CN108113926A - A kind of Herba Saussureae Involueratae ultrasonic extraction process - Google Patents
A kind of Herba Saussureae Involueratae ultrasonic extraction process Download PDFInfo
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- CN108113926A CN108113926A CN201711500278.5A CN201711500278A CN108113926A CN 108113926 A CN108113926 A CN 108113926A CN 201711500278 A CN201711500278 A CN 201711500278A CN 108113926 A CN108113926 A CN 108113926A
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- herba saussureae
- saussureae involueratae
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- general flavone
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
Abstract
The invention discloses a kind of Herba Saussureae Involueratae ultrasonic extraction process, include the following steps：S1, by Herba Saussureae Involueratae using solid-liquid ratio be respectively 1: 50 be immersed in volume fraction as 80% ethanol solution in 15min, with ultrasonic power 60w extract 15min to get Herba Saussureae Involueratae general flavone；S2, the Herba Saussureae Involueratae general flavone of gained is evaporated ethyl alcohol, obtains alcohol extract, carried out alkali extraction and acid precipitation purifying, measure the absorbance of its 320nm on ultraviolet specrophotometer afterwards, Herba Saussureae Involueratae general flavone preventing ultraviolet effect, the results show are judged according to absorbance：Absorbance is 1.742, it is seen that the Herba Saussureae Involueratae alcohol extract obtained by above-mentioned Herba Saussureae Involueratae ultrasonic extraction process can be used for ultraviolet light sun-proof.Apparatus of the present invention are relatively simple, operation is more convenient, have the characteristics that efficient, expense is low and at low cost.
Description
Technical field
The present invention relates to field of plant extraction, and in particular to a kind of Herba Saussureae Involueratae ultrasonic extraction process.
Background technology
Herba Saussureae Involueratae (Saussurea involucrata (Kar.et Kir.) Sch.-Bip.) is that composite family Genus Saussurea is planted
Object, while also known as Xinjiang Saussurea involucrate, snow lotus of big bud, it is the conventional crude drugs of the Uygur nationality, Uighur name " tower since time immemorial power this ", with
And Tibetan language name " proper fruit Soviet Union bar ".Herba Saussureae Involueratae is distributed mainly on Xinjiang of China Tianshan Mountains, Altai Mountains, this band of Kun Lun Mountain, growth
In the rock seam, cliff and ice debt ruckle of height above sea level 2600-4000m.It is not only exotic flowers and rare herbs rare to see and lifts
The well-known Tibetan medicine being of great rarity of generation.Herba Saussureae Involueratae can make it is medicinal, while also have certain ornamental value.Among the people with complete
Grass is used as medicine, first appeared in Tibetan medicine document《Month king's drug》, afterwards in the Qing Dynasty《A Supplement to the Compendium of Materia Medica》Also it is on the books, be Tibet, Xinjiang,
The common ethnic drug and Folk medicine of the plateaus such as Qinghai, Gansu, Yunnan, Sichuan refrigerant latitudes.Described in its traditional Chinese medical science, Herba Saussureae Involueratae
Its is warm-natured, mildly bitter flavor, enters liver,spleen,kidney three and passes through, and has warming kidney and enhancing yang, expelling wind and eliminating dampness, the primary efficacy of clearing and activating the channels and collaterals, while can
For treating the symptoms such as rheumatism numbness pain, rheumatoid arthritis, cold and pain in the lower abdomen, irregular menstruation.In its Uygur medicine, Tianshan Mountains snow
Its property two level of lotus is damp and hot, have the function of invigorating kidney, promoting blood circulation, strengthening the bones and muscles, trophic nerve, adjust body fluid exception and available for rheumatic
The symptoms such as arthritis, arthralgia, lung cold cough, kidney and cold and pain in the lower abdomen, leukorrhea.
Just because of this, Herba Saussureae Involueratae becomes a kind of a kind of Chinese herbal medicine among the people for having and developing and utilizing value, together
When, many scientists have medically done it more research.According to research and development, show to contain flavones in Herba Saussureae Involueratae
The Multiple components such as class, alkaloid, lactones, sterol, polysaccharide and volatile oil, it is pregnant with anti-inflammatory, antirheumatic, analgesia, cardiac stimulant, termination
It is pregnent, anticancer, anti-oxidant, radioresistance, spasmolysis, is depressured and a variety of pharmacological activity such as relieving asthma.
The content of the invention
To solve the above problems, the present invention provides a kind of Herba Saussureae Involueratae ultrasonic extraction process.
To achieve the above object, the technical solution taken of the present invention is：
A kind of Herba Saussureae Involueratae ultrasonic extraction process, includes the following steps：
S1, by Herba Saussureae Involueratae using solid-liquid ratio be respectively 1: 50 be immersed in volume fraction as 80% ethanol solution in 15min,
15min is extracted to get Herba Saussureae Involueratae general flavone with ultrasonic power 60w；
S2, the Herba Saussureae Involueratae general flavone of gained is evaporated ethyl alcohol, obtains alcohol extract, carry out alkali extraction and acid precipitation purifying, it
The absorbance of its 320nm is measured on ultraviolet specrophotometer afterwards, Herba Saussureae Involueratae general flavone antiultraviolet is judged according to absorbance
Effect, the results show：Absorbance is 1.742, it is seen that the Herba Saussureae Involueratae alcohol extract obtained by above-mentioned Herba Saussureae Involueratae ultrasonic extraction process
It is sun-proof available for ultraviolet light.
Apparatus of the present invention are relatively simple, operation is more convenient, have the characteristics that efficient, expense is low and at low cost.
Description of the drawings
Fig. 1 is rutin standard curve.
Influence line chart of Fig. 2 concentration of alcohol to recovery rate.
Fig. 3 is influence line chart of the soaking time to recovery rate.
Fig. 4 is influence line chart of the solid-liquid ratio to recovery rate.
Fig. 5 is influence line chart of the ultrasonic power to recovery rate.
Specific embodiment
In order to which objects and advantages of the present invention are more clearly understood, the present invention is carried out with reference to embodiments further
It is described in detail.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to limit this hair
It is bright.
Herba Saussureae Involueratae used in following embodiment is Herba Saussureae Involueratae bouquet, the mineral water of natural drying
The present embodiment Jilin agricultural science and technology institute Chemistry for Chinese Traditional Medicine laboratory complete, by single factor experiment (concentration of alcohol,
Soaking time, solid-liquid ratio, ultrasonic time, ultrasonic power) in determine 4 factors being affected to extraction efficiency (concentration of alcohol,
Soaking time, solid-liquid ratio, ultrasonic time) orthogonal test is carried out, it is preferred so as to carry out ultrasonic extraction process.Then optimised process is used
The Herba Saussureae Involueratae general flavone of extraction is evaporated, i.e. Herba Saussureae Involueratae alcohol extract, using its absorbance of determined by ultraviolet spectrophotometry,
Judge whether Herba Saussureae Involueratae alcohol extract has the effect of antiultraviolet according to the evaluation of 5 sun-proof result of table.
The drafting of rutin standard curve
The preparation of standard solution
Precision weighs control substance of Rutin 5mg, adds in the dissolving of 95% ethyl alcohol, and constant volume obtains in 25mL volumetric flasks
The standard solution of 0.2mg/mL.
The formulation of standard curve：
Precision draws rutin titer 0.0,1.0,2.0,3.0,4.0,5.0mL, puts respectively in 25mL volumetric flasks, adds 5%
NaNO2Solution 1mL, shakes up, and places 6min, adds 10%AL (NO)3Solution 1mL, shakes up, and places 6min, and finally plus 4%NaOH is tried
Liquid 10mL, then distilled water is added to be diluted to scale, it shakes up, 15min is placed, using determined by ultraviolet spectrophotometry, with the first reagent
Solution is blank control, in 200-800nm range scans, determines maximum absorption band.Absorbance is measured in maximum, with absorbance
For ordinate, concentration is abscissa, draws standard curve, obtains equation：Y=12.554x-0.003 (R2=0.9982), the result is shown in
Fig. 1.The result shows that concentration linear relationship in the range of 8-48ug/ml is good.
Concentration of alcohol influences to measure
Precision weighs Herba Saussureae Involueratae 0.3g or so respectively, is immersed in 15min, solid-liquid ratio 1: 80, and concentration of alcohol is respectively：
70%th, 75%, 80%, 85%, 90%, with ultrasonic power 60w, extract 1 time, ultrasonic time 15min in triplicate, is prepared into
Test solution measures its absorbance, investigates influence of the soaking time to its content, determines optimal soaking time.
The measure that the soaking time of saussurea involucrata in organic solvent influences
Precision weighs Herba Saussureae Involueratae 0.3g or so respectively, is immersed in 80% ethanol solution, solid-liquid ratio 1: 80, the time point
It is not：5min, 10min, 15min, 20min, 25min with ultrasonic power 60w, extract 1 time, ultrasonic time 15min, repeat three
It is secondary, test solution is prepared into, measures its absorbance, influence of the soaking time to its content is investigated, determines optimal immersion
Time.
The measure that solid-liquid ratio influences
Precision weighs Herba Saussureae Involueratae 0.3g or so respectively, and solid-liquid ratio is respectively 1: 60,1: 70,1: 80,1: 90,1: 100, leaching
Bubble 15min in 80% ethanol solution with ultrasonic power 60w, is extracted 1 time, ultrasonic extraction 15min is repeated, is prepared into three times
Test solution measures its absorbance, investigates the influence that liquid ratio carries Herba Saussureae Involueratae component content, determines optimal material
Liquor ratio.
The measure that ultrasonic time influences
Precision weighs Herba Saussureae Involueratae 0.3g or so respectively, and solid-liquid ratio 1: 80 is immersed in 15min in 80% ethanol solution,
It with ultrasonic power 60w, extracts 1 time, ultrasonic extraction 5min, 10min, 15min, 20min, 25min, repeat three times respectively, prepare
Into test solution, its absorbance is measured, investigates the influence that ultrasonic time carries Herba Saussureae Involueratae component content, is determined most preferably
Ultrasonic time.
The measure that ultrasonic power influences
Precision weighs Herba Saussureae Involueratae 0.3g or so respectively, and solid-liquid ratio 1: 100 is immersed in 15min in 80% ethanol solution,
Respectively with ultrasonic power 40w, 50w, 60w, 70w, 80w, extract 1 time, ultrasonic extraction 15min repeats, is prepared into test sample three times
Solution measures its absorbance, investigates the influence that ultrasonic power carries Herba Saussureae Involueratae component content, determines optimal ultrasonic work(
Rate.
Orthogonal Experiment and Design
Herba Saussureae Involueratae coarse powder about 5g is taken, it is accurately weighed, it puts in round-bottomed flask, further according to single factor experiment the results show that reality
It tests using the extraction of 80w power ultrasonics once, selectes concentration of alcohol soaking time, ultrasonic time, four factor of solid-liquid ratio, three level, choosing
With L9 (34) orthogonal arrage tested.After ethanol evaporation, then the NaOH adjusting pH value for adding in 4-6 drops 1mol/L shakes up for 10
The HCl to pH of 1mol/L is added dropwise as 2-3, stands 20min, 10 000r/min centrifugation 20min obtain tan precipitate.Collect supernatant
Liquid (continues to concentrate, repeat above-mentioned settling step), and residual liquid is blotted with filter paper, adds in 70% ethyl alcohol of 20mL volume fractions and adjusts
Section pH is 10-11, is completely dissolved precipitation, and dropwise addition HCl to pH is 1-2, stands 20min, and repeated centrifugation collects precipitation.With steaming
Distilled water repeated washing precipitation 3 times, removes water-solubility impurity and salt.Using water as blank, existed with ultraviolet-visible spectrophotometer
Absorbance is measured at 510nm wavelength, the concentration (g/mL) of general flavone in test solution is drawn from standard curve, calculates day
The recovery rate of mountain Saussurea involucrate general flavone, is shown in Table 1.Variance analysis is carried out with orthogonal test, is shown in Table 2.
1 factor of table and water-glass
Note：A is concentration of alcohol, and B is soaking time, and C is solid-liquid ratio, and D is ultrasonic time
As a result with analysis
Influence of the concentration of alcohol to recovery rate
Influence of 2 concentration of alcohol of table to recovery rate
Analysis：Concentration of alcohol is in 70%-90%, with the increase of concentration of alcohol, Herba Saussureae Involueratae general flavone recovery rate by
Gradually rise and then decline and rise again, maximum is at 80%.As concentration of alcohol ＞ 80%, general flavone recovery rate first under
Rise after drop, this may be since concentration of alcohol is at 85%, and other materials are easier to be dissolved in ethyl alcohol, cause impurity in leaching liquor
Content is higher, finally general flavone content is caused to reduce.
Variance analysis
3 analysis of variance table of table
Analysis：Variance analysis learns, F0.01(4,10)=5.99 are learnt by table 2, F value ＞ critical values 5.99, therefore notable
Property the refusal of level 0.01 time H0, illustrate that its yield has marked difference under 5 kinds of different ethanol concentrations of Herba Saussureae Involueratae general flavone.
Conclusion：When concentration of alcohol is 70%, 75%, 80%, 85%, 90%, corresponding Herba Saussureae Involueratae general flavone is received
Yield is respectively 2.431%, 2.732%, 3.690%, 1.918%, 2.487%.Therefore according to table 3, selected three water
It puts down as 75%, 80%, 85%.
Influence of the soaking time to recovery rate
Data analysis
Influence of 4 soaking time of table to recovery rate
Variance analysis
5 analysis of variance table of table
Analysis：Variance analysis learns, F0.01(4,10)=5.99 are learnt by table 5, F value ＞ critical values 5.99, therefore notable
Property the refusal of level 0.01 time H0, illustrate that its yield has marked difference under 5 kinds of different soaking times of Herba Saussureae Involueratae general flavone.
Conclusion：When its soaking time is 5min, 10min, 15min, 20min, 25min, corresponding Herba Saussureae Involueratae is always yellow
Ketone recovery rate is respectively 2.138%, 2.274%, 3.368%, 2.751%, 2.837%.According to table 5, selected three water
It puts down as 10min, 15min, 20min.
Influence of the solid-liquid ratio to recovery rate
Data analysis
Influence of 6 solid-liquid ratio of table to recovery rate
Variance analysis
7 analysis of variance table of table
Analysis：Variance analysis learns, F0.01(4,10)=5.99 are learnt by table 7, F value ＞ critical values 5.99, therefore notable
Property the refusal of level 0.01 time H0, illustrate 5 kinds of different feed liquids of Herba Saussureae Involueratae general flavone has marked difference than its lower yield.
Conclusion：When solid-liquid ratio is 1: 20,1: 30,1: 40,1: 50,1: 60, corresponding Herba Saussureae Involueratae general flavone is received
Rate is respectively 2.657%, 2.715%, 3.411%, 2.397%, 2.503%.According to table 7, selected three levels are 1:
30、 1∶40、1∶50。
Influence of the ultrasonic time to recovery rate
Data analysis
Influence of 8 ultrasonic time of table to recovery rate
Variance analysis
9 analysis of variance table of table
Analysis：Variance analysis learns, F0.01(4,10)=5.99 are learnt by table 9, F value ＞ critical values 5.99, therefore notable
Property the refusal of level 0.01 time H0, illustrate that its yield has marked difference under 5 kinds of different ultrasonic times of Herba Saussureae Involueratae general flavone.
Conclusion：When ultrasonic time is 5min, 10min, 15min, 20min, 25min, corresponding Herba Saussureae Involueratae is always yellow
Ketone recovery rate is respectively 2.009%, 2.252%, 3.367%, 2.717%, 2.719%.According to table 9, selected three water
It puts down as 10min, 15min, 20min.
Influence of the ultrasonic power to recovery rate
Data analysis
Influence of 10 ultrasonic power of table to recovery rate
Variance analysis
11 analysis of variance table of table
Analysis：Variance analysis learns, F0.01(4,10)=5.99 are learnt by table 11, F value ＜ critical values 5.99, therefore aobvious
Receive H under work property level 0.010, illustrate that its recovery rate does not have marked difference under 5 kinds of ultrasonic powers of Herba Saussureae Involueratae general flavone.
Conclusion：When ultrasonic power is 60w, 70w, 80w, 90w, 100w, corresponding Herba Saussureae Involueratae general flavone recovery rate
Respectively 1.629%, 1.682%, 1.909%, 1.791%, 1.795%.Because the influence of its factor is not notable, choose
Level is peak 80w.
Orthogonal test determines optimum extraction process
According to single factor experiment the results show that experiment is extracted once using 80w power ultrasonics, select concentration of alcohol, impregnate
Time, solid-liquid ratio, four factor of ultrasonic time three are horizontal, select L9 (34) orthogonal arrage tested.Factor is determined with horizontal, calculating
Recovery rate carries out variance analysis with orthogonal test, is shown in Table 12.
The orthogonal test scheme and result of 12 general flavone extraction conditions of table
Conclusion：By the R values of four factors in 12 tables, show that influence degree is followed successively by solid-liquid ratio (C) ＞ ultrasonic times (D) ＞
Soaking time (B) ＞ concentration of alcohol (A)；The K values of four factors, the influence degree for showing each factor level are A1＞ A2＞ A3, B2
＞ B3＞ B1, C2＞ C3＞ C1, D2＞ D3＞ D1It is A that i.e. optimal extraction conditions, which are,1B2C2D2.I.e. concentration of alcohol is 75%, impregnates
The yield highest of Herba Saussureae Involueratae general flavone when time is 15min, solid-liquid ratio 1: 40, ultrasonic time are 15min.
Supercritical ultrasonics technology is compared with heating alcohol extracting method
The Contrast on effect of 13 ultrasonic extraction of table and water extraction method
Conclusion：During using ultrasonic extraction, the yield of Herba Saussureae Involueratae general flavone is 8.45%, molten compared to alcohol compared with alcohol extracting method
Property general flavone yield add 3.46%, shorten 105min on extraction time, extraction time is reduced 1 time.Therefore using ultrasound
Ripple extraction method has apparent advantage.
VB areas sun-proof result evaluation (absorbance)
General flavone in Herba Saussureae Involueratae is extracted using ultrasonic wave optimised process, is waved under the action of Rotary Evaporators
Hair, after ethyl alcohol is volatilized, obtains alcohol extract, and the NaOH adjusting pH value for adding 4-6 drops 1mol/L shakes up and then is added dropwise for 10
The HCl of 1mol/L to pH is 2-3, stands 20min, and 10 000r/min centrifugation 20min obtain tan precipitate.It collects supernatant
(continuing to concentrate, repeat above-mentioned settling step), residual liquid is blotted with filter paper, is added in 70% ethyl alcohol of 20mL volume fractions and is adjusted
PH is 10-11, is completely dissolved precipitation, and dropwise addition HCl to pH is 1-2, stands 20min, and repeated centrifugation collects precipitation.With distillation
Water repeated washing precipitation 3 times, remove water-solubility impurity and salt to get to purified T1.It is surveyed on ultraviolet specrophotometer
Determine the absorbance of 320nm.It may determine that Herba Saussureae Involueratae extract has sun-proof result according to the evaluation of 14 sun-proof result of table：
14 sun-proof result of table is evaluated
And conclusion as a result：The absorbance measured is 1.742, is judged according to table 14, the sun-proof result of Herba Saussureae Involueratae alcohol extract
It is irradiated for high efficient protected ultraviolet light, use condition is irradiated for summer sunburst.
This specific implementation extracts Herba Saussureae Involueratae general flavone by supercritical ultrasonics technology from Herba Saussureae Involueratae, preferably its extraction process.
5 large effect factors are determined, are respectively concentration of alcohol, soaking time, solid-liquid ratio, ultrasonic time, ultrasonic power.It is right
This five factors carry out experiment of single factor, and 4 factors being affected therefrom are determined, and carry out orthogonal examination for its four factor
It tests, show that optimum extraction process is A by analysis1B2C2D2, i.e., concentration of alcohol be 75%, soaking time 15min, solid-liquid ratio
For 1: 80, ultrasonic time 15min.The recovery rate highest of Herba Saussureae Involueratae general flavone under optimum process condition is 8.45%.China
The heating alcohol extracting method that pharmacopeia (two 2010 editions) uses the extraction of sophora bud general flavone extracts, and operation is loaded down with trivial details, time-consuming, Tianshan Mountains snow
Lotus general flavone uses similar approach to measure its yield as 4.99%, less than ultrasonic method yield.Therefore ultrasonic method and forefathers' experiment pair
It is simpler than device, operation is more convenient, have the characteristics that efficient, expense is low and at low cost.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, without departing from the principle of the present invention, several improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (2)
1. a kind of Herba Saussureae Involueratae ultrasonic extraction process, which is characterized in that include the following steps：
S1, by Herba Saussureae Involueratae using solid-liquid ratio be respectively 1: 50 be immersed in volume fraction as 80% ethanol solution in 15min, with super
Acoustical power 60w extracts 15min to get Herba Saussureae Involueratae general flavone；
S2, the Herba Saussureae Involueratae general flavone of gained is evaporated ethyl alcohol to get alcohol extract.
2. the application of the Herba Saussureae Involueratae alcohol extract obtained by Herba Saussureae Involueratae ultrasonic extraction process as described in claim 1, feature
It is, it is sun-proof available for ultraviolet light.
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Cited By (3)
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| CN111374936A (en) * | 2020-04-02 | 2020-07-07 | 安赛搏(重庆)生物技术有限公司 | Method for preparing saussurea involucrate cell extract by using multi-component liquid system and application thereof |
| CN112675121A (en) * | 2020-12-28 | 2021-04-20 | 大连普瑞康生物技术有限公司 | Method for extracting functional active substances of saussurea involucrate cells and application thereof |
| CN116712473A (en) * | 2023-06-20 | 2023-09-08 | 齐鲁理工学院 | Method for extracting flavone from snow lotus herb |
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| CN112675121A (en) * | 2020-12-28 | 2021-04-20 | 大连普瑞康生物技术有限公司 | Method for extracting functional active substances of saussurea involucrate cells and application thereof |
| CN112675121B (en) * | 2020-12-28 | 2022-07-12 | 大连普瑞康生物技术有限公司 | Method for extracting functional active substances of saussurea involucrate cells and application thereof |
| CN116712473A (en) * | 2023-06-20 | 2023-09-08 | 齐鲁理工学院 | Method for extracting flavone from snow lotus herb |
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