CN106916065A - The method that high-purity chlorogenic acid is prepared from radix bardanae - Google Patents
The method that high-purity chlorogenic acid is prepared from radix bardanae Download PDFInfo
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- C07C67/00—Preparation of carboxylic acid esters
- C07C67/28—Preparation of carboxylic acid esters by modifying the hydroxylic moiety of the ester, such modification not being an introduction of an ester group
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- C07—ORGANIC CHEMISTRY
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- C07C67/48—Separation; Purification; Stabilisation; Use of additives
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Abstract
A kind of method that chlorogenic acid is isolated and purified from radix bardanae, it is characterized in that it comprises the following steps：（1）The preparation of radix bardanae extract solution；Dry radix bardanae is taken, is crushed, use alkaline soak 30min, extracted using ultrasound or microwave, extract solution is adjusted between pH to 11 12；（2）Purification enrichment；Will（1）Gained extract solution loading anion-exchange column, column packing in anion-exchange column is weak anion resin, the impurity such as carbohydrate are removed using distillation water elution successively, eluted with alkali lye again, eluent adjusts pH to 25 with sour water, isometric ethyl acetate is extracted three times, combined ethyl acetate phase, and be concentrated under reduced pressure dry Burdok root P.E；（3）High-speed countercurrent chromatography is used to isolate and purify the solubility that the chlorogenic acid present invention greatly improves chlorogenic acid with buck as Extraction solvent, without organic solvent extracting and enriching, separation process need not be using immobilization carrier and without dead absorption, process is simple, rapidly and efficiently, reproducible, gained target compound purity is high.
Description
Technical field
Method the present invention relates to being extracted from food, being enriched with and separate its active ingredient, it is more particularly to a kind of from burdock
The extracting method of high-purity chlorogenic acid is prepared in root.
Background technology
Burdock（ArctiumlappaL.）It is the biennial herbaceous plant of composite family, medicine-food two-purpose, from incoming Japan of China and quilt
Quality vegetables are modified to, a large amount of plantations in the area such as Cangshan of Feng County, Pei County and Shandong Province in China Jiangsu.Radix bardanae is burdock
Main Leaf-feeding insects, with splendid health value,《Compendium of Materia Medica》Record its can " logical the twelve regular channels, except the five internal organs bad odor ",
" long term usage is made light of one's life by commiting suicide resistance to old ".Medical research proves, the effects such as radix bardanae has strengthening the spleen and stomach, clearing heat and detoxicating and protection blood vessel, to hypertension,
The diseases such as diabetes, artery sclerosis have therapeutic action.
Radix bardanae rich in the composition such as synanthrin, organic acid and sterols, its Content of Chlorogenic Acid be organic acid compound it is main into
Point, carry out quality evaluation frequently as index composition important in radix bardanae at present.Chlorogenic acid has extensive bioactivity, tool
There are antibacterial, antiviral, antitumor, lowering blood pressure and blood fat, remove the effect such as free radical and stimulating central nervous system system, modern section
Learn and the multiple fields such as food, health care, medicine and daily-use chemical industry have been deep into the research of chlorogenic acid bioactivity.
Literature search to prior art finds：
Chinese Patent Application No. 200510000484.0 discloses the technique of the chlorogenic acid extracting from burdock leaf, and the technique needs chlorine
Imitative degreasing, the sample sour water after degreasing needs to again pass by the extracting and enriching of ethyl acetate after extracting, and then passes twice through AB-8
Resin column such as is eluted, concentrated and is recrystallized at the operating procedure that method obtains chlorogenic acid, early stage enrichment and the enrichment of the step of later stage two
Cumbersome, energy consumption is big and consumes a large amount of organic chemical reagents.
Chinese Patent Application No. CN03111946.8 also discloses that a kind of technique of the chlorogenic acid extracting from burdock leaf, and what is used carries
Solvent is taken for acidic organic solvent, the burdock Leave extract for obtaining crosses polyamide column, eluted with water and methyl alcohol, eluent is through dense
Contract, refine, obtain chlorogenic acid pure product.
Chinese Patent Application No. has been 201310726468.4 disclosure of the invention, and a kind of breaking-wall cell low temperature extraction bark of eucommia is green
The method of ortho acid, the cuticula destroyed on blade with the alkaline soak that pH is 9, then with the pure water dynamic that 7 times of pH is 2 often
Temperature extracts 2 h, is repeated to extract with sour water, then prepares bark of eucommia chlorogenic acid through techniques such as film process, counter-infiltration and resin wash-outs.
Chinese Patent Application No. has been 201010542548.0 disclosure of the invention, and one kind isolates and purifies green original from folium cortex eucommiae
The method of acid, high speed adverse current chromatogram point is carried out using the microwave thing of folium cortex eucommiae by ethyl acetate/n-butanol/water as dicyandiamide solution
From purifying 200min, high-purity chlorogenic acid is obtained.
A kind of utilization high speed adverse current chromatogram of the disclosure of the invention of Chinese Patent Application No. 201510316229.0 is isolated and purified
The method of monomeric compound in Chinese mahonia leaf, by the use of n-hexane-ethyl acetate-methanol-water as dicyandiamide solution, by a high speed
Counter current chromatography purifies 100min, can obtain high-purity chlorogenic acid from the ethyl acetate phase of Gaulis Mahoniae ethanol extract
18.32mg。
Lou is in auspicious academic dissertation《Burdock functional components and its anti-oxidant, antibacterial activity research》（Southern Yangtze University, 2010：
34-36）It is Extraction solvent to use 70% ethanol solution, is distinguished by petroleum ether, ethyl acetate, n-butanol, water after extract concentration
Four parts are obtained after extraction, wherein ethyl acetate component passes through high speed adverse current chromatogram n-butanol-second again after being pre-processed through column chromatography
Acid-water（The ︰ 5 of 4 ︰ 1）System is prepared for the chlorogenic acid that purity is respectively 98.3%.
Chen Dejing etc. exists《Food Science》（2008,29 (11):298~299）" high-speed countercurrent chromatography isolates and purifies green original
Find to utilize n-butanol-glacial acetic acid-water in acid research "（The ︰ 5, V/V of 4 ︰ 1）System, being prepared through 6h can be from Honegsukle flower P.E
Obtain the mg of chlorogenic acid 40 of purity 98.1%.
Rosy clouds etc. are opened to exist《Pharmaceutical Analysis magazine》（2010,30 (1)：106~109）" high-speed countercurrent chromatography isolates and purifies gold and silver
Find to utilize with n-butanol-acetic acid-water in chlorogenic acid in spending "（4∶1∶5）It is dicyandiamide solution, can obtain purity through 4h preparations is
98.2% chlorogenic acid 11.2mg.
Shibo's sun is waited《Food Science》（2013,34 (13):87~90）" the height of purple sweetpotato Content of Chlorogenic Acid and isochlorogenic acid
Fast adverse current chromatogram is separated " find to utilize acetate-methanol-water（The ︰ 5 of 3 ︰ 1）For dicyandiamide solution through 4h preparative separations compound from
200mg purple sweetpotato extracts obtain the chlorogenic acid 28mg that purity is 98.5%.
The solvent of above method extraction process is the neutrality such as water, sour water, water-alcohol, acidity alcohol solution or acidic extraction is molten
Agent, gained extract solution Content of Chlorogenic Acid keeps molecular state substantially；It is extract obtained by ethyl acetate extracting and enriching or soda acid
Treatment carries out preliminary purification, and pre-treatment operating procedure is relatively complicated.It is green that the breaking-wall cell low temperature that existing patent is related to extracts the bark of eucommia
Although having carried out pre-treatment in the method for ortho acid with buck, target is to destroy the cuticula of blade, and the later stage is entered using sour water
Row is extracted, and is still fallen within acid and is carried category.Additionally, high-speed counter-current separation process involved in early-stage Study has a preparation amount low, system
For the problem that the time is long, the most 40mg of preparation amount, required time is substantially in more than 100min.
The content of the invention
The present invention provides a kind of method that separating chlorogenic acid is quickly and efficiently extracted from radix bardanae, and the method passes through buck
Extract, be enriched with, high speed adverse current chromatogram is isolated and purified, so as to obtain the chlorogenic acid of high-purity（Purity>90% ）.The program overcomes
The problems such as preparation amount of previous purification method is low, preparation time system solvent species long and required is more, substantially increases production
Efficiency, reduce production cost.
In order to realize foregoing invention purpose, the technical solution adopted by the present invention is：
A kind of method that chlorogenic acid is isolated and purified from radix bardanae, its step is as follows：
（1）The preparation of radix bardanae extract solution；
Dry radix bardanae is taken, is crushed, with the min of alkaline soak 30, extracted using ultrasound or microwave, extract solution adjusts pH
To between 11-12；
（2）Purification enrichment；
Will（1）Gained extract solution loading anion-exchange column, the column packing in anion-exchange column is weak anion resin,
The impurity such as carbohydrate are removed using distillation water elution successively, then is eluted with alkali lye, eluent adjusts pH to 2-5, isometric second with sour water
Acetoacetic ester is extracted three times, combined ethyl acetate phase, and be concentrated under reduced pressure dry Burdok root P.E.
（3）Chlorogenic acid is isolated and purified using high-speed countercurrent chromatography：By ethyl acetate, first alcohol and water by volume 4 ~ 5：
0.1~1：4 ~ 5 are made into mixed solution is placed in static 30min in separatory funnel, takes upper strata as fixing phase, and lower floor is used as mobile phase；
Fixing phase is first pumped into, phase to be fixed is full of high speed adverse current chromatogram post, opens speed regulator, pumps into mobile phase, treats mobile phase stream
Go out and UV-detector reading relatively stablize when, take（2）Middle gained Burdok root P.E is dissolved in the fixing phase and mobile phase of equivalent simultaneously
Sample introduction, observes chromatogram, and cut is collected when target peak occurs, and spin concentration removal organic solvent, freeze-drying obtains high-purity
Chlorogenic acid.
Specific feature of the invention also has, step（1）Described in alkali in buck be potassium hydroxide, NaOH, carbonic acid
The basic sample solutions such as sodium, potassium carbonate, sodium methoxide and caustic alcohol, concentration is 0.1mol/L-1mol/L.
Step（2）In, weak anion resin is diethylamine anion exchange resin, macroreticular weakly base polystyrene the moon
Ion exchange resin or weak-base anion-exchange resin D301.
Step（2）In, the sour water for adjusting eluent pH value is hydrochloric acid, sulfuric acid, phosphoric acid, the formic acid of 1mol/L-5mol/L.
Step（3）In, high-speed counter-current chromatograph is same field TBE-3000C, flow rate of mobile phase 5-12mL/min.
The beneficial effects of the invention are as follows：With it is existing with the chlorogenic acid extracting technology of neutral and acidic extraction agent compared with, this hair
It is bright initiative by the use of buck as Extraction solvent so that chlorogenic acid reacts forming salt with alkali, can greatly improve chlorogenic acid and exist
Solubility in extractant, chlorogenic acid solubility can improve more than 50% compared with neutral and acidic extraction agent in prior document；
Green original in the enrichment means such as ethyl acetate extraction after anion exchange resin and tune pH can greatly improve sample to be prepared
The purity of acid；The high speed adverse current chromatogram separation method flow velocity set up is fast, and preparation time is short, and preparation efficiency is the 5-10 in document
Times, with prominent effect.
Brief description of the drawings
Fig. 1 is high speed adverse current chromatogram figure.
Specific embodiment
With reference to specific embodiment, the present invention is described further.
Embodiment 1：
A kind of method that high-purity chlorogenic acid is prepared from radix bardanae, comprises the following steps：
（1）The preparation of radix bardanae extract solution
The dry medicine materical crude slice of radix bardanae was crushed into 40 mesh sieves, 100g samples were taken, was entered with the NaOH of 0.1mol/L by the ︰ 10 of solid-liquid ratio 1
Row ultrasonic extraction three times, the time is respectively 1.5h, 1h and 0.5h, merges extract solution；PH to 11- is adjusted with the hydrochloric acid of 0.1mol/L
12。
（2）Purification enrichment
Take radix bardanae extract solution to be loaded to diethylamine anion exchange resin chromatographic column, loading speed is 3BV/h, first uses distilled water
Wash-out 2h is carried out with 5BV/h, then 1h is eluted with 3BV/h with the sodium hydroxide solution of 0.1mol/L, 500mL connects as a unit
Receive, merge eluent；PH to 4-5, isometric ethyl acetate is adjusted to extract three times with the hydrochloric acid of 1mol/L, combined ethyl acetate phase is simultaneously
Dry Burdok root P.E 3.6g concentrated under reduced pressure.
（3）Chlorogenic acid is isolated and purified using high-speed countercurrent chromatography
By ethyl acetate, first alcohol and water according to the ︰ 5 of 5 ︰ 1 volume ratio, i.e. ethyl acetate 1000mL, methyl alcohol 200mL and water 1000mL
In separatory funnel, concussion is uniform, and static 30min is standby.Upper strata is taken as fixing phase, it is lower as mobile phase.Take（2）In
Obtained Burdok root P.E 450mg, is dissolved in the dissolution system of the upper and lower phase composition of 15mL equivalent, ultrasonic dissolution, by same respectively
Field TBE-3000C high speed adverse current chromatograms are separated.Constant temperature circulator temperature is set as 25 DEG C, opens UV-detector, with
40mL/min pumps into fixing phase, and phase to be fixed turns speed to 800rpm full of main frame after pipeline, is opened, with 10mL/min flow pumps
Enter mobile phase.After thering is mobile phase to flow out and the limit is steady, by sample introduction valve injection and adverse current chromatogram figure is recorded, collect cut
（10mL/pipe）, cut is detected with high performance liquid chromatograph, spin concentration removal organic solvent, freeze-drying obtains the green original of high-purity
Acid.
（4）The purity detecting of target compound
HPLC testing conditions are as follows：Gemini NX-C18（250 × 4.6 mm, 5 μm）, 25 DEG C of column temperature；Mobile phase：Methyl alcohol-
0.1% formic acid water（27：73）；Flow velocity 1mL/min；The μ L of sample size 10；Detection wavelength 330nm.
The collection of target compound：Detected by the cut collected, find target compound（Chlorogenic acid）In height
Flowed out between 23 ~ 38min of fast adverse current chromatogram.
The purity testing of target compound, collects the cut between 26 ~ 35min of high speed adverse current chromatogram, removes organic molten
Agent, obtains sample 108.3mg after freezing, it is 93.1% that high performance liquid chromatography area normalization method measures purity.
Embodiment 2：
The present embodiment part same as Example 1 is repeated no more, and difference is：It is green that one kind prepares high-purity from radix bardanae
The method of ortho acid, step（3）Carry out isolating and purifying chlorogenic acid using same field TBE-3000C high speed adverse current chromatograms, by ethyl acetate,
First alcohol and water according to the ︰ 4 of 4 ︰ 0.1 volume ratio, i.e. ethyl acetate 1000mL, methyl alcohol 200mL and water 1000mL fall separatory funnel
In, concussion is uniform, and static 30min is standby.Upper strata is taken as fixing phase, it is lower as mobile phase.
The collection of target compound：Detected by the cut collected, find target compound（Chlorogenic acid）In height
Flowed out between 25 ~ 43min of fast adverse current chromatogram.
The purity testing of target compound, collects the cut between 28 ~ 37min of high speed adverse current chromatogram, except organic solvent
Sample 80.1mg is obtained after lyophilized, it is 94.2% that high performance liquid chromatography area normalization method measures purity.
Embodiment 3：
The present embodiment part same as Example 1 is repeated no more, and difference is：
（1）The preparation of radix bardanae crude extract：
The exocuticle discarded after radix bardanae is processed, drying and crushing crosses 40 mesh sieves, 50g is sampled, with the 1mol/L hydroxides of 500mL
The h of potassium solution ultrasonic extraction 1；Repeat to extract once, merge extract solution, pH to 3-4, isometric acetic acid second are adjusted with 1mol/L hydrochloric acid
Ester is extracted three times, and be concentrated under reduced pressure dry Burdok root P.E 2.1g；
（2）High speed adverse current chromatogram is separated：
By ethyl acetate, first alcohol and water according to the ratio of the ︰ 4 of 5 ︰ 0.5, take（1）In obtained radix bardanae ethyl acetate extract
240mg, is dissolved in the dissolution system of the upper and lower phase composition of 5mL equivalent, ultrasonic dissolution respectively.By same field TBE-3000C high-speed counter-currents
Chromatogram is separated, and preparation process and method for detecting purity are evaporated off obtaining sample after organic solvent is lyophilized with embodiment 1
59.4mg, it is 93.6% that high performance liquid chromatography area normalization method measures purity.
Embodiment 4：
A kind of method that high-purity chlorogenic acid is prepared from radix bardanae, it is mainly comprised the following steps：The radix bardanae 100g of drying and crushing is taken,
The 2000 mL potassium hydroxide immersion 30min of 0.1mol/L, using ultrasonic extraction 60min, extract solution adjusts pH to 12；By extract solution
Upper diethylamine anion exchange resin exchange column after filtering, removes the impurity such as carbohydrate using 1500mL distillation water elutions successively, then
Eluted with the 0.1mol/L potassium hydroxide of 1500 mL, eluent adjusts pH to 3 with hydrochloric acid；Merge the isometric acetic acid second of eluent
Ester is extracted three times, combined ethyl acetate phase, the Burdok root P.E being concentrated to dryness；By ethyl acetate, first alcohol and water by volume
The ︰ 4 of 5 ︰ 1 are made into mixed solution and are placed in static 30min in separatory funnel, and upper strata uses together as fixing phase, lower floor as mobile phase
Field TBE-3000C high-speed counter-current chromatographs are separated；Observation adverse current chromatogram figure, collects the mL/ of cut 10 when target peak occurs
Bottle, liquid phase monitoring purity, the sample by purity more than 90% merges, and spin concentration removal organic solvent, freeze-drying obtains high-purity
Chlorogenic acid.
Dicyandiamide solution of the selection with suitable allocation coefficient, it is considered that distribution coefficient K values separating effect between 0.5 ~ 2
Preferably.From preparation time, on the premise of purity requirement is met, the time is shorter, and preparation efficiency is higher, and the unit interval produces
Value is higher.
K values are determined：Separation system is reached into balance in advance, mutually each 1mL up and down is taken respectively and is placed in during 5mL connects lid centrifuge tube,
Add the refined thing of radix bardanae（About 1-3mg）, shaking dissolves it.After standing, upper and lower Xiang Gequ 0.5mL are respectively placed in 2mL and even cover
In centrifuge tube, nitrogen drying, 1mL methyl alcohol dissolving carries out HPLC detections, and the peak area of phase is designated as A respectively up and down1And A2, K=A1/
A2, the results are shown in Table 1.
Separative efficiency is investigated：Pumped into lower according to different flow velocitys, lower chlorogenic acid different in flow rate is investigated respectively
Separating effect, the results are shown in Table 2.
As can be known from Table 1, in petroleum ether-ethyl acetate-methanol-water solution, the distribution coefficient for measuring is less than normal, it is seen that solid
The solubility determined in phase is too small, when being separated using same field TBE-3000C high speed adverse current chromatograms, target compound wash-out speed
Degree is too fast, it is difficult to efficiently separated, in actual high-speed counter-current separation test, it was demonstrated that the dissolubility of sample it is insufficient and
Target compound is not isolated and purified well, therefore unsuitable as separate dissolution system.As can be known from Table 2, flow velocity
For 10 mL/min can also efficiently separate target compound, the high speed adverse current chromatogram figure of preparation is shown in Fig. 1, collects component to be measured
Through liquid-phase chromatographic analysis, purity is more than 90%.
It is a discovery of the invention that in the separation system of acetate-methanol-water and ethyl acetate, alcohol and water, what is measured divides
Distribution coefficient is more suitable, therefore selects the system as the solvent system of separating chlorogenic acid.By testing, acetate-methanol-
Water（4~5：0~1：4~5）The system of other ratios is compared, is prepared under flow velocity in the height of 10 mL/min, purity can be isolated and purified
Target compound higher.
The appearance time for preparing target component different in flow rate of table 2
Comparative example one：
According to Chinese Patent Application No. CN03111946.8, by the radix bardanae 100g of drying and crushing, 2000mL ethanol is used（pH2）Point
2 are inferior to 78 DEG C of difference refluxing extraction 2h and 1h, every time merging filtrate, radix bardanae crude extract concentrated under reduced pressure after extracting.By burdock
Polyamide column on root crude extract, is eluted with water and methyl alcohol, and eluent is concentrated refined, obtains high-purity chlorogenic acid.
Comparative example two：
According to Chinese Patent Application No. CN200510000484.0,1300mL chloroforms are added to return the radix bardanae 100g of drying and crushing
Stream extracts 80 min, and filtering discards chloroform layer；The sulfuric acid water of 1200 mLpH 3.5 will be added by the sample after chloroform recovery
Solution refluxing extraction 2h, is concentrated into the crude extract of 100 mL；By crude extract 4 extractions of 2000mL ethyl acetate point, second is discarded
Acetoacetic ester part；Surplus solution crosses AB-8 macroreticular resins, respectively with the sour waters of pH 3.5 and 30% ethanol elution；Ethanol eluate
Loading AB-8 macroreticular resins again after concentration, respectively with the sour waters of pH 3.5 and 30% ethanol elution, concentration recrystallize high-purity is green
Ortho acid.
Comparative example three：
According to article, " high-speed countercurrent chromatography isolates and purifies chlorogenic acid and studies（《Food Science》, Chen Dejing etc., 2008）", by ox
Burdock root accurately weighs 50g after crushing, and uses 70% ethanol solution, the ︰ 10, pH4 of solid-liquid ratio 1, extraction 2 times in 70 DEG C of water-baths, every time
1.5h is extracted, merging filtrate is concentrated under reduced pressure into 100ml or so at 60 DEG C, is extracted respectively with petroleum ether, ethyl acetate successively
Three times, be concentrated under reduced pressure ethyl acetate layer at 60 DEG C.Select n-butanol-glacial acetic acid-water (︰ 5, V/V of 4 ︰ 1) system to separate, obtain
High-purity chlorogenic acid.
Shown through many experimental results, only technical solution of the present invention can obtain purity in 60min from radix bardanae
Chlorogenic acid more than 90%.
Claims (5)
1. a kind of method that chlorogenic acid is isolated and purified from radix bardanae, it is characterized in that it comprises the following steps：
（1）The preparation of radix bardanae extract solution；
Dry radix bardanae is taken, is crushed, use alkaline soak 30min, extracted using ultrasound or microwave, extract solution adjusts pH extremely
Between 11-12；
（2）Purification enrichment；
Will（1）Gained extract solution loading anion-exchange column, the column packing in anion-exchange column is weak anion resin,
The impurity such as carbohydrate are removed using distillation water elution successively, then is eluted with alkali lye, eluent adjusts pH to 2-5, isometric second with sour water
Acetoacetic ester is extracted three times, combined ethyl acetate phase, and be concentrated under reduced pressure dry Burdok root P.E；
（3）Chlorogenic acid is isolated and purified using high-speed countercurrent chromatography：By ethyl acetate, first alcohol and water by volume 4 ~ 5：0.1~1：
4 ~ 5 are made into mixed solution is placed in static 30min in separatory funnel, takes upper strata as fixing phase, and lower floor is used as mobile phase；First pump into
Fixing phase, phase to be fixed is full of high speed adverse current chromatogram post, opens speed regulator, pumps into mobile phase, treats mobile phase outflow and purple
When external detector reading is relatively stablized, take（2）Middle gained Burdok root P.E is dissolved in the fixing phase and mobile phase and sample introduction of equivalent, sees
Chromatogram is examined, cut is collected when target peak occurs, spin concentration removal organic solvent, freeze-drying obtains high-purity chlorogenic acid.
2. the method that chlorogenic acid is isolated and purified from radix bardanae according to claim 1, it is characterized in that step（1）Described in
Alkali in buck is the basic sample solutions such as potassium hydroxide, NaOH, sodium carbonate, potassium carbonate, sodium methoxide and caustic alcohol, and concentration is
0.1mol/L-1mol/L。
3. the method that chlorogenic acid is isolated and purified from radix bardanae according to claim 1, it is characterized in that step（2）Middle weak base
Property resin anion (R.A.) be diethylamine anion exchange resin, macroreticular weakly base styrene series anion exchange resin or alkalescent
Anion exchange resin D301.
4. the method that chlorogenic acid is isolated and purified from radix bardanae according to claim 1, it is characterized in that step（2）In, adjust
The sour water for saving eluent pH value is hydrochloric acid, sulfuric acid, phosphoric acid, the formic acid of 1mol/L-5mol/L.
5. the method that chlorogenic acid is isolated and purified from radix bardanae according to claim 1, it is characterized in that step（3）In, it is high
Fast counter-current chromatograph is same field TBE-3000C, flow rate of mobile phase 5-12mL/min.
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| CN107879937A (en) * | 2017-10-24 | 2018-04-06 | 四川九章生物科技有限公司 | A kind of new crystal of chlorogenic acid and preparation method thereof |
| CN109293509A (en) * | 2018-11-30 | 2019-02-01 | 浙江科技学院 | A method of preparing high-purity chlorogenic acid from bamboo extractive |
| CN109293509B (en) * | 2018-11-30 | 2021-08-03 | 浙江科技学院 | Method for preparing high-purity chlorogenic acid from bamboo leaf extract |
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