A method for extracting and enriching camptothecin and 10-hydroxycamptothecin

Abstract

The present application relates to the technical field of natural product extraction and enrichment, and particularly relates to a method for extracting and enriching camptothecin (CPT) and 10-hydroxyl camptothecin (HCPT). The method solves the problems of low efficiency, poor safety, high cost and the like of common technologies. Dry camptothecin tissue powder and non-ionic surfactant Genapol X-080 are mixed according to a certain solid-liquid ratio, and then are placed in an extraction device for extraction. After extraction, CPT and HCPT extraction liquid is obtained through centrifugation. Inorganic salt is added to the extraction liquid at a certain temperature, and is uniformly stirred. The solution is subjected to directional enrichment treatment for a certain time, and is subjected to centrifugation or standing to separate the solution into layers. The upper surfactant-rich phase is taken, and the enrichment product of the CPT and HCPT extraction liquid is obtained. The extraction and enrichment process of the present application does not use any organic solvent, and is low in cost, safe and reliable, has high economic and environmental benefits, and has a good popularization and application prospect. The method has important significance for the industrialized safe production of CPT and HCPT.

Description

Technical Field

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

[0002] Camptotheca acuminata (Camptotheca acuminata) Camptotheca acuminata Camptotheca acuminata is a perennial deciduous tree belonging to the genus Camptotheca in the family Nephrolepisaceae. It is endemic to China and widely distributed in regions south of the Yangtze River. The entire plant (including seeds, fruits, leaves, flowers, branches, bark, and roots) contains camptothecin (CPT), an alkaloid with antitumor activity, which exerts its unique anticancer effect by inhibiting topoisomerase I. Screening for new compounds with the same anticancer mechanism as camptothecin but with high efficacy and low toxicity from camptothecin derivatives has become an important method for obtaining novel anticancer drugs. 10-Hydroxycamptothecin (HCPT) is a derivative of CPT, a natural alkaloid isolated from Camptotheca acuminata, with low toxicity and good anticancer activity. Camptothecin is an important precursor compound for developing novel anticancer pharmaceutical intermediates. Currently, extracting camptothecin and its derivatives from Camptotheca acuminata seeds, fruits, branches, and leaves remains one of the most important methods.

[0003] Since CPT and HCPT are poorly soluble in water, the main methods for extracting and enriching camptothecin and hydroxycamptothecin from Camptotheca acuminata seeds currently include: organic solvent extraction (ethanol, methanol, ethyl acetate, chloroform, etc.) (Chinese Patent CN02132721.1: High-efficiency production process of camptothecin; Chinese Patent CN201010529441.2: An extraction method for camptothecin; Chinese Patent CN200810046161.9: A high-efficiency separation and purification method for camptothecin), alkaline water extraction (Chinese Patent CN201010517601.1: An extraction and preparation method for camptothecin; Zhang Zhengxiang, Mao Gaoxiang, Zhang Maoxiang, Zhang Wenjin. A new alkaline water extraction process for camptothecin, Natural Product Research and Development, 2006, 18:302-303), and supercritical extraction. Extraction (Zhang Yuhong, Zhou Zhiqiang, Wang Yang. Supercritical carbon dioxide extraction of camptothecin from Camptotheca acuminata seeds, Fine Chemicals, 2012, 29, 8, 757-760), etc. Organic solvent extraction methods have drawbacks such as poor safety, high cost, and significant environmental pollution. Alkaline water extraction easily damages the camptothecin structure and is difficult to treat wastewater. Supercritical carbon dioxide extraction… Extraction methods have drawbacks such as high extraction costs and expensive equipment.

[0004] To solve the problems of low efficiency, poor safety, high cost and so on in the traditional extraction and enrichment methods of CPT and HCPT, in recent years, the environmentally friendly extraction and enrichment technology based on surfactant water-based micelles has been increasingly concerned. Compared with the traditional solvent extraction, the extraction and enrichment method based on the cloud point characteristics of non-ionic surfactants is a technology for extracting and enriching liposoluble natural products by using the solubilization effect and cloud point characteristics of surfactant solution. The process does not need organic solvents, has little impact on the environment, can better protect the original properties of the extracted natural products, and can improve the extraction and enrichment efficiency. At present, there is no report on the use of non-ionic surfactant Genapol X-080 aqueous system to extract and enrich CPT and HCPT. Therefore, it is urgent to propose a method for extracting and enriching CPT and HCPT to solve the above technical problems. SUMMARY

[0005] The present application is to solve the problems of low efficiency, poor safety, high cost and so on in the conventional technology, and a brief summary of the present application is given below.

[0006] The present application provides a new method for extracting and enriching CPT and HCPT in camptotheca acuminata tissues. The method is safe, simple to operate, low in running cost, and has good extraction and enrichment effects on CPT and HCPT. It not only improves the industrialized safe production of CPT and HCPT, but also has great significance

[0007] The technical scheme of the present application is as follows:

[0008] A method for extracting and enriching CPT and HCPT, comprising the following steps:

[0009] Step one: dry camptotheca acuminata tissues are crushed and sieved to obtain camptotheca acuminata tissue powder;

[0010] Step two: the camptotheca acuminata tissue powder and non-ionic surfactant Genapol X-080 are mixed and placed in an extraction device for extraction. After extraction, CPT and HCPT extract is obtained by centrifugation;

[0011] Step three: inorganic salt is added to the CPT and HCPT extract, stirred uniformly, and treated at a certain temperature. After reaction treatment, the solution is separated by centrifugation or standing, and the upper surfactant-rich phase is taken as the enrichment product of the CPT and HCPT extract.

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

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

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

[0015] The extraction method comprises one or more of ultrasonic extraction, microwave extraction, ultrahigh pressure extraction, hot reflux extraction, Soxhlet extraction, homogenate extraction, and negative pressure cavitation extraction, preferably 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 extraction solution.

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

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

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

[0021] Compared with the prior extraction and enrichment method, the present application provides a new environment-friendly method for extracting and enriching CPT and HCPT by using a non-ionic surfactant Genapol X-080 aqueous solution.

[0022] The present application grinds Camptotheca acuminata seeds to a certain particle size, thereby expanding the contact area of the extraction solvent and accelerating the dissolution rate of CPT and HCPT; the non-ionic surfactant Genapol X-080 is used to extract CPT and HCPT from Camptotheca acuminata seed (or other tissues of Camptotheca acuminata) powder; in the extraction process, the hydrophobic interaction and hydrogen bond interaction 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, CPT and HCPT in the extraction solution are directionally enriched by adding salt and changing temperature.

[0023] Genapol X-080 is a kind of alkyl polyethylene glycol ether non-ionic surfactant, which has excellent solubilizing effect and emulsifying performance, can reduce the surface tension of the solution during the extraction process, and can interact with camptothecin and 10-hydroxycamptothecin through the hydrophobic group in its molecular structure, while the hydrophilic group interacts with water, thereby increasing the solubility of camptothecin and 10-hydroxycamptothecin in micelles, making it easier to transfer from the camptothecin tissue to the extraction solvent, and promoting the release of camptothecin and 10-hydroxycamptothecin from the plant cells. Compared with some cationic or anionic surfactants, Genapol X-080 has higher biodegradability and environmental friendliness, and can complete the extraction under relatively mild conditions, so it has advantages in environmental protection. In addition, Genapol X-080 can also be used in combination with other surfactants, such as polyether surfactants, to improve the extraction selectivity and efficiency through the interaction between surfactants.

[0024] The present application uses Genapol X-080 aqueous solution to extract and enrich CPT and HCPT, and the extraction solvent is environmentally friendly, safe and less harmful to human body, and the treatment temperature during enrichment is relatively low, thereby saving energy consumption; the present application avoids the defects of traditional organic solvent extraction, such as large solvent consumption, high extraction cost, serious environmental pollution and safety hazards; and the extraction and enrichment method provided by the present application is simple in operation, high in extraction and enrichment efficiency, low in energy consumption, free of any organic solvent during the extraction process, safe and reliable in the whole process, and has high economic and environmental benefits, and is suitable for industrial production.

[0025] The extraction method of the present application can achieve an extraction rate of 96.64% and 95.59% for CPT and HCPT in camptothecin seeds respectively, and an enrichment rate of 99.32% and 99.18% for CPT and HCPT in the extraction solution respectively. BRIEF DESCRIPTION OF DRAWINGS

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

[0027] Figure 2 is the comparison diagram of camptothecin and 10-hydroxycamptothecin extraction solution before enrichment (A) and after enrichment (B);

[0028] Figure 3 is the HPLC chromatogram of camptothecin and 10-hydroxycamptothecin of standard and sample; in the figure, (A) is mixed standard, (B) is Genapol X 080 extraction solution of camptothecin seeds, (C) is the enrichment phase of Genapol X 080 extraction solution;

[0029] Figure 4 High performance liquid chromatography standard curve of CPT and HCPT; (A) standard curve of CPT, (B) standard curve of HCPT. DETAILED DESCRIPTION

[0030] The application will be described in detail below with reference to the embodiments and the accompanying drawings. However, the following embodiments should not be considered as limiting the scope of the application.

[0031] High performance liquid chromatography detection: the extraction rate and enrichment rate of the product were detected by high performance liquid chromatography in the following examples. The specific detection conditions are as follows:

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

[0033] Detection wavelength: 254 nm;

[0034] Injection volume: 10 μL;

[0035]

[0036]

[0037] Determination of CPT or HCPT content in samples by high performance liquid chromatography standard curve: take an appropriate amount of CPT and HCPT standard, and prepare a mixed standard mother liquor containing 114 μg / mL of CPT and 187 μg / mL of HCPT with chromatographic methanol; dilute the mixed standard solution to 1 / 2, 1 / 4, 1 / 8, 1 / 16, 1 / 32 of the mother liquor concentration; take the dilution concentration of the standard solution as the abscissa, and the chromatographic peak area of CPT and HCPT as the ordinate, respectively draw the standard curve of CPT and HCPT, and the obtained standard curve is shown in Figure 4 The standard curve of CPT is y=544.65x-1560.8 ( =0.9999), and the standard curve of HCPT is y=137.82x-154.77 ( =0.9999), wherein x is the concentration of CPT and HCPT respectively, and y is the corresponding peak area. According to the standard curve and the corresponding peak area in the chromatogram, the content of CPT and HCPT in the sample is calculated.

[0038] Example 1

[0039] A method for extracting and enriching CPT and HCPT, comprising the following steps:

[0040] Step one: dry Camptotheca acuminata seeds were crushed through a 60 mesh sieve to obtain dry Camptotheca acuminata seed powder for later use;

[0041] Step two: 20 g of the crushed seed powder and 50 times the amount of 10% (v / v) non-ionic surfactant Genapol X-080 solution were placed in an ultrasonic extraction device and extracted for 50 min at a power of 480 W, with 3 extractions. After each extraction, the mixture was centrifuged at 6000 rpm for 10 min, and the supernatant was collected, which was the CPT and HCPT extraction solution. The contents of CPT and HCPT in the seed powder and the extraction solution were determined by high performance liquid chromatography (the average of 3 experiments), and the extraction rates of CPT and HCPT were calculated to be 96.09% and 93.2%, respectively;

[0042] Step three: 20% (w / v) sodium chloride was added to the CPT and HCPT extraction solution, and the mixture was stirred for 15 min to fully mix, and then the extraction solution was enriched at 40°C for 30 min. After enrichment, the mixture was centrifuged at 6000 rpm for 10 min to separate the layers, and the upper surfactant-rich phase was collected, which was the enrichment product of the CPT and HCPT extraction solution. The contents of CPT and HCPT in the extraction solution before and after enrichment were determined by high performance liquid chromatography, and the enrichment rates of CPT and HCPT were calculated to be 99.32% and 99.12%, respectively.

[0043] Example 2

[0044] A method for extracting and enriching CPT and HCPT, comprising the following steps:

[0045] Step one: dry Camptotheca acuminata seeds were crushed through a 60 mesh sieve to obtain dry Camptotheca acuminata seed powder for later use;

[0046] Step two: 20 g of the crushed seed powder and 40 times the amount of 12% (v / v) non-ionic surfactant Genapol X-080 solution were placed in an ultrasonic extraction device and extracted for 30 min at a power of 540 W, with 3 extractions. After each extraction, the mixture was centrifuged at 8000 rpm for 6 min, and the supernatant was collected, which was the CPT and HCPT extraction solution. The contents of CPT and HCPT in the seed powder and the extraction solution were determined by high performance liquid chromatography (the average of 3 experiments), and the extraction rates of CPT and HCPT were calculated to be 94.33% and 92.93%, respectively;

[0047] Step three: add 15% (w / v) of sodium sulfate to the CPT and HCPT extract, stir for 15 min to make it fully mixed, and enrich the extract at 25°C for 30 min. After enrichment, centrifuge at 6500 rpm for 8 min to separate the layers, and take the upper surfactant-rich phase, to obtain the enrichment product of CPT and HCPT. Detect the contents of CPT and HCPT in the extract before and after enrichment by high performance liquid chromatography, and calculate the enrichment rates of CPT and HCPT to be 99.08% and 99.24%, respectively.

[0048] Example 3

[0049] A method for extracting and enriching CPT and HCPT, comprising the following steps:

[0050] Step one: crush the dried camptotheca acuminata seeds through an 80-mesh sieve to obtain dried camptotheca acuminata seed powder for later use;

[0051] Step two: take 20 g of the crushed seed powder and 60 times the amount of 14% (v / v) non-ionic surfactant Genapol X-080 solution, and place them in an ultrasonic extraction device to extract for 80 min at a power of 420 W, for 3 times. After each extraction, centrifuge at 7500 rpm for 10 min, and take the supernatant, which is the CPT and HCPT extract. Detect the contents of CPT and HCPT in the seed powder and the extract (take the average of 3 experiments) by high performance liquid chromatography, and calculate the extraction rates of CPT and HCPT to be 95.36% and 93.47%, respectively.

[0052] Step three: add 4% (w / v) of ammonium sulfate to the CPT and HCPT extract, stir for 15 min to make it fully mixed, and enrich the extract at 45°C for 15 min. After enrichment, centrifuge at 4000 rpm for 5 min to separate the layers, and take the upper surfactant-rich phase, to obtain the enrichment product of the CPT and HCPT extract. Detect the contents of CPT and HCPT in the extract before and after enrichment by high performance liquid chromatography, and calculate the enrichment rates of CPT and HCPT to be 98.88% and 97.98%, respectively.

[0053] Example 4

[0054] A method for extracting and enriching CPT and HCPT, comprising the following steps:

[0055] Step one: crush the dried camptotheca acuminata seeds through a 60-mesh sieve to obtain dried camptotheca acuminata seed powder for later use;

[0056] Step two: take 20 g of the crushed and sieved CPT and HCPT seed powder and 70 times the amount of 10% (v / v) non-ionic surfactant Genapol X-080 solution, place them in an ultrasonic extraction device, and extract them for 50 min at a power of 600 W, three times. After each extraction, centrifuge the mixture at a speed of 4500 rpm for 10 min, and take the supernatant, which is the CPT and HCPT extraction solution. Determine the CPT and HCPT contents in the seed powder and the CPT and HCPT contents in the extraction solution (take the average of three experiments) by high-performance liquid chromatography, and calculate the CPT and HCPT extraction rates to be 96.46% and 95.21%, respectively.

[0057] Step three: add sodium chloride to the CPT and HCPT extraction solution at a volume of 18% (w / v) of the extraction solution, stir for 15 min to fully mix, and enrich the extraction solution at 50°C for 10 min. After enrichment, centrifuge the mixture at a speed of 5500 rpm for 6 min to separate the layers, and take the upper surfactant-rich phase, which is the enrichment product of the CPT and HCPT extraction solution. Determine the CPT and HCPT contents in the extraction solution before and after enrichment by high-performance liquid chromatography, and calculate the CPT and HCPT enrichment rates to be 99.55% and 99.28%, respectively.

[0058] Example 5

[0059] A method for extracting and enriching CPT and HCPT, comprising the following steps:

[0060] Step one: crush dry CPT and HCPT seed through a 30-mesh sieve to obtain dry CPT and HCPT seed powder for later use;

[0061] Step two: take 20 g of the crushed and sieved CPT and HCPT seed powder and 15 times the amount of 12% (v / v) non-ionic surfactant Genapol X-080 solution, place them in an ultrasonic extraction device, and extract them for 40 min at a power of 360 W, three times. After each extraction, centrifuge the mixture at a speed of 6500 rpm for 8 min, and take the supernatant, which is the CPT and HCPT extraction solution. Determine the CPT and HCPT contents in the seed powder and the CPT and HCPT contents in the extraction solution (take the average of three experiments) by high-performance liquid chromatography, and calculate the CPT and HCPT extraction rates to be 90.10% and 91.35%, respectively.

[0062] Step three: add 16% (w / v) of potassium chloride to the CPT and HCPT extract solution, stir for 15 min to make it fully mixed, and enrich the extract solution at 45°C for 20 min. After enrichment, stand for 10 min at room temperature to make it stratify, and take the upper surfactant-rich phase, to obtain the enrichment product of the CPT and HCPT extract solution. Detect the CPT and HCPT contents in the extract solution before and after enrichment by high performance liquid chromatography, and calculate the CPT and HCPT enrichment rates as 99.32% and 99.18%, respectively.

[0063] Example 6

[0064] A method for extracting and enriching CPT and HCPT, comprising the following steps:

[0065] Step one: crush the dried CPT and HCPT extract solution, and obtain the dried CPT and HCPT extract solution for use.

[0066] Step two: take 20 g of the crushed and sieved CPT and HCPT extract solution and 25 times of 10% (v / v) non-ionic surfactant Genapol X-080 solution, and place them in an ultrasonic extraction device to extract for 80 min at a power of 420 W, for 3 times. After each time, centrifuge at a speed of 1100 rpm for 5 min, and take the supernatant, to obtain the CPT and HCPT extract solution. Detect the CPT and HCPT contents in the seed powder and the CPT and HCPT contents in the extract solution by high performance liquid chromatography (take the average of 3 experiments), and calculate the CPT and HCPT extraction rates as 96.64% and 95.59%, respectively.

[0067] Step three: add 16% (w / v) of potassium chloride to the CPT and HCPT extract solution, stir for 15 min to make it fully mixed, and enrich the extract solution at 45°C for 20 min. After enrichment, stand for 10 min at room temperature to make it stratify, and take the upper surfactant-rich phase, to obtain the enrichment product of the CPT and HCPT extract solution. Detect the CPT and HCPT contents in the extract solution before and after enrichment by high performance liquid chromatography, and calculate the CPT and HCPT enrichment rates as 99.32% and 99.18%, respectively.

[0068] Example 7

[0069] A method for extracting and enriching CPT and HCPT, comprising the following steps:

[0070] Step one: crush the dried CPT and HCPT extract solution, and obtain the dried CPT and HCPT extract solution for use.

[0071] Step two: 20 g of the pulverized and sieved Camptotheca acuminata seed powder and 30 times the amount of 6% (v / v) non-ionic surfactant Genapol X-080 solution were placed in a polyethylene bag, which was sealed and placed in a high-pressure cylinder of a pressurized extraction device. The extraction pressure was set to 50 MPa, and the pressure was maintained for 5 min, repeated 3 times. After the extraction, the supernatant was obtained by centrifugation at 8000 rpm for 15 min. The CPT and HCPT extraction solution was obtained. The contents of CPT and HCPT in the seed powder and the extraction solution were determined by high performance liquid chromatography (the average of 3 experiments). The extraction rates of CPT and HCPT were 89.09% and 87.31%, respectively.

[0072] Step three: 10% (w / v) of sodium chloride was added to the CPT and HCPT extraction solution, and the mixture was stirred for 15 min to fully mix. The extraction solution was enriched at 35°C for 20 min. After enrichment, the extraction solution was centrifuged at 10000 rpm for 8 min to separate the layers. The upper surfactant-rich phase was obtained, which was the enrichment product of CPT and HCPT. The contents of CPT and HCPT in the extraction solution before and after enrichment were detected by high performance liquid chromatography. The enrichment rates of CPT and HCPT were 99.11% and 99.17%, respectively.

[0073] Example 8

[0074] A method for extracting and enriching CPT and HCPT, comprising the following steps:

[0075] Step one: dry Camptotheca acuminata seeds were pulverized and sieved through a 20-mesh sieve to obtain dry Camptotheca acuminata seed powder for later use.

[0076] Step two: 20 g of the pulverized and sieved Camptotheca acuminata seed powder and 20 times the amount of 8% (v / v) non-ionic surfactant Genapol X-080 solution were placed in a microwave extraction device. The extraction was carried out at a microwave temperature of 25°C and a microwave power of 420 W for 15 min, repeated 3 times. After the extraction, the supernatant was obtained by centrifugation at 4000 rpm for 12 min. The CPT and HCPT extraction solution was obtained. The contents of CPT and HCPT in the seed powder and the extraction solution were determined by high performance liquid chromatography (the average of 3 experiments). The extraction rates of CPT and HCPT were 88.10% and 85.39%, respectively.

[0077] Step three: 8% (w / v) sodium chloride of the volume of the extract was added into the CPT and HCPT extract, and stirred for 15 min to make it fully mixed, and the extract was enriched at 30°C for 15 min. After enrichment, it was centrifuged at 10000 rpm for 5 min to make it layered, and the upper surfactant-rich phase was taken, and the enrichment product of the CPT and HCPT extract was obtained. The content of CPT and HCPT in the extract before and after enrichment was detected by high performance liquid chromatography, and the enrichment rates of CPT and HCPT were 99.07% and 99.04%, respectively.

[0078] It should be noted that in the above examples, as long as the technical solutions are not contradictory, they can be arranged and combined. Those skilled in the art can exhaust all possibilities according to the mathematical knowledge of arrangement and combination, so the technical solutions after arrangement and combination will not be described one by one, but it should be understood that the technical solutions after arrangement and combination have been disclosed by the present application.

[0079] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Those skilled in the art can make various changes and modifications to the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A method for the extraction and enrichment of camptothecin (CPT) and 10-hydroxycamptothecin (HCPT), characterized in that, Includes the following steps: Step 1: Crush and sieve 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 after extraction, and take the supernatant to obtain CPT and HCPT extracts; Step 3: Add inorganic salts to the CPT and HCPT extracts, stir well, and treat the extracts at a certain temperature for a certain time. After the reaction treatment, the solution is separated into layers by centrifugation or standing. The upper surfactant-rich phase is the enriched product of the CPT and HCPT extracts.

2. The method for extraction and enrichment of CPT and HCPT according to claim 1, characterized in that: The Camptotheca tissue is one or more of the following: Camptotheca seeds, Camptotheca fruits, Camptotheca leaves, Camptotheca branches, and Camptotheca root bark.

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

4. The method for extraction and 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 mass ratio of the Camptotheca acuminata tissue powder to the volume ratio of the nonionic surfactant Genapol X-080 is 1 g: 10-80 mL.

5. The method for extraction and enrichment of CPT and HCPT according to claim 1, characterized in that: The extraction method includes one or more of the following: ultrasonic extraction, microwave extraction, ultra-high pressure extraction, hot reflux extraction, Soxhlet extraction, homogenization extraction, and negative pressure cavitation extraction.

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

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

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

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