CN111253345A - Method for extracting paclitaxel from branches and leaves of taxus chinensis - Google Patents
Method for extracting paclitaxel from branches and leaves of taxus chinensis Download PDFInfo
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Abstract
The invention provides a method for extracting taxol from taxus chinensis branches and leaves, belonging to the field of extraction of fat-soluble chemical components. The invention realizes the high-efficiency extraction of the paclitaxel in the branches and leaves of the taxus chinensis by the ultrahigh pressure auxiliary micelle extraction technology, N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride is a green and environment-friendly surfactant, under the conditions of normal temperature and high pressure, the micelle extraction solvent and the taxus chinensis branch and leaf powder are subjected to the action of strong shearing force, impact force, turbulent flow, cavity explosive force generated by sudden rising and falling of static pressure and the like when passing through a narrow gap, so that the original suspension of coarse particles is processed into suspension in which fine particles stably exist, the paclitaxel is quickly diffused into the solvent, thereby realizing the quick low-temperature extraction, and the operation is simple, the extraction efficiency is high, plays a certain role in protecting heat-sensitive active ingredients and is suitable for large-scale industrial extraction production.
Description
Technical Field
The invention relates to the technical field of fat-soluble chemical component extraction, in particular to a method for extracting taxol from taxus chinensis branches and leaves.
Background
Taxus cuspidata is a taxaceae, and extracts of fruits, barks, branches and leaves of the taxus cuspidata have various active ingredients with medicinal values, wherein secondary metabolites of taxane diterpenoids such as paclitaxel and the like have an anticancer effect with a unique mechanism, and the active ingredients of the paclitaxel can also be used for treating chronic inflammatory diseases. The injection prepared from paclitaxel has well-known breast cancer resisting effect, and can be used for treating ovarian cancer and non-small cell lung cancer.
The traditional extraction methods include immersion, percolation, soxhlet extraction, etc. The dipping method needs simple equipment and is easy to operate, but the dipping time is long, the working strength is high, and the extraction of effective components is incomplete. The percolation method is suitable for extracting components which are unstable and easy to decompose, is simple, has high extraction efficiency, and causes unnecessary waste due to large solvent consumption. The Soxhlet extraction method is simple to operate, but the extraction time is too long, volatile organic solvents such as ethanol and the like are required to extract effective components in the extraction process, the effective components are obtained, and meanwhile, various impurity components are contained, so that the later separation is difficult, and the environmental-friendly development is not facilitated.
The traditional extraction method not only has complex operation on the extraction process, but also has the defect of low extraction efficiency.
Disclosure of Invention
In view of the above, the present invention aims to provide a method for extracting paclitaxel from branches and leaves of taxus chinensis. The method for extracting the taxol components in the branches and leaves of the taxus chinensis through the ultrahigh pressure auxiliary micelle is simple to operate and high in extraction efficiency.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a method for extracting paclitaxel from branches and leaves of taxus chinensis, which comprises the following steps:
pulverizing branches and leaves of taxus chinensis to obtain taxus chinensis branch and leaf powder;
mixing the taxus chinensis branch and leaf powder with a micelle extraction solvent, and soaking to obtain a mixed solution, wherein the micelle extraction solvent is an N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution;
and carrying out ultrahigh pressure crushing extraction on the mixed solution to obtain a paclitaxel extract, wherein the ultrahigh pressure crushing extraction pressure is 25-300 MPa.
Preferably, the micelle extraction solvent contains 0.4-2% of N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride by volume fraction.
Preferably, the granularity of the taxus chinensis branch and leaf powder is 40-200 meshes.
Preferably, the dosage ratio of the taxus chinensis branch and leaf powder to the micelle extraction solvent is 1g: 10-50 mL.
Preferably, the soaking time is 6-24 h.
Preferably, the mixed solution is placed in a vacuum packaging bag for sealing and then is subjected to ultrahigh pressure crushing extraction.
Preferably, the time for ultrahigh pressure crushing and extraction is 1-15 min.
Preferably, the temperature of the ultrahigh pressure crushing extraction is not higher than room temperature.
Preferably, the ultrahigh pressure crushing extraction is followed by centrifuging the obtained extract to obtain the paclitaxel extract.
Preferably, the rotation speed of the centrifugation is 5000rpm, and the time is 10 min.
The invention provides a method for extracting paclitaxel from branches and leaves of taxus chinensis, which comprises the following steps: pulverizing branches and leaves of taxus chinensis to obtain taxus chinensis branch and leaf powder; mixing the taxus chinensis branch and leaf powder with a micelle extraction solvent, and soaking to obtain a mixed solution, wherein the micelle extraction solvent is an N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution; and carrying out ultrahigh pressure crushing extraction on the mixed solution to obtain a paclitaxel extract, wherein the ultrahigh pressure crushing extraction pressure is 25-300 MPa. The invention realizes the high-efficiency extraction of the paclitaxel in the branches and leaves of the taxus chinensis by the ultrahigh pressure auxiliary micelle extraction technology, N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride is a green and environment-friendly surfactant, under the conditions of normal temperature and high pressure, the micelle extraction solvent and the taxus chinensis branch and leaf powder are subjected to the action of strong shearing force, impact force, turbulent flow, cavity explosive force generated by sudden rising and falling of static pressure and the like when passing through a narrow gap, so that the original suspension of coarse particles is processed into suspension in which fine particles stably exist, the paclitaxel is quickly diffused into the solvent, thereby realizing the quick low-temperature extraction, and the operation is simple, the extraction efficiency is high, plays a certain role in protecting heat-sensitive active ingredients and is suitable for large-scale industrial extraction production. The data of the examples show that the extraction yield of the extraction method of the invention is as high as 88.3%.
Detailed Description
The invention provides a method for extracting paclitaxel from branches and leaves of taxus chinensis, which comprises the following steps:
pulverizing branches and leaves of taxus chinensis to obtain taxus chinensis branch and leaf powder;
mixing the taxus chinensis branch and leaf powder with a micelle extraction solvent, and soaking to obtain a mixed solution, wherein the micelle extraction solvent is an N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution;
and carrying out ultrahigh pressure crushing extraction on the mixed solution to obtain a paclitaxel extract, wherein the ultrahigh pressure crushing extraction pressure is 25-300 MPa.
The invention pulverizes the taxus chinensis branches and leaves to obtain taxus chinensis branch and leaf powder. In the invention, the granularity of the taxus chinensis branch and leaf powder is preferably 40-200 meshes, and more preferably 80-100 meshes.
After the taxus chinensis branch and leaf powder is obtained, the taxus chinensis branch and leaf powder and a micelle extraction solvent are mixed and soaked to obtain a mixed solution.
In the invention, the volume fraction of the N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride in the micelle extraction solvent is preferably 0.4-2%, more preferably 0.8-1.6%, and most preferably 1.2%. The source of the N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution is not particularly limited, and the N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution can be prepared by a preparation method well known to a person skilled in the art.
In the invention, the dosage ratio of the taxus chinensis branch and leaf powder to the micelle extraction solvent is preferably 1g: 10-50 mL, more preferably 1g: 20-40 mL, and most preferably 1g:30 mL.
In the invention, the soaking time is preferably 6-24 h.
After the mixed solution is obtained, the mixed solution is subjected to ultrahigh pressure crushing extraction to obtain a paclitaxel extract, wherein the ultrahigh pressure crushing extraction pressure is 25-300 MPa.
In the invention, the mixed solution is preferably placed in a vacuum packaging bag for sealing and then is subjected to ultrahigh pressure crushing extraction.
In the invention, the time for the ultrahigh-pressure crushing extraction is preferably 1-15 min, more preferably 5-7 min, and the pressure is preferably 100-150 MPa.
In the present invention, the temperature of the ultrahigh pressure crushing extraction is preferably not higher than room temperature. In the present invention, the number of times of the ultrahigh-pressure crushing extraction is preferably 1.
In the present invention, after the ultrahigh pressure disruption extraction, the method preferably further comprises centrifuging the obtained extract to obtain the paclitaxel extract.
In the present invention, the rotation speed of the centrifugation is preferably 5000rpm, and the time is preferably 10 min.
In order to further illustrate the present invention, the following will describe the method for extracting paclitaxel from the branches and leaves of taxus chinensis in detail with reference to the examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Firstly, dried branches and leaves of taxus chinensis are taken as raw materials, crushed and sieved by a 100-mesh sieve, and 5g of branch and leaf powder of taxus chinensis is weighed. Adding micelle extraction solvent (N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution) with volume fraction of 0.8% into the raw material powder, wherein the addition volume (mL) of the micelle extraction solvent is 20 times of the mass (g) of the branch and leaf powder of the taxus chinensis. Then, the suspension is placed in a vacuum packaging bag and hermetically put into a high-pressure crushing extraction device, the extraction pressure is 150MPa, the extraction times are 1 time, and the extraction time is 7 min. And finally, centrifuging the extracting solution after high-pressure crushing at the rotating speed of 5000rpm for 10min, and separating filtrate from filter residue to obtain filtrate, namely the paclitaxel extracting solution. By high performance liquid chromatography detection and analysis, the extraction rate of the paclitaxel is calculated to be 66.7% according to the content of the paclitaxel in the branches and leaves of the taxus chinensis and the content of the paclitaxel in the extracting solution.
Example 2
Firstly, dried branches and leaves of taxus chinensis are taken as raw materials, crushed, sieved by a 80-mesh sieve, and 5g of branch and leaf powder of taxus chinensis is weighed. Adding a micelle extraction solvent (N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution) with the volume fraction of 1.2% into the raw material powder, wherein the addition volume (mL) of the micelle extraction solvent is 30 times of the mass (g) of the branch and leaf powder of the taxus chinensis. Then, the suspension is placed in a vacuum packaging bag and hermetically put into a high-pressure crushing and extracting device, the extracting pressure is 100MPa, the extracting times are 1 time, and the extracting time is 5 min. And finally, centrifuging the extracting solution after high-pressure crushing at the rotating speed of 5000rpm for 10min, and separating filtrate from filter residue to obtain filtrate, namely the paclitaxel extracting solution. Through high performance liquid chromatography detection and analysis, the extraction rate of the paclitaxel is calculated to be 88.3 percent according to the content of the paclitaxel in the branches and leaves of the taxus chinensis and the content of the paclitaxel in the extracting solution.
Example 3
Firstly, dried branches and leaves of taxus chinensis are taken as raw materials, crushed, sieved by a 40-mesh sieve, and 5g of branch and leaf powder of taxus chinensis is weighed. Adding a micelle extraction solvent (N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution) with the volume fraction of 1.6% into the raw material powder, wherein the addition volume (mL) of the micelle extraction solvent is 40 times of the mass (g) of the branch and leaf powder of the taxus chinensis. Then, the suspension is placed in a vacuum packaging bag and hermetically put into a high-pressure crushing extraction device, the extraction pressure is 150MPa, the extraction times are 1 time, and the extraction time is 7 min. And finally, centrifuging the extracting solution after high-pressure crushing at the rotating speed of 5000rpm for 10min, and separating filtrate from filter residue to obtain filtrate, namely the paclitaxel extracting solution. Through high performance liquid chromatography detection and analysis, the extraction rate of the paclitaxel is calculated to be 83.3 percent according to the content of the paclitaxel in the branches and leaves of the taxus chinensis and the content of the paclitaxel in the extracting solution.
Example 4
Firstly, dried branches and leaves of taxus chinensis are taken as raw materials, crushed and sieved by a 100-mesh sieve, and 5g of branch and leaf powder of taxus chinensis is weighed. Adding a micelle extraction solvent (N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution) with the volume fraction of 1.2% into the raw material powder, wherein the addition volume (mL) of the micelle extraction solvent is 50 times of the mass (g) of the branch and leaf powder of the taxus chinensis. Then, the suspension is placed in a vacuum packaging bag and hermetically put into a high-pressure crushing and extracting device, the extracting pressure is 100MPa, the extracting times are 1 time, and the extracting time is 5 min. And finally, centrifuging the extracting solution after high-pressure crushing at the rotating speed of 5000rpm for 10min, and separating filtrate from filter residue to obtain filtrate, namely the paclitaxel extracting solution. Through high performance liquid chromatography detection and analysis, the extraction rate of the paclitaxel is calculated to be 56.7 percent according to the content of the paclitaxel in the branches and leaves of the taxus chinensis and the content of the paclitaxel in the extracting solution.
Example 5
Firstly, dried branches and leaves of taxus chinensis are taken as raw materials, crushed, sieved by a 80-mesh sieve, and 5g of branch and leaf powder of taxus chinensis is weighed. Adding a micelle extraction solvent (N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution) with the volume fraction of 1.2% into the raw material powder, wherein the addition volume (mL) of the micelle extraction solvent is 30 times of the mass (g) of the branch and leaf powder of the taxus chinensis. Then, the suspension is placed in a vacuum packaging bag and hermetically put into a high-pressure crushing and extracting device, the extracting pressure is 100MPa, the extracting times are 1 time, and the extracting time is 5 min. And finally, centrifuging the extracting solution after high-pressure crushing at the rotating speed of 5000rpm for 10min, and separating filtrate from filter residue to obtain filtrate, namely the paclitaxel extracting solution. Through high performance liquid chromatography detection and analysis, the extraction rate of the paclitaxel is calculated to be 80% according to the content of the paclitaxel in the branches and leaves of the taxus chinensis and the content of the paclitaxel in the extracting solution.
Example 6
Firstly, dried branches and leaves of taxus chinensis are taken as raw materials, crushed and sieved by a 100-mesh sieve, and 5g of branch and leaf powder of taxus chinensis is weighed. Adding a micelle extraction solvent (N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution) with the volume fraction of 1.6% into the raw material powder, wherein the addition volume (mL) of the micelle extraction solvent is 20 times of the mass (g) of the branch and leaf powder of the taxus chinensis. Then, the suspension is placed in a vacuum packaging bag and hermetically put into a high-pressure crushing extraction device, the extraction pressure is 150MPa, the extraction times are 1 time, and the extraction time is 7 min. And finally, centrifuging the extracting solution after high-pressure crushing at the rotating speed of 5000rpm for 10min, and separating filtrate from filter residue to obtain filtrate, namely the paclitaxel extracting solution. Through high performance liquid chromatography detection and analysis, the extraction rate of the paclitaxel is calculated to be 70.6 percent according to the content of the paclitaxel in the branches and leaves of the taxus chinensis and the content of the paclitaxel in the extracting solution.
Example 7
Firstly, dried branches and leaves of taxus chinensis are taken as raw materials, crushed, sieved by a 80-mesh sieve, and 5g of branch and leaf powder of taxus chinensis is weighed. Adding a micelle extraction solvent (N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution) with the volume fraction of 1.6% into the raw material powder, wherein the addition volume (mL) of the micelle extraction solvent is 20 times of the mass (g) of the branch and leaf powder of the taxus chinensis. Then, the suspension is placed in a vacuum packaging bag and hermetically put into a high-pressure crushing extraction device, the extraction pressure is 150MPa, the extraction times are 1 time, and the extraction time is 3 min. And finally, centrifuging the extracting solution after high-pressure crushing at the rotating speed of 5000rpm for 10min, and separating filtrate from filter residue to obtain filtrate, namely the paclitaxel extracting solution. By high performance liquid chromatography detection and analysis, the extraction rate of the paclitaxel is calculated to be 55.6% according to the content of the paclitaxel in the branches and leaves of the taxus chinensis and the content of the paclitaxel in the extracting solution.
Example 8
Firstly, dried branches and leaves of taxus chinensis are taken as raw materials, crushed, sieved by a 40-mesh sieve, and 5g of branch and leaf powder of taxus chinensis is weighed. Adding a micelle extraction solvent (N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution) with the volume fraction of 1.6% into the raw material powder, wherein the addition volume (mL) of the micelle extraction solvent is 40 times of the mass (g) of the branch and leaf powder of the taxus chinensis. Then, the suspension is placed in a vacuum packaging bag and hermetically put into a high-pressure crushing and extracting device, the extracting pressure is 50MPa, the extracting times are 1 time, and the extracting time is 3 min. And finally, centrifuging the extracting solution after high-pressure crushing at the rotating speed of 5000rpm for 10min, and separating filtrate from filter residue to obtain filtrate, namely the paclitaxel extracting solution. Through high performance liquid chromatography detection and analysis, the extraction rate of the paclitaxel is calculated to be 67.8% according to the content of the paclitaxel in the branches and leaves of the taxus chinensis and the content of the paclitaxel in the extracting solution.
Example 9
Firstly, dried branches and leaves of taxus chinensis are taken as raw materials, crushed and sieved by a 100-mesh sieve, and 5g of branch and leaf powder of taxus chinensis is weighed. Adding a micelle extraction solvent (N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanolammonium chloride solution) with the volume fraction of 2% into the raw material powder, wherein the addition volume (mL) of the micelle extraction solvent is 10 times of the mass (g) of the branch and leaf powder of the taxus chinensis. Then, the suspension is placed in a vacuum packaging bag and hermetically put into a high-pressure crushing and extracting device, the extracting pressure is 25MPa, the extracting times are 1 time, and the extracting time is 7 min. And finally, centrifuging the extracting solution after high-pressure crushing at the rotating speed of 5000rpm for 10min, and separating filtrate from filter residue to obtain filtrate, namely the paclitaxel extracting solution. By high performance liquid chromatography detection and analysis, the extraction rate of the paclitaxel is calculated to be 43.7% according to the content of the paclitaxel in the branches and leaves of the taxus chinensis and the content of the paclitaxel in the extracting solution.
Example 10
Firstly, dried branches and leaves of taxus chinensis are taken as raw materials, crushed and sieved by a 100-mesh sieve, and 5g of branch and leaf powder of taxus chinensis is weighed. Adding micelle extraction solvent (N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution) with volume fraction of 0.4% into the raw material powder, wherein the addition volume (mL) of the micelle extraction solvent is 50 times of the mass (g) of the branch and leaf powder of the taxus chinensis. Then, the suspension is placed in a vacuum packaging bag and hermetically put into a high-pressure crushing and extracting device, the extracting pressure is 300MPa, the extracting times are 1 time, and the extracting time is 7 min. And finally, centrifuging the extracting solution after high-pressure crushing at the rotating speed of 5000rpm for 10min, and separating filtrate from filter residue to obtain filtrate, namely the paclitaxel extracting solution. Through high performance liquid chromatography detection and analysis, the extraction rate of the paclitaxel is calculated to be 69.3 percent according to the content of the paclitaxel in the branches and leaves of the taxus chinensis and the content of the paclitaxel in the extracting solution.
The foregoing is merely a preferred embodiment of the invention and is not intended to limit the invention in any manner. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be construed as the protection scope of the present invention.
Claims (10)
1. A method for extracting paclitaxel from branches and leaves of taxus chinensis is characterized by comprising the following steps:
pulverizing branches and leaves of taxus chinensis to obtain taxus chinensis branch and leaf powder;
mixing the taxus chinensis branch and leaf powder with a micelle extraction solvent, and soaking to obtain a mixed solution, wherein the micelle extraction solvent is an N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride solution;
and carrying out ultrahigh pressure crushing extraction on the mixed solution to obtain a paclitaxel extract, wherein the ultrahigh pressure crushing extraction pressure is 25-300 MPa.
2. The extraction method according to claim 1, wherein the micelle extraction solvent contains 0.4 to 2% by volume of N- (3-hydrogenated abietic acid acyl-2-hydroxy) propyl-N, N, N-triethanol ammonium chloride.
3. The extraction method according to claim 1, wherein the particle size of the Taxus chinensis shoot and leaf powder is 40-200 mesh.
4. The extraction method according to claim 1, 2 or 3, wherein the ratio of the taxus chinensis branch and leaf powder to the micelle extraction solvent is 1g: 10-50 mL.
5. The extraction method according to claim 1, wherein the soaking time is 6-24 hours.
6. The extraction method according to claim 1, wherein the mixed solution is subjected to ultrahigh pressure crushing extraction after being sealed in a vacuum packaging bag.
7. The extraction method according to claim 1, wherein the time for the ultra-high pressure crushing extraction is 1 to 15 min.
8. The extraction method according to claim 1, wherein the temperature of the ultra-high pressure crushing extraction is not higher than room temperature.
9. The extraction method according to claim 1, further comprising centrifuging the obtained extract after the ultra-high pressure disruption extraction to obtain the paclitaxel extract.
10. The extraction method according to claim 9, wherein the centrifugation is performed at 5000rpm for 10 min.
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