• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    The invention provides a process for extracting total flavone from seeds of fragrant cedarel, which comprises the following steps: mixing the powder of the seeds of fragrant cedarel in a NADES eutectic solvent, performing the homogenization, centrifugation and filtering , purify the filtrate using an AB-8 resin, elute the total flavone from the seeds of fragrant cedarella using ethanol at a gradient of 70% to 95%. The method of the invention improves the efficiency of the extraction of total flavone from the seeds of sweet cedar.
    公开号:BE1027352B1
    申请号:E20205091
    申请日:2020-02-14
    公开日:2021-01-20
    发明作者:Changqing Qu；Shaoyun Yue；Jingxia Yang；Xinyue Dong；Yuntao Ji；Xin Jin
    申请人:Univ Fuyang Normal；
    IPC主号:
    专利说明:

    Process for extracting total flavone from seeds of fragrant cedarella Technical Field The invention belongs to the field of medicinal chemistry, particularly to the process for extracting total flavone using a eutectic solvent.
    State of the art Sweet cedar is a traditional Chinese medicinal material in China, also known as Toona sinensis.
    Its root bark, bark, germ, leaves and fruits can be used as the medicine to commonly treat enteritis, dysentery and inflammation of the skin in the field of traditional Chinese medicine.
    Their tissues from various parts have been used to treat various diseases.
    According to specific studies, the seeds of sweet cedar are rich in flavone.
    Bioflavonoids have a variety of pharmacological activities, such as antibacterial effect, antiviral effect, anti-tumor effect, antioxidant effect, anti-inflammatory effect, analgesic effect, etc., especially the anti-free radical effect, anti-cancer effect, etc.
    Therefore, the application of bioflavonoids has broad prospects in the field of medicine, food, etc.
    Regarding the method of extracting the total flavone from the seeds of sweet cedar, it is generally the traditional methods of extraction, such as extraction in water, extraction in alcohol, precipitation in alcohol, extraction of acetone-ethyl acetate, etc.
    However, these traditional processes have problems such as large loss of raw materials, low yield, etc.
    And the extraction of total flavone from the seeds of sweet cedar is usually a crude extraction.
    Therefore, there is a need to further study methods of extracting total flavone from the seeds of sweet cedar.
    Disclosure of the Invention The object of the invention is to provide a method of extracting total flavone from seeds of sweet cedarel to improve the efficiency of extracting total flavone from seeds of sweet cedar.
    The present invention provides a process for extracting the total flavone from the seeds of odoriferous cedarel comprising the following steps: mixing the powder of the seeds of odoriferous cedarel in a NADES eutectic solvent, carrying out the homogenization, centrifugation and filtering, purifying the filtrate using an AB-8 resin, elute the total flavone from the seeds of odoriferous cedarella using ethanol at a gradient of 70% to 95%.
    Among them, the eutectic solvent NADES is composed of choline chloride having a water content of 30% and 1,2-propanediol, whose optimum molar ratio between choline chloride and 1,2-propanediol is 1 / 4.
    Among them, the mass ratio between the seed powder of odoriferous cedarel and said eutectic solvent NADES is 1/50.
    Among them, the duration of said ultrasound is 15 min.
    Among them, the rotational speed of said centrifugation is 4000 rpm, while the centrifugation time is 25 min.
    The invention has chosen an ecological soluble eutectic solvent from several different polyhydric alcohol hydrogen donors to obtain a process of extracting total flovan from the seeds of fragrant cedarella in order to obtain the best process conditions of auxiliary ultrasonic extraction (UAE) of total flovan from seeds of fragrant cedarella: choline chloride having the water content of 30% and 1,2-propanediol were used as the extractant by adopting the solid ratio -liquid of 1/50 and the extraction time of 35 min to extract the total flovane from the seeds of fragrant cedarella. Compared to traditional extraction processes (with 0.0020 water and 75% ethanol 0.0027), the efficiency will be higher for NADES dedicated to extracting total flovan from the seeds of sweet cedar. Brief description of the figures Figure 1 shows the standard curve between rutin concentration and absorbance. Figure 2 presents the results of comparison of the total concentrations of flavone in the seeds of sweet cedar extracted in the NADES, the waters, and the different ethanols. Figure 3 shows an infrared spectrum diagram with A: choline chloride, B: 1,2-propanediol; C: Chcl: 1,2-propanediol. Figure 4 shows a diagram of the relationship between the different parameters of the extraction processes and the concentrations of the total flavone, with: À: molar ratio of the hydrogen donor; B: water content; C: solid-liquid ratio; D: extraction time.
    Detailed Description The following examples may enable those skilled in the art to fully understand the present invention, but do not limit the present invention in any way. Unless otherwise indicated, the test methods in the following examples are conventional methods. Unless otherwise noted, the test materials used in the following examples were purchased from conventional biochemical reagent companies. NADES is a eutectic mixture composed of a hydrogen bond acceptor (Hydrogen Bond Acceptor, HBA) and a hydrogen bond donor (Hydrogen Bond Donnors) of a solid halide salt with a certain stoichiometric ratio, whose point melting rate is significantly lower than that of substance of all groups. As a new advanced solvent, NADES is a fully biodegradable and biocompatible material. By combining the characteristics of ionic liquids, NADES is inexpensive and non-toxic, adopts the simple preparation process which only adopts mechanical heating and tempering. It is a new ecological solvent. The invention provides different environmentally friendly eutectic methods for extracting total flavone from seeds of sweet cedar. After comparison and study with traditional test methods, he is able to finish the preliminary screening of polyhydric alcohol hydrogen donors and optimize the extraction of total flavone parameters from cedar seed. odoriferous using soluble NADES. 1 Material and process
    1.1 Material The seeds of sweet cedar were purchased in Taihe County of Fuyang Town. 1,2-Propanediol, Glycerol, 1,4-Propanediol were purchased from Wuxi Prospect Chemical Reagent Co., Ltd. Choline chloride was purchased from Sinopharm Chemical Reagent Co., Ltd. Rutin was purchased from Wuxi Prospect Chemical Reagent Co., Ltd. Other reagents such as ethanol (> 95%) are of analytical grade.
    1.2 Instrument Ultraviolet spectrophotometer: made in Germany; electronic balance: manufactured by Shanghai Precision Scientific Instrument Co., Ltd .; Constant Temperature Intelligent Band Crucible: Manufactured by Beijing Dongfang Jingrui Technology Development Co., Ltd .; rotary evaporator: manufactured by Anhui Water Science Instrument Co., Ltd .; ultrasonic cleaner KQ-50B; Fourier infrared spectrometer: manufactured by Agilent Technologies.
    1.3 Process
    1.3.1 Preparation of different NADES soluvants Place the NADES with different alcoholic hydrogen donors indicated in the following table in a conical flask, carry out the water bath at 65 ° C and stir until a liquid forms. homogeneous. After the uniform liquid has formed, add the water to the NADES Solvent in proportion and leave it for use.
    Table 1
    1.3.2 Extraction of the total flavone from the seeds of odoriferous cedarella Mix the powder of the seeds of odoriferous cedarella of 0.2g in the aforementioned NADES solvent 5 of 5mL, activate the ultrasound for 15 min, carry out the centrifugation for 25min at 4000 rpm and filtering, mix the solvent and the aqueous hydrochloric acid solution (mass concentration: 50%), carry out the magnetic stir of the mixed solution for 20min at 65 ° C, and filter it, remove the deposit, and then use AB-8 resin to carry out the purification, and finally recover the total flavones with ethanol at a gradient of 70% to 95% ethanol. Carry out the decompression and the distillation of the total flavone from the seeds of odoriferous cedarella purified and recovered to the minimum volume. Store it for later testing. At the same time, use water and 75% ethanol to extract the total flavone from the seeds of sweet cedar, adopt the same extraction conditions according to section 1.3.2.
    1.3.3 Drawing a standard curve Carry out the precise weighing of the 50 mg rutin reference product and place it in a 50 ml flask, dissolve in 0.95 ethanol and add it up to the gradient, precisely aspirate a 10ml solvent in a 100ml flask, dissolve it up to the gradient using 0.60 ethanol, pipette 0.5, 1.0, 2.0, 3.0, 4.0, 5 respectively , 0 ml of rutin solution as reference (0.1 mg / ml) in a 10 ml flask, add the 0.3 ml sodium nitrite solution (with mass concentration of 5%), shake it well and shake, put deposit for 6min at room temperature, and then add the 0.3ml aluminum nitrite solution (with mass concentration of 10%), shake it well and shake, deposit for 6min at room temperature, finally add the solution of sodium hydroxide of 4ml (with mass concentration of 4%), calibrate the volume up to the gradient using 60% ethanol, shake it well and shake, put e n deposit for 12 min at room temperature, take the ethanol reagent as the virgin reference liquid, measure all the absorbances of all the groups (OD value) at 510nm using an ultraviolet spectrophotometer.
    1.3.4 Determination of the total flavone from the seeds of sweet cedarella Take respectively the ImL solution of the total flavone from the seeds of sweet cedar, determine the total flavone content from the seeds of sweet cedar under different conditions according to the process indicated in article 1.3.3.
    1.3.5 Infrared characterization of NADES NADES, synthesized by different hydrogen donors and hydrogen acceptors, has different physical properties, such as electrical conductivity, density, viscosity, surface tension, etc. Through the primary screening of NADES, the present assay determined 1,2-propanediol as the hydrogen acceptor and indicated infrared characterization, determined Chcl + 1 and 1,2-propanediol to synthesize the environmentally friendly eutectic solvent ecological dedicated to participate in the optimization test of the extraction parameters.
    1.3.6 Parameter optimization Based on polyhydric alcohol hydrogen donors, select the molar ratio of HBA and HDA (1/1, 1/2, 1/3, 1/4, 1/5), the water content of NADES (10%, 20%, 30%, 40%, 50%), the liquid-solid ratio between NADES and odoriferous cedar seed powder (from 1/25, 1/50, 1/100 , 1 / 200mg ml) as well as the extraction time (15 min, 25 min, 35 min, 45 min) to determine the optimal extraction parameters.
    1.3.7 Data analysis Adopt statistical methods to analyze data. Express all the data in the text as the result of (mean +/- standard difference), adopt SPSS 19.0 to perform the significant difference analysis for the different NADES and the total flavone concentrations from the seeds of sweet cedar in water and in ethanol.
    2 results
    2.1 Drawing of standard curve According to the standard curve drawn according to the method indicated in article 1.3.3, the regression equation is: Y = 11.035x + 0.0047, while the correlation coefficient is R2 = 0 , 9987. This means that the concentration of rutin shows a good linear relationship with the absorbance in the range of 0 to 0.06 mg / mL, the standard curve of which is given in Figure 1.
    2.2 Concentrations of total flavone from seeds of sweet cedarella obtained from different NADES, water and ethanol The results of comparison of the concentrations of total flavone from seeds of sweet cedar obtained from different NADES, water and ethanol are shown in Figure 2. When extracting the active flavone components from the drug using aqueous soluble NADES, the highest extraction efficiency is obtained from polyhydric alcohol donors. The invention adopts the typical polyhydric alcohol of hydrogen ions to compare with the traditional extraction process to finish the choice of the conditions of primary choice.
    After choosing different hydrogen donors (polyhydric alcohol, polyacid, etc.), it is able to synthesize different NADES. Different hydrogen donors will obtain different physical properties (viscosity, density, surface tension, conductivity, etc.). These adjustable different physical properties are the prerequisites for exceptional application of NADES in all fields. For the extraction of the active flavone components using NADES, the choice of polyhydric alcohols as the hydrogen donor has been shown to have better excellence than other hydrogen donors. Referring to figure 2, the average content of total flavone extracted from the eutectic solvent of choline chloride and 1,2-propanediol is 0.0151mg / ml, forming a significant difference with that of 0.0027mg / ml obtained from 75% trational ethanol and that of 0.0020 mg / ml obtained from water. At the same time, compared to the other two NADES polyhydric alcohols, choline chloride and 1,2-propanediol will have a higher extraction concentration due to the formation of more hydrogen bonds under combination force. stronger hydrogen bonds between 1,2-propanediol and choline chloride.
    2.3 Infrared characterization scheme The infrared characterization scheme is shown in figure 3.
    After choosing the different polyhydric alcohol donors, the present test provides the infrared characterization of better NADES to optimize subsequent extraction parameters. From Example A in Figure 3, the absorption peak of the characterization of choline chloride is 3255 ° ® "! Of these, 10851 is caused by OH vibration and CH vibration. Example B of Figure 3, the OH- vibration peaks are 3384 ° "- !, 297 0e"! 2932 '"- !. Compared to the ordinary vibration of OH-, the peak width is possibly caused by hydrogen bonding. In Example C of Figure 3, the Chel: 1, 2-propanediol CH3 vibration peak is 1402 ° "" !. Therefore, the infrared characterization is successful for Chel: 1, 2-propanediol (1/3).
    2.4 Optimization of NADES extraction parameters
    2.4.1 Molar ratio of hydrogen donor As an environmentally friendly eutectic solvent, the choice of different hydrogen donors is very important for the formation of NADES and the physical properties. The number of hydrogen donors determines the efficiency and stability of the extraction of NADES. Generally speaking, the number of hydrogen donors determines the viscosity of NADES. As the hydrogen donor increases, more hydrogen bonds will be formed with the hydrogen acceptors and waters improving the stability of the solvent. The invention compares the efficiency of extraction of NADES hydrogen donors at different mole ratios. Referring to Example C of Figure 4, as the different molar ratios (1/1 to 1/3) increase, the extraction efficiency will be increased gradually, the maximum will be obtained at molar ratio of 1/3, and will be reduced in case of extraction efficiency of 1/4 to 1/5 due to the decrease in the binding capacity of choline chloride and total flavones of odoriferous cedarel due to the donor excessive hydrogen.
    2.4.2 Water content After formation of aqueous soluble NADES, many donor hydroxyl chemical groups will be exposed. Therefore, an appropriate amount of water is added to form more hydrogen bonds with these groups to make NADES more stable. The surface tension of the NADES solvent is the most important physical property of the water content. However, the excessive water content reduces the concentration of NADES after group saturation, which will affect the efficiency of extraction. In this way, it is necessary to study the different water content of NADES in the extraction. Referring to Example B in Fig. 4, the water content of 30% will make the highest extraction efficiency of total flavone from the seeds of sweet cedar, while the water content of 50% and that of 70% will make the extraction efficiency reduced, which is caused by the reduction in NADES concentration in case of reduction in surface tension.
    2.4.3 Solid-liquid ratio During the extraction of the active components of the drug, the solid-liquid ratio
    different liquid exhibits the efficiency of extracting total flavone from the seeds of different fragrant cedarella. During extraction of total flavone from the seeds of sweet cedar in aqueous soluble NADES, the too low solid-liquid ratio will cause the formation of hydrogen bond between the NADES molecules causing insufficient extraction of the flavone from the seeds of Cedarwood, while too large a solid-liquid ratio will cause sufficient combination not possible with the total flavone of the seeds of Cedarwood. From Example C in Figure 4, the extraction efficiency will be highest at 1 / 50mg / ml "!.
    2.44 Extraction Time Referring to Example D in Figure 4, water soluble NADES exhibits a high total flavone extraction rate over a period of 35 minutes. As the time passes from 5 to 35 minutes, the extraction efficiency is increased, and will be reduced from 45 ° "° minute, which is caused by the increase in temperature and by the evaporation of NADES. The invention has chosen an ecological soluble eutectic solvent from several different polyhydric alcohol hydrogen donors to obtain a process of extracting total flovan from the seeds of fragrant cedarella in order to obtain the best process conditions of the auxiliary ultrasonic extraction (UAE) of total flovan from the seeds of sweet cedar: choline chloride having the water content of 30% and 1,2-propanediol were used as the extractant in adopting the solid-liquid ratio of 1/50 and the extraction time of 35 min to extract the total flovan from the seeds of odoriferous cedarel. Compared to traditional extraction methods (with water of 0.0020 and 75% of ethanol 0.0027), the efficiency is ra higher for NADES dedicated to extracting the total flovan from the seeds of sweet cedar. Those skilled in the art should understand that the aforementioned exemplary embodiments are only exemplary embodiments, and that various changes, substitutions and changes may be made without departing from the spirit and scope of the present invention.
    权利要求:
    Claims (5)
    [1]
    Claims 1 Process for extracting the total flavone from the seeds of odoriferous cedarel, comprising the following steps: mixing the powder of the seeds of odoriferous cedarel in a NADES eutectic solvent, carrying out the homogenization, centrifugation and filtering, purifying the filtrate Using an AB-8 resin, elute the total flavone from the seeds of sweet cedar with ethanol at a gradient of 70% to 95%.
    [2]
    2 Method according to claim 1, characterized in that the eutectic solvent NADES is composed of choline chloride having a water content of 30% and 1,2-propanediol, of which the optimum molar ratio between the choline chloride and the 1 , 2-propanediol is 1/3.
    [3]
    3 Method according to claim 1, characterized in that the mass ratio between the seed powder of odoriferous cedarella and the eutectic solvent NADES is 1/50.
    [4]
    4 The method of claim 1, characterized in that the duration of said ultrasound is 15 min.
    [5]
    5 The method of claim 1, characterized in that the rotational speed of said centrifugation is 4000 rpm, while the centrifugation time is 25 min.
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    同族专利:
    公开号 | 公开日
    BE1027352A1|2021-01-13|
    CN110302248A|2019-10-08|
    CN110302248B|2021-10-26|
    引用文献:
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    法律状态:
    2021-04-09| FG| Patent granted|Effective date: 20210120 |
    优先权:
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