CN111253345A - A kind of extraction method of paclitaxel in yew branches and leaves - Google Patents

Abstract

The invention provides a method for extracting paclitaxel from branches and leaves of Taxus chinensis, which belongs to the field of extraction of fat-soluble chemical components. The invention realizes the efficient extraction of paclitaxel from the branches and leaves of Taxus chinensis by the ultra-high pressure assisted micelle extraction technology, and N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanolammonium chloride It is a green and environmentally friendly surfactant. When the micelle extraction solvent and the yew branch and leaf powder pass through a narrow gap under normal temperature and high pressure conditions, they are subjected to strong shearing force, impact force and turbulent flow, as well as sudden rise and fall of static pressure. The resulting hole explosion force, etc., makes the suspension of the original coarse particles process into a suspension with stable existence of fine particles, so that the paclitaxel diffuses into the solvent quickly, so as to achieve rapid low-temperature extraction, and the operation is simple and the extraction efficiency is high. , has a certain protective effect on heat-sensitive active ingredients, and is suitable for large-scale industrial extraction and production.

Description

A kind of extraction method of paclitaxel in yew branches and leaves

technical field

The invention relates to the technical field of extraction of fat-soluble chemical components, in particular to a method for extracting paclitaxel from branches and leaves of Taxus chinensis.

Background technique

Taxus chinensis belongs to the Taxus family, and the extracts of its fruit, bark and branches and leaves have a variety of active ingredients of medicinal value, among which the taxane diterpenoid secondary metabolites such as paclitaxel have anti-cancer effects with a unique mechanism. The active ingredient of paclitaxel can also be used to treat chronic inflammatory diseases. The injection made of paclitaxel has a very well-known anti-breast cancer effect, and has a therapeutic effect on ovarian cancer and non-small cell lung cancer.

Traditional extraction methods include dipping, percolation, and Soxhlet extraction. The equipment required for the dipping method is simple and easy to operate, but the leaching time is long, the work intensity is high, and the extraction of active ingredients is incomplete. The percolation method is suitable for the extraction of thermally unstable and easily decomposed components. The method is simple and the extraction efficiency is high, but the large amount of solvent causes unnecessary waste. The Soxhlet extraction method is simple to operate, but the extraction time is too long, and volatile organic solvents such as ethanol are required to extract the active ingredients during the extraction process. Conducive to environment-friendly development.

The traditional extraction method is not only complicated in extraction process operation, but also has the disadvantage of low extraction efficiency.

SUMMARY OF THE INVENTION

In view of this, the purpose of the present invention is to provide a method for extracting paclitaxel from branches and leaves of Taxus chinensis. The invention provides a method for extracting paclitaxel components from the branches and leaves of Taxus chinensis by ultra-high pressure-assisted micelles, which is simple in operation and high in extraction efficiency.

In order to achieve the above-mentioned purpose of the invention, the present invention provides the following technical solutions:

The invention provides a method for extracting paclitaxel from branches and leaves of Taxus chinensis, comprising the following steps:

The branches and leaves of the yew are pulverized to obtain the powder of the branches and leaves of the yew;

The yew branch and leaf powder is mixed with a micellar extraction solvent for soaking to obtain a mixed solution, and the micellar extraction solvent is N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N- triethanol ammonium chloride solution;

The mixed solution is subjected to ultra-high pressure crushing and extraction to obtain a paclitaxel extract, and the pressure of the ultra-high pressure crushing and extraction is 25-300 MPa.

Preferably, the volume fraction of N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanolammonium chloride in the micelle extraction solvent is 0.4-2%.

Preferably, the particle size of the yew branch and leaf powder is 40-200 meshes.

Preferably, the dosage ratio of the yew branch and leaf powder to the micellar extraction solvent is 1 g: 10-50 mL.

Preferably, the soaking time is 6-24 hours.

Preferably, the mixed solution is placed in a vacuum packaging bag to be sealed and then subjected to ultra-high pressure crushing and extraction.

Preferably, the time for the ultra-high pressure crushing and extraction is 1-15 min.

Preferably, the temperature of the ultra-high pressure crushing and extraction is not higher than room temperature.

Preferably, the ultra-high pressure crushing and extraction further comprises centrifuging the obtained extract to obtain the paclitaxel extract.

Preferably, the rotational speed of the centrifugation is 5000 rpm, and the time is 10 min.

The invention provides a method for extracting paclitaxel from branches and leaves of Taxus chinensis, comprising the following steps: pulverizing the branches and leaves of Taxus chinensis to obtain branch and leaf powder of Taxus chinensis; mixing the branch and leaf powder of Taxus chinensis with a micellar extraction solvent for soaking to obtain a mixed solution , the micelle extraction solvent is N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanol ammonium chloride solution; the mixed solution is subjected to ultra-high pressure crushing and extraction, The paclitaxel extract is obtained, and the pressure of the ultra-high pressure crushing and extraction is 25-300 MPa. The invention realizes the efficient extraction of paclitaxel from the branches and leaves of Taxus chinensis by the ultra-high pressure assisted micelle extraction technology, and N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanol ammonium chloride It is a green and environmentally friendly surfactant. When the micelle extraction solvent and the yew branch and leaf powder pass through a narrow gap under normal temperature and high pressure conditions, they are subjected to strong shearing force, impact force and turbulent flow, as well as sudden rise and fall of static pressure. The resulting hole explosion force, etc., makes the suspension of the original coarse particles process into a suspension with stable existence of fine particles, so that the paclitaxel diffuses into the solvent quickly, so as to achieve rapid low-temperature extraction, and the operation is simple and the extraction efficiency is high. , has a certain protective effect on heat-sensitive active ingredients, and is suitable for large-scale industrial extraction and production. The data of the examples show that the extraction rate of the extraction method of the present invention is as high as 88.3%.

Detailed ways

The invention provides a method for extracting paclitaxel from branches and leaves of Taxus chinensis, comprising the following steps:

The branches and leaves of the yew are pulverized to obtain the powder of the branches and leaves of the yew;

The yew branch and leaf powder is mixed with a micellar extraction solvent for soaking to obtain a mixed solution, and the micellar extraction solvent is N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N- triethanol ammonium chloride solution;

The mixed solution is subjected to ultra-high pressure crushing and extraction to obtain a paclitaxel extract, and the pressure of the ultra-high pressure crushing and extraction is 25-300 MPa.

In the present invention, the branches and leaves of Taxus chinensis are pulverized to obtain the branches and leaves of Taxus chinensis powder. In the present invention, the particle size of the yew branch and leaf powder is preferably 40-200 mesh, more preferably 80-100 mesh.

After the yew branch and leaf powder is obtained, the present invention mixes the yew branch and leaf powder with a micellar extraction solvent for soaking to obtain a mixed solution.

In the present invention, the volume fraction of N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanolammonium chloride in the micelle extraction solvent is preferably 0.4-2% , more preferably 0.8 to 1.6%, and most preferably 1.2%. The present invention has no special limitation on the source of the N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanol ammonium chloride solution, and adopts the preparation method well known to those skilled in the art. method can be obtained.

In the present invention, the dosage ratio of the yew branch and leaf powder to the micelle extraction solvent is preferably 1g:10-50mL, more preferably 1g:20-40mL, and most preferably 1g:30mL.

In the present invention, the soaking time is preferably 6-24 hours.

After the mixed solution is obtained, in the present invention, the mixed solution is subjected to ultra-high pressure crushing and extraction to obtain a paclitaxel extract, and the pressure of the ultra-high pressure crushing and extraction is 25-300 MPa.

In the present invention, the mixed solution is preferably placed in a vacuum packaging bag to be sealed and then subjected to ultra-high pressure crushing and extraction.

In the present invention, the ultra-high pressure crushing and extraction time 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 ultra-high pressure crushing and extraction is preferably not higher than room temperature. In the present invention, the number of times of ultra-high pressure crushing and extraction is preferably 1 time.

In the present invention, the ultra-high pressure crushing and extraction preferably further comprises centrifuging the obtained extract to obtain the paclitaxel extract.

In the present invention, the rotational speed of the centrifugation is preferably 5000 rpm, and the time is preferably 10 min.

In order to further illustrate the present invention, the method for extracting paclitaxel from the branches and leaves of Taxus chinensis provided by the present invention is described in detail below with reference to examples, but they should not be construed as limiting the protection scope of the present invention.

Example 1

First, the dried branches and leaves of Taxus chinensis were used as raw materials, pulverized, passed through a 100-mesh sieve, and 5 g of the branches and leaves of Taxus chinensis powder were weighed. Add 0.8% volume fraction of micellar extraction solvent (N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanolammonium chloride solution) to the raw material powder, micellar extraction The added volume (mL) of the solvent is 20 times the mass (g) of the leaf powder of Taxus chinensis. Then, the above suspension was placed in a vacuum packaging bag, sealed and put into a high-pressure crushing and extraction device. The extraction pressure was 150 MPa, the extraction frequency was 1 time, and the extraction time was 7 min. Finally, the high-pressure crushed extract was centrifuged at 5000 rpm for 10 min, the filtrate was separated from the filter residue, and the obtained filtrate was the paclitaxel extract. Through high performance liquid chromatography detection and analysis, according to the content of paclitaxel in the branches and leaves of Taxus chinensis and the content of paclitaxel in the extract, the extraction rate of paclitaxel was calculated to be 66.7%.

Example 2

First, the dried branches and leaves of Taxus chinensis were used as raw materials, pulverized, passed through an 80-mesh sieve, and 5 g of the branches and leaves of Taxus chinensis powder were weighed. Add 1.2% volume fraction of micellar extraction solvent (N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanolammonium chloride solution) to the raw material powder, micellar extraction The added volume (mL) of the solvent is 30 times the mass (g) of the leaf powder of Taxus chinensis. Then, the above suspension was placed in a vacuum packaging bag, sealed and put into a high-pressure crushing and extraction device. The extraction pressure was 100 MPa, the extraction frequency was 1 time, and the extraction time was 5 min. Finally, the high-pressure crushed extract was centrifuged at 5000 rpm for 10 min, the filtrate was separated from the filter residue, and the obtained filtrate was the paclitaxel extract. Through high performance liquid chromatography detection and analysis, according to the content of paclitaxel in the branches and leaves of Taxus chinensis and the content of paclitaxel in the extract, the extraction rate of paclitaxel was calculated to be 88.3%.

Example 3

First, the dried branches and leaves of Taxus chinensis were used as raw materials, pulverized, passed through a 40-mesh sieve, and 5 g of the branches and leaves of Taxus chinensis powder were weighed. Add 1.6% volume fraction of micellar extraction solvent (N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanolammonium chloride solution) to the raw material powder, micellar extraction The added volume (mL) of the solvent is 40 times the mass (g) of the leaf powder of Taxus chinensis. Then, the above suspension was placed in a vacuum packaging bag, sealed and put into a high-pressure crushing and extraction device. The extraction pressure was 150 MPa, the extraction frequency was 1 time, and the extraction time was 7 min. Finally, the high-pressure crushed extract was centrifuged at 5000 rpm for 10 min, the filtrate was separated from the filter residue, and the obtained filtrate was the paclitaxel extract. Through the detection and analysis of high performance liquid chromatography, according to the content of paclitaxel in the branches and leaves of Taxus chinensis and the content of paclitaxel in the extract, the extraction rate of paclitaxel was calculated to be 83.3%.

Example 4

First, the dried branches and leaves of Taxus chinensis were used as raw materials, pulverized, passed through a 100-mesh sieve, and 5 g of the branches and leaves of Taxus chinensis powder were weighed. Add 1.2% volume fraction of micellar extraction solvent (N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanolammonium chloride solution) to the raw material powder, micellar extraction The added volume (mL) of the solvent is 50 times the mass (g) of the leaf powder of Taxus chinensis. Then, the above suspension was placed in a vacuum packaging bag, sealed and put into a high-pressure crushing and extraction device. The extraction pressure was 100 MPa, the extraction frequency was 1 time, and the extraction time was 5 min. Finally, the high-pressure crushed extract was centrifuged at 5000 rpm for 10 min, the filtrate was separated from the filter residue, and the obtained filtrate was the paclitaxel extract. Through high performance liquid chromatography detection and analysis, according to the content of paclitaxel in the branches and leaves of Taxus chinensis and the content of paclitaxel in the extract, the extraction rate of paclitaxel was calculated to be 56.7%.

Example 5

First, the dried branches and leaves of Taxus chinensis were used as raw materials, pulverized, passed through an 80-mesh sieve, and 5 g of the branches and leaves of Taxus chinensis powder were weighed. Add 1.2% volume fraction of micellar extraction solvent (N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanolammonium chloride solution) to the raw material powder, micellar extraction The added volume (mL) of the solvent is 30 times the mass (g) of the leaf powder of Taxus chinensis. Then, the above suspension was placed in a vacuum packaging bag, sealed and put into a high-pressure crushing and extraction device. The extraction pressure was 100 MPa, the extraction frequency was 1 time, and the extraction time was 5 min. Finally, the high-pressure crushed extract was centrifuged at 5000 rpm for 10 min, the filtrate was separated from the filter residue, and the obtained filtrate was the paclitaxel extract. Through high performance liquid chromatography detection and analysis, according to the content of paclitaxel in the branches and leaves of Taxus chinensis and the content of paclitaxel in the extract, the extraction rate of paclitaxel was calculated to be 80%.

Example 6

First, the dried branches and leaves of Taxus chinensis were used as raw materials, pulverized, passed through a 100-mesh sieve, and 5 g of the branches and leaves of Taxus chinensis powder were weighed. Add 1.6% volume fraction of micellar extraction solvent (N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanolammonium chloride solution) to the raw material powder, micellar extraction The added volume (mL) of the solvent is 20 times the mass (g) of the leaf powder of Taxus chinensis. Then, the above suspension was placed in a vacuum packaging bag, sealed and put into a high-pressure crushing and extraction device. The extraction pressure was 150 MPa, the extraction frequency was 1 time, and the extraction time was 7 min. Finally, the high-pressure crushed extract was centrifuged at 5000 rpm for 10 min, the filtrate was separated from the filter residue, and the obtained filtrate was the paclitaxel extract. Through high performance liquid chromatography detection and analysis, according to the content of paclitaxel in the branches and leaves of Taxus chinensis and the content of paclitaxel in the extract, the extraction rate of paclitaxel was calculated to be 70.6%.

Example 7

First, the dried branches and leaves of Taxus chinensis were used as raw materials, pulverized, passed through an 80-mesh sieve, and 5 g of the branches and leaves of Taxus chinensis powder were weighed. Add 1.6% volume fraction of micellar extraction solvent (N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanolammonium chloride solution) to the raw material powder, micellar extraction The added volume (mL) of the solvent is 20 times the mass (g) of the leaf powder of Taxus chinensis. Then, the above suspension was placed in a vacuum packaging bag, sealed, and put into a high-pressure crushing and extraction device. The extraction pressure was 150 MPa, the extraction frequency was 1 time, and the extraction time was 3 min. Finally, the high-pressure crushed extract was centrifuged at 5000 rpm for 10 min, the filtrate was separated from the filter residue, and the obtained filtrate was the paclitaxel extract. Through high performance liquid chromatography detection and analysis, according to the content of paclitaxel in the branches and leaves of Taxus chinensis and the content of paclitaxel in the extract, the extraction rate of paclitaxel was calculated to be 55.6%.

Example 8

First, the dried branches and leaves of Taxus chinensis were used as raw materials, pulverized, passed through a 40-mesh sieve, and 5 g of the branches and leaves of Taxus chinensis powder were weighed. Add 1.6% volume fraction of micellar extraction solvent (N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanolammonium chloride solution) to the raw material powder, micellar extraction The added volume (mL) of the solvent is 40 times the mass (g) of the leaf powder of Taxus chinensis. Then, the above suspension was placed in a vacuum packaging bag, sealed, and put into a high-pressure crushing and extraction device. The extraction pressure was 50 MPa, the extraction frequency was 1 time, and the extraction time was 3 min. Finally, the high-pressure crushed extract was centrifuged at 5000 rpm for 10 min, the filtrate was separated from the filter residue, and the obtained filtrate was the paclitaxel extract. Through high performance liquid chromatography detection and analysis, according to the content of paclitaxel in the branches and leaves of Taxus chinensis and the content of paclitaxel in the extract, the extraction rate of paclitaxel was calculated to be 67.8%.

Example 9

First, the dried branches and leaves of Taxus chinensis were used as raw materials, pulverized, passed through a 100-mesh sieve, and 5 g of the branches and leaves of Taxus chinensis powder were weighed. Add 2% volume fraction of micelle extraction solvent (N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanol ammonium chloride solution) to the raw material powder, micelle extraction The added volume (mL) of the solvent is 10 times the mass (g) of the leaf powder of Taxus chinensis. Then, the above suspension was placed in a vacuum packaging bag, sealed and put into a high-pressure crushing and extraction device. The extraction pressure was 25 MPa, the extraction frequency was 1 time, and the extraction time was 7 min. Finally, the high-pressure crushed extract was centrifuged at 5000 rpm for 10 min, the filtrate was separated from the filter residue, and the obtained filtrate was the paclitaxel extract. Through high performance liquid chromatography detection and analysis, according to the content of paclitaxel in the branches and leaves of Taxus chinensis and the content of paclitaxel in the extract, the extraction rate of paclitaxel was calculated to be 43.7%.

Example 10

First, the dried branches and leaves of Taxus chinensis were used as raw materials, pulverized, passed through a 100-mesh sieve, and 5 g of the branches and leaves of Taxus chinensis powder were weighed. Add 0.4% volume fraction of micelle extraction solvent (N-(3-hydroabietic acid acyl-2-hydroxy)propyl-N,N,N-triethanolammonium chloride solution) to the raw material powder to extract micelles The added volume (mL) of the solvent is 50 times the mass (g) of the leaf powder of Taxus chinensis. Then, the above suspension was placed in a vacuum packaging bag, sealed and put into a high-pressure crushing and extraction device. The extraction pressure was 300 MPa, the extraction frequency was 1 time, and the extraction time was 7 min. Finally, the high-pressure crushed extract was centrifuged at 5000 rpm for 10 min, the filtrate was separated from the filter residue, and the obtained filtrate was the paclitaxel extract. Through high performance liquid chromatography detection and analysis, according to the content of paclitaxel in the branches and leaves of Taxus chinensis and the content of paclitaxel in the extract, the extraction rate of paclitaxel was calculated to be 69.3%.

The above descriptions are only preferred embodiments of the present invention, and do not limit the present invention in any form. It should be pointed out that for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can also be made, and these improvements and modifications should also be regarded 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.