Green extraction and efficient enrichment method of camptothecin and 10-hydroxycamptothecin

Abstract

The invention relates to the technical field of extraction and enrichment of natural products, in particular to a green extraction and efficient enrichment method of camptothecin (CPT) and 10-hydroxycamptothecin (HCPT). The problems of low efficiency, poor safety, high cost and the like in the common technology are solved. The method comprises the following steps: mixing dried camptotheca acuminate tissue powder with a nonionic surfactant Genapol X-080 according to a certain solid-to-liquid ratio, extracting in extraction equipment, and centrifuging after extraction to obtain CPT and HCPT extracting solutions; adding inorganic salt into the extracting solution at a certain temperature, uniformly stirring, carrying out directional enrichment treatment for a certain time, layering the solution by centrifuging or standing, and taking an upper-layer surfactant-rich phase, so as to obtain an enriched product of the CPT and HCPT extracting solution. According to the method, no organic solvent is used in the extraction and enrichment process, the cost is low, safety and reliability are achieved, high economic benefits and environmental benefits are achieved, good application and popularization prospects are achieved, and the method has important significance on industrial safety production of CPT and HCPT.

Description

Technical Field

[0001] The invention relates to the technical field of natural product extraction and enrichment, and in particular to a method for green extraction and efficient enrichment of camptothecin and 10-hydroxycamptothecin.

[0002] Camptotheca acuminata Decne is a perennial deciduous tree of the family Camptotheca and the genus Camptotheca. It is endemic to China and widely distributed in various regions south of the Yangtze River. The whole plant of Camptotheca acuminata (such as seeds, fruits, leaves, flowers, branches, bark, and roots) contains antitumor active alkaloids, Camptothecin (CPT), which exerts a unique anticancer effect of cytotoxicity by inhibiting topoisomerase I. Screening new compounds with the same anticancer mechanism as Camptothecin and high efficiency and low toxicity from Camptothecin derivatives has become one of the important methods for obtaining new anticancer drugs. 10-Hydroxycamptothecin (HCPT) is a derivative of CPT and a natural alkaloid isolated from Camptotheca acuminata. It has low toxicity and good anticancer activity. Camptothecin is an important precursor compound for the development of new anticancer pharmaceutical intermediates. At present, extracting Camptothecin and its derivatives from Camptotheca acuminata seeds, fruits, branches and leaves is still one of the most important ways to obtain Camptothecin and its derivatives.

[0003] Since CPT and HCPT are insoluble in water, the main methods for extracting and enriching camptothecin and hydroxycamptothecin from Camptotheca acuminata seeds are: organic solvent method (ethanol, methanol, ethyl acetate, chloroform, etc.) (Chinese patent CN02132721.1: High-efficiency production process of camptothecin; Chinese patent CN201010529441.2: A method for extracting camptothecin; Chinese patent CN200810046161.9: A method for efficiently separating and purifying camptothecin) , alkaline water extraction (Chinese patent CN201010517601.1: A method for extracting and preparing camptothecin; Zhang Zhengxiang, Mao Gaoxiang, Zhang Maoxiang, Zhang Wenjin. New process for alkaline water extraction of camptothecin, Natural Product Research and Development, 2006, 18: 302-303), supercritical CO2 extraction (Zhang Yuhong, Zhou Zhiqiang, Wang Yang. Extraction of camptothecin from camptothecin seeds by supercritical carbon dioxide, Fine Chemicals, 2012, 29, 8, 757-760), etc. The organic solvent extraction method has the disadvantages of poor safety, high cost, and great environmental pollution. The alkaline water extraction method easily destroys the structure of camptothecin and is difficult to treat wastewater. The supercritical CO2 extraction has the disadvantages of high extraction cost and expensive equipment.

[0004] In order to solve the problems of low efficiency, poor safety and high cost in the traditional extraction and enrichment methods of CPT and HCPT, green extraction and enrichment technology based on surfactant water-based micelles has received increasing attention in recent years. Compared with traditional solvent extraction, the extraction and enrichment method based on the turbidity characteristics of non-ionic surfactants is a green extraction and enrichment technology for fat-soluble natural products that utilizes the solubilization effect and turbidity characteristics of surfactant solutions. This process does not require organic solvents and has little impact on the environment. At the same time, it can better protect the original properties of the extracted natural products and improve their extraction and enrichment efficiency. At present, there is no research report on the extraction and enrichment of CPT and HCPT using the non-ionic surfactant Genapol X-080 aqueous system. Therefore, it is urgent to propose a method for green extraction and efficient enrichment of CPT and HCPT to solve the above technical problems. Summary of the invention

[0005] The present invention aims to solve the problems of low efficiency, poor safety and high cost existing in the conventional technologies. A brief overview of the present invention is given below.

[0006] The present invention provides a new method for green extraction and efficient enrichment of camptothecin and hydroxycamptothecin in acuminata tissue. The method is safe, simple to operate, low in running cost, and can fully extract camptothecin and hydroxycamptothecin and achieve good enrichment effect. It is of great significance to improve the industrial safety production of camptothecin and hydroxycamptothecin.

[0007] The technical solution of the present invention:

[0008] A method for green extraction and efficient enrichment of CPT and HCPT comprises the following steps:

[0009] Step 1: crushing and sieving the dried Camptotheca acuminata tissue to obtain Camptotheca acuminata tissue powder;

[0010] Step 2: Mix the Camptotheca acuminata tissue powder and the nonionic surfactant Genapol X-080, place them in an extraction device for extraction, and obtain CPT and HCPT extracts by centrifugation after extraction;

[0011] Step 3: Add inorganic salts to the CPT and HCPT extracts, stir evenly, and treat at a certain temperature. After the reaction treatment, separate the solution by centrifugation or standing, and take the upper surfactant-rich phase to obtain the enriched product of the CPT and HCPT extracts.

[0012] The acuminata tissue is one or more of acuminata seeds, acuminata fruits, acuminata leaves, acuminata branches, and acuminata root bark. The preferred tissue material is acuminata seeds or acuminata fruits.

[0013] Preferably, the particle size of the tissue powder is 20-80 mesh.

[0014] Preferably, the volume concentration of the nonionic surfactant Genapol X-080 is 1-20% v / v; the volume ratio of the Camptotheca acuminata tissue powder to the nonionic surfactant Genapol X-080 is 1 g:10-80 mL.

[0015] The extraction method comprises one or more of ultrasonic extraction, microwave extraction, ultra-high pressure extraction, thermal reflux extraction, Soxhlet extraction, homogenate extraction, and negative pressure cavitation extraction. The preferred extraction method is ultrasonic extraction.

[0016] The inorganic salt is one of sodium chloride, sodium sulfate, potassium chloride, potassium sulfate, magnesium sulfate, calcium nitrate and ammonium sulfate, preferably sodium chloride.

[0017] Preferably, the mass percentage of the added inorganic salt is 2-20% w / v of the extract.

[0018] Preferably, in step three, the treatment temperature is 20-65° C. and the treatment time is 5-60 min.

[0019] Preferably, the centrifugal speed is 4000-12000 rpm, and the centrifugal time is 5-30 min.

[0020] The present invention has the following beneficial effects:

[0021] Compared with the existing extraction and enrichment methods, the present invention provides a new method for efficiently extracting and enriching CPT and HCPT using an aqueous solution of a nonionic surfactant GenapolX-080;

[0022] The invention crushes the seeds of Camptotheca acuminata to a certain particle size, thereby expanding the contact area of ​​the extraction solvent and accelerating the dissolution rate of CPT and HCPT; adopts the nonionic surfactant Genapol X-080 to carry out green extraction of CPT and HCPT in the powder of the seeds of Camptotheca acuminata (or other tissues of Camptotheca acuminata); during the extraction process, the hydrophobic interaction and hydrogen bonding between Genapol X-080 and CPT and HCPT can wrap CPT and HCPT in the hydrophobic core to form a supramolecular system; finally, according to the cloud point characteristics of Genapol X-080, salt is added and the temperature is changed to carry out directional and efficient enrichment of CPT and HCPT in the extract.

[0023] Genapol X-080 is an alkyl polyglycol ether nonionic surfactant with excellent solubilizing and emulsifying properties. It can reduce the surface tension of the solution during the extraction process. By penetrating the plant cell membrane, the hydrophobic groups in its molecular structure can interact with camptothecin and 10-hydroxycamptothecin, while the hydrophilic groups interact with water, increasing the solubility of camptothecin and 10-hydroxycamptothecin in micelles, making it easier to transfer from camptothecin tissue to the extraction solvent, and promoting the release of camptothecin and 10-hydroxycamptothecin from plant cells. Compared with some cationic or anionic surfactants, Genapol X-080 has higher biodegradability and environmental friendliness, and can complete the extraction under milder conditions, so it has advantages in environmental protection. In addition, Genapol X-080 can also be mixed with other surfactants, such as polyether surfactants, to improve the extraction selectivity and extraction efficiency through the interaction between surfactants.

[0024] The present invention uses Genapol X-080 aqueous solution to extract and enrich CPT and HCPT. The extraction solvent is green, safe, and less harmful to the human body. The processing temperature during enrichment is low, which saves energy consumption. It avoids the shortcomings of traditional organic solvent extraction, such as large solvent usage, high extraction cost, environmental unfriendliness, and potential safety hazards. In summary, the extraction and enrichment method provided by the present invention is simple to operate, has high extraction and enrichment efficiency, low energy consumption, does not use any organic solvents during the extraction process, and the entire process is safe and reliable, has high economic and environmental benefits, and is suitable for industrial production.

[0025] The extraction method of the invention can achieve extraction rates of CPT and HCPT in Camptotheca acuminata seeds of 96.64% and 95.59% respectively, and enrichment rates of CPT and HCPT in the extract of 99.32% and 99.18% respectively. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is the structural diagram of camptothecin and 10-hydroxycamptothecin;

[0027] Figure 2 It is a comparison diagram of camptothecin and 10-hydroxycamptothecin extracts before enrichment (A) and after enrichment (B);

[0028] Figure 3 HPLC chromatograms of camptothecin and 10-hydroxycamptothecin of standard and sample; (A) mixed standard, (B) Genapol X-080 extract of camptothecin seeds, (C) enriched phase of Genapol X-080 extract; in the chromatogram, 1. 10-hydroxycamptothecin, 2. camptothecin;

[0029] Figure 4These are the HPLC standard curves of camptothecin and 10-hydroxycamptothecin; (A) in the figure is the standard curve of camptothecin, and (B) is the standard curve of 10-hydroxycamptothecin. DETAILED DESCRIPTION

[0030] The present invention is described in detail below in conjunction with the embodiments and the accompanying drawings. However, the following embodiments should not be considered as limiting the scope of the present invention.

[0031] High performance liquid chromatography detection: In the following examples, high performance liquid chromatography was used to detect the extraction rate and enrichment rate of the product. The specific detection conditions are as follows:

[0032] Mobile phase conditions: The ratio of pure water (A) to methanol (B) is: 0-11min, 45-55%B; 11-15min, 55-45%B.

[0033] Detection wavelength: 254nm;

[0034] Injection volume: 10 μL;

[0035]

[0036] Determine the content of CPT or HCPT in the sample using a high performance liquid chromatography standard curve: take an appropriate amount of CPT and HCPT standards, and use chromatographic methanol to prepare a mixed standard stock solution containing 114 μg / mL CPT and 187 μg / mL HCPT; dilute the mixed standard solution to 1 / 2, 1 / 4, 1 / 8, 1 / 16, and 1 / 32 of the mother solution concentration; use the concentration of the diluted standard solution as the horizontal axis and the corresponding chromatographic peak area of ​​CPT and HCPT as the vertical axis to draw the standard curves of CPT and HCPT, respectively. The obtained standard curves are as follows: Figure 4 As shown, the standard curve of CPT obtained by fitting is y=544.65x-1560.8 (R 2 =0.9999), the standard curve of HCPT is y=137.82x-154.77 (R 2 =0.9999), where x is the concentration of CPT and HCPT, and y is the corresponding peak area. The contents of CPT and HCPT in the sample were calculated based on the corresponding peak areas in the standard curve and the chromatogram.

[0037] Example 1

[0038] A method for green extraction and efficient enrichment of CPT and HCPT comprises the following steps:

[0039] Step 1: Grind the dried Camptotheca acuminata seeds through a 40-mesh sieve to obtain dried Camptotheca acuminata seed powder for later use;

[0040] Step 2: Take 20g of Camptotheca acuminata seed powder obtained by crushing and sieving and 30 times the amount of 6% (v / v) non-ionic surfactant Genapol X-080 solution, put them into a polyethylene bag and seal them, place them in a high-pressure cylinder of a pressurized extraction equipment, set the extraction pressure to 50MPa, hold the pressure for 5min, repeat 3 times, centrifuge at 8000rpm for 15min after the extraction, take the supernatant, which is the extract of CPT and HCPT, and determine the contents of CPT and HCPT in the seed powder and the contents of CPT and HCPT in the extract by high performance liquid chromatography (taking the average value of 3 experiments), and calculate the extraction rates of CPT and HCPT to be 89.09% and 87.31%, respectively;

[0041] Step 3: adding sodium chloride in an amount of 10% (w / v) of the volume of the extract to the CPT and HCPT extracts, stirring for 15 minutes to fully mix the extracts, and enriching the extracts at 35° C. for 20 minutes; after the enrichment of the extracts, centrifuging at 10,000 rpm for 8 minutes to separate the layers, taking the upper surfactant-rich phase to obtain the enriched products of CPT and HCPT; detecting the contents of CPT and HCPT in the extracts before and after enrichment by high performance liquid chromatography, and calculating the enrichment rates of CPT and HCPT to be 99.11% and 99.17%, respectively.

[0042] Example 2

[0043] A method for green extraction and efficient enrichment of CPT and HCPT comprises the following steps:

[0044] Step 1: Grind the dried Camptotheca acuminata seeds through a 20-mesh sieve to obtain dried Camptotheca acuminata seed powder for later use;

[0045] Step 2: Take 20g of Camptotheca acuminata seed powder obtained by crushing and sieving and 20 times the amount of 8% (v / v) non-ionic surfactant Genapol X-080 solution, place them in a microwave extraction device, extract for 15 minutes at a microwave temperature of 25°C and a microwave power of 420W, and extract three times. After the extraction, centrifuge at 4000rpm for 12 minutes, and take the supernatant, which is the extract of CPT and HCPT. The contents of CPT and HCPT in the seed powder and the contents of CPT and HCPT in the extract are determined by high performance liquid chromatography (taking the average value of 3 experiments), and the extraction rates of CPT and HCPT are calculated to be 88.10% and 85.39%, respectively;

[0046] Step 3: Add 8% (w / v) sodium chloride to the CPT and HCPT extracts, stir for 15 minutes to fully mix, and enrich the extracts at 30°C for 15 minutes. After the enrichment, centrifuge at 10,000 rpm for 5 minutes to separate the layers, and take the upper surfactant-rich phase to obtain the enriched product of the CPT and HCPT extracts. The contents of CPT and HCPT in the extracts before and after enrichment were detected by high performance liquid chromatography, and the enrichment rates of CPT and HCPT were calculated to be 99.07% and 99.04%, respectively.

[0047] Example 3

[0048] A method for green extraction and efficient enrichment of CPT and HCPT comprises the following steps:

[0049] Step 1: Grind the dried Camptotheca acuminata seeds through a 60-mesh sieve to obtain dried Camptotheca acuminata seed powder for later use;

[0050] Step 2: Take 20g of Camptotheca acuminata seed powder obtained by crushing and sieving and 50 times the amount of 10% (v / v) non-ionic surfactant Genapol X-080 solution, place them in an ultrasonic extraction device, extract at 480W power for 50 minutes, and extract 3 times. After each extraction, centrifuge at 6000rpm for 10 minutes, and take the supernatant, which is the extract of CPT and HCPT. The content of CPTHCPT in the seed powder and the content of CPT and 10-hydroxyCPT in the extract were determined by high performance liquid chromatography (taking the average value of 3 experiments), and the extraction rates of CPT and HCPT were calculated to be 96.09% and 93.2%, respectively;

[0051] Step 3: Add 20% (w / v) sodium chloride to the CPT and HCPT extracts, stir for 15 minutes to fully mix, and enrich the extracts at 40°C for 30 minutes. After the enrichment, centrifuge at 6000 rpm for 10 minutes to separate the layers, and take the upper surfactant-rich phase to obtain the enriched product of the CPT and HCPT extracts. The contents of CPT and HCPT in the extracts before and after enrichment were detected by high performance liquid chromatography, and the enrichment rates of CPT and HCPT were calculated to be 99.32% and 99.12%, respectively.

[0052] Example 4

[0053] A method for green extraction and efficient enrichment of CPT and HCPT comprises the following steps:

[0054] Step 1: Grind the dried Camptotheca acuminata seeds through a 60-mesh sieve to obtain dried Camptotheca acuminata seed powder for later use;

[0055] Step 2: Take 20g of Camptotheca acuminata seed powder obtained by crushing and sieving and 40 times the amount of 12% (v / v) non-ionic surfactant Genapol X-080 solution, place them in an ultrasonic extraction device, extract at 540W power for 30 minutes, and extract 3 times. After each extraction, centrifuge at 8000rpm for 6 minutes, and take the supernatant, which is the extract of CPT and HCPT. The contents of CPT and HCPT in the seed powder and the contents of CPT and HCPT in the extract are determined by high performance liquid chromatography (taking the average value of 3 experiments), and the extraction rates of CPT and HCPT are calculated to be 94.33% and 92.93%, respectively;

[0056] Step 3: Add sodium sulfate at 15% (w / v) of the volume of the extract to the CPT and HCPT extracts, stir for 15 minutes to fully mix, and enrich the extract at 25°C for 30 minutes. After the enrichment, centrifuge at 6500 rpm for 8 minutes to separate the layers, and take the upper surfactant-rich phase to obtain the enriched products of CPT and HCPT. The contents of CPT and HCPT in the extracts before and after enrichment were detected by high performance liquid chromatography, and the enrichment rates of CPT and HCPT were calculated to be 99.08% and 99.24%, respectively.

[0057] Example 5

[0058] A method for green extraction and efficient enrichment of CPT and HCPT comprises the following steps:

[0059] Step 1: Grind the dried Camptotheca acuminata seeds through an 80-mesh sieve to obtain dried Camptotheca acuminata seed powder for later use;

[0060] Step 2: Take 20g of Camptotheca acuminata seed powder obtained by crushing and sieving and 60 times the amount of 14% (v / v) non-ionic surfactant Genapol X-080 solution, place them in an ultrasonic extraction device, extract at 420W power for 80 minutes, and extract 3 times. After each extraction, centrifuge at 7500rpm for 10 minutes, and take the supernatant, which is the extract of CPT and HCPT. The contents of CPT and HCPT in the seed powder and the contents of CPT and HCPT in the extract are determined by high performance liquid chromatography (taking the average value of 3 experiments), and the extraction rates of CPT and HCPT are calculated to be 95.36% and 93.47%, respectively;

[0061] Step 3: Add 4% (w / v) ammonium sulfate to the CPT and HCPT extracts, stir for 15 minutes to fully mix, and enrich the extracts at 45°C for 15 minutes. After the enrichment, centrifuge at 4000 rpm for 5 minutes to separate the layers, and take the upper surfactant-rich phase to obtain the enriched product of the CPT and HCPT extracts. The contents of CPT and HCPT in the extracts before and after enrichment were detected by high performance liquid chromatography, and the enrichment rates of CPT and HCPT were calculated to be 98.88% and 97.98%, respectively.

[0062] Example 6

[0063] A method for green extraction and efficient enrichment of CPT and HCPT comprises the following steps:

[0064] Step 1: Grind the dried Camptotheca acuminata seeds through a 60-mesh sieve to obtain dried Camptotheca acuminata seed powder for later use;

[0065] Step 2: Take 20g of Camptotheca acuminata seed powder obtained by crushing and sieving and 70 times the amount of 10% (v / v) non-ionic surfactant Genapol X-080 solution, place them in an ultrasonic extraction device, extract at 600W power for 50 minutes, and extract 3 times. After each extraction, centrifuge at 4500rpm for 10 minutes, and take the supernatant, which is the extract of CPT and HCPT. The contents of CPT and HCPT in the seed powder and the contents of CPT and HCPT in the extract are determined by high performance liquid chromatography (taking the average value of 3 experiments), and the extraction rates of CPT and HCPT are calculated to be 96.46% and 95.21%, respectively;

[0066] Step 3: Add 18% (w / v) sodium chloride to the CPT and HCPT extracts, stir for 15 minutes to fully mix, and enrich the extracts at 50°C for 10 minutes. After the enrichment, centrifuge at 5500 rpm for 6 minutes to separate the layers, and take the upper surfactant-rich phase to obtain the enriched product of the CPT and HCPT extracts. The contents of CPT and HCPT in the extracts before and after enrichment were detected by high performance liquid chromatography, and the enrichment rates of CPT and HCPT were calculated to be 99.55% and 99.28%, respectively.

[0067] Example 7

[0068] A method for green extraction and efficient enrichment of CPT and HCPT comprises the following steps:

[0069] Step 1: Grind the dried Camptotheca acuminata seeds through a 30-mesh sieve to obtain dried Camptotheca acuminata seed powder for later use;

[0070] Step 2: Take 20g of Camptotheca acuminata seed powder obtained by crushing and sieving and 15 times the amount of 12% (v / v) non-ionic surfactant Genapol X-080 solution, place them in an ultrasonic extraction device, extract at 360W power for 40 minutes, and extract 3 times. After each extraction, centrifuge at 6500rpm for 8 minutes, and take the supernatant, which is the extract of CPT and HCPT. The contents of CPT and HCPT in the seed powder and the contents of CPT and HCPT in the extract are determined by high performance liquid chromatography (taking the average value of 3 experiments), and the extraction rates of CPT and HCPT are calculated to be 90.10% and 91.35%, respectively.

[0071] Step 3: Add 14% (w / v) magnesium sulfate to the CPT and HCPT extracts, stir for 15 minutes to fully mix, and enrich the extracts at 50°C for 15 minutes. After the enrichment, stand at room temperature for 10 minutes to separate the layers, and take the upper surfactant-rich phase to obtain the enriched product of the CPT and HCPT extracts. The contents of CPT and HCPT in the extracts before and after enrichment were detected by high performance liquid chromatography, and the enrichment rates of CPT and HCPT were calculated to be 98.25% and 98.88%, respectively.

[0072] Example 8

[0073] A method for green extraction and efficient enrichment of CPT and HCPT comprises the following steps:

[0074] Step 1: Grind the dried Camptotheca acuminata seeds through a 50-mesh sieve to obtain dried Camptotheca acuminata seed powder for later use;

[0075] Step 2: Take 20g of Camptotheca acuminata seed powder obtained by crushing and sieving and 25 times the amount of 10% (v / v) non-ionic surfactant Genapol X-080 solution, place them in an ultrasonic extraction device, extract at 420W power for 80 minutes, and extract 3 times. After each extraction, centrifuge at 1100rpm for 5 minutes, and take the supernatant, which is the extract of CPT and HCPT. The contents of CPT and HCPT in the seed powder and the contents of CPT and HCPT in the extract are determined by high performance liquid chromatography (taking the average value of 3 experiments), and the extraction rates of CPT and HCPT are calculated to be 96.64% and 95.59%, respectively;

[0076] Step 3: Add 16% (w / v) potassium chloride to the CPT and HCPT extracts, stir for 15 minutes to fully mix, and enrich the extracts at 45°C for 20 minutes. After the enrichment, stand at room temperature for 10 minutes to separate the layers, and take the upper surfactant-rich phase to obtain the enriched product of the CPT and HCPT extracts. The contents of CPT and HCPT in the extracts before and after enrichment were detected by high performance liquid chromatography, and the enrichment rates of CPT and HCPT were calculated to be 99.32% and 99.18%, respectively.

[0077] It should be noted that in the above embodiments, as long as the technical solutions are not contradictory, they can be arranged and combined, and those skilled in the art can exhaust all possibilities based on the mathematical knowledge of arrangement and combination. Therefore, the present invention will no longer describe the technical solutions after arrangement and combination one by one, but it should be understood that the technical solutions after arrangement and combination have been disclosed by the present invention.

[0078] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for green extraction and efficient enrichment of camptothecin (CPT) and 10-hydroxycamptothecin (HCPT), characterized in that: The steps include: Step 1: crushing and sieving the dried Camptotheca acuminata tissue to obtain Camptotheca acuminata tissue powder; Step 2: Mix the Camptotheca acuminata tissue powder and the nonionic surfactant Genapol X-080, place them in an extraction device for extraction, centrifuge them after extraction, and take the supernatant to obtain CPT and HCPT extracts; Step 3: Add inorganic salts to the CPT and HCPT extracts, stir evenly, treat the extracts at a certain temperature for a certain time, and separate the solutions by centrifugation or standing after the reaction treatment. The upper surfactant-rich phase is the enriched product of the CPT and HCPT extracts.

2. The method for green extraction and efficient enrichment of CPT and HCPT according to claim 1, characterized in that: The acuminata tissue is one or more of acuminata seed, acuminata fruit, acuminata leaf, acuminata branch, and acuminata root bark.

3. The method for green extraction and efficient enrichment of CPT and HCPT according to claim 1, characterized in that: The particle size of the Camptotheca acuminata tissue powder is 20-80 meshes.

4. The method for green extraction and efficient enrichment of CPT and HCPT according to claim 1, characterized in that: The volume concentration of the nonionic surfactant Genapol X-080 is 1-20% v / v; the volume ratio of the mass of the camptotheca acuminata tissue powder to the nonionic surfactant Genapol X-080 is 1g:10-80mL.

5. The method for green extraction and efficient enrichment of CPT and HCPT according to claim 1, characterized in that: The extraction method comprises one or more of ultrasonic extraction, microwave extraction, ultra-high pressure extraction, thermal reflux extraction, Soxhlet extraction, homogenate extraction, and negative pressure cavitation extraction.

6. The method for green extraction and efficient enrichment of CPT and HCPT according to claim 1, characterized in that: The inorganic salt is one or more of sodium chloride, sodium sulfate, potassium chloride, potassium sulfate, magnesium sulfate, calcium nitrate and ammonium sulfate.

7. The method for green extraction and efficient enrichment of CPT and HCPT according to claim 1, characterized in that: The mass percentage of the added inorganic salt is 2-20% w / v of the extract.

8. The method for green extraction and efficient enrichment of CPT and HCPT according to claim 1, characterized in that: In step 3, the treatment temperature is 20-65° C. and the treatment time is 5-60 min.

9. The method for green extraction and efficient enrichment of CPT and HCPT according to claim 1, characterized in that: The centrifugal speed is 4000-12000 rpm, and the centrifugal time is 5-30 min.

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