CN106138240A - A kind of extracting method of buckwheat total flavone - Google Patents
A kind of extracting method of buckwheat total flavone Download PDFInfo
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
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- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/0261—Solvent extraction of solids comprising vibrating mechanisms, e.g. mechanical, acoustical
- B01D11/0265—Applying ultrasound
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- 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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- 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
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Abstract
The invention discloses the extracting method of a kind of buckwheat total flavone, comprise the steps: to take Semen Fagopyri Esculenti seed, pulverize, defat, use propanol sulfuric acid ammonium double-aqueous phase system to extract, stratification, take supernatant, to obtain final product.Using extraction process of the present invention to extract the Flavonoid substances in Semen Fagopyri Esculenti, can obtain the extraction ratio higher than 0.3%, high extraction reaches 0.57%.
Description
Technical field
The present invention relates to the extracting method of a kind of buckwheat total flavone, belong to field of medicaments.
Background technology
Semen Fagopyri Esculenti (Fagopyrum esculentum Moench) belongs to the dicotyledonous annual crop of buckwheat.Produce
Upper cultivation mainly have two kinds, i.e. sweet cherry roots (F.esculentumMoench) and Radix Et Rhizoma Fagopyri Tatarici (F.esculentum (L.)
Gaerth).Modern medicine study shows, it is many that Semen Fagopyri Esculenti has antioxidation, anti-inflammatory, antitumor, blood sugar lowering, blood fat reducing etc.
Pharmacological action, nutrition, health care and medical value are higher.Along with the raising of people's living standard, the goods such as buckwheat foods, medicine are just
It is increasingly subject to the favor of people.
Semen Fagopyri Esculenti does not contain only rich in protein, carbohydrate, unsaturated fatty acid and vitamin, possibly together with abundant
The flavone compounds such as bioactive ingredients rutin, kaempferol and Quercetin.Many studies have shown that flavone compound has
Have multiple biological activity, in addition to having the effects such as antibacterial, antiinflammatory, blood pressure lowering, antioxidation, anticancer, give protection against cancer, suppress the aspects such as lipase
Also there is remarkable result.Owing to flavone compound has biological activity so the most widely to its research, develop and utilize
Cause increasing concern.For better profiting from Semen Fagopyri Esculenti resource, offer one is provided badly and can effectively extract wherein flavonoid
The method of material.
The extracting method of traditional buckwheat flavonoids is mainly ethanol immersion, supercritical ultrasonics technology, cellulose enzyme process etc..These sides
Although flavones ingredient can be proposed from Semen Fagopyri Esculenti by method, but there is various defect, such as, a large amount of of organic solvent make
Obtaining dissolvent residual to be difficult to avoid that, operation is complicated, higher to equipment requirements, causes production cost increase etc., strongly limit this
The application industrially of class extractive technique.
10, double-aqueous phase system is typically by two kinds of polymer, polymer and salt, ionic liquid and salt, little molecule hydrophilic organic
Solvent and salt composition.After becoming phase material fully to dissolve in water when two kinds, in intermolecular mutually exclusive power and steric restriction effect
Under the influence of, balance through after a while, form mutual impervious two-phase system.The principle that aqueous two-phase extracts and conventional liquid/
Liquid-solution solvent extraction principle is similar, is in accordance with material and distributes at two alternate selectivitys, but its more conventional liquid liquid solvent extraction
Mode has a clear superiority in: 1, operating condition is gentle, and content of water in system is high (generally 75%~90%), close to physiological environment
System extracts, bioactive substance inactivation or degeneration will not be caused;2, the residue problem of organic solvent it is generally not present;
3, being prone to continuous operation, equipment is simple, it is not necessary to carries out special handling, is suitable to industrialized production.But, use Two-phase system
Extract the effective ingredient not a duck soup in plant, generally require the physicochemical property according to material to be extracted and carry out lot of experiments
Just can determine that the most suitable extraction conditions, thus obtain higher extraction ratio;The selection especially becoming phase material is particularly important,
Generally target substance had appreciable impact in the distribution behavior of aqueous two-phase system.
Therefore it provides a kind of aqueous two-phase extraction process that can effectively extract total flavones from Semen Fagopyri Esculenti, become one urgently
Problem to be solved.
Summary of the invention
It is an object of the invention to provide the extracting method of a kind of buckwheat total flavone.
The invention provides the extracting method of a kind of buckwheat total flavone, comprise the steps: to take Semen Fagopyri Esculenti seed, pulverize, de-
Fat, uses propanol-ammonium sulfate double-aqueous phase system to extract, and stratification takes supernatant, to obtain final product.
Further, described extraction conditions is: propanol is 3:10~9:10 with the volume ratio of water, and ammonium sulfate concentrations is 0.27
~0.45g/ml, solid-liquid ratio is 1:10~1:40, and extraction time is 15~45min, and Extracting temperature is 30~60 DEG C.
Preferably, described extraction conditions is: propanol is 8:10 with the volume ratio of water, and ammonium sulfate concentrations is 0.39~0.42g/
Ml, solid-liquid ratio is 1:20~1:25, and extraction time is 25~30min, and Extracting temperature is 50~55 DEG C.
It is further preferred that described extraction conditions is: propanol is 8:10 with the volume ratio of water, and ammonium sulphate concentration is 0.39g/
Ml, solid-liquid ratio is 1:20, and extraction time is 30min, and Extracting temperature is 55 DEG C.
Further, using ultrasonic assistant propanol-ammonium sulfate double-aqueous phase system to extract, ultrasonic power is 250W.
Further, described Semen Fagopyri Esculenti is sweet buckwheat.
Further, described degreasing method is: take Fagopyrum esculentum Moench powder, adds petroleum ether, ultrasonic extraction, filters, collects filter
Slag, to obtain final product.
Further, adding solid-liquid ratio is the petroleum ether of 1:10, in 30 DEG C~60 DEG C of ultrasonic extraction 30min, ultrasonic merit
Rate is 250W.
The invention provides a kind of buckwheat total flavone prepared according to described extracting method.
The invention provides the extracting method of a kind of buckwheat total flavone, have the advantages that
1, the present invention uses aqueous two-phase extractive technique to extract the Flavonoid substances in Semen Fagopyri Esculenti, and determines the one-tenth of aqueous two-phase system
Phase material is propanol-ammonium sulfate, and compared to using polymer as becoming phase material in traditional double-aqueous phase system, the present invention has
Low cost, easy and simple to handle, be prone to the advantages such as subsequent treatment.
2, the invention provides aqueous two-phase and extract the actual conditions parameter of buckwheat flavonoids, the extraction higher than 0.3% can be obtained
Rate, high extraction can reach 0.57%.
3, the present invention uses ultrasonic assistant aqueous two-phase to extract further, utilizes the cavitation of ultrasound wave, machinery
Effect and heat effect etc. are accelerated the release of Flavonoid substances in plant cell, are spread and dissolves, thus improve extraction efficiency, shortening
Extraction time, save solvent, and avoid the high temperature destruction to flavone compound.
Obviously, according to the foregoing of the present invention, according to ordinary technical knowledge and the customary means of this area, without departing from
Under the present invention above-mentioned basic fundamental thought premise, it is also possible to make the amendment of other various ways, replace or change.
The detailed description of the invention of form by the following examples, remakes the most specifically the foregoing of the present invention
Bright.But this should not being interpreted as, the scope of the above-mentioned theme of the present invention is only limitted to Examples below.All based on foregoing of the present invention
The technology realized belongs to the scope of the present invention.
Accompanying drawing explanation
Fig. 1 is rutin standard curve figure;
Fig. 2 be alcohol water comparison extract yield affect figure;
Fig. 3 is that ammonium sulfate concentrations affects figure to extract yield;
Fig. 4 is that solid-liquid ratio affects figure to extract yield;
Fig. 5 is extraction time to affect figure to extract yield;
Fig. 6 is that Extracting temperature affects figure to extract yield.
Detailed description of the invention
The raw material, the equipment that use in the specific embodiment of the invention are known product, obtain by buying commercially available prod.
1, experimental raw
Sweet cherry roots seed (the raw grain place of production: Chaoyang;China Oil and Food Import and Export Corporation's rice industry (huge rock) company limited produces, and gold is full of board)
It is standby that Fagopyrum esculentum Moench powder is made in pretreatment: after being pulverized by sweet cherry roots, excessively 100 mesh sieves.
2, primary drug reagent
Rutin standard substance (Tianjin Sigma science and technology);Petroleum ether (boiling range: 60-90 DEG C);Propanol, aluminum chloride, potassium acetate, first
Alcohol, dehydrated alcohol, ammonium sulfate etc..
3, key instrument equipment
UV-1200 type ultraviolet spectrophotometer: Shanghai Mei Puda Instrument Ltd.;Pulverizer;SB-5200DTDN is ultrasonic
Washer (ultrasonic power 250W);JA1203 electronic analytical balance: Shanghai Yue Ping tech equipment company limited;SZ-93 type is automatic
Dual pure water distillator: Shanghai Yarong Biochemical Instrument Plant etc..
The extracting method of embodiment 1 buckwheat total flavone of the present invention
Weigh 2.00g Fagopyrum esculentum Moench powder and fill in conical flask in 100ml tool, add 20ml petroleum ether, in 60 DEG C of ultrasonic extraction
30min, ultrasonic power 250W, remove liposoluble constituent, filters, and filtering residue removes wherein remaining oil in room temperature volatile dry
Ether.According to 1:20 solid-liquid ratio (m/v), adding alcohol water ratio (v/v) is 8:10, and ammonium sulfate concentrations is the aqueous two-phase of 0.39g/ml preparation
Solution, arranges supersound extraction temperature 55 DEG C, supersound extraction 30min, ultrasonic power 250W.In separatory funnel after stratification,
Take supernatant, to obtain final product.Total flavones yield is 0.57%.
Beneficial effects of the present invention is proved below by way of experimental example.
The content assaying method of total flavones
Method according to NY/T1295-2007 " mensuration of general flavone content in Semen Fagopyri Esculenti and goods thereof " measures containing of total flavones
Amount.
First, prepare 0.1mol/L aluminum chloride, 0.1mol/L potassium acetate solution, methanol aqueous solution (7+3), then, claim
Take rutin standard substance 0.0573g in beaker, dissolve with methanol, move in 1000mL volumetric flask, by methanol constant volume to scale, i.e.
Obtain the rutin standard solution of 0.0500mg/mL.Rutin standard curve is drawn: draw respectively accurately 0.25 with pipet, 0.50,
1.0,2.0,3.0,4.0mL rutin standard solution be placed in 10mL volumetric flask, be separately added into 0.1mol/L aluminum chloride 2mL,
0.1mol/L potassium acetate solution 3mL, adds methanol solution and is settled to scale, rock uniformly, and ambient temperatare puts 30min.Make simultaneously
Blank.In standard curve, rutin content is respectively 0.00125mg/mL, 0.00250mg/mL, 0.00500mg/mL, 0.0100mg/
mL、0.0150mg/mL、0.0200mg/mL.Absorbance is measured at 420nm wavelength.Think the horizontal seat of rutin concentration of standard solution
Mark, absorbance are vertical coordinate, draw standard curve, and result is shown in Fig. 1.
Determination of total flavonoids: accurate measuring buckwheat total flavone liquid to be measured 1.0mL, in 10mL volumetric flask, adds 0.1mol/L
Aluminum chloride 2mL, 0.1mol/L potassium acetate solution 3mL, adds methanol solution and is settled to scale, shake up, and ambient temperatare is put
30min.Then at 420nm wavelength, measure absorbance, the absorbance that will obtain, substitute into the regression equation of rutin standard curve,
Calculate concentration and the total flavones quality of total flavones.Quality × 100% of total flavones quality/Semen Fagopyri Esculenti in total flavones yield=Semen Fagopyri Esculenti.
The impact of yield is extracted in experimental example 1 alcohol water comparison
Weigh 2.00g Fagopyrum esculentum Moench powder and fill in conical flask in 100ml tool, add 20ml petroleum ether, in 60 DEG C of ultrasonic extraction
30min, ultrasonic power is 250W, removes liposoluble constituent, filters, and filtering residue removes wherein remaining oil in room temperature volatile dry
Ether.According to 1:25 solid-liquid ratio (m/v), be separately added into alcohol water ratio (v/v) be 3:10,4:10,5:10,6:10,7:10,8:10,9:
10, concentration be 0.35g/ml ammonium sulfate preparation aqueous two-phase solution (propanol-ammonium sulfate double-aqueous phase system is according to list of references system
Standby: 1, Ma Hongfei, Han Qiuju, Li Wei. propanol-ammonium sulfate double-aqueous phase system and Application research [J]. aminoacid and living resources
.2011,33(4):67-69;2, high clouds, Wu Lisheng, Wang Wei. propanol-ammonium sulfate-water liquid-liquid system extract and separate platinum, palladium,
Rhodium and gold [J]. analytical chemistry .2001,29 (8): 901-903), supersound extraction temperature 60 C, supersound extraction 25min are set, surpass
Acoustical power 250W.In separatory funnel after stratification, take supernatant and be placed in 100mL volumetric flask, water-soluble with 30%v/v ethanol
Liquid constant volume, this solution is total flavones liquid to be measured, measures general flavone content, and result is shown in Fig. 2.
Experimental result: along with alcohol water than increase total flavones extract to take the lead in increase after reduce.Along with alcohol water is than increasing, third
Determining alcohol increases, and makes extractant strengthen the penetrating power of material, improves the dissolubility of total flavones, to alcohol water ratio (v/v) be
During 7:10, gained extraction ratio is the highest;But alcohol water is than continuing to increase, and can affect the two-phase partitioning of aqueous two-phase, and total flavones dissolubility drops
Low, extract yield and decline.
The impact on extracting yield of experimental example 2 ammonium sulfate concentrations
Weigh 2.00g Fagopyrum esculentum Moench powder and fill in conical flask in 100ml tool, add 20ml petroleum ether, in 60 DEG C of ultrasonic extraction
30min, ultrasonic power is 250W, removes liposoluble constituent, filters, and filtering residue removes wherein remaining oil in room temperature volatile dry
Ether.According to 1:25 solid-liquid ratio (m/v), being separately added into alcohol water ratio (v/v) is 6:10, ammonium sulfate concentrations is 0.27,0.30,0.33,
0.36,0.39,0.42, the aqueous two-phase solution of 0.45g/ml preparation, supersound extraction temperature 60 C is set, supersound extraction 25min, super
Acoustical power 250W.In separatory funnel after stratification, take supernatant and be placed in 100mL volumetric flask, water-soluble with 30%v/v ethanol
Liquid constant volume, this solution is total flavones liquid to be measured, measures general flavone content, and result is shown in Fig. 3.
Experimental result: along with the increase of ammonium sulfate concentrations, total flavones extract to take the lead in increase after reduce.Due to ammonium sulfate
Concentration increase, aqueous two-phase more tends towards stability, to ammonium sulfate concentrations be 0.4g/ml time aqueous two-phase the most stable, extract yield the highest;But
Be as ammonium sulfate concentrations continue increase siphon away large quantity of moisture so that on mutually be organic facies moisture reduce, some are water miscible
Flavone compound dissolubility reduces, and extracts yield and decreases.
The impact on extracting yield of experimental example 3 solid-liquid ratio
Weigh 2.00g Fagopyrum esculentum Moench powder and fill in conical flask in 100ml tool, add 20ml petroleum ether, in 60 DEG C of ultrasonic extraction
30min, ultrasonic power is 250W, removes liposoluble constituent, filters, and filtering residue removes wherein remaining oil in room temperature volatile dry
Ether.Respectively according to 1:10,1:15,1:20,1:25,1:30,1:35,1:40 solid-liquid ratio (m/v), adding alcohol water ratio (v/v) is 6:
10, concentration is the aqueous two-phase solution of the ammonium sulfate preparation of 0.35g/ml, in supersound extraction temperature 60 C, supersound extraction 25min, surpasses
Acoustical power 250W.In separatory funnel after stratification, take supernatant and be placed in 100mL volumetric flask, water-soluble with 30%v/v ethanol
Liquid constant volume, this solution is total flavones liquid to be measured, measures general flavone content, and result is shown in Fig. 4.
Experimental result: along with the increase of solid-liquid ratio, total flavones extracts and takes the lead in increasing, reaches maximum when arriving about 1:30,
Then begin to decline.Solid-liquid ratio is little, and the speed that flavone compound is dissolved in solvent is slow;Solid-liquid ratio is too big, and solvent load increases,
Relatively the consumption of ammonium sulfate increases, and affects stablizing of aqueous two-phase so that extracting flavonoids yield reduces.
The impact on extracting yield of experimental example 4 extraction time
Weigh 2.00g Fagopyrum esculentum Moench powder and fill in conical flask in 100ml tool, add 20ml petroleum ether, in 60 DEG C of ultrasonic extraction
30min, ultrasonic power is 250W, removes liposoluble constituent, filters, and filtering residue removes wherein remaining oil in room temperature volatile dry
Ether.According to 1:25 solid-liquid ratio (m/v), adding alcohol water ratio (v/v) is 6:10, and concentration is double water of the ammonium sulfate preparation of 0.35g/ml
Phase solution, under conditions of supersound extraction temperature 60 C, ultrasonic power 250W, respectively supersound extraction 15,20,25,30,35,40,
45min.In separatory funnel after stratification, take supernatant and be placed in 100mL volumetric flask, fixed with 30%v/v ethanol water
Holding, this solution is total flavones liquid to be measured, measures general flavone content, and result is shown in Fig. 5.
Experimental result: extraction time increases extracts yield increase, when extraction time is 30min, extracts yield and reaches maximum,
Increasing along with the time extracted afterwards, the oxidized decomposition of flavone compound can be made, beginning to decline so extracting yield.
The impact on extracting yield of experimental example 5 Extracting temperature
Weigh 2.00g Fagopyrum esculentum Moench powder and fill in conical flask in 100ml tool, add 20ml petroleum ether, in 60 DEG C of ultrasonic extraction
30min, ultrasonic power is 250W, removes liposoluble constituent, filters, and filtering residue removes wherein remaining oil in room temperature volatile dry
Ether.According to 1:25 solid-liquid ratio (m/v), adding alcohol water ratio (v/v) is 6:10, and concentration is double water of the ammonium sulfate preparation of 0.35g/ml
Phase solution, under conditions of supersound extraction temperature is respectively 30,35,40,45,50,55,60 DEG C, supersound extraction 25min, ultrasonic
Power 250W.In separatory funnel after stratification, take supernatant and be placed in 100mL volumetric flask, use 30%v/v ethanol water
Constant volume, this solution is total flavones liquid to be measured, obtains total flavones liquid to be measured, measures general flavone content, and result is shown in Fig. 6.
Experimental result: total flavones extracts yield and raises with temperature and increase, because flavone compound dissolves in propanol
Degree increases with increased temperature, and temperature increases, and flavone compound diffusion coefficient increases so that extracts yield and increases the most therewith;
But after temperature increases to 50 DEG C, extract yield and just begin to decline, be primarily due to flavonoid at relatively high temperatures
Thing can occur the reactions such as oxidation Decomposition to be destroyed.
Experimental example 6 uses the comparison of various extracting conditions total flavones extraction ratio
Being divided into 16 test group, often group weighs 2.00g Fagopyrum esculentum Moench powder in 100ml tool plug conical flask, adds 20ml oil
Ether, in 60 DEG C of ultrasonic extraction 30min, ultrasonic power is 250W, removes liposoluble constituent, filters, and filtering residue is dry in room temperature volatilization
The petroleum ether that dry removing is wherein remaining.Remaining each step conditional parameter is the most as shown in table 1.
Table 1 uses the comparison of various extracting conditions total flavones extraction ratio
Experimental result: use extraction conditions of the present invention that the extraction yield of buckwheat total flavone can be made to reach more than 0.3%;Its
In, the extraction yield of test group 11 and 12 buckwheat total flavone is more than 0.5%, and relatively other test group yield significantly improves, and shows
Volume ratio at propanol Yu water is 8:10, and ammonium sulfate concentrations is 0.39~0.42g/ml, and solid-liquid ratio is 1:20~1:25, during extraction
Between be 25~30min, Extracting temperature is under the extraction conditions of 50~55 DEG C, i.e. can reach higher extraction ratio;Above-mentioned all
In test group, the extraction yield of test group 11 buckwheat total flavone is the highest, can reach 0.57%, shows optimum extraction condition of the present invention
For: alcohol water than be 0.39g/ml for 8:10, ammonium sulfate concentrations, solid-liquid ratio be 1:20, extraction time be 30min, Extracting temperature be
55℃。
The checking test of experimental example 7 extraction process of the present invention
Take with a collection of Semen Fagopyri Esculenti, use to enter according to optimum extraction condition described in experimental example 6 (i.e. the extraction conditions of test group 11)
The extraction of row total flavones, is repeated 3 times, and total flavones extracts yield and is respectively 0.53%, 0.57%, 0.59%, average out to 0.56%,
RSD0.03%.
The above results shows that extraction process of the present invention repeatability, stability are high, is suitable to industrialized production.
Claims (9)
1. an extracting method for buckwheat total flavone, is characterized in that: comprise the steps: to take Semen Fagopyri Esculenti seed, pulverizes, defat, adopts
Extract with propanol-ammonium sulfate double-aqueous phase system, stratification, take supernatant, to obtain final product.
2. extracting method as claimed in claim 1, is characterized in that: described extraction conditions is: propanol is 3 with the volume ratio of water:
10~9:10, ammonium sulfate concentrations is 0.27~0.45g/ml, and solid-liquid ratio is 1:10~1:40, and extraction time is 15~45min, carries
Taking temperature is 30~60 DEG C.
3. extracting method as claimed in claim 2, is characterized in that: described extraction conditions is: propanol is 8 with the volume ratio of water:
10, ammonium sulfate concentrations is 0.39~0.42g/ml, and solid-liquid ratio is 1:20~1:25, and extraction time is 25~30min, Extracting temperature
It it is 50~55 DEG C.
4. extracting method as claimed in claim 3, is characterized in that: described extraction conditions is: propanol is 8 with the volume ratio of water:
10, ammonium sulphate concentration is 0.39g/ml, and solid-liquid ratio is 1:20, and extraction time is 30min, and Extracting temperature is 55 DEG C.
5. the extracting method as described in Claims 1 to 4 any one, is characterized in that: use ultrasonic assistant propanol-ammonium sulfate
Double-aqueous phase system extracts, and ultrasonic power is 250W.
6. extracting method as claimed in claim 1, is characterized in that: described Semen Fagopyri Esculenti is sweet buckwheat.
7. extracting method as claimed in claim 1, is characterized in that: described degreasing method is: take Fagopyrum esculentum Moench powder, adds oil
Ether, ultrasonic extraction, filters, collects filtering residue, to obtain final product.
8. extracting method as claimed in claim 7, is characterized in that: adding solid-liquid ratio is the petroleum ether of 1:10, in 30 DEG C~60
DEG C ultrasonic extraction 30min, ultrasonic power is 250W.
9. the buckwheat total flavone prepared according to extracting method described in claim 1~8 any one.
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CN107137466A (en) * | 2017-05-19 | 2017-09-08 | 内蒙古医科大学 | The preparation method of liver-protection health-care composition |
CN111150087A (en) * | 2019-12-31 | 2020-05-15 | 西昌市正中食品有限公司 | Embedding wall material and microcapsule tartary buckwheat extract and preparation process thereof |
CN111150087B (en) * | 2019-12-31 | 2022-03-18 | 西昌市正中食品有限公司 | Microcapsule containing tartary buckwheat extract and preparation process thereof |
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