethylamine into the reaction kettle, then raising the temperature in the reaction kettle to 50 ℃, reacting at the constant temperature for 6 hours, after the reaction is finished, naturally cooling the mixture in the kettle to room temperature, filtering, then sequentially washing the mixture with absolute ethyl alcohol and deionized water, and finally placing the obtained product in a vacuum drying oven to dry to constant weight to obtain a finished product of the modified polystyrene resin HPS-355;
wherein the dosage of the ethylene glycol is 6 times of that of the polystyrene resin HPS-355, and the ratio of the trihydroxy triethylamine to the polystyrene resin HPS-355 is 4: 1.
The flow rate of the eluent at the time of elution in step S5 was 10min per fraction.
In S6, when separation and purification are carried out, the selected mobile phase group is prepared by mixing acetonitrile and water according to the volume ratio of 2: 3.
In S6, when separation and purification are performed, the flow rate of the mobile phase is 1mL/min, the column temperature is 30 ℃, and the detection wavelength is 234 nm.
Example 2
A preparation process of high-purity paclitaxel comprises the following steps:
s1, cleaning the branches and leaves of the taxus chinensis with clear water, and then placing the branches and leaves of the taxus chinensis in a constant-temperature drying box for drying treatment; then crushing and grinding the dried taxus chinensis branches and leaves until the granularity is 70 meshes; storing the obtained fine powder of Taxus chinensis for use;
s2, accurately weighing 90g of the fine taxus chinensis powder prepared in the previous step, soaking the fine taxus chinensis powder in a proper amount of extracting solution at the temperature of 45 ℃, carrying out ultrasonic extraction for 3 times, wherein the ultrasonic extraction time is 18 hours each time, and carrying out ultrasonic treatment on the extracting solution during extraction; then collecting and combining the leaching liquor obtained each time; storing and standby;
s3, pumping the leaching liquor obtained in the step S2 into supercritical extraction equipment equipped with an enhanced ultrasonic assembly, adding an organic extractant, setting the temperature of supercritical extraction to 40 ℃, the pressure of supercritical extraction to 4MPa and the time of supercritical extraction to 5 hours; after extraction is finished, collecting extract liquor; storing for later use;
s4, continuously inputting ozone into the extraction liquid obtained in the step S3; wherein, the ozone amount per liter of reaction system per hour is 5g, the temperature in the extraction liquid is set to be 25 ℃, and the ozone continuous introduction time is 2 h; concentrating the extractive solution under normal pressure until no solvent overflows, concentrating under reduced pressure under vacuum, and drying under reduced pressure to obtain paclitaxel extract;
s5, mixing the mixed components into macroporous resin with the same quantity as the paclitaxel extract, and loading the mixture onto a column by a sample dry method; then adding an eluent to elute the paclitaxel extract, and controlling gradient elution to separate each fraction by 40mL during elution; after obtaining the corresponding elution fraction, recovering the solvent from the elution fraction at normal pressure; then pumping into a concentration tank for reduced pressure concentration to obtain a concentrated solution;
s6, adding dichloromethane into the concentrated solution obtained in the step S5 to extract the concentrated solution, pumping the extract phase into a concentration tank to concentrate the extract to obtain an extract, namely a taxol extract crude product, dissolving the taxol crude extract obtained after chromatographic separation by using acetone, and putting the taxol crude extract into a preparative high performance liquid chromatograph to separate and purify the taxol crude extract; preparing paclitaxel fraction, concentrating under reduced pressure, and pumping to obtain paclitaxel product.
The extract used in step S2 is prepared by mixing ethyl acetate, acetone and the like in mass.
The ultrasonic leaching frequency in the step S2 is 28kHz, and the ultrasonic power is 600W.
Used in step S3The organic extractant is methane; and the dosage standard of the extracting agent is V(organic extractant)：m(Leaching liquor)=30ml/g。
The mixed components in the step S5 are prepared by mixing chloroform and methanol according to the volume ratio of 95: 5.
The macroporous resin used in the step S5 is modified polystyrene resin HPS-355, and the preparation process comprises the following steps: placing the polystyrene resin HPS-355 in a reaction kettle, then respectively adding ethylene glycol and trihydroxy triethylamine into the reaction kettle, then raising the temperature in the reaction kettle to 60 ℃, reacting at the constant temperature for 7 hours, after the reaction is finished, naturally cooling the mixture in the kettle to room temperature, filtering, then sequentially washing the mixture with absolute ethyl alcohol and deionized water, and finally placing the obtained product in a vacuum drying oven to dry to constant weight to obtain a finished product of the modified polystyrene resin HPS-355;
wherein the dosage of the ethylene glycol is 7 times of that of the polystyrene resin HPS-355, and the ratio of the trihydroxy triethylamine to the polystyrene resin HPS-355 is 4: 1.
The flow rate of the eluent at the time of elution in step S5 was 12min per fraction.
In S6, when separation and purification are carried out, the selected mobile phase group is prepared by mixing acetonitrile and water according to the volume ratio of 2: 3.
In S6, when separation and purification are performed, the flow rate of the mobile phase is 2mL/min, the column temperature is 33 ℃, and the detection wavelength is 234 nm.
Example 3
A preparation process of high-purity paclitaxel comprises the following steps:
s1, cleaning the branches and leaves of the taxus chinensis with clear water, and then placing the branches and leaves of the taxus chinensis in a constant-temperature drying box for drying treatment; then crushing and grinding the dried taxus chinensis branches and leaves until the granularity is 90 meshes; storing the obtained fine powder of Taxus chinensis for use;
s2, accurately weighing 100g of the fine taxus chinensis powder prepared in the previous step, soaking the fine taxus chinensis powder in a proper amount of extracting solution at the temperature of 50 ℃, carrying out ultrasonic extraction for 4 times, wherein the ultrasonic extraction time is 20 hours each time, and carrying out ultrasonic treatment on the extracting solution during extraction; then collecting and combining the leaching liquor obtained each time; storing and standby;
s3, pumping the leaching liquor obtained in the step S2 into supercritical extraction equipment equipped with an enhanced ultrasonic assembly, adding an organic extractant, setting the temperature of the supercritical extraction to be 45 ℃, the pressure of the supercritical extraction to be 5MPa and the time of the supercritical extraction to be 6 hours; after extraction is finished, collecting extract liquor; storing for later use;
s4, continuously inputting ozone into the extraction liquid obtained in the step S3; wherein, the ozone amount per liter of reaction system per hour is 7g, the temperature in the extraction liquid is set to be 40 ℃, and the ozone continuous introduction time is 3 h; concentrating the extractive solution under normal pressure until no solvent overflows, concentrating under reduced pressure under vacuum, and drying under reduced pressure to obtain paclitaxel extract;
s5, mixing the mixed components into macroporous resin with the same quantity as the paclitaxel extract, and loading the mixture onto a column by a sample dry method; then adding an eluent to elute the paclitaxel extract, and controlling each fraction to be 50mL during elution; after obtaining the corresponding elution fraction, recovering the solvent from the elution fraction at normal pressure; then pumping into a concentration tank for reduced pressure concentration to obtain a concentrated solution;
s6, adding dichloromethane into the concentrated solution obtained in the step S5 to extract the concentrated solution, pumping the extract phase into a concentration tank to concentrate the extract to obtain an extract, namely a taxol extract crude product, dissolving the taxol crude extract obtained after chromatographic separation by using acetone, and putting the taxol crude extract into a preparative high performance liquid chromatograph to separate and purify the taxol crude extract; preparing paclitaxel fraction, concentrating under reduced pressure, and pumping to obtain paclitaxel product.
The extract used in step S2 is prepared by mixing ethyl acetate, acetone and the like in mass.
The ultrasonic leaching frequency in the step S2 was 30kHz, and the ultrasonic power was 700W.
The organic extractant used in the step S3 is ethylene; and the dosage standard of the extracting agent is V(organic extractant)：m(Leaching liquor)＝35ml/g。
The mixed components in the step S5 are prepared by mixing chloroform and methanol according to the volume ratio of 95: 5.
The macroporous resin used in the step S5 is modified polystyrene resin HPS-355, and the preparation process comprises the following steps: placing the polystyrene resin HPS-355 in a reaction kettle, then respectively adding ethylene glycol and trihydroxy triethylamine into the reaction kettle, then raising the temperature in the reaction kettle to 65 ℃, reacting at the constant temperature for 8 hours, after the reaction is finished, naturally cooling the mixture in the kettle to room temperature, filtering, then sequentially washing the mixture with absolute ethyl alcohol and deionized water, and finally placing the obtained product in a vacuum drying oven to dry to constant weight to obtain a finished product of the modified polystyrene resin HPS-355;
wherein the dosage of the ethylene glycol is 8 times of that of the polystyrene resin HPS-355, and the ratio of the trihydroxy triethylamine to the polystyrene resin HPS-355 is 4: 1.
The flow rate of the eluent at the time of elution in step S5 was 15min per fraction.
In S6, when separation and purification are carried out, the selected mobile phase group is prepared by mixing acetonitrile and water according to the volume ratio of 2: 3.
In S6, when separation and purification are performed, the flow rate of the mobile phase is 3mL/min, the column temperature is 35 ℃, and the detection wavelength is 234 nm.
Performance testing
Comparative example: a paclitaxel product extracted according to a method for extracting paclitaxel from roots of yew trees provided in patent document CN 201510492051.5;
example (b): paclitaxel products prepared by examples 1 to 3 of the present invention (referred to as examples 1 to 3);
HPLC detection is respectively carried out on the paclitaxel products prepared in the comparative example and the examples 1-3, and the obtained data are recorded in the following table:
| percent content of paclitaxel% | Purity/% of paclitaxel | Percent of cephalomannine/%) | Percent of 7-epitaxol/%) | |
| Example 1 | 97.8 | 99.8 | 0.086 | 0.067 |
| Example 2 | 98.6 | 99.4 | 0.092 | 0.034 |
| Example 3 | 99.4 | 99.6 | 0.084 | 0.031 |
| Comparative example | 82.7 | 86.3 | 2.63 | 3.86 |
As can be seen from the relevant data in the above table, the paclitaxel prepared by the present invention is not only higher in content but also higher in purity, compared to the method for extracting paclitaxel provided in the comparative example. The paclitaxel preparation process provided by the invention has wider market prospect and is more suitable for popularization.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; such modifications and substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Claims (9)
1. A preparation process of high-purity paclitaxel is characterized by comprising the following steps:
s1, cleaning the branches and leaves of the taxus chinensis with clear water, and then placing the branches and leaves of the taxus chinensis in a constant-temperature drying box for drying treatment; then crushing and grinding the dried taxus chinensis branches and leaves until the granularity is 60-90 meshes; storing the obtained fine powder of Taxus chinensis for use;
s2, accurately weighing 80-100 g of the fine taxus chinensis powder prepared in the step, soaking the fine taxus chinensis powder in a proper amount of extracting solution at the temperature of 40-50 ℃, performing ultrasonic extraction for 3-4 times, wherein the ultrasonic extraction time is 15-20 hours each time, and performing ultrasonic treatment on the extracting solution during extraction; then collecting and combining the leaching liquor obtained each time; storing and standby;
s3, pumping the leaching liquor obtained in the step S2 into supercritical extraction equipment equipped with an enhanced ultrasonic assembly, adding an organic extractant, setting the temperature of supercritical extraction to be 30-45 ℃, the pressure of supercritical extraction to be 3-5 MPa, and setting the time of supercritical extraction to be 2-6 h; after extraction is finished, collecting extract liquor; storing for later use;
s4, continuously inputting ozone into the extraction liquid obtained in the step S3; wherein, the amount of ozone introduced into each liter of reaction system per hour is 3-7 g, the temperature in the extraction liquid is set to be 10-40 ℃, and the continuous ozone introduction time is 2-3 h; concentrating the extractive solution under normal pressure until no solvent overflows, concentrating under reduced pressure under vacuum, and drying under reduced pressure to obtain paclitaxel extract;
s5, mixing the mixed components into macroporous resin with the same quantity as the paclitaxel extract, and loading the mixture onto a column by a sample dry method; then adding an eluent to elute the paclitaxel extract, and controlling each fraction to be 30-50 mL during elution; after obtaining the corresponding elution fraction, recovering the solvent from the elution fraction at normal pressure; then pumping into a concentration tank for reduced pressure concentration to obtain a concentrated solution;
s6, adding dichloromethane into the concentrated solution obtained in the step S5 to extract the concentrated solution, pumping the extract phase into a concentration tank to concentrate the extract to obtain an extract, namely a taxol extract crude product, dissolving the taxol crude extract obtained after chromatographic separation by using acetone, and putting the taxol crude extract into a preparative high performance liquid chromatograph to separate and purify the taxol crude extract; preparing paclitaxel fraction, concentrating under reduced pressure, and pumping to obtain paclitaxel product.
2. The process according to claim 1, wherein the paclitaxel is prepared by the following steps: the extracting solution used in the step S2 is prepared by mixing ethyl acetate, acetone and the like in mass.
3. The process according to claim 1, wherein the paclitaxel is prepared by the following steps: in the step S2, the ultrasonic leaching frequency is 25-30 kHz, and the ultrasonic power is 500-700W.
4. The process according to claim 1, wherein the paclitaxel is prepared by the following steps: the organic extracting agent used in the step S3 is any one of carbon dioxide, methane or ethylene; and the dosage standard of the extracting agent is V(organic extractant)：m(Leaching liquor)=25～35ml/g。
5. The process according to claim 1, wherein the paclitaxel is prepared by the following steps: the mixed components in the step S5 are prepared by mixing chloroform and methanol according to a volume ratio of 95: 5.
6. The process according to claim 1, wherein the paclitaxel is prepared by the following steps: the macroporous resin used in the step S5 is modified polystyrene resin HPS-355, and the preparation process comprises the following steps: placing the polystyrene resin HPS-355 in a reaction kettle, then respectively adding ethylene glycol and trihydroxy triethylamine into the reaction kettle, raising the temperature in the reaction kettle to 50-65 ℃, reacting at the constant temperature for 6-8 h, after the reaction is finished, naturally cooling the mixture in the kettle to room temperature, filtering, then sequentially washing the mixture with absolute ethyl alcohol and deionized water, and finally placing the obtained product in a vacuum drying oven to dry to constant weight to obtain a modified polystyrene resin HPS-355 finished product;
wherein the dosage of the ethylene glycol is 6-8 times of that of the polystyrene resin HPS-355, and the ratio of the trihydroxy triethylamine to the polystyrene resin HPS-355 is 4: 1.
7. The process according to claim 1, wherein the paclitaxel is prepared by the following steps: and the flow rate of the eluent in the step S5 is 10-15 min per fraction.
8. The process according to claim 1, wherein the paclitaxel is prepared by the following steps: in the S6, when separation and purification are carried out, the selected mobile phase group is prepared by mixing acetonitrile and water according to the volume ratio of 2: 3.
9. The process according to claim 1, wherein the paclitaxel is prepared by the following steps: in the S6, when separation and purification are carried out, the flow velocity of a mobile phase is 1-3 mL/min, the column temperature is 30-35 ℃, and the detection wavelength is 234 nm.
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| CN112409300A (en) * | 2020-11-06 | 2021-02-26 | 巢湖学院 | Extraction process of paclitaxel |
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| CN112409300A (en) * | 2020-11-06 | 2021-02-26 | 巢湖学院 | Extraction process of paclitaxel |
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