CN104496760B - A kind of preparation method of high purity tyrosol - Google Patents
A kind of preparation method of high purity tyrosol Download PDFInfo
- Publication number
- CN104496760B CN104496760B CN201510004953.XA CN201510004953A CN104496760B CN 104496760 B CN104496760 B CN 104496760B CN 201510004953 A CN201510004953 A CN 201510004953A CN 104496760 B CN104496760 B CN 104496760B
- Authority
- CN
- China
- Prior art keywords
- tyrosol
- concentrated solution
- extract concentrated
- extraction
- extract
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C37/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring
- C07C37/004—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring by obtaining phenols from plant material or from animal material
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C37/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring
- C07C37/68—Purification; separation; Use of additives, e.g. for stabilisation
- C07C37/70—Purification; separation; Use of additives, e.g. for stabilisation by physical treatment
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C37/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring
- C07C37/68—Purification; separation; Use of additives, e.g. for stabilisation
- C07C37/70—Purification; separation; Use of additives, e.g. for stabilisation by physical treatment
- C07C37/82—Purification; separation; Use of additives, e.g. for stabilisation by physical treatment by solid-liquid treatment; by chemisorption
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C37/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring
- C07C37/68—Purification; separation; Use of additives, e.g. for stabilisation
- C07C37/70—Purification; separation; Use of additives, e.g. for stabilisation by physical treatment
- C07C37/84—Purification; separation; Use of additives, e.g. for stabilisation by physical treatment by crystallisation
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/10—Process efficiency
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Botany (AREA)
- Crystallography & Structural Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention relates to a kind of preparation method of high purity tyrosol, comprise the following steps: extraction step, get Root of Kirilow Rhodiola medicinal material, add water carry out decocting, ultrasonic or microwave extraction, extracting solution carries out concentrating under reduced pressure, obtains the first extract concentrated solution; Purifying, adopts alcohol deposition method to carry out ethanol precipitation twice the first extract concentrated solution, filters acquisition second extract concentrated solution, upper polyamide column carries out water elution, obtains water elution liquid and concentrates, obtain the 3rd extract concentrated solution; Refine, macroporous resin column on the 3rd extract concentrated solution is carried out wash-out, collect elutriant, after concentrated, obtain the 4th extract concentrated solution, through the tyrosol that extractive crystallization acquisition purity is greater than 80%.It is higher that the inventive method obtains tyrosol purity; Operational process of craft is simple, is applicable to suitability for industrialized production; Dead absorption is less, and the tyrosol rate of transform is high, and material can reuse, energy-conserving and environment-protective; Only use water, ethanol and ethyl acetate, all belong to low toxicity or innocuous agents, little to operator's body harm.
Description
Technical field
The present invention relates to a kind of preparation method of high purity tyrosol, belong to tyrosol extraction and separation method field.
Background technology
Tyrosol is a kind of natural antioxidant, is one of main active ingredient in Root of Kirilow Rhodiola medicinal material, is a kind of derivative of phenylethyl alcohol.It has extremely strong to resist myocardial ischemia, reducing myocardial infarction area, reduce apoptosis of cardiac muscle number, reduce the incidence and severity of ischemia-reperfusion animal ventricular tachycardia and room quivering.The resistance of oxidation knocking out low density lipoprotein receptor mouse of feeding high cholesterol diet can also be improved simultaneously, and reduce upborne blood cholesterol and triglyceride levels.
Tyrosol can significantly improve superoxide-dismutase in coxsackie myocarditis mouse heart and activity of glutathione peroxidase; reduce serum lactic dehydrogenase (SLDH) activity and LPO; there is good provide protection to heart, be mainly used in clinically treating cardiovascular aspect disease.
The method of current extraction and isolation tyrosol mainly contains:
Rhodioside in Radix Rhodiolae, tyrosol, production technique (the Wang Huatian of tonka bean camphor, Northeast Agricultural University's Master's thesis) disclose tyrosol extracting method in a kind of Root of Kirilow Rhodiola, specifically: adopt the granulated slag after supercritical extraction, extract by 40% alcohol immersion, extracting liquid filtering concentrates, macroporous resin on concentrated solution, after ethanol-dichloromethane (3:7) wash-out, be concentrated into dry, a small amount of distilled water dissolves, filter, be concentrated into small volume, vacuum-freeze-dry, obtain the extract of rhodioside and tyrosol, wherein rhodioside and tyrosol content sum are about 80%, yield is 0.27%.Employ methylene dichloride in a large number in the method leaching process, its toxicity is comparatively large, cause product to there is potential safety hazard, and tyrosol yield is lower, wastes medicinal material greatly.
In olive polyphenol the composition such as tyrosol separation and purification research (yellow Beichuan. Northwest University's Master's thesis) disclose a kind of tyrosol extraction and separation method, specifically: have employed supersound extraction, extraction conditions is: temperature is 60, and the time is 20min, and solid-liquid ratio is 1:25.Extracting solution adopts silica gel column chromatography to be separated tyrosol, and the tyrosol purity separated is 85.81%.Being separated tyrosol purity by alumina adsorption method is 40%.It is adopt silica gel column chromatography to be separated that the method extracts tyrosol, because silicagel column extremely adsorbs comparatively serious, causes tyrosol to lose more, and can use a large amount of organic solvents in industrialization process, unfavorable to operator's health; Due to a large amount of with an organic solvent and the silicagel column that sepn process uses cannot reuse, cause cost in junket preparation process higher.
Preparation method's research of rhodioside and Root of Kirilow Rhodiola aglycon standard substance (Ding Chaowu. analytical test journal, 1997,16 (1): 38-41) preparation method of tyrosol standard substance is disclosed, it specifically have employed supersound extraction, extracting solution adds plumbic acetate solution except tannin, flavones etc., after use charcoal absorption depigmentation, be separated with preparative chromatography post finally by after membrane filtration, the tyrosol purity obtained is 97%.The method adopts preparative chromatography to be separated, and can only adapt to the separation in laboratory, cannot realize industrialization.
Rhodioside and tyrosol Study on extraction (Li Chen in Root of Kirilow Rhodiola, Chen Dongsheng, Chen Juan etc. Chinese medicinal materials, 2006,29 (11): 1239-1241) disclose tyrosol extraction process, it have employed 80% alcohol reflux, and solvent load is 6 times, extract 2 times, each 2 hours.Extraction Technology of Rhodiola Sacra research (Chinese Journal of Modern Applied Pharmacy, 2012 that He Meiyan, Zhang little Long. Tang Gute delivers, 29 (6): 495-497) tyrosol extracting method is disclosed, it is refluxing extraction when have employed 8 times amount 50% ethanol 80 DEG C, extracts 2 times, each 1 hour.Ma Tao, Xiong Chunmei, open the rhodioside in the orthogonal experiment extraction Rhodiola Crenulata of loyal grey hair table and tyrosol (Kunming Normal Higher Traiing School's journal, 2006,28 (4): 14-16) extracting method of tyrosol is disclosed, by 90% methyl alcohol or anhydrous methanol supersound extraction when in fact have employed 65 DEG C, extraction time is 15 minutes, and solid-liquid ratio is 1:20.The tyrosol content obtained is 0.015%.The tyrosol CO2 supercritical extraction method of patent name to be a kind of ethanol be entrainment agent has opened a kind of method extracting tyrosol, and in fact adopt the method for supercritical extraction to extract tyrosol, the extraction yield obtaining tyrosol is 1.637%.These methods are all preliminary extraction research, and it have employed refluxing extraction, supersound extraction and supercritical extraction respectively, and do not have further separation and purification tyrosol, therefore purity is all lower.
Patent name name is called that a kind of method of purifying tyrosol from coarse product of olive polyphenol discloses the another kind of method extracting tyrosol, specifically olive polyphenol meal aqueous acid is dissolved, sodium-chlor is added in above-mentioned olive polyphenol solution, the olive polyphenol solution that obtained salt is saturated; Be extracted with ethyl acetate the tyrosol in the saturated olive polyphenol solution of salt; Removing extraction into ethyl acetate mutually in moisture.Liquid phase method record ethyl acetate mutually in tyrosol content be 70%-90%.The tyrosol purity that the method obtains is lower, and 70%-90% described in it is for adopting areas of peak normalization method gained, and true purity should be 50%-70%.
Patent name is that tyrosol and tyrosol by product salidroside plant extraction and preparation thereof and purposes disclose the method that another extracts tyrosol, specifically by starting material alcohol reflux, dipping or seepage are extracted, collect extracting solution, be concentrated into without alcohol taste, add water refrigeration, go precipitation, aqueous solution acidic solution adjusts PH to 2-4, refrigeration, go precipitation, low-pole macroporous resin on clear liquor, first wash with water to effluent liquid in neutral, again with basic solution wash-out macroporous resin, and then be washed till neutrality with deionized water, again with acidic solution wash-out, partial neutral is eluted to again with deionized water, above elutriant all discards, again with 5%-30% ethanol elution, collect elutriant, elutriant concentrates, and dry rear organic solvent dissolution, removes insolubles, clear liquor concentrates, and adds precipitation organic solvent, fully stirs, refrigeration, goes precipitation, get clear liquor concentrated after, add chromatographic silica gel and carry out mixing sample, evaporate to dryness, upper silica gel column chromatography, adopt organic solvent or mixed organic solvents to carry out removal of impurities and wash-out or gradient elution, adopt thin layer to detect, collecting principal spot is the elutriant of tyrosol, reclaims elutriant, obtains solid matter and carry out recrystallization with recrystallization organic solvent, to obtain final product.The method complicated operation is loaded down with trivial details, and the separation and purification time is long, and silicagel column extremely adsorbs seriously, and tyrosol loss is more; Elutriant thin layer monitors tyrosol, collects the elutriant merging that principal spot is tyrosol, concentrated, in the inapplicable and large-scale production of this mode; Because silicagel column cannot reuse, cost is higher, and is unfavorable for environmental protection and energy-saving and emission-reduction.
Summary of the invention
In order to overcome above-mentioned defect of the prior art, technical problem to be solved by this invention is to provide a kind of preparation method of high purity tyrosol.
The technical scheme that the present invention solves the problems of the technologies described above is as follows: a kind of preparation method of high purity tyrosol, comprise the following steps: step (1): extract, get Root of Kirilow Rhodiola medicinal material, add water carry out decocting, ultrasonic or microwave extraction, extracting solution carries out concentrating under reduced pressure, obtains the first extract concentrated solution; Step (2): purifying, the first extract concentrated solution step (1) prepared adopts alcohol deposition method to carry out ethanol precipitation twice, filter acquisition second extract concentrated solution, again polyamide column on the second extract concentrated solution is carried out water elution, obtain water elution liquid to concentrate, obtain the 3rd extract concentrated solution; Step (3) is refined, on the 3rd extract concentrated solution step (2) obtained, macroporous resin column carries out wash-out, collect elutriant, after concentrated, obtain the 4th extract concentrated solution, the tyrosol that described 4th extract concentrated solution is greater than 80% through extractive crystallization acquisition purity.
On the basis of technique scheme, the present invention can also do following improvement.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, Root of Kirilow Rhodiola medicinal material described in step (1) is the fine powder according to Chinese materia medica classifying and dividing.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, described decoction, ultrasonic or microwave extraction extraction conditions are: extract 1 ~ 3 time, amount of water is 5 ~ 20 times, and extraction time is 2 ~ 60 minutes.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, described decoction extraction conditions is mix after extraction three times, and each amount of water is 10 times, and decocting time is 2 hours, and temperature is 100 DEG C; Described ultrasonic extraction conditions is: mix after extracting three times, each amount of water is 10 times, ultrasonic 45 minutes at every turn; Described microwave extraction condition is mix after extraction three times, and each extraction adds 10 times of water yields, each microwave 5 minutes, and wherein microwave power is low and middle-grade to high-grade.Be preferably after microwave extraction extracts three times and mix, each extraction adds 10 times of water yields, each microwave 5 minutes, and microwave power is 435w ~ 450w, and the tyrosol efficiency in microwave extraction Root of Kirilow Rhodiola is high, and method is simple to operation and extraction time is short, shortened process.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, the relative density of described first extract concentrated solution 50 DEG C of step (2) is 1.05-1.35; Described first extract concentrated solution adopts alcohol deposition method to carry out ethanol precipitation twice, and wherein alcohol precipitation ethanol content reaches 50%-70wt% for the first time; Second time alcohol precipitation ethanol content reaches 75%-85wt%; The alcohol precipitation time is 12-48 hour.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, described first time alcohol precipitation ethanol content reach 50%-70wt%, leave standstill refrigeration 24 hours; Second time alcohol precipitation ethanol content reaches 75%-85wt%, leaves standstill refrigeration 24 hours.First time, alcohol precipitation can make vegetable-protein, polysaccharide, starch, and tannin etc. form precipitation; Second time alcohol precipitation can remove the macromolecule impurities such as polysaccharide further, can improve the clarity of solution after the removing of these impurity.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, on described second extract concentrated solution of step (2), polyamide column carries out water elution is that employing 10 times of cylinder ponding carry out wash-out; Silon particle diameter in wherein said polyamide column is 14 ~ 120 orders; Part by weight 2:1 ~ the 1:4 of the Silon in Root of Kirilow Rhodiola medicinal material and polyamide column.Use the technical process of above-mentioned acid amides post effectively can remove tannin.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, the Silon particle diameter in described polyamide column is 60 ~ 80 orders.The particle diameter of polymeric amide is excessive, and the amount of the polymeric amide of use is more.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, described in step (3), macroporous resin column is SP825.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, the part by weight of described SP825 and Root of Kirilow Rhodiola medicinal material is more than 1:1.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, the part by weight 2:1 of described SP825 and Root of Kirilow Rhodiola medicinal material.The part by weight adopting SP825 and Root of Kirilow Rhodiola medicinal material is the loss that 2:1 can greatly reduce tyrosol, improves macroporous resin to the adsorptive capacity of tyrosol.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, on 3rd extract concentrated solution described in step (3), macroporous resin column is carried out the concrete steps of wash-out and is: macroporous resin dress post after pre-treatment, the 3rd extract concentrated solution is added according to the ratio 2:1 of macroporous resin and Root of Kirilow Rhodiola medicinal material, first wash with water to elutriant near colorless, again with being greater than 5% ethanol elution, collect the elutriant of ethanol elution; The relative density of wherein said 3rd extract concentrated solution 50 DEG C is 1.15-1.2.Preferred, after pre-treatment, macroporous resin dress post, adds the 3rd extract concentrated solution according to the ratio 2:1 of macroporous resin and Root of Kirilow Rhodiola medicinal material, first washes with water to elutriant near colorless, then use 10% ~ 25wt% ethanol elution; The consumption of the ethanol of described 10 ~ 25wt% is 21 ~ 25 times of column volumes.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, in step (3), the condition of macroporous resin wash-out is: the liquor strength in described 3rd extract concentrated solution is 0.5-2.0g/ml, blade diameter length ratio 1 ~ the 1:15 of macroporous resin, wherein said concentration is Root of Kirilow Rhodiola raw medicinal herbs/the 3rd extract concentrated solution volume ratio, during elution flow rate 1-8BV/h, the liquor strength in described 3rd extract concentrated solution is the quality of Root of Kirilow Rhodiola and the volume ratio of described 3rd extract concentrated solution.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, blade diameter length ratio is 1:10, and the liquor strength in the 3rd extract concentrated solution is 1.0g/ml, and elution speed is 4BV/h.Blade diameter length ratio has a significant impact tyrosol purity, adopts above-mentioned blade diameter length ratio and liquor strength and elution speed can greatly improve tyrosol purity.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, liquor strength in described 4th extract concentrated solution of step (3) is 50g-1g/ml, and the liquor strength in described 4th extract concentrated solution is that every 1ml the 4th extract concentrated solution contains 1g Root of Kirilow Rhodiola medicinal material containing 50g Root of Kirilow Rhodiola medicinal material to every 1ml the 4th extract concentrated solution.
The preparation method of the present invention's a kind of high purity tyrosol described above, further, described extractive crystallization is: add the ethyl acetate with the corresponding amount of described 4th extract concentrated solution to described 4th extract concentrated solution, extract 1-10 time, combining extraction liquid, decompression and solvent recovery, refrigerates to every ml containing tyrosol 20mg to 200mg, crystallization, obtains the tyrosol that purity is greater than 80%.The extractive crystallization step being separated this step of tyrosol process obtained through macroporous resin effectively can improve the purity of tyrosol.
The present invention, a preferred specifically embodiment, specifically comprises the following steps:
Step (1) is extracted, and get Root of Kirilow Rhodiola medicinal material, grind to form fine powder according to pharmacopoeial requirements, add water and carry out microwave extraction, extracting solution carries out concentrating under reduced pressure, obtains the first extract concentrated solution.The preferred power of microwave extraction is 100 ~ 800w power draw, and preferred fine powder particle diameter is 30 object powder, and the microwave extraction time is 5 ~ 20 minutes, is preferably 10 minutes, amount of water is 5 ~ 20 times, and be preferably 10 times, extraction time is 1 ~ 3 time, is preferably 3 times, and the relative density obtaining 50 DEG C is first extract concentrated solution of 1.05-1.35, step (2): purifying, alcohol deposition method is adopted to carry out first time alcohol precipitation step (1), namely in the first extract concentrated solution, the amount adding the ethanol of 95% makes ethanol content in the first extract concentrated solution reach 50%-70wt%, be preferably 70%, more than 12 hours are left standstill under conventional refrigeration conditions, preferably leave standstill 24 hours, then filter, filtrate recycling ethanol is also concentrated, concentrated solution carries out second time alcohol precipitation, add 95% ethanol again, its content is made to reach 75%-85%, preferred content is to 80%, carry out secondary alcohol precipitation, leave standstill refrigeration 24 hours, then filter, obtain the second extract concentrated solution,
After 60-80 order Silon is soaked 24 hours, load the glass column of 3 same inner diameter, not aobvious muddy to effluent liquid with water treatment, again polyamide column on the second extract concentrated solution is carried out water elution, obtain water elution liquid to concentrate, obtain the 3rd extract concentrated solution, the part by weight 2:1 ~ 1:4 of the Silon wherein in Root of Kirilow Rhodiola medicinal material and polyamide column, be preferably 2:1, step (3) is refined, liquor strength in the 3rd extract concentrated solution step (2) obtained is 0.5-2.0g/ml, preferred liquor strength is 1.0g/ml, blade diameter length ratio 1 ~ the 1:15 of macroporous resin, preferred blade diameter length ratio is 1:10, wherein said concentration is raw medicinal herbs/medicine liquid volume ratio, elution flow rate 1-8BV/h, preferred elution speed is that on 4BV/h, macroporous resin column carries out wash-out, collect elutriant, the 4th extract concentrated solution is obtained after concentrated, described 4th extract concentrated solution herb liquid concentration is 50g-1g/ml (namely every 1ml solution contains 1g raw medicinal herbs containing 50g raw medicinal herbs to every 1ml), add the ethyl acetate of corresponding amount, extract 1-10 time, combining extraction liquid, decompression and solvent recovery contains tyrosol 200mg to 20mg to every ml, let cool, crystallization, namely the tyrosol that purity is greater than 92% is obtained.
The Root of Kirilow Rhodiola that the inventive method uses can select Radix Rhodiolae, the one or several arbitrarily in Radix Rhodiolae, Radix Rhodiolae, Rhodiola kirilowii (Regel) Maxim., Rhodiola fastigita, Rhodiola rosea, Rhodiola algida Var. tangutica, large purplish red red-spotted stonecrop, Rhodiola quadrifida, rhodiola kirilowii Regel.
The present invention compared with prior art has following beneficial effect:
(1) obtain the inventive method acquisition tyrosol purity higher, can more than 95% be reached, can directly with the medicine for the treatment of cardiovascular aspect disease;
(2) operational process of craft is simple, is applicable to suitability for industrialized production;
(3) the inventive method is used owing to have employed polymeric amide and macroporous resin as parting material, and dead absorption is less, and the tyrosol rate of transform is high, and material can reuse, energy-conserving and environment-protective;
(4) this technique only uses water, ethanol and ethyl acetate, all belongs to low toxicity or innocuous agents, little to operator's body harm.
Embodiment
Be described principle of the present invention and feature with the following Examples, example, only for explaining the present invention, is not intended to limit scope of the present invention.
The raw material used in following specific embodiment is Radix Rhodiolae, rhodiola kirilowii Regel, Radix Rhodiolae, and wherein in Radix Rhodiolae, tyrosol content is 0.73%, and in rhodiola kirilowii Regel, tyrosol content is 0.65%, and in Radix Rhodiolae, tyrosol content is 0.47%.。Tyrosol content detection mainly uses HPLC method, and design parameter is as follows:
Chromatographic condition and system suitability test take octadecylsilane chemically bonded silica as weighting agent; With acetonitrile-water (10:90) for moving phase; Determined wavelength is 275nm.Theoretical plate number should lower than 2000 by tyrosol peak.
The preparation of reference substance solution
Get tyrosol reference substance appropriate, accurately weighed, add water and make the solution of every 1ml containing 0.4mg, to obtain final product.
The preparation of need testing solution
Get this product powder (crossing No. three sieves) about 0.5g, accurately weighed, put in tool plug Erlenmeyer flask, precision adds methyl alcohol 10ml, close plug, weighed weight, supersound process 30 minutes, lets cool, more accurately weighed, supplies the weight of less loss with methyl alcohol, shake up, filter, get subsequent filtrate, to obtain final product.
Assay method is accurate respectively draws reference substance solution and each 10um of need testing solution, injection liquid chromatography, measures, to obtain final product.
Embodiment 1
Root of Kirilow Rhodiola medicinal material granularity, extracting method, extraction conditions etc. in tyrosol extracting method in Radix Rhodiolae are investigated respectively
1, the investigation of Root of Kirilow Rhodiola medicinal material granularity
Get Radix Rhodiolae medicinal material 5 parts, every part of 100g, wherein 4 parts shatter into most meal, meal, middle powder, fine powder by " Chinese Pharmacopoeia " requirement respectively, add 10 times of water gaging microwave extraction 3 times, each 10 minutes, united extraction liquid, detect wherein tyrosol content, the results are shown in following table 1:
Table 1. Root of Kirilow Rhodiola shatters the impact of degree on extraction yield
As can be seen from Table 1, the extraction yield impact of medicinal material granularity on tyrosol is larger, fine powder extraction yield is the highest, if but particle diameter is meticulous, pulverize comparatively difficulty, and be difficult to after extracting filter, for industrial mass production brings the problem significantly improved of cost, therefore more preferably raw material particle size is that meal can better improve tyrosol extraction yield and easily be separated, and is conducive to the production of industrial-scale.
2, the investigation of extracting mode
Get Radix Rhodiolae medicinal material coarse powder 3 parts, every part of 100g, extract by decoction, ultrasonic, microwave three kinds of modes respectively, extract 3 times, amount of water is 10 times, and decocting time is 2 hours at every turn, ultrasonic each 45 minutes, each 10 minutes of microwave time.Tyrosol content in the extracting solution that comparison three kinds of modes obtain.The results are shown in following table 2:
Tyrosol content in the extracting solution that table 2. three kinds of modes obtain
| Extracting mode | Decoct | Ultrasonic | Microwave |
| Tyrosol content (mg) | 368.7 | 397.8 | 446.3 |
| Extraction yield (%) | 50.5 | 54.5 | 61.1 |
As can be seen from Table 2, three kinds of modes all have taller and bigger extraction yield to the tyrosol in Root of Kirilow Rhodiola, are more preferably microwave extraction, and the extracted amount of microwave extraction tyrosol is significantly higher than decoction and ultrasonic extracting method.
3, the investigation of extraction conditions
According to general Chinese medicinal materials extraction conditions, solubilization dosage, extraction time and extraction time are the principal element affecting effective component extraction rate in raw medicinal herbs, determination solvent of the present invention is water, to this Three factors of amount of water, extraction time and extraction time, select L9 (34) orthogonal experiment to be optimized, select three conventional levels in each factor and consider.Level of factor and experimental result are in table 3.
Factors Factor in table 3. extraction conditions and water-glass
Take 9 parts of medicinal materials, every part of 100g, grinds to form fine powder, adopts microwave extraction mode and extracts according to the extraction conditions of table 3, measures the content of tyrosol in extracting solution.
Table 4 extraction conditions experimental establishment and result
Table 5 tyrosol extraction conditions investigates experimental result analysis of variance table
F
0.05(2,2)＝19，F
0.01(2,2)＝99
According to the extreme difference shown in table 5 and variance analysis, the extracted amount that result shows tyrosol with add water consumption and extraction time has substantial connection.And Extraction solvent consumption at 5-20 doubly, extraction time, extraction time all had higher extraction yield to the tyrosol in Root of Kirilow Rhodiola at 1-3 time at 2-60 minute.The preferred optimum production process condition of the present invention is chosen to be A
3b
2c
3, namely amount of water is 10 times, extracts 3 times, each extraction 5 minutes.
4, three process certifications test
Take 9 parts of medicinal materials, every part of 100g, grinds to form fine powder, and adopt microwave extraction mode, extraction conditions is: amount of water is 10 times, extracts 3 times, and each extraction 5 minutes, measures tyrosol content in extracting solution, the results are shown in Table 6.
Table 6. three process certification test-results
Show from table 6 result, this process stabilizing is feasible.
Embodiment 2
To the investigation of purification condition in tyrosol extracting method in Radix Rhodiolae
Rhodiola extract is obtained more muddy through above-mentioned extracting method, adopting alcohol deposition method to carry out ethanol precipitation twice can make it clarify, alcohol precipitation can make vegetable-protein for the first time, polysaccharide, starch, tannins etc. form precipitation to remove, and second time alcohol precipitation can remove the macromolecule impurities such as polysaccharide further, can improve the clarity of solution after the removing of these impurity.
1, investigate the relative density of first time alcohol precipitation extracting solution, alcohol precipitation concentration, the alcohol precipitation time is the principal element affecting purifying process, to this Three factors, select L9 (34) orthogonal experiment to be optimized, select three conventional levels in each factor and consider, level of factor is in table 7.
Get 900g Root of Kirilow Rhodiola fine medicinal material powder, extract according to the preferred version of embodiment 1, extracting solution filters, and filtrate is equally divided into 9 parts, tests according to the empirical factor shown in table 7 and method.Experimental result is in table 8, table 9 and table 10.
Table 7. first time alcohol precipitation factor and water-glass
Table 8. test arrangement and experimental result
Table 9. tyrosol analysis of variance table
F
0.05(2,2)＝19，F
0.01(2,2)＝99
Table 10. precipitation capacity analysis of variance table
F
0.05(2,2)＝19，F
0.01(2,2)＝99
Consider in conjunction with the principal constituent rate of transform and precipitation capacity, extracting solution relative density has highly significant to affect on the tyrosol rate of transform, and alcohol precipitation concentration has a significant effect to the tyrosol rate of transform.Adopt the parameter of following scope, suitable alcohol precipitation product can be obtained.Extracting solution relative density 1.05-1.35 (50 DEG C of surveys), during alcohol precipitation, alcohol concn is 50-85%, and the alcohol precipitation time is 12-48 hour; Preferred condition is: 50 DEG C of relative densities are 1.15-1.20, and alcohol precipitation concentration is 70%, and the alcohol precipitation time is 24 hours.
Get 900g Root of Kirilow Rhodiola fine medicinal material powder, extract according to the preferred version of embodiment 1, extracting solution filters, is evaporated to 50 DEG C of relative densities is 1.15-1.20, and alcohol precipitation concentration is 70%, and the alcohol precipitation time is 24 hours, measures tyrosol content.
Three times confirmatory experiment shows, according to identical technique alcohol precipitation, the weight of three gained precipitations and the content of tyrosol are respectively:
Table 11. three proof test results
Above result shows, this purifying process is feasible.
2, second time alcohol precipitation
After above-mentioned first time, alcohol precipitation process carried out first time purifying, majority of plant albumen in extracting solution, polysaccharide, starch, tannins etc. form precipitation, then filter, ethanol is added again in filtrate, make its content reach 75% ~ 85%, carry out secondary alcohol precipitation, second time alcohol precipitation can remove the macromolecule impurities such as polysaccharide further.
3, in polyamide column, Silon granularity is investigated
Take 300g Root of Kirilow Rhodiola medicinal material and obtain extracting solution according to the selected technique of embodiment 1 and above-mentioned alcohol precipitation process, concentrating under reduced pressure, concentrated solution is divided into 3 parts, for subsequent use.Separately get 3 parts of Silons, every part of 100g, granularity is respectively 14-30 order, 30-60 order, 60-80 order, after soaking 24 hours, load the glass column of 3 same inner diameter, not aobvious muddy to effluent liquid with water treatment, by above-mentioned ready 3 parts of concentrated solution loadings, wash with water, collect 10 times of column volume elutriants, whether check in effluent liquid containing tannin.The results are shown in following table 12.
Table 12. Silon globule size and tannin detection result
As can be seen from Table 12, in equal quality situation, 60-80 order Silon is better than the Silon of other 2 particle diameters except tannin effect, therefore selects 60-80 object Silon.When particle diameter is thinner time, filter more difficult.
Granularity 60-120 order can reach the effect except complete tannin, and when particle diameter is larger time, Silon consumption is more.
4, maximal absorptive capacity is investigated
Take 500g Root of Kirilow Rhodiola medicinal material, according to above-mentioned Optimization Technology extraction purification, concentrated, concentrated solution is divided into 5 parts, and every part is equivalent to Root of Kirilow Rhodiola raw medicinal herbs 100g.Root of Kirilow Rhodiola raw medicinal herbs and Silon are crossed polyamide column in following ratio, is respectively 4:1,2:1,1:1,1:2,1:4, carry out wash-out with water, collect 10 times of column volume elutriants, check whether containing tannin in elutriant, table 13.
The part by weight of table 13. Root of Kirilow Rhodiola raw medicinal herbs and Silon and wash-out result
As can be seen from the above results, when raw medicinal herbs and Silon weight ratio are for being no more than 2:1, tannin is not checked out in effluent liquid, therefore all suitable in the scope that raw medicinal herbs and Silon mass ratio (w/w) are 2:1 to 1:4, preferred raw medicinal herbs and Silon weight ratio are 2:1, and the consumption effectively can saving polymeric amide can reach again the effect removing tannin preferably.
Embodiment 3
To the investigation that the macroporous resin kind used in process for refining in Root of Kirilow Rhodiola tyrosol extracting method, maximum applied sample amount, eluent-liquor strength, flow velocity and blade diameter length ratio are selected.
1, choose 3 parts of macroporous resins, model is respectively D101, D301, SP825.The specifications and characteristics of three kinds of resins is as following table 14.
The character of table 14 three kinds of macroporous resins and water content
Take 300g Root of Kirilow Rhodiola meal, adopt embodiment 1 and the selected processing parameter of embodiment 2 to carry out extracting, concentrate, mistake polyamide column, elutriant is concentrated into relative density 1.15-1.20 (50 DEG C), is divided into 3 parts, for subsequent use; Take the processed good macroporous resin in 3 parts of upper tables again, every part of 100g, dress post, by above-mentioned ready 3 parts of concentrated solution upper props.Wash 5 times of column volumes with water, collect elutriant, detect tyrosol content in elutriant; Carry out wash-out with 70% ethanol afterwards, collect 8 times of column volume elutriants, detect rhodioside and tyrosol content in elutriant.Result is as following table 15.
The absorption of table 15 tyrosol on 3 kinds of resins and desorption investigate tyrosol result
As can be seen from the result of table 15, the adsorption rate reconciliation adsorption rate of SP825 type macroporous resin to tyrosol is all larger, illustrate that SP825 type macroporous resin is comparatively large to the adsorptive capacity of tyrosol, and dead adsorption rate is less, is very suitable for the separation of tyrosol.
2, the selection of maximum applied sample amount
Get each 50g of SP825 type macroporous resin that 5 parts of pre-treatment are good, filling post respectively, is 1:0.5 according to resin and raw medicinal herbs weight ratio, 1:1,1:2,1:3, the ratio of 1:4, by dynamic for rhodiola extract loading, collects debris, water elution 8BV, merge debris and water elution liquid, after concentrated constant volume, measure the content of tyrosol, observe the relation between the number of dropouts of tyrosol and applied sample amount, experimental result is in table 16.
The investigation of table 16 tyrosol turnover rate and raw medicinal herbs applied sample amount relation
As can be known from Table 16, when resin and raw medicinal herbs weight ratio are 2:1, tyrosol chance is not lost, and therefore selects resin and raw medicinal herbs weight ratio to be greater than the ratio of 2:1, is preferably 2:1.
3, the investigation of eluant strength
Get the good SP825 type macroporous resin 200g of pre-treatment and fill post, add rhodiola extract according to the ratio 2:1 of macroporous resin and raw medicinal herbs, first wash with water to elutriant near colorless, collect elutriant, and record water consumption, then add 5% successively, 10%, 15%, 20%, 25% ethanol, 15 times of column volumes all collected by the ethanol of each concentration, collect successively, the ethanol eluate of same concentrations merges, and measures tyrosol content and medicinal extract amount after concentrated constant volume.Experimental result is in table 17.
The selection of table 17 macroporous resin eluant strength and eluting rate result
As can be known from the results of Table 17, most tyrosol can elute by 10% ethanol, the tyrosol overwhelming majority also can elute by 15% ethanolic soln, but impurity also elutes more, consider the reason of cost and impurity aspect, more excellent selection water and 5% ethanolic soln wash-out removing impurity, 10% ~ 15% ethanol, as the eluting solvent of macroporous resin column, collects this part solution.
4, the selection of eluting agent
Get the good macroporous resin 200g of pre-treatment and fill post, be that 2:1 adds rhodiola extract in the ratio of resin and raw medicinal herbs, first wash with water to elutriant colourless, use 5% ethanol elution colourless to effluent liquid again, then wash-out is carried out with the ethanol of 10%, every 5 times of column volumes combine, and measure rhodioside and tyrosol content after concentrated constant volume, measurement result is in table 18.
Table 18 eluting agent investigates result
| Column volume | 1-5 | 6-10 | 11-15 | 16-20 | 21-25 | 26 |
| Tyrosol eluting rate (%) | 8 | 25 | 40 | 15 | 6 | 2 |
As can be seen from Table 18, during 21-25 times of column volume, the tyrosol amount eluted seldom, does not then almost have during 26 times of column volumes, therefore, selects eluting solvent to be 25 times of column volumes.
5, the selection of liquor strength, flow velocity and blade diameter length ratio
Liquor strength, flow velocity, blade diameter length ratio are three important factors affecting purifying process, and to these three factors, we select L9 (34) orthogonal test to be optimized, and each factor is selected three levels and investigated, and factor is in table 19.
Get the macroporous resin of 9 parts of 200gSP825 models, post is filled respectively according to following blade diameter length ratio, get 100g Root of Kirilow Rhodiola medicinal material 9 parts respectively, extraction purification is carried out according to aforementioned fixed extraction and purification process, be condensed into following liquor strength respectively, dynamic upper prop, carries out wash-out according to fixed washing and dehydrating integrated machine concentration, and HPLC method measures tyrosol content and purity in elutriant.
Table 19. liquor strength, flow velocity, blade diameter length ratio Three factors and level
Table 20. test arrangement and experimental result
The variance analysis of table 21. Three factors test-results
F
0.05(2,2)＝19，F
0.01(2,2)＝99
As can be seen from Table 21, blade diameter length ratio has a significant impact tyrosol purity, and in conjunction with range analysis, when technique is A3B3C2, tyrosol purity is the highest.In conjunction with industrial production practical situation, select A
3b
2c
3, namely blade diameter length ratio is 1:10, and liquor strength is 1.0g/ml, and elution speed is 4BV/h.
6, three verification experimental verifications
Get 3 parts of SP825 type macroporous resins, resin and Root of Kirilow Rhodiola raw medicinal herbs weight ratio 2:1, (resin 200g, Root of Kirilow Rhodiola medicinal material 100g) be that 1:10 fills post according to blade diameter length ratio, purify according to the above-mentioned purifying process determined, dynamic upper prop, liquor strength is 1.0g/ml, elution speed is 4BV/h, merges elutriant, measures tyrosol content and purity.
Three process certifications experiment shows, extracts 3 times, the content of tyrosol and purity table 22 respectively according to identical condition.
The tyrosol content that table 22. three proof tests obtain and purity
Table 22 shows result and shows, this feasible process.
7, extractive crystallization
Above-mentioned resin elution liquid is concentrated into 50g-1g/ml (every 1ml solution contains 1g raw medicinal herbs containing 50g raw medicinal herbs to every 1ml), add the ethyl acetate of corresponding amount, extract 1-10 time, combining extraction liquid, decompression and solvent recovery contains tyrosol 200mg to 20mg to every ml, let cool, crystallization, obtain the tyrosol that purity is greater than 80%.
Embodiment 4
The preparation method of a kind of high purity tyrosol of the present invention, comprises the following steps (1) and extracts, get and take Root of Kirilow Rhodiola medicinal material 1kg, meal is ground to form according to pharmacopoeial requirements, add water and carry out microwave extraction, extracting solution carries out concentrating under reduced pressure, obtains the first extract concentrated solution.The preferred power of microwave extraction is 100 ~ 800w power draw, and the microwave extraction time is 5 minutes; Amount of water is 10 times, and extraction time is 3 times, and the relative density obtaining 50 DEG C is first extract concentrated solution of 1.15; Step (2): purifying, alcohol deposition method is adopted to carry out first time alcohol precipitation step (1), namely in the first extract concentrated solution, the amount adding the ethanol of 95% makes ethanol content in the first extract concentrated solution reach 70%, 24 hours are left standstill under conventional refrigeration conditions, then filter, filtrate recycling ethanol is also concentrated, concentrated solution carries out second time alcohol precipitation, then adds 95% ethanol, makes its content reach 85%, carry out secondary alcohol precipitation, leave standstill refrigeration 24 hours, then filter, obtain the second extract concentrated solution;
After 80 order Silons are soaked 24 hours, load the glass column of 3 same inner diameter, not aobvious muddy to effluent liquid with water treatment, again polyamide column on the second extract concentrated solution is carried out water elution, obtain water elution liquid to concentrate, obtain the 3rd extract concentrated solution, the part by weight of the Silon wherein in Root of Kirilow Rhodiola medicinal material and polyamide column is 2:1, step (3) is refined, liquor strength in the 3rd extract concentrated solution step (2) obtained is 1.0g/ml, the blade diameter length ratio blade diameter length ratio of macroporous resin is 1:10, wherein said concentration is raw medicinal herbs/medicine liquid volume ratio, elution flow rate 1 carries out wash-out for macroporous resin column on 4BV/h, collect elutriant, the 4th extract concentrated solution is obtained after concentrated, described 4th extract concentrated solution herb liquid concentration is 50g-1g/ml (namely every 1ml solution contains 1g raw medicinal herbs containing 50g raw medicinal herbs to every 1ml), add the ethyl acetate of corresponding amount, extract 5-6 time, combining extraction liquid, decompression and solvent recovery contains tyrosol 5mg to 10mg to every ml, let cool, crystallization, namely the tyrosol of purity 95% is obtained.
Embodiment 5
Take Root of Kirilow Rhodiola medicinal material coarse powder 1kg, microwave extraction 3 times, each 5 times of water gagings, extract 5 minutes, extracting solution merges, be evaporated to relative density 1.1 (50 DEG C of surveys), adding 95% ethanol makes alcohol content reach 60%, leave standstill after 24 hours under 2-8 DEG C of condition, filter, filtrate adds 95% ethanol makes alcohol content reach 85%, leave standstill after 24 hours under 2-8 DEG C of condition, filter, filtrate is concentrated into without alcohol taste, upper polyamide column, Silon granularity wherein in polyamide column is 60 orders, soak 24 little fashionable dress posts, raw medicinal herbs and Silon weight ratio are 2:1, water elution 10 times of column volumes, macroporous resin column on elutriant, raw medicinal herbs is 1:1 with the ratio of the consumption of resin column, liquor strength is 1.0g/ml, blade diameter length ratio is 1:6, after water elution is extremely colourless, with 5% ethanol elution, 12 times of column volumes, wash-out is carried out afterwards with 20% ethanol, flow velocity is 2BV/h, after elutriant reclaim under reduced pressure to 100ml, use 50ml extraction into ethyl acetate, extract 5 times, combining extraction liquid.Extraction liquid is evaporated to small volume, and refrigeration, crystallization, obtains tyrosol crystallization 2.9g, and purity is 90.7%.
Embodiment 6
Take powder 1kg in Root of Kirilow Rhodiola medicinal material, decoct extraction 3 times, each 10 times of water gagings, extract 2 hours, extracting solution merges, be evaporated to relative density 1.15 (50 DEG C of surveys), adding 95% ethanol makes alcohol content reach 60%, leave standstill after 24 hours under 2-8 DEG C of condition, filter, filtrate adds 95% ethanol makes alcohol content reach 85%, leave standstill after 12 hours under 2-8 DEG C of condition, filter, filtrate is concentrated into without alcohol taste, upper polyamide column, Silon granularity wherein in polyamide column is 60-80 order, soak 24 little fashionable dress posts, when raw medicinal herbs and Silon weight ratio are 1:1, water elution 12 times of column volumes, macroporous resin column on elutriant, resin is 3:1 with the ratio of the consumption of raw medicinal herbs, liquor strength is 1.5g/ml, blade diameter length ratio is 1:10, after water elution is extremely colourless, with 5% ethanol elution, 10 times of column volumes, wash-out is carried out afterwards with 10% ethanol, eluent flow rate is 3BV/h, after elutriant reclaim under reduced pressure to 200ml, use 100ml extraction into ethyl acetate, extract 5 times, combining extraction liquid.Extraction liquid is evaporated to small volume, and refrigeration, crystallization, obtains tyrosol crystallization 2.3g, and purity is 88.5%.
Embodiment 7
Take Root of Kirilow Rhodiola fine medicinal material powder 1kg, supersound extraction 3 times, each 10 times of water gagings, extract 30 minutes, extracting solution merges, be evaporated to relative density 1.10 (50 DEG C of surveys), adding 95% ethanol makes alcohol content reach 50%, leave standstill after 24 hours under 2-8 DEG C of condition, filter, filtrate adds 95% ethanol makes alcohol content reach 80%, leave standstill after 18 hours under 2-8 DEG C of condition, filter, filtrate is concentrated into without alcohol taste, upper polyamide column, Silon granularity wherein in polyamide column is 70 orders, soak 24 little fashionable dress posts, raw medicinal herbs and Silon weight ratio are 1:2, water elution 10 times of column volumes, macroporous resin column on elutriant, resin is 2:1 with the ratio of the consumption of Root of Kirilow Rhodiola raw medicinal herbs, liquor strength is 2.0g/ml, blade diameter length ratio is 1:15, after water elution is extremely colourless, with 5% ethanol elution, 8 times of column volumes, wash-out is carried out afterwards with 20% ethanol, flow velocity is 4BV/h, after elutriant reclaim under reduced pressure to 50ml, use 50ml extraction into ethyl acetate, extract 5 times, combining extraction liquid.Extraction liquid is evaporated to small volume, and refrigeration, crystallization, obtains tyrosol crystallization 2.8g, and purity is 92.5%.
Embodiment 8
Take Root of Kirilow Rhodiola medicinal material coarse powder 1kg, microwave extraction 2 times, each 10 times of water gagings, extract 10 minutes, extracting solution merges, be evaporated to relative density 1.15 (50 DEG C of surveys), adding 95% ethanol makes alcohol content reach 70%, leave standstill after 12 hours under 2-8 DEG C of condition, filter, filtrate adds 95% ethanol makes alcohol content reach 85%, leave standstill after 24 hours under 4-8 DEG C of condition, filter, filtrate is concentrated into without alcohol taste, upper polyamide column, Silon granularity wherein in polyamide column is 60 orders, soak 24 little fashionable dress posts, raw medicinal herbs and Silon weight ratio are 1:4, water elution 15 times of column volumes, macroporous resin column on elutriant, resin is 2:1 with the ratio of the consumption of raw medicinal herbs, liquor strength is 1.0g/ml, blade diameter length ratio is 1:10, after water elution is extremely colourless, with 5% ethanol elution, 10 times of column volumes, wash-out is carried out afterwards with 25% ethanol, flow velocity is 5BV/h, after elutriant reclaim under reduced pressure to 400ml, use 100ml extraction into ethyl acetate, extract 5 times, combining extraction liquid.Extraction liquid is evaporated to small volume, and refrigeration, crystallization, obtains tyrosol crystallization 3.2g, and purity is 94.3%.
Embodiment 9
Take powder 1kg in Root of Kirilow Rhodiola medicinal material, decoct extraction 1 time, 20 times of water gagings, extract 15 minutes, extracting solution merges, be evaporated to relative density 1.15-1.25 (50 DEG C of surveys), adding 95% ethanol makes alcohol content reach 70%, leave standstill after 12 hours under 2-8 DEG C of condition, filter, filtrate adds 95% ethanol makes alcohol content reach 85%, leave standstill after 48 hours under 4 DEG C of conditions, filter, filtrate is concentrated into without alcohol taste, upper polyamide column, Silon granularity wherein in polyamide column is 80 orders, soak 24 little fashionable dress posts, raw medicinal herbs and Silon weight ratio are 2:1, water elution 15 times of column volumes, macroporous resin column on elutriant, resin is 2:1 with the ratio of the consumption of raw medicinal herbs, liquor strength is 1.0g/ml, blade diameter length ratio is 1:10, after water elution is extremely colourless, with 5% ethanol elution, 8 times of column volumes, wash-out is carried out afterwards with 20% ethanol, flow velocity is 4BV/h, after elutriant reclaim under reduced pressure to 100ml, use 80ml extraction into ethyl acetate, extract 5 times, combining extraction liquid.Extraction liquid is evaporated to small volume, and refrigeration, crystallization, obtains tyrosol crystallization 2.7g, and purity is 92.8%.
Embodiment 10
Take Root of Kirilow Rhodiola medicinal material coarse powder 1kg, supersound extraction 2 times, 8 times of water gagings, extract 45 minutes, extracting solution merges, be evaporated to relative density 1.25-1.35 (50 DEG C of surveys), adding 95% ethanol makes alcohol content reach 70%, leave standstill after 24 hours under 2-8 DEG C of condition, filter, filtrate adds 95% ethanol makes alcohol content reach 85%, leave standstill after 12 hours under 8 DEG C of conditions, filter, filtrate is concentrated into without alcohol taste, upper polyamide column, Silon granularity wherein in polyamide column is 80 orders, soak 24 little fashionable dress posts, raw medicinal herbs and Silon weight ratio are 2:1, water elution 10 times of column volumes, macroporous resin column on elutriant, resin is 3:1 with the ratio of the consumption of raw medicinal herbs, liquor strength is 1.0g/ml, blade diameter length ratio is 1:8, after water elution is extremely colourless, with 5% ethanol elution, 10 times of column volumes, wash-out is carried out afterwards with 15% ethanol, flow velocity is 6BV/h, after elutriant reclaim under reduced pressure to 200ml, use 80ml extraction into ethyl acetate, extract 3 times, combining extraction liquid.Extraction liquid is evaporated to small volume, and refrigeration, crystallization, obtains tyrosol crystallization 2.9g, and purity is 90.3%.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (5)
1. a preparation method for tyrosol, is characterized in that, comprises the following steps:
Step (1): extract, get Root of Kirilow Rhodiola medicinal material, add water carry out decocting, ultrasonic or microwave extraction, extracting solution carries out concentrating under reduced pressure, obtains the first extract concentrated solution; The relative density of described first extract concentrated solution 50 DEG C is 1.05-1.35;
Described decoction extraction conditions is mix after extraction three times, and each amount of water is 10 times, and decocting time is 2 hours, and temperature is 100 DEG C; Described ultrasonic extraction conditions is: mix after extracting three times, each amount of water is 10 times, ultrasonic 45 minutes at every turn; Described microwave extraction condition is mix after extraction three times, and each extraction adds 10 times of water yields, each microwave 5 minutes;
Step (2): purifying, the first extract concentrated solution step (1) prepared adopts alcohol deposition method to carry out ethanol precipitation twice, filter acquisition second extract concentrated solution, again polyamide column on the second extract concentrated solution is carried out water elution, obtain water elution liquid to concentrate, obtain the 3rd extract concentrated solution;
Described first extract concentrated solution adopts alcohol deposition method to carry out ethanol precipitation twice, and wherein alcohol precipitation ethanol content reaches 50%-70% for the first time; Second time alcohol precipitation ethanol content reaches 75%-85%; The alcohol precipitation time is 12-48 hour;
On described second extract concentrated solution, polyamide column carries out water elution is that employing 10 times of cylinder ponding carry out wash-out; Silon particle diameter in wherein said polyamide column is 60 ~ 120 orders; Part by weight 2:1 ~ the 1:4 of the Silon in Root of Kirilow Rhodiola medicinal material and polyamide column;
Step (3) is refined, on the 3rd extract concentrated solution step (2) obtained, macroporous resin column carries out wash-out, collect elutriant, after concentrated, obtain the 4th extract concentrated solution, the tyrosol that described 4th extract concentrated solution is greater than 80% through extractive crystallization acquisition purity;
Described macroporous resin column is SP825; The part by weight 2:1 of described SP825 and Root of Kirilow Rhodiola medicinal material;
On described 3rd extract concentrated solution, macroporous resin column is carried out the concrete steps of wash-out and is: macroporous resin dress post after pre-treatment, the 3rd extract concentrated solution is added according to the ratio 2:1 of macroporous resin and Root of Kirilow Rhodiola medicinal material, first wash with water to elutriant near colorless, use 5% ethanol elution again, with 10 ~ 25% ethanol elutions; Collect the elutriant of 10 ~ 25% ethanol elutions; The relative density of wherein said 3rd extract concentrated solution 50 DEG C is 1.15-1.2, and the consumption of the ethanol of described 10 ~ 25% is 21 ~ 25 times of column volumes;
The condition of macroporous resin wash-out is: the liquor strength in described 3rd extract concentrated solution is 0.5-2.0g/ml, blade diameter length ratio 1:1 ~ 15 of macroporous resin, elution flow rate 1-8BV/h;
Liquor strength in described 4th extract concentrated solution is 50g-1g/ml.
2. the preparation method of a kind of tyrosol according to claim 1, is characterized in that, described first time alcohol precipitation ethanol content reach 50%-70%, leave standstill refrigeration 24 hours; Second time alcohol precipitation ethanol content reaches 75%-85%, leaves standstill refrigeration 24 hours.
3. the preparation method of a kind of tyrosol according to claim 1, it is characterized in that, the Silon particle diameter in described polyamide column is 60 ~ 80 orders.
4. the preparation method of a kind of tyrosol according to claim 1, it is characterized in that, described 3rd extract concentrated solution herb liquid concentration is 1.0g/ml, and the blade diameter length ratio of macroporous resin is 1:10, and elution speed is 4BV/h.
5. the preparation method of a kind of tyrosol according to claim 1, it is characterized in that, described extractive crystallization is: add the ethyl acetate with the corresponding amount of described 4th extract concentrated solution to described 4th extract concentrated solution, extract 1-10 time, combining extraction liquid, decompression and solvent recovery, refrigerates to every ml containing tyrosol 20mg to 200mg, crystallization, obtains the tyrosol that purity is greater than 80%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510004953.XA CN104496760B (en) | 2015-01-06 | 2015-01-06 | A kind of preparation method of high purity tyrosol |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510004953.XA CN104496760B (en) | 2015-01-06 | 2015-01-06 | A kind of preparation method of high purity tyrosol |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104496760A CN104496760A (en) | 2015-04-08 |
| CN104496760B true CN104496760B (en) | 2016-04-27 |
Family
ID=52938236
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510004953.XA Active CN104496760B (en) | 2015-01-06 | 2015-01-06 | A kind of preparation method of high purity tyrosol |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104496760B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107661219A (en) * | 2017-07-31 | 2018-02-06 | 芜湖杨燕制药有限公司 | A kind of production technology of traditional Chinese medicine |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1130994C (en) * | 1998-04-21 | 2003-12-17 | 沈阳药科大学 | Health rhodiola root beverage |
| CN100496520C (en) * | 2006-06-28 | 2009-06-10 | 南京蓝本科技有限公司 | Chinese-medicinal extract for treating cardiovascular disease, its preparation and use |
| CN101143157A (en) * | 2006-09-11 | 2008-03-19 | 江苏中康药物科技有限公司 | Tyrosol and tyrosol bypass salidroside plant extraction and preparation and use thereof |
-
2015
- 2015-01-06 CN CN201510004953.XA patent/CN104496760B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN104496760A (en) | 2015-04-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1919222B (en) | Process for preparing notoginsen triterpenes | |
| CN101974045B (en) | Method for preparing salidroside | |
| CN102423329B (en) | A kind of discoloration method of panax notoginsenoside extract | |
| CN104725450B (en) | A kind of method that high-purity oleuropein is extracted from jasmine | |
| CN102631414B (en) | SepHaniadelavayi Diels total alkaloid extraction and purification technology | |
| CN102875450B (en) | Technological method for extracting 1-deoxynojirimycin from mulberry leaf | |
| CN106242959B (en) | A kind of extracting method of giant knotweed bioactive ingredients | |
| CN101721516B (en) | Preparation method of gardenia extract | |
| CN102924467B (en) | Preparation method for extracting and purifying bruceine D form brucea javanica | |
| CN105566409B (en) | A method of the extraction separation glucorphanin from broccoli seed | |
| CN103006721B (en) | Method for extracting phenylethanoid glycoside and flavonoid C-glycoside from corallodiscus flabellata | |
| CN103494858A (en) | Method for enriching total flavonoids in taraxacum mongolicum with macroporous resins | |
| CN104496760B (en) | A kind of preparation method of high purity tyrosol | |
| CN102659903A (en) | Method for extracting and purifying Chinese thorowax root saponin a or Chinese thorowax root saponin d | |
| CN104610401A (en) | Method for simultaneously extracting baicalin, baicalein and wogonin from scutellaria baicalensis | |
| CN112159451A (en) | Gynostemma pentaphylla saponin extract and preparation method thereof | |
| CN109776515A (en) | The method of mangiferin is extracted from myrica rubra leaf | |
| CN104189073B (en) | The preparation method of salvianolic acid | |
| CN109912582A (en) | The method of mangiferin is extracted from mango leaf | |
| CN102327324B (en) | Mulberry leaf total alkali extract and preparation method and application thereof | |
| CN101445544B (en) | Method for preparing ginsenoside Rb<1> | |
| CN106916162A (en) | A kind of preparation method of rock root of Beijing euphorbia lactone B bulk drugs | |
| CN106336440A (en) | Method for extracting and separating out oleanolic acid from olive leaves | |
| CN108191933B (en) | Method for preparing new astilbin by taking rhizoma smilacis glabrae as raw material | |
| CN102078400B (en) | Preparation method of extract of total triterpene acid of loquat leaf |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |