CN108042618A - A kind of method using Subcritical water chromotagraphy total glucosides of paeony - Google Patents
A kind of method using Subcritical water chromotagraphy total glucosides of paeony Download PDFInfo
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- 235000019441 ethanol Nutrition 0.000 claims abstract description 26
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- YKRGDOXKVOZESV-WRJNSLSBSA-N Paeoniflorin Chemical compound C([C@]12[C@H]3O[C@]4(O)C[C@](O3)([C@]1(C[C@@H]42)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)C)OC(=O)C1=CC=CC=C1 YKRGDOXKVOZESV-WRJNSLSBSA-N 0.000 description 11
- YKRGDOXKVOZESV-UHFFFAOYSA-N paeoniflorin Natural products O1C(C)(C2(CC34)OC5C(C(O)C(O)C(CO)O5)O)CC3(O)OC1C24COC(=O)C1=CC=CC=C1 YKRGDOXKVOZESV-UHFFFAOYSA-N 0.000 description 11
- VYQNWZOUAUKGHI-UHFFFAOYSA-N monobenzone Chemical compound C1=CC(O)=CC=C1OCC1=CC=CC=C1 VYQNWZOUAUKGHI-UHFFFAOYSA-N 0.000 description 10
- 239000003814 drug Substances 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 8
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- 235000010233 benzoic acid Nutrition 0.000 description 7
- 238000004128 high performance liquid chromatography Methods 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- QQUHMASGPODSIW-UHFFFAOYSA-N Albiflorin Natural products C=1C=CC=CC=1C(=O)OCC12C(=O)OC3(C)CC(O)C1CC32OC1OC(CO)C(O)C(O)C1O QQUHMASGPODSIW-UHFFFAOYSA-N 0.000 description 5
- QQUHMASGPODSIW-ICECTASOSA-N albiflorin Chemical compound O([C@@]12C[C@H]3[C@H](O)C[C@@]1(OC(=O)[C@]32COC(=O)C=1C=CC=CC=1)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O QQUHMASGPODSIW-ICECTASOSA-N 0.000 description 5
- 238000010828 elution Methods 0.000 description 5
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 4
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- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
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- 229930014626 natural product Natural products 0.000 description 3
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- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
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- 244000236658 Paeonia lactiflora Species 0.000 description 1
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- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/65—Paeoniaceae (Peony family), e.g. Chinese peony
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The present invention provides a kind of method using Subcritical water chromotagraphy total glucosides of paeony, step is as follows：A, Chinese herbaceous peony powder is taken, by liquid ratio 20~40：It after 1 (mL/g) is soaked, is placed in Subcritical water chromotagraphy device, controlled at 158~180 DEG C, keeps the temperature 20~30min, obtain extracting solution；B, ethyl alcohol is added in the extracting solution of step a, mixing filters, concentration, upper macroporous resin adsorption column, after washed with impurities, is eluted with 20~40% ethanol solutions, collects eluent, concentration, drying to get total glucosides of paeony.The Subcritical water chromotagraphy of total glucosides of paeony can be realized using simple device by the present invention, use extractant as water, environmentally protective, and obtained total glucosides of paeony recovery rate is high, purity is high, and low energy consumption, extraction cost is low, and suitable for industrialized production, application prospect is good.
Description
Technical field
The invention belongs to Chinese material medicine resource field of comprehensive utilization, and in particular to a kind of to utilize Subcritical water chromotagraphy total glucosides of paeony
Method.
Background technology
Chinese herbaceous peony (Paeonia lactiflora Pall.) is Paeoniaceae (Paeoniaceae) Paeonia (Paeonia L.)
Herbaceos perennial is used as medicine with root, is the rare medicinal herbs in Chinese medicine, and medicinal history is long, and medical value is high, modern medicine
Reason research shows that the main active of Chinese herbaceous peony for Paeoniflorin, in addition there is the monoterpene glycosides such as albiflorin, Hydroxy peoniflorin
Close object, abbreviation total glucosides of paeony (TGP).Total glucosides of paeony has the pharmacological actions such as anti-inflammatory, antiulcer, antithrombotic and immunological regulation.Root
According to the difference of the place of production and processing method etc., the dry root of commodity Chinese herbaceous peony is divided into Radix Paeoniae Alba and the radix paeoniae rubrathe, and the two is main using Paeoniflorin
One of active ingredient.Wherein, Radix Paeoniae Alba is in the significant curative effect to virus hepatitis, rheumatoid arthritis and gynaecology's class disease, with
Radix Paeoniae Alba for primary raw material Chinese patent drug (Xiaoyao San, small Green Dragon particle, peony root total glycosides capsules, nourishing lung and activating blood capsule etc.) demand with
Day all increasings, the market prospects of Radix Paeoniae Alba are more and more wide.
At present, the extracting method of total glucosides of paeony is more, mainly there is decocting cooking method, Diluted Alcohol reflux extraction, ultrasonic extraction
Method, microwave loss mechanisms etc..However, the complexity due to natural products ingredient so that these conventional methods mostly there are efficiency it is low,
Yield is low and the shortcomings of energy consumption is big, and mostly using inflammable, explosive, toxic organic solvent such as：The extractions such as methanol, ethyl alcohol, chloroform,
Environmental pollution and dissolvent residual are easily caused, and endangers health.With the development of science and technology, some new extraction and separation technology quilts
For the extraction of active ingredient of natural product, as supercritical extract, physical field strengthen extraction, homogenate extraction, aqueous two-phase extraction and
Inverse micelle abstraction etc., but new technology again mostly it is of high cost, equipment is complicated, it is difficult to realize practical application in the industry.It is subcritical
Water extraction method is a kind of new extractive technique to grow up in the latest 20 years, with green non-pollution, quick, low cost
Advantage, be subject to Natural products research, person is favored, and becomes a kind of popular extraction means.
Subcritical water chromotagraphy technology is using subcritical water as extractant, by controlling temperature and pressure, accelerates the diffusion of water
Efficiency reduces its surface tension and viscosity, reaches the green extraction of extraction polarity or Nonpolar Solute.Faced at present using Asia
Boundary's water extractive technique extraction Chinese herbaceous peony active ingredient has a small number of reports, but all not fully up to expectations to the recovery rate of total glucosides of paeony, still needs to
It is further improved.
There are two reports using Subcritical water chromotagraphy technology extraction Chinese herbaceous peony active ingredient at present, wherein opening the extraction of fine jade report
Method【Open fine jade etc., the comparative studies of Paeoniflorin extracting method in Radix Paeoniae Alba, modern traditional Chinese medicine, 2013,33 (1)：84-93】, it is only right
Paeoniflorin is extracted in radix paeoniae alba decoction pieces and has carried out rough temperature change experiment, as a result this method is relatively low to the recovery rate of Paeoniflorin.
The extracting method of Wang Qian reports【Wang Qian etc., accelerated solvent extraction extraction the radix paeoniae rubrathe in Paeoniflorin, drug inspection, 2006,21
(2)：184-186】, the extractant used is ethanol solution, at high temperature using ethyl alcohol with certain security risk；And it carries
Complicated equipment need to be used when taking, operates comparatively laborious, easy failure, it is very high to extraction raw material particle size requirement (2~3mm compared with
It is suitable), it is unfavorable for industrial applications.
The content of the invention
To solve the above-mentioned problems, it is an object of the invention to provide a kind of Chinese herbaceous peonies environmentally protective, recovery rate is high, purity is high
The total glycosides extracting method of medicine.
The present invention provides a kind of method of Subcritical water chromotagraphy total glucosides of paeony, step is as follows：
A, Chinese herbaceous peony powder is taken, by liquid ratio 20~40：After 1 (mL/g) is soaked, it is placed in Subcritical water chromotagraphy device,
Controlled at 158~180 DEG C, 20~30min is kept the temperature, obtains extracting solution；
B, ethyl alcohol is added in the extracting solution of step a, mixing filters, concentration, upper macroporous resin adsorption column, washed with impurities
Afterwards, eluted with 20~40% ethanol solutions, collect eluent, concentration, drying to get total glucosides of paeony.
Wherein, the Subcritical water chromotagraphy device is made of high-pressure hydrothermal reaction kettle and constant temperature high temperature oven.
Wherein, in step a, the time soaked is 10min.
Wherein, in step a, after preheating 2min at 158~180 DEG C, then 20~30min is kept the temperature.
Wherein, in step a, the liquid ratio is 20~21：1 (mL/g), temperature are 158~160 DEG C, and the time of heat preservation is
20~22min.
Wherein, the liquid ratio is 21：1 (mL/g), temperature are 158 DEG C, and the time of heat preservation is 22min.
Wherein, in step b, isometric ethyl alcohol is added in the extracting solution of step a.
Wherein, in step b, the macroreticular resin is D101.
Wherein, in step b, during upper macroporous resin adsorption column, the amount ratio of applied sample amount and macroreticular resin is 1.5：1(mL/
g)。
Wherein, in step b, the concentration of the ethanol solution is 20%.
Sichuan Zhong Jiang is exactly one of Genuine producing area of Chinese herbaceous peony since ancient times.This patent is using middle river Radix Paeoniae Alba as raw material, by right
Influencing the factors of Subcritical water chromotagraphy total glucosides of paeony includes：After Extracting temperature, time, liquid ratio optimize, obtain
A kind of total glucosides of paeony extracting method environmentally protective, recovery rate is high, purity is high.And Extraction solvent of the present invention is water, uses peace
Entirely, low energy consumption；Extraction equipment is simple in structure, at low cost；To sample without particular/special requirement, it is only necessary to which crushing is particularly suitable for industry
Metaplasia is produced, and new technological means is provided for the reasonable development of Chinese herbaceous peony resource.
Obviously, the above according to the present invention according to the ordinary technical knowledge and customary means of this field, is not departing from
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.
The specific embodiment of form by the following examples remakes further specifically the above of the present invention
It is bright.But the scope that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to following example.It is all to be based on the above of the present invention
The technology realized all belongs to the scope of the present invention.
Description of the drawings
Fig. 1 simplifies Subcritical water chromotagraphy device
The total glycosides recovery rates of Fig. 2 measure HPLC chromatogram
Fig. 3 Subcritical water chromotagraphies sample and macroporous resin purification sample HPLC chromatogram
Specific embodiment
It is described further below with embodiment, but the present invention is not limited to these embodiments.
Experiment reagent and instrument used in the present invention is as follows：
1. material and reagent
Middle river herbaceous peony powder；Absolute ethyl alcohol；Sodium hydroxide；Concentrated hydrochloric acid；Acetonitrile (chromatographically pure, Fisher)；Benzoic acid (>=
99.5%)；Paeoniflorin, albiflorin standard items (>=98%)；D101 macroreticular resins, in addition to indicating, other reagents are point
It analyses pure.
2. laboratory apparatus
ZX 101-1AB-C high temperature drying casees, 1260 high performance liquid chromatographs of Agilent Technologies, BT
124s type electronic balances, high-pressure hydrothermal reaction kettle, B-260 Rotary Evaporators, WP-Z-UV types laboratory ultrapure water machine, recirculated water
Formula vacuum pump.
The extracting method of 1 total glucosides of paeony of the present invention of embodiment
Extraction equipment：Simple subcritical water static state extraction element (see Fig. 1), the device is simple, at low cost, easy to operate.It should
Device is made of constant temperature oven (controllable temperature >=250 DEG C) and high-pressure hydrothermal reaction kettle.First high temperature constant temperature baking oven is set using preceding
Surely assigned temperature is arrived, preheats 30min.
Extracting method：Dry middle river herbaceous peony powder is taken, adds in the distilled water of 21 times of amounts, impregnates 10min；It stirs evenly, fills
Enter autoclave, and the preheating 2min in constant temperature oven (158 DEG C), 158 DEG C of heat preservation extraction 21min；Take out reaction kettle cooling
To≤50 DEG C, isometric ethyl alcohol is added in into extracting solution and is stirred evenly, filter to obtain total glycosides extracting solution；Extracting solution is concentrated into about 1/10
Volume is loaded on D101 macroreticular resins, and it is not muddy that pure water is eluted to eluent, then using 20% ethyl alcohol as eluant, eluent, 1BV
h-1Speed elution, collect eluent；HPLC is analyzed, and is merged same composition, is concentrated under reduced pressure and is evaporated, obtain unformed powder Chinese herbaceous peony
Total glycosides.
The extracting method of 2 total glucosides of paeony of the present invention of embodiment
Dry middle river herbaceous peony powder is taken, adds in the distilled water (liquid ratio 21mL/g) of 21 times of amounts, impregnates 10min；Stirring is equal
It is even, it is fitted into Subcritical water chromotagraphy device, and the preheating 2min in constant temperature oven (158 DEG C), 158 DEG C of heat preservation extraction 22min；It takes
Go out reaction kettle and be cooled to≤50 DEG C, isometric ethyl alcohol is added in into extracting solution and is stirred evenly, filter to obtain total glycosides extracting solution；Extracting solution
Be concentrated into about 1/10 volume, be loaded on D101 macroreticular resins, it is not muddy that pure water is eluted to eluent, then using 20% ethyl alcohol as
Eluant, eluent, 1BVh-1Speed elution, collect eluent；HPLC is analyzed, and is merged same composition, is concentrated under reduced pressure and is evaporated, obtain nothing
Sizing powder total glucosides of paeony.Total glycosides recovery rate is measured as 7.08%.
Beneficial effects of the present invention are illustrated below by way of test example：
1 extraction process of test example is investigated
First, experimental method
1st, middle river total glucoside extractive technique route
River herbaceous peony powder in a certain amount of drying → a certain amount of distilled water of addition is weighed, immersion 10min → stir evenly is filled
Enter autoclave → in constant temperature oven preheat 2min → heat preservation is extracted under certain temperature → be cooled to≤50 DEG C → to extraction
Isometric ethyl alcohol is added in liquid to stir evenly → filtering to obtain total glycosides extracting solution → to be concentrated into 1/10 volume and be loaded to D101 macropore trees
On fat → pure water is eluted to transparent (sugar-free) → 20% ethanol elution → HPLC is analyzed → and merges same composition, be concentrated under reduced pressure and be evaporated
→ obtain the total glycosides of unformed powder.
2nd, Testing index
The extraction process of river Radix Paeoniae Alba in being investigated using total glycosides recovery rate as index, total glycosides recovery rate are measured using high-efficient liquid phase color
Spectrum analysis method, specific method are as follows：
Sample hydrolyzes：Subcritical water chromotagraphy liquid is taken, isometric ethyl alcohol mixing, pipette, extract 1mL are added in into extracting solution
Extracting solution, 50 DEG C are evaporated, and add in 2mL 1%NaOH and shake up, are transferred to 25mL volumetric flasks, with each 5mL washings flask of 50% ethyl alcohol
Twice, cleaning solution moves into volumetric flask, and it is 3-6 then to adjust pH with 5%HCl, and 50% ethyl alcohol constant volume is spare.
Chromatographic condition：Chromatographic column：InertSustain C18 chromatographic columns (4.6 × 250mm, 5 μm)；Flow visualizing：
0.1% phosphoric acid water (A)-acetonitrile (B), condition of gradient elution：0~10min 30%B, 10~12min 30%~90%B, 12~
15min 90%B, run 5min afterwards；Detection wavelength 230nm；40 DEG C of column temperature, sample size 10 μ L, flow velocity 1mL/min.
3rd, the making of standard curve
It is appropriate that benzoic acid, Paeoniflorin and albiflorin standard items are weighed respectively, respectively at the volumetric flask of 3 10.00mL
In, scale is settled to ethyl alcohol, ultrasonic dissolution is to get corresponding standard solution.Concentration of benzoic acid is 0.04mg/mL, Paeoniflorin
For 0.47mg/mL, albiflorin 0.20mg/mL.
The corresponding mark solution injection HPLC of 1,2,5,10,15 μ L is drawn respectively, with concentration of standard solution (X) for abscissa,
Peak area (Y) draws standard curve for ordinate, and calibration curve equation is drawn by linear regression.
Concentration of benzoic acid in extracting solution is calculated with standard curve, is then calculated as follows benzoic acid content：
In formula, C represents the concentration of benzoic acid (mg/mL) that establishing criteria curve calculates, and V represents body after sample hydrolysis constant volume
Product (mL), n represent specific extension rate, and m is the sample quality (g) weighed.
Total glycosides (in terms of Paeoniflorin) recovery rate is calculated finally by benzoic acid content：
4th, the investigation of extraction conditions
(1) Subcritical water chromotagraphy Study of operational conditions
Using total glycosides recovery rate as index, to influencing the principal element of Subcritical water chromotagraphy method extraction efficiency：Extracting temperature carries
Time and liquid ratio is taken to be screened, draws the optimal extraction process of river total glucoside in optimal subcritical water-swollen squid.
(2) total glycosides purifying
River Radix Paeoniae Alba Subcritical water chromotagraphy liquid in being obtained with optimum extraction process, is concentrated under reduced pressure to the 1/10 of total volume, passed through
Screen macroreticular resin species (H103, D101 and S-8), applied sample amount: macroreticular resin dosage (3: 1,2: 1,1.5: 1) (i.e. applied sample amount
3-1.5ml, corresponding 1g macroreticular resins), eluent (water, 20% ethyl alcohol, 40% ethyl alcohol), obtain optimal separation macroreticular resin and wash
De- liquid composition.
2nd, experimental result
1st, standard curve
1 standard curve of table
As it can be seen that during three kinds of compound determinations, R20.99 is all higher than, standard curve is linearly good.
2nd, the optimization of extraction conditions
Dry middle river herbaceous peony powder (different liquid ratio test groups are weighed the herbaceous peony powder of respective amount by corresponding liquid ratio) is weighed,
The distilled water of same volume is separately added into, is extracted under different temperatures, time, the recovery rate of total glucosides of paeony in Detection and Extraction liquid.
The selection result is shown in Table 2.
Influence of the different extracting parameters of table 2 to total glycosides recovery rate
| Extraction time/min | Extracting temperature/DEG C | Liquid ratio/mL/g | Total glycosides recovery rate (%) | |
| 1 | 10.00 | 160 | 10.00 | 6.48 |
| 2 | 10.00 | 120 | 20.00 | 6.06 |
| 3 | 30.00 | 160 | 10.00 | 6.56 |
| 4 | 20.00 | 160 | 20.00 | 7.03 |
| 5 | 20.00 | 120 | 30.00 | 6.24 |
| 6 | 20.00 | 200 | 30.00 | 6.21 |
| 7 | 20.00 | 200 | 10.00 | 6.12 |
| 8 | 10.00 | 200 | 20.00 | 6.06 |
| 9 | 30.00 | 120 | 20.00 | 6.37 |
| 10 | 30.00 | 160 | 30.00 | 6.77 |
| 11 | 20.00 | 120 | 10.00 | 6.25 |
| 12 | 30.00 | 200 | 20.00 | 6.12 |
| 13 | 10.00 | 160 | 30.00 | 6.43 |
| 14 | 10.00 | 180 | 20.00 | 5.90 |
| 15 | 20.00 | 180 | 20.00 | 6.93 |
| 16 | 30.00 | 180 | 20.00 | 6.83 |
| 17 | 40.00 | 180 | 20.00 | 6.72 |
| 18 | 20.00 | 120 | 20.00 | 6.29 |
| 19 | 20.00 | 140 | 20.00 | 6.56 |
| 20 | 20.00 | 200 | 20.00 | 6.06 |
| 21 | 20.00 | 160 | 10.00 | 5.83 |
| 22 | 20.00 | 160 | 30.00 | 6.80 |
| 23 | 20.00 | 160 | 40.00 | 6.91 |
| 24 | 22.00 | 158 | 21.00 | 7.08 |
As shown in Table 2, optimal extraction process is number 24, i.e.,：Extraction time 22min, 158 DEG C of Extracting temperature, liquid ratio
21mL/g, it is 7.08 ± 0.20% that checking test, which measures total glycosides recovery rate, three times.
3rd, the purifying of total glycosides
It the results are shown in Table 3 and Fig. 3.
3 macroporous resin adsorption of table screens (Dynamic Adsorption and elution)
| Resin model | Maximal absorptive capacity/mg/g | Eluting rate/% | Total glycosides content/% |
| H103 | 54.2 | 51.3 | 82.6 |
| D101 | 82.3 | 78.3 | 88.6 |
| HP-20 | 62.6 | 93.1 | 73.6 |
As shown in Table 3, D101 macroreticular resins are best to the enrichment of total glycosides, purification effect, have the higher rate of recovery (>=
88%), optimal applied sample amount: macroreticular resin dosage answers≤1.5, and optimal eluant, eluent is 20% ethyl alcohol.Measured through HPLC, purifying it is total
Paeoniflorin content ＞ 63% in glycosides, albiflorin content ＞ 25%.
1 extraction process of comparative example is investigated
Compare different extraction process by water：
Water extracting method 1：Extraction time 22min, liquid ratio 21mL/g, 90 DEG C of Extracting temperature, checking test measure is total three times
Glycosides recovery rate is only 5.91 ± 0.15%；
Water extracting method 2：Ultrasonic method, ultrasonic time 22min, liquid ratio 21mL/g, ultrasonic power 400W, room temperature extraction, three
It is 5.51 ± 0.07% that secondary checking test, which measures total glycosides recovery rate,.
Subcritical water chromotagraphy method of the present invention：Extraction time 22min, 158 DEG C of Extracting temperature, liquid ratio 21mL/g are tested three times
It is 7.08 ± 0.20% that confirmatory test, which measures total glycosides recovery rate,.
As it can be seen that under the conditions of identical extraction time and liquid ratio, the method for the present invention is to the extraction efficiency of total glycosides in the Radix Paeoniae Alba of river
Most preferably, compared with 2 kinds of control methods, 19.8% and 28.5% is respectively increased in recovery rate, is more suitable for total glucosides of paeony extraction.
To sum up, the present invention has been obtained the total glucosides of paeony that a kind of recovery rate is high, purity is high and has been carried by the optimization to extracting parameter
Take method；And low energy consumption, operating cost is low for the method for the present invention, is particularly suitable for industrialized production, and application prospect is good.
Claims (10)
- A kind of 1. method using Subcritical water chromotagraphy total glucosides of paeony, it is characterised in that：Step is as follows：A, Chinese herbaceous peony powder is taken, by liquid ratio 20~40：It after 1 (mL/g) is soaked, is placed in Subcritical water chromotagraphy device, controls Temperature is 158~180 DEG C, keeps the temperature 20~30min, obtains extracting solution；B, ethyl alcohol is added in the extracting solution of step a, mixing filters, concentration, upper macroporous resin adsorption column, after washed with impurities, uses 20~40% ethanol solutions elute, and collect eluent, concentration, drying to get total glucosides of paeony.
- 2. according to the method described in claim 1, it is characterized in that：The Subcritical water chromotagraphy device is by high-pressure hydrothermal reaction kettle It is formed with constant temperature high temperature oven.
- 3. according to the method described in claim 1, it is characterized in that：In step a, the time soaked is 10min.
- 4. according to the method described in claim 1, it is characterized in that：In step a, after preheating 2min at 158~180 DEG C, then Keep the temperature 20~30min.
- 5. according to the method described in any of claim 1 to 4, it is characterised in that：In step a, the liquid ratio for 20~ 21：1 (mL/g), temperature are 158~160 DEG C, and the time of heat preservation is 20~22min.
- 6. according to the method described in claim 5, it is characterized in that：The liquid ratio is 21：1 (mL/g), temperature are 158 DEG C, The time of heat preservation is 22min.
- 7. according to the method described in claim 1, it is characterized in that：In step b, added in the extracting solution of step a isometric Ethyl alcohol.
- 8. according to the method described in claim 1, it is characterized in that：In step b, the macroreticular resin is D101.
- 9. according to the method described in claim 1, it is characterized in that：In step b, during upper macroporous resin adsorption column, applied sample amount with The amount ratio of macroreticular resin is 1.5：1(mL/g).
- 10. according to the method described in claim 1, it is characterized in that：In step b, the concentration of the ethanol solution is 20%.
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| CN112791165A (en) * | 2020-12-31 | 2021-05-14 | 杭州电子科技大学 | Chinese herbal medicine extract, preparation method thereof and application thereof in fruit fly anti-aging |
| CN115677796A (en) * | 2022-11-23 | 2023-02-03 | 四川聚元药业集团有限公司 | Method for extracting paeoniflorin from white paeony root |
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| CN112791165A (en) * | 2020-12-31 | 2021-05-14 | 杭州电子科技大学 | Chinese herbal medicine extract, preparation method thereof and application thereof in fruit fly anti-aging |
| CN115677796A (en) * | 2022-11-23 | 2023-02-03 | 四川聚元药业集团有限公司 | Method for extracting paeoniflorin from white paeony root |
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