CN105497169A - Method for extracting phenolic acid effective constituents of rhizome cimicifugae - Google Patents
Method for extracting phenolic acid effective constituents of rhizome cimicifugae Download PDFInfo
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- CN105497169A CN105497169A CN201610010292.6A CN201610010292A CN105497169A CN 105497169 A CN105497169 A CN 105497169A CN 201610010292 A CN201610010292 A CN 201610010292A CN 105497169 A CN105497169 A CN 105497169A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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Abstract
The invention discloses a method for extracting phenolic acid effective constituents of rhizome cimicifugae. The method includes the steps of taking and smashing rhizome-cimicifugae medicinal materials, adding a 55%-85%-ethyl alcohol solvent with the volume 6-14 times the weight of the rhizome-cimicifugae medicinal materials, and carrying out extracting 1-3 times, wherein each time of extracting lasts for 50-300 min; combining extracted solutions, and carrying out concentrating to obtain rhizome-cimicifugae extract containing the phenolic acid effective constituents. According to the extracting method, as factors such as the dissolvent ratio, the ethyl alcohol volume fraction, extracting time and the number of extracting times are analyzed and screened through a system, and the interaction relationships of the main influence factors such as the dissolvent ratio, the ethyl alcohol volume fraction and the extracting time are analyzed with a response surface analysis method on the basis of the screening results, and an optimal combination is screened out. An experimental result is verified to prove that according to the method, a process route is stable, operability is high, the content of the phenolic acid effective constituents in the extract obtained through preparing is high, and great application significance is achieved.
Description
Technical field
The present invention relates to a kind of extracting method of Effective Component of Chinese Medicine, be specifically related to the method that a kind of aggregative weighted average response face method optimizes Rhizoma Cimicifugae extraction process.
Background technology
Rhizoma Cimicifugae is the Chinese medicine of China, has and delivers rash, heat-clearing and toxic substances removing, effect of elevate a turnable ladder yang-energy.For headache due to pathogenic wind-heat, toothache, aphtha, laryngopharynx swelling and pain, measles without adequate eruption, YANG toxin syndrome sends out speckle, proctoptosis, uterine prolapse.Clinical practice is extensive, aboundresources.Wherein liposoluble ingredient has antioxidation through modern pharmacological research, antiinflammatory, anti-platelet aggregation, antitumor, and reduce the effects such as blood glucose, clinical drug effect is remarkable.
But the processing of current Rhizoma Cimicifugae extract, do not have unified standard, still adopt outmoded technique, cause the waste of resource, crudy is unstable, inferior quality.
Summary of the invention:
Goal of the invention: technical problem to be solved by this invention overcomes the deficiencies in the prior art, adopt the optimization of aggregative weighted average response face method, preferably obtain a kind of extraction yield high, and phenolic acids active constituent content is high, process stabilizing is reliable, the extracting method of workable Rhizoma Cimicifugae phenolic acids effective ingredient.
Technical scheme: in order to realize above object, the technical solution used in the present invention is:
An extracting method for Rhizoma Cimicifugae phenolic acids effective ingredient, is characterized in that, comprise the following steps:
Get Rhizoma Cimicifugae medical material, pulverize, add 55 ~ 85% alcohol solvents of Rhizoma Cimicifugae medical material weight 6 ~ 14 times amount volume, extract 1 ~ 3 time, each 50 ~ 300min, merge extractive liquid, the concentrated Rhizoma Cimicifugae extract obtained containing liposoluble ingredient.
Preferably, the extracting method of above-described Rhizoma Cimicifugae phenolic acids effective ingredient, comprises the following steps:
Get Rhizoma Cimicifugae medical material, pulverize, add 70% alcohol solvent of Rhizoma Cimicifugae medical material weight 8 times amount volume, extract 2 times, each 220min, merge extractive liquid, the concentrated Rhizoma Cimicifugae extract obtained containing liposoluble ingredient.
Preferably, the extracting method of above-described Rhizoma Cimicifugae phenolic acids effective ingredient, containing caffeic acid, ferulic acid and Hesperetic acid in Rhizoma Cimicifugae extract.
Preferably, the extracting method of above-described Rhizoma Cimicifugae phenolic acids effective ingredient, in Rhizoma Cimicifugae extract containing the caffeic acid of percentage by weight 0.015 ~ 0.018%, the ferulic acid of 0.033 ~ 0.041% and 0.229 ~ 0.263% Hesperetic acid.
Preferably, the extracting method of above-described Rhizoma Cimicifugae phenolic acids effective ingredient, Rhizoma Cimicifugae extract yield is 0.18 ~ 0.21%.
Craft screening is tested:
1, the assay method of index composition:
The formulation of standard curve: accurately weighed caffeic acid, ferulic acid, Hesperetic acid reference substance is appropriate, adds methanol and is mixed with concentration and is respectively 47.20,144.80,247.40mgL
-1mixing reference substance.Mixing reference substance solution 2,5,8,10,15,18 μ L, measure by under chromatographic condition, and carry out statistical regression with peak area (Y) to sample size (X), obtain caffeic acid, ferulic acid, the regression equation of Hesperetic acid is respectively
Y＝273287X-1149084650(r＝0.9999)；
Y＝745351X+38143(r＝0.9999)
Y＝1000000X+283628(r＝0.9995)。
According to above regression equation, HPLC is adopted to measure the content of liposoluble ingredient in Rhizoma Cimicifugae.Chromatographic condition is: chromatographic column: YMC-Pack-ODS-C4 (250 × 4.6mm, 5 μm); Flowing acetonitrile-0.1% formic acid solution (0 ~ 12min, 5% ~ 13%A; 12 ~ 23min, 13% ~ 18%A; 23 ~ 30min, 18% ~ 5%A); Flow velocity: 1.0mLmin
-1; Column temperature: 30 DEG C; Determined wavelength: 320nm; Sample size: 10 μ L.
2, the extraction of index composition:
The extraction of Rhizoma Cimicifugae liposoluble ingredient: get Rhizoma Cimicifugae and cross No. 2 sieves after crushed, accurately take 10.0g Largetrifoliolious Bugbane Rhizome in 250mL boiling flask, add ethanol, reflux, extract, in water-bath, filter and get final product.
3, single factor test and response surface experimental result as follows:
3.1 different ethanol concentration are on the impact of Rhizoma Cimicifugae liposoluble ingredient extraction ratio
The present invention adopts the ethanol extraction Rhizoma Cimicifugae of 20% ~ 80% different volumes concentration, as shown in Figure 1, solvent compares 10:1, under the condition of extraction time 120min, along with ethanol contend concentration increases, the peak area of total phenolics slowly rises, when ethanol contend concentration reaches 70%, peak area reaches maximum, obviously declines later, shows to adopt liposoluble ingredient religion in volumetric concentration 70% ethanol extraction Rhizoma Cimicifugae good.
3.2 Different solution ratios (mL/g) are on the impact of Rhizoma Cimicifugae liposoluble ingredient extraction ratio
The present invention adopts ethanol contend and Rhizoma Cimicifugae medical material weight ratio to be respectively 5 ~ 16 times to carry out extraction Rhizoma Cimicifugae, as shown in Figure 2, be 70% in ethanol contend concentration, under the condition of extraction time 120min, along with solvent is than increasing, the peak area of total phenolics raises, when ethanol contend (mL) and Rhizoma Cimicifugae medical material weight (g) are than when being 8, the peak area of total phenolics reaches maximum, and peak area obviously declines afterwards.Above result shows: when phenolic acid components unsaturation, increase along with the increase of quantity of solvent, when preventing solution to be in supersaturation, then prevents the stripping of effective ingredient.
3.3 different extraction times are on the impact of Rhizoma Cimicifugae liposoluble ingredient extraction ratio
The present invention extracted Rhizoma Cimicifugae by 50 ~ 300 minutes different extraction times, Fig. 3 is known, and at ethanol contend 70%, solvent is than under the condition of 8:1, along with the increase of extraction time, the extraction ratio of total phenolics raises gradually, and when the time is 240min, extraction rate reached is to maximum, but during overlong time, extract cell membrane fragmentation also to strengthen, also stripping is too much for impurity thereupon, affects the extraction of effective ingredient.
4, response phase method optimization experiment:
On single factor experiment basis, the present invention chooses the factor larger on liposoluble ingredient impact, according to Box-Benhnken EXPERIMENTAL DESIGN principle, the response surface analysis test of design Three factors-levels, research volume fraction of ethanol, liquid ratio and extraction time and reciprocal action thereof are on the impact of Rhizoma Cimicifugae index constituents extraction rate;
Single factor test and response surface experiment comprise as follows:
A, volume fraction of ethanol (55-85%) are on the impact of Rhizoma Cimicifugae index composition
B, liquid ratio (6:1-10:1) are on the impact of Rhizoma Cimicifugae index composition
C, reflux extracting time (180-270min) are on the impact of Rhizoma Cimicifugae index composition.
By consulting literatures and trial test, set comprehensive scoring full marks are 100 points, according to active constituent content in Rhizoma Cimicifugae and drug effect impact, caffeic acid, ferulic acid, Hesperetic acid, extractum weight coefficient is respectively 20,30,40,10, carry out total weighted scoring on this basis, comprehensive grading value (P) is utilized to analyze result of the test: the measured value representing index components in test with Y, with the maximum (Y of each index
max) as reference, standardization being carried out to each data of same index, C represents the standardized data (C=Y/Y of the measured value of this index components in test
max), comprehensive grading P=C
caffeic acid× 20+C
ferulic acid× 30+C
hesperetic acid× 40+C
extractum× 10.According to Box-Behnken center combination EXPERIMENTAL DESIGN principle, comprehensive single-factor influence result of the test, chooses solvent ratio (X
1), concentration of alcohol (X
2) and extraction time (X
3) as investigating object, adopt the Responds Surface Methodology arrangement test of DesignExpert8.0 statistical analysis software, to obtain the suitableeest technological parameter.
The present invention adopts table 1 aggregative weighted average response face method to optimize EXPERIMENTAL DESIGN and the result of Rhizoma Cimicifugae extraction process
Table 1: response phase method optimizes EXPERIMENTAL DESIGN and the result of Rhizoma Cimicifugae extraction process
The results are shown in Table 1 by the response surface, utilize Design-Expert software to carry out regression statistical analysis to experimental result, the regression equation obtained is as follows:
P＝-16.82+5.95X1+1.64X2+0.31X3+0.10X1X2+0.0067X1X3-0.00022X2X3-0.94X12-0.018X22-0.0008432(r＝0.9671)
Regression equation once in item the size of every absolute coefficient directly reflect the influence degree of each factor to response value, the positive and negative direction reflecting impact of coefficient.From equation, the primary and secondary order affecting the factor of Rhizoma Cimicifugae index composition is X1>X2>X3, carries out analysis of variance in regression further to equation.
By the variance analysis of response phase method in table 2, in regression equation, each variable judges the significance F inspection that response value affects, probability P value is less, then the significance degree of relevant variable is higher, during P < 0.01, impact is highly significant, and P < 0.05 affects significantly.As can be seen from the analysis result of table 2, model P < 0.01, illustrates that the multinomial model of secondary has the significance of height, model coefficient of determination R2=0.9671, shows that model-fitting degree is good, the success of quadratic term models fitting.
Table 2: the results of analysis of variance
From response phase method optimization experiment result, ethanol contend, solvent ratio, the reciprocal action of extraction time all has appreciable impact, optimizing Rhizoma Cimicifugae liposoluble ingredient extraction process optimum condition according to aggregative weighted average response face method is: liquid ratio is 8, concentration of alcohol 70%, extraction time 200min, extraction time is 2 times.
5, demonstration test
Whether consistently with truth predicting the outcome to check, with the theoretical optimum condition of bend plane conversion for operating condition, carrying out the demonstration test of three groups of repetitions.The weighed 3 parts of Largetrifoliolious Bugbane Rhizome 10.0g of precision parallel, 2 times are extracted according to prediction optimum extraction process, liquid ratio is 8, ethanol contend concentration 70%, extraction time is that 200min carries out demonstration test, result average aggregate scoring 93.26, compared with theoretical expectation values, relative error is respectively 2.20%, is less than 5%, within range of error.Therefore, Comparison of experiment results is stablized at the process conditions, and repeatability is good, also proves that the present invention selects Design-ExpertSoftware to design the reliability of screening each influence factor simultaneously.
Therefore, the optimum extraction process of the Rhizoma Cimicifugae that the present invention preferably obtains is: volume 70% alcohol solvent adding Rhizoma Cimicifugae medical material weight 8 times amount, extracts 2 times, each 200min.The present invention has investigated solvent ratio through great many of experiments screening, volume fraction of ethanol, extraction time 3 factors, take comprehensive grading as reference index, rational weighted coefficient distribution is carried out to selected index components, makes to evaluate extraction process with comprehensive grading more reasonable, the information of extract effective ingredient can be reflected comprehensively, optimize the extracting factor of Rhizoma Cimicifugae, to its industrialized great production, there is important directive significance.
Beneficial effect: compared to the prior art the extracting method of Rhizoma Cimicifugae phenolic acids effective ingredient provided by the invention has the following advantages:
The extracting method of Rhizoma Cimicifugae phenolic acids effective ingredient provided by the invention, by system to solvent ratio, volume fraction of ethanol, the factor such as extraction time and extraction time carries out analyzing and screening, and on the result basis of screening, recycling Response surface methodology analyzes solvent ratio, volume fraction of ethanol, the reciprocal effect relation of the major influence factors such as extraction time, and filter out optimum combination.Shown by confirmatory experiment result, the extracting method of Rhizoma Cimicifugae phenolic acids effective ingredient provided by the invention, process route is stable, feasible, workable, prepares in extract, phenolic acids active constituent content is high, can reach more than 0.32%, and impurity content is low, the optimum extracting method that the present invention preferably obtains, extraction conditions accurately and reliably, extraction efficiency is high, can provide scientific basis as Rhizoma Cimicifugae extraction process, can ensure that stablizing of Rhizoma Cimicifugae extract is significant.
Accompanying drawing explanation
Fig. 1 different ethanol concentration is on the impact of Rhizoma Cimicifugae liposoluble ingredient extraction ratio;
The impact of Fig. 2 Different solution comparison Rhizoma Cimicifugae liposoluble ingredient extraction ratio;
Fig. 3 different extraction time is on the impact of Rhizoma Cimicifugae liposoluble ingredient extraction ratio;
The response surface figure that Fig. 4 volume fraction of ethanol, ultrasonic temperature and sonication times reciprocal action affect extraction ratio.
Detailed description of the invention
According to following embodiment, the present invention may be better understood.But those skilled in the art will readily understand, concrete material proportion, process conditions and result thereof described by embodiment only for illustration of the present invention, and should can not limit the present invention described in detail in claims yet.
Embodiment 1
Accurately weighed Rhizoma Cimicifugae medical material, pulverizes, adds 70% alcohol solvent of Rhizoma Cimicifugae medical material weight 8 times amount volume, extract 2 times, each 200min, merge extractive liquid, the concentrated Rhizoma Cimicifugae extract obtained containing liposoluble ingredient.
And adopt above-mentioned high-efficient liquid phase chromatogram condition and regression equation, calculating extract yield is 0.21%, the caffeic acid of content percentage by weight 0.018% in extract, the ferulic acid of 0.041% and, the Hesperetic acid of 0.263%.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (5)
1. an extracting method for Rhizoma Cimicifugae phenolic acids effective ingredient, is characterized in that, comprises the following steps:
Get Rhizoma Cimicifugae medical material, pulverize, add 55 ~ 85% alcohol solvents of Rhizoma Cimicifugae medical material weight 6 ~ 14 times amount volume, extract 1 ~ 3 time, each 50 ~ 300min, merge extractive liquid, the concentrated Rhizoma Cimicifugae extract obtained containing liposoluble ingredient.
2. the extracting method of Rhizoma Cimicifugae phenolic acids effective ingredient according to claim 1, is characterized in that, comprise the following steps:
Get Rhizoma Cimicifugae medical material, pulverize, add 70% alcohol solvent of Rhizoma Cimicifugae medical material weight 8 times amount volume, extract 2 times, each 220min, merge extractive liquid, the concentrated Rhizoma Cimicifugae extract obtained containing liposoluble ingredient.
3. the extracting method of Rhizoma Cimicifugae phenolic acids effective ingredient according to claim 1 and 2, is characterized in that, containing caffeic acid, ferulic acid and Hesperetic acid in Rhizoma Cimicifugae extract.
4. the extracting method of Rhizoma Cimicifugae phenolic acids effective ingredient according to claim 3, it is characterized in that, in Rhizoma Cimicifugae extract containing the caffeic acid of percentage by weight 0.015 ~ 0.018%, the ferulic acid of 0.033 ~ 0.041% and 0.229 ~ 0.263% Hesperetic acid.
5. the extracting method of Rhizoma Cimicifugae phenolic acids effective ingredient according to claim 1 and 2, is characterized in that, Rhizoma Cimicifugae extract yield is 0.18 ~ 0.21%.
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Citations (4)
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|---|---|---|---|---|
| CN1580062A (en) * | 2004-03-23 | 2005-02-16 | 肖培根 | Method for extracting total aglycone of Macrotys and its antitumour action |
| CN101411753A (en) * | 2008-12-04 | 2009-04-22 | 闫明 | Broomrape extract, broomrape total glycosides, broomrape polysaccharide as well as preparation method and application thereof |
| CN102060693A (en) * | 2010-12-20 | 2011-05-18 | 大兴安岭嘉迪欧营养原料有限公司 | Method for enriching and purifying ferulic acid in cimicifugae foetidae |
| CN102180933A (en) * | 2011-03-28 | 2011-09-14 | 南京泽朗农业发展有限公司 | Preparation method of skunk bugbane monomer components |
-
2016
- 2016-01-06 CN CN201610010292.6A patent/CN105497169A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1580062A (en) * | 2004-03-23 | 2005-02-16 | 肖培根 | Method for extracting total aglycone of Macrotys and its antitumour action |
| CN101411753A (en) * | 2008-12-04 | 2009-04-22 | 闫明 | Broomrape extract, broomrape total glycosides, broomrape polysaccharide as well as preparation method and application thereof |
| CN102060693A (en) * | 2010-12-20 | 2011-05-18 | 大兴安岭嘉迪欧营养原料有限公司 | Method for enriching and purifying ferulic acid in cimicifugae foetidae |
| CN102180933A (en) * | 2011-03-28 | 2011-09-14 | 南京泽朗农业发展有限公司 | Preparation method of skunk bugbane monomer components |
Non-Patent Citations (2)
| Title |
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| 沈保家: "升麻质量评价及药代动力学研究", 《医药卫生科技辑》 * |
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