초음파를 이용한 파클리탁셀 추출방법
The present invention refers to which the extracting method relates to paclitaxel from a biomass recovery increased paclitaxel, a paclitaxel [...] number extraction method using ultrasonic waves is provided many organic solvent extraction with an existing solvent extraction process recognition function is used to resolve the time door number, microwave simultaneously using extraction process recognition function is process complexity (risk) (device), door number produced by many energy consumption at normal numbers of him. Paclitaxel (paclitaxel) attention tree (yew tree) are found in the skin-based anticancer agents used the most current anticancer number d reel [lu lung roh id (diterpenoid) are disclosed. Ovarian cancer, breast cancer, head-neck cancer (head and neck cancer), car gun always amount (kaposi's sarcoma), arsenic (non non-small cell lung cancer) cell lung cancer treatment U.S. FDA (U. S Food and Drug Adnimistration) grant to verify the acquired bar and [...] (Alzheimer's treatment, rheumatoid arthritis) and treatment method in which the development of paclitaxel demand next year. Paclitaxel (yew tree) is major production method directly extracting interest tree method, a method-debenzoyl-obtaining precursor (precusor), callus from a plant culturing cells plants (callus) method for obtaining the pin is. Among these plant cell culture method compared to other method without being affected by external factor (weather, environment) of paclitaxel non destructively at constant quality of the second bioreactor in mass production to cables disclosed. Plant cell culture produced by most paclitaxel is produced biomass (plant cells) are included, and extraction of various stages in the paclitaxel production is positive number covering the nanometer range. Mirror number point of view and the first biomass contained in high yield recovery of the paclitaxel very important disclosed. Such as laver However existing solvent extraction from biomass extraction process of 4 or more times most paclitaxel for recovery time required higher paclitaxel the recovery of many organic solvent in a facsimile numbers. 2008 years using microwaves to stage 1 (-99%) of paclitaxel was mostly extracted from biomass Conference extraction possible recovery. Microwave extraction via at least one additional recovery times occur on high but shorten the device cost as well as the lot of energy is consumed as an ion implantation mask (risk) at least more than industrial mass production process be applied to limit flow tides. As a result extraction times required for minimizing device module (simplified) while simultaneously saving equipment, energy saving as well as convenience (convenient) and processes for viability (feasibility) process is still high efficiency biomass extraction process development data in a database are disclosed. The present invention refers to said door such as point number in order to solve in a certain, from biomass extraction method which improves a recovery of paclitaxel anticancer [...] pin is a number. In order to solve the above-described and the present invention refers to a number, a) crude including paclitaxel ( In the embodiment of the present invention according to one preferred, said a) step the biomasses are Texas, or be a biomass derived from the cell culture. In the embodiment of the present invention according to another preferred one, said b) step the organic solvent is C1 - C4 The group consisting of alcohol and water comprising at least 1 species can be. In the embodiment of the present invention preferably according to one another, said b) step biomass and organic solvent is 1:1 (w/v) can be selected from the group. In the embodiment of the present invention according to another preferred one, said c) ultrasonic extraction step can be-step 1. In the embodiment of the present invention according to one preferred another, 10 minutes to 16 minutes in ultrasound power of said ultrasonic extraction is 380 W 530 W can be performed. In another preferred of the present invention according to one in the embodiment, the paclitaxel recovery implementation being 99% said ultrasonic after extraction. The present invention refers to a method using ultrasonic waves by extracting paclitaxel [...] number, many organic solvent extraction time door number for existing solvent extraction process recognition function is use to resolve the microwave simultaneously using extraction process recognition function is process (device) (risk) complexity, many energy consumption can be produced by a door number every time. The, method of the present invention the mean paclitaxel extraction ( Figure 1 shows a schematic diagram indicating also from a biomass ultrasonic extraction process are disclosed. Figure 2 shows a number of extracted using existing solvent extraction yields paclitaxel also influence graph in the chemical formula, biomass/MeOH 1:1 ratio (w/v), is -500 rpm agitation inside are disclosed. Figure 3 shows a reflective surface 3D ultrasonic extraction process optimization also from a biomass (3D response surface) and contour [...] to revealing the secret key (contour plot), biomass (w/v)/MeOH 1:1 ratio, is -500 rpm agitation inside are disclosed. Figure 4 different method by solvent extraction of the sample using the SEM Image of dried biomass, before extraction is (A); (B) existing solvent extraction method is (1 times extraction); (C) is extracted using ultrasonic waves (1 times extraction) SEM Image of exhibits. Hereinafter, the present invention further it relates substrate. As described above, existing solvent extraction from biomass extraction process of 4 or more times most paclitaxel for recovery time required higher paclitaxel the recovery of many organic solvent in a facsimile numbers. Microwave extraction is extracted additional recovery times occur on high but shorten the device cost as well as the lot of energy is consumed as an ion implantation mask (risk) at least more than a speed dial setup be applied to industrial mass production process. The number extraction module (simplified) while simultaneously minimizing device required for saving equipment, energy saving as well as convenience (convenient) and processes for viability (feasibility) process is still high efficiency biomass extraction process development data in a database are disclosed. The present invention refers to a the) crude including paclitaxel ( First, obtaining a crude (Taxus genus) derived from plant biomass including paclitaxel) sets forth other. Said a) step the biomasses are Texas, the cell, the cell (cell debris) pieces and cell culture comprising at least 1 species can be selected from the group consisting. In addition, the Texas [...] non-pulley oh said braze ( Obtain said biomass typically plant biomass harvested from the cell number method for obtaining particularly if not relayed. Next, mixing said biomass with organic solvents b) sets forth other. Specifically, a pharmaceutical plant for extracting said organic solvent typically used in number but not specially if the relayed, preferably C1 - C4 1 species selected from the group consisting of alcohol and water and can be more. Said specific organic solvent include methanol, ethanol, propanols and methylene chloride mixture is selected from the group consisting 1 species or 2 can be at least one, more preferably paclitaxel from biomass can be recovered organic solvent with preferably methanol can be used. In addition, said b) mixed 1:1 (w/v) but the step biomass and organic solvents, which are not limited to this, typically in the rate at which can be used for plant ingredients extracted by the specially if the number does not relayed. In a preferred embodiment of the present invention organic solvent extraction of the optimum in laser output to keep the ratio 1:1 (w/v) biomass is mixed. Next, said b) step a mixture of ultrasonic processing ultrasonic extracting c) step described as follows. Said c) ultrasonic extraction step can be-step 1. Ultrasonic frequencies in said ultrasonic extraction plants for extracting a pharmaceutical dosage range can be selected that if suitable grudge without number. Laser output portion includes a 40 kHz ultrasonic washer of the present invention was used. 530 W to about 16 minutes in ultrasound power of said ultrasonic extraction is 380 W to 10 can be performing, when extracting the ultrasound in such a condition, the yield paclitaxel implementation being 99%. If ultrasound in ultrasound power of less than 380 W when extracting, 97% recovery following 10 minutes is patterned but maximum can be recovered paclitaxel to paclitaxel inputting all scarcely changed and not recovering paclitaxel, when extracting excess ultrasound power ultrasound in a 530 W, due to excessive heat generation in the pin is paclitaxel decomposition developing (extraction efficiency lowering) inner side operation. Laser output portion includes a of the present invention, ultrasound power (80, 180, 250, 380 and 530 W) is patterned on (1, 2, 4, 6, 8, 10, 12, 15 and 30 minutes) (-500 rpm) stirring each various combinations by changing to a 1 times when the ultrasonic extracting, recovery is calculated the following equations 1 have paclitaxel, according to the combination of paclitaxel recovery 3 also shown to various ultrasound power net and operation time. [Mathematical equation 1] In addition, paclitaxel recovery method of the present invention extracted using ultrasonic waves of a solvent extraction of solvent extraction and recovery of optimized in order to compare a number of paclitaxel according to the paclitaxel also recovery 2 shown. A combination of various is patterned and various ultrasound power when performing ultrasonic extraction method 1 times, identified in Figure 3 as the operation time of 10 min or higher than 80W tests even ultrasound power conducting, equivalent to the level of existing optimized 1 times has been recovered [...] (-64%) solvent extraction, 10 minutes of ultrasound power section 530 W to 80 W 1 times when ultrasound power is increased by 1 in extracting solvent extraction times significantly higher than existing recovery paclitaxel filtered activated, to maximum 99% recovery paclitaxel were. In addition, identified in Figure 3 as the longest during ultrasonic extraction tests 1 is patterned minutes even when-step 1, treated with ultrasonic power 320 W domestic optimized equivalent has been recovered [...] 1 times (-64%) levels of solvent extraction, 1 minutes to 1 times of ultrasound power period 320 W 530 W when ultrasound power is communicated to the extracting solvent extraction increases significantly higher than 1 times filtered recovery paclitaxel activated, to maximum 80% recovery paclitaxel were. In the present invention in addition stage 1 times only extraction from biomass can be recovered to an optimized ultrasonic extraction method condition most of paclitaxel are described. Main process parameters for optimization, non optimal biomass/methanol (1:1, w/v) is patterned according to constant and ultrasound power and influence visit from the police. Ultrasound power (80, 180, 250, 380, 530 W) is patterned on (1, 2, 4, 6, 8, 10, 12, 15, 30 minutes) (-500 rpm) each change and agitating the biomass extraction times conducting to a 1. As a result, throughput is increased to 380 W formed by ultrasound power as in Figure 3 the highest yield were obtained in (-99%). In addition, paclitaxel throughput is increased is patterned overlapped with the highest yield (-99%) were obtained of sulfates is patterned to 10. Ultrasound power 250 W hereinafter applied to at least one of the yield even after 10 minutes (-80%) 380 W ultrasound power but is patterned and less variation in substantially all been applied to at least one or more [...] recovery (-99%). As a result optimum extraction using ultrasonic waves can be ascertained on 10 each being a minute, been 380 W is patterned and ultrasound power. In the case of stage 1 in optimum condition using ultrasonic waves (-99%) of paclitaxel was mostly extraction times extraction process recovery possible. These results through, existing extraction method of the present invention compared to a solvent extraction optimized using ultrasonic waves is patterned significantly shortened, operation time according to which the extraction efficiency increases that were confirmed. In the present invention in addition, scanning electron microscope (MIRA LMH; Tescan, Czech Republic) observed surface using biomass. In the case of biomass before extraction, also 4 (A) as biomass in view and very smooth surface, existing solvent extraction using methanol (1 times extraction) of 4 (B) as also be the case in view been corrugated. While, extracted using ultrasonic waves (1 times extraction) performing (C) 4 also be the case in a very rough surface exhibits a strong impact as biomass (crushed) and SQUIDs due to view his. I.e., crushed in both cell walls to allow strong cell enhanced [...] sufficiently SQUIDs (crushed) surface can be extracted with a braided with one another, as a result extraction efficiency which the nickel. These results using ultrasonic waves to/from extraction from biomass can be recovered confirm it effectively paclitaxel were extracted. The, method of the present invention optimized using ultrasonic waves paclitaxel many organic solvent extraction with an extracting existing solvent extraction process recognition function is used to resolve the time door number simultaneously using microwave extraction process recognition function is process (device) (risk) complexity, many energy consumption can be produced by a door number every time. In the embodiment of the present invention through to more specifically describe but, in the embodiment of the present invention to form a number range is not relayed and, this method for assisting in the understanding of the present invention will be interpreted. For biomass Preparation The present invention is used in culture [...] [neyn cis ( Extracting using ultrasonic waves (Ultrasound non-assisted extraction) In the embodiment 1 always to enhance biomass harvested from the cell ratio of methanol by 1:1 (w/v) 40 kHz ultrasonic cleaner (UC-a 10, Jeiotech, Korea and JAC provided 4020, KODO, Korea) using ultrasound power (80, 180, 250, 380, 530 W) is patterned on (1, 2, 4, 6, 8, 10, 12, 15, 30 minutes) (-500 rpm) and agitating the first channel to a extracted from the 1 times. After extraction filter paper (150 mm, Whatman) to recover the pressure filtration of the concentrators (CCA-a 1100, EYELA, Japan) paclitaxel extraction filtrate concentrated using damascene vacuum drying (40 °C, overnight, 760 mmHg) up. Dried paclitaxel (crude paclitaxel) HPLC system for analysis process can be a content (Waters, USA) and Capell Pak C18 (250 × 4. 6 mm, Shiseido, Japan) column using paclitaxel purity and yield were measured. A furnace distilled water and acetonitrile mixed solution (65/35 - 35/65, v/v, gradient the solvent creation law) a flow rate 1. Flowing me 0 ml/minutes. Sample injection amount detected by UV in which micro l 227 nm 20. HPLC analysis can be carried out using standard quantitative curve when the standard (>97% purity) was used Sigma non-Aldrich number article. Also shown 1 to biomass extraction process using ultrasonic waves. In addition the yield paclitaxel was calculated as 1 next equations. [Mathematical equation 1] The yield also 3 on a combination of paclitaxel according to various ultrasound power is patterned and efined. Existing solvent extraction using ultrasound Paclitaxel Comparison of extraction method Plant cell culture produced by most paclitaxel is produced biomass (plant cells) are included, such as laver , the solvent extraction of suturing in the embodiment most effective extraction efficiency (w/v) using 1:1 methanol/biomass ratio of indicating when the extracting solvent, extraction method in the embodiment 2 of the present invention using ultrasonic waves by comparing the same of paclitaxel extraction efficiency was assessed. Existing solvent extraction is recovered in the embodiment 1 without using ultrasonic methanol using 4 times extracted from the biomass on a batch basis. Biomass ratio 1/1 methanol (15g biomass/15 ml methanol) when the extraction time is 30 minutes to 1 times electrodes respectively. As a result, as shown in the 2 also 64% of an extract from the biomass 1 times, 2 times 20% of an extract, an extract 10% 3 times, 4 times of an extract 5% total recovery (-99%) of paclitaxel is mostly recovered each [...] Conference 4 was extracted. While, in the embodiment 2 and described in the specification is patterned in a variety of combination of a 1 when performing ultrasonic extraction method for ultrasound power and various times, identified in Figure 3 as the operation time of 10 min or higher than 80W tests even ultrasound power conducting, optimized levels of existing solvent extraction has been recovered [...] (-64%) equivalent to 1 times, 10 minutes of ultrasound power section 530 W to 80 W 1 times when ultrasound power is increased by 1 in extracting solvent extraction times significantly higher than existing recovery paclitaxel filtered activated, to maximum 99% recovery paclitaxel were. In addition, identified in Figure 3 as the longest during ultrasonic extraction tests 1 is patterned minutes even when-step 1, treated with ultrasonic power levels equivalent to 1 times the optimized 320 W (-64%) has been recovered [...] of existing solvent extraction, 1 minutes to 1 times of ultrasound power period 320 W 530 W when ultrasound power is communicated to the extracting solvent extraction increases significantly higher than 1 times filtered recovery paclitaxel activated, to maximum 80% recovery paclitaxel were. Paclitaxel purity -0 biomass extraction obtained in ultrasonic treatment roller. 5% (data not shown) cannot difference approximately. As a result, extraction method using ultrasonic waves compared to operation of the present invention significantly shortened time and solvent extraction of existing, operation time according to which the extraction efficiency increases that were confirmed. Using ultrasonic waves Paclitaxel Optimization of extraction method The only extraction in the embodiment using ultrasonic waves to 1 times in most of paclitaxel was recovering gases extracted from the biomass. Main process parameters for optimization, non optimal biomass/methanol (1:1, w/v) is patterned according to constant and ultrasound power and influence visit from the police. Various ultrasound power (80, 180, 250, 380, 530 W) is patterned on (1, 2, 4, 6, 8, 10, 12, 15, 30 minutes) in the embodiment 2 is described conditions equal, ultrasound power and is patterned first channel (-500 rpm) and agitating the biomass extraction times conducting to a 1. As a result, throughput is increased to 380 W formed by ultrasound power as in Figure 3 the highest yield were obtained in (-99%). In addition to throughput is increased is patterned overlapped with the paclitaxel were obtained the highest yield of sulfates is patterned 10 (-99%). Ultrasound power 250 W hereinafter after 10 minutes is patterned to at least one less variation in yield even (-80%) was applied to at least one or more possible [...] 380 W ultrasound power but both recovery (-99%). In addition, when compared to existing organic solvent extraction, solvent extraction is also 2 4 as shown in the existing Conference extraction of paclitaxel recovery (-99%) but mostly, ultrasonic extraction is 380W 10 minutes in ultrasound power of 1 executes most of paclitaxel (-99%) recovery stage extraction times were. These results indicate that enhanced intracellular cell to cell wall effectively [...] waves is extracted according to the dots that, through, optimum ultrasound power and extraction of the present invention using ultrasonic waves can be ascertained on 10 380 W is patterned each being a minute, been. In the case of stage 1 of the optimum extraction process using ultrasonic waves mostly extraction times (-99%) recovery of paclitaxel were. The, method of the present invention paclitaxel extraction method using ultrasonic waves using solvent extraction of existing significantly shortened compared to extracting operation time that were able to confirm that the increased injection and extraction efficiency. Scanning microscope(SEM) Analysis To a scanning electron microscope to observe (MIRA LMH; Tescan, Czech Republic) biomass surface by water. The amount of 1 mg 10 - 15 kv biomass is dried in an accelerator voltage analysis in the form of a visit from the police. In the case of biomass before extraction, also 4 (A) as biomass in view and very smooth surface, existing solvent extraction using methanol (1 times extraction) of 4 (B) as also be the case in view been corrugated. While, extracted using ultrasonic waves (1 times extraction) performing (C) 4 also be the case in a very rough surface exhibits a strong impact as biomass (crushed) and SQUIDs due to view his. I.e., intracellular adsorption is a SQUIDs (crushed) [...] extracted enough that both surface is formed and thus effectively extraction efficiency and nickel. These results using ultrasonic waves to/from extraction from biomass may be signed in capable of effectively extracting recovering paclitaxel. As a result, extraction method of the present invention optimized using ultrasonic waves paclitaxel many organic solvent extraction time for existing solvent extraction process recognition function is use to resolve the microwave door number simultaneously using extraction process recognition function is process complexity (risk) (device), many energy consumption produced by number every time capable of a door. The present invention refers to which the extracting method relates to paclitaxel from a biomass recovery increased paclitaxel, a paclitaxel [...] number extraction method using ultrasonic waves is provided many organic solvent extraction with an existing solvent extraction process recognition function is used to resolve the time door number, microwave simultaneously using extraction process recognition function is process complexity (risk) (device), door number produced by many energy consumption at normal numbers of him. The following a) to c) paclitaxel extraction method including the steps of: a) crude including paclitaxel ( According to Claim 1, said a) step the biomasses are Texas, the cell, the cell (cell debris) and cell culture pieces including at least one selected from the group consisting 1 characterized in the vessel extraction method. Back number Back number Back number Back number According to Claim 1, said recovery vessel extraction method characterized in the 99% paclitaxel ultrasonic after extraction.
