CN109316501A - A method of extracting triterpenoid saponin from gyro fruit seed meal - Google Patents
A method of extracting triterpenoid saponin from gyro fruit seed meal Download PDFInfo
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- CN109316501A CN109316501A CN201811457822.7A CN201811457822A CN109316501A CN 109316501 A CN109316501 A CN 109316501A CN 201811457822 A CN201811457822 A CN 201811457822A CN 109316501 A CN109316501 A CN 109316501A
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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)
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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
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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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, 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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The method that the present invention relates to a kind of to extract triterpenoid saponin from gyro fruit seed meal, cellulase, beta-glucosidase and citric acid solution are sequentially added in gyro fruit seed meal after drying, granularity is 20-120 mesh, it is subsequently placed on shaking table, 0.5-2.5h is reacted under conditions of 45-55 DEG C, 100-200r/min, reaction solution is filtered into the dissolution fluid to get triterpenoid saponin, macroporous absorbent resin is then added and adsorbs 1-5h, filtering, obtains the macroporous absorbent resin for being adsorbed with triterpenoid saponin；Ethanol solution is added to be desorbed, after removing ethanol solution, finally obtains triterpenoid saponin sterling.The present invention is using the gyro fruit seed meal generated after extracting oil as raw material, provide a kind of method that triterpenoid saponin is extracted from gyro fruit seed meal, the present invention makes full use of the gyro fruit seed meal generated after oil expression, and the extracting method is simple, low energy consumption, triterpenoid saponin extraction efficiency is high.
Description
Technical field
The method that the present invention relates to a kind of to extract triterpenoid saponin from gyro fruit seed meal, belongs to natural drug extractive technique
Field.
Background technique
Gyro fruit (Melliodendron xylocarpum Hand.-Mazz.) fruit likeness in form gyro, therefore named gyro fruit,
For Styracaceae (Styracaceae) gyro fruit category (Melliodendron) plant, rosy jasmin, water are also referred to as civil
Wax gourd, crow head pears, produce southeastern Yunnan, South Sichuan, Guizhou, Guangxi (except the southeast), Hunan, in the middle part of Guangdong to the north of, river
West and Fujian are China's endemic plant.Gyro fruit tree shape is beautiful, and trunk is logical straight, first spends rear leaf, trees are in full blossom when spending in full bloom seemingly
Snow, can do garden ornamental tree and shade tree.The tree lumber is yellow-white, and it is good commerical tree species that material is close, is suitble to system
Make furniture, tool etc..Gyro fruit seed is rich in grease, and main oil-producing position is kernel, and oil content is up to 49.6%, and edible
With being first-class oilseed plant, also containing there are also a large amount of such as triterpenoid saponin multiple compounds in seed.Gyro fruit seed by with
After extracting oil, the gyro fruit seed meal (gyro fruit seed oil expression waste matter) of generation usually can be all discarded, due to gyro fruit seed
Also containing multiple compounds such as triterpenoid saponins in the dregs of rice, and triterpene saponin componds have extensive pharmacological action and important life
Object activity, especially antibacterial, anti-inflammatory, liver protection, anticancer and in terms of expression effect it is prominent, therefore there are resources
The problem of waste.
Currently, the method for extracting triterpenoid saponin mainstream has water extract method, ultrasonic extraction, organic solvent method, chemistry pure
Change method etc., but there is the problems such as complex process, condition requirement is high, energy consumption is high, these are all constrained rests in peace including gyro fruit etc.
The utilization rate of triterpene saponin componds natural products etc. in savoury herb.
Summary of the invention
Present invention aims to solve the deficiencies of the prior art, and provides a kind of one kind, and triterpene soap is extracted from gyro fruit seed meal
The method of glycosides makes full use of the gyro fruit seed meal generated after oil expression, and the method for extracting triterpenoid saponin it is simple,
Low energy consumption, extraction efficiency is high.
Technical solution
A method of it extracting triterpenoid saponin from gyro fruit seed meal, includes the following steps:
(1) dissolve out: toward after dry, granularity for sequentially added in the 2g gyro fruit seed meal of 20-120 mesh cellulase,
Beta-glucosidase and citric acid-sodium citrate buffer, be subsequently placed on shaking table, in 45-55 DEG C, 100-200r/min
Under the conditions of react 0.5-2.5h, by reaction solution filter to get triterpenoid saponin dissolution fluid；
(2) it adsorbs: 2g macroporous absorbent resin being added into the triterpenoid saponin dissolution fluid of step (1), in 45-55 DEG C, 100-
1-5h is adsorbed under conditions of 200r/min, is filtered, is obtained the macroporous absorbent resin for being adsorbed with triterpenoid saponin；
(3) desorb: the addition ethanol solution in the macroporous absorbent resin of triterpenoid saponin that is adsorbed with toward step (2) is solved
It inhales, removes ethanol solution then to get triterpenoid saponin sterling.
Further, in step (1), the granularity of gyro fruit seed meal is 80-100 mesh.
Further, in step (1), the dosage of the cellulase and beta-glucosidase is 1-2.5%.
Further, in step (1), the citric acid-sodium citrate buffer by 9.2mL 0.1mol/L citric acid
The sodium citrate solution of solution and 10.8mL 0.1mol/L mix.
Further, in step (1), the solid-to-liquid ratio of gyro fruit seed meal and citric acid solution is 1:5-30, more preferably
For 1:25.
Further, in step (1), the reaction temperature is 50 DEG C, revolving speed 150r/min, reaction time 1h.
Further, in step (3), the volumetric concentration of the ethanol solution is 80-100%, more preferably 90%.Work as ethyl alcohol
Volume fraction when ever-increasing, the desorption efficiency of triterpenoid saponin can also become larger, when volume fraction reaches 90%,
The desorption efficiency of triterpenoid saponin starts to stablize, and from economic angle, desorbs triterpenoid saponin using the ethyl alcohol of 90% volume fraction
Most preferably.
Beneficial effects of the present invention: the present invention using the gyro fruit seed meal generated after extracting oil as raw material, provide it is a kind of from
The method that triterpenoid saponin is extracted in gyro fruit seed meal, the present invention have carried out abundant benefit to the gyro fruit seed meal generated after oil expression
With, and the extracting method is simple, low energy consumption, triterpenoid saponin extraction efficiency is high, is the similar Styracaceae plant resources in China
It efficiently utilizes and provides technical support.
Detailed description of the invention
Fig. 1 is the dissolution standard curve of triterpenoid saponin；
Fig. 2 is the ultraviolet spectrogram for the triterpenoid saponin that embodiment 1 is extracted；
Fig. 3 is that the triterpenoid saponin that embodiment 1 is extracted eliminates free radical activity curve.
Specific embodiment
The present invention will be further explained below with reference to the attached drawings and specific examples.
In following embodiments, gyro fruit seed, place of production Guangxi Gold show, it is big to pick up from Guangxi Gold show triterpenoid saponin content 16%
Yao Shanlinchang；Cellulase, the trade mark are especially your L-100, filter paper enzyme activity 188.54U, by Hunan especially your biochemical limited public affairs
Department provides；The enzyme activity of beta-glucosidase, trade mark BG-188, beta-glucosidase is 237.54U, is mentioned by Beijing Novi letter
For；Macroporous absorbent resin, trade mark AB-8, is provided by Sangon Biotech (Shanghai) Co., Ltd..But it is not limited to this.
The gyro fruit seed meal generated after oil expression is subjected to constituent analysis, the results are shown in Table 1:
1 gyro fruit seed meal main component of table
Embodiment 1
A method of it extracting triterpenoid saponin from gyro fruit seed meal, includes the following steps:
(1) it dissolves out: successively adding in the gyro fruit seed meal (gyro fruit seed meal granularity is 80-100 mesh) after 2g drying
Enter cellulase (1%), beta-glucosidase (1%) and 50mL citric acid-sodium citrate (by the lemon of 9.2mL 0.1mol/L
The sodium citrate solution of acid solution and 10.8mL 0.1mol/L mix) buffer solution, gyro fruit seed meal and citric acid are slow
The solid-to-liquid ratio for rushing solution is 1:25, is subsequently placed on shaking table, reacts 1h under conditions of 50 DEG C, 150r/min, by reaction solution mistake
Filter is to get triterpenoid saponin dissolution fluid；
(2) it adsorbs: 2g macroporous absorbent resin being added into the triterpenoid saponin dissolution fluid of step (1), in 50 DEG C, 150r/min
Under conditions of adsorb 4h, filter, obtain the macroporous absorbent resin for being adsorbed with triterpenoid saponin；
(3) desorb: toward step (2) to be adsorbed in the macroporous absorbent resin of triterpenoid saponin addition 90% ethyl alcohol of 10mL molten
Liquid is desorbed, and ethanol solution is then removed, and obtains triterpenoid saponin sterling.The ultraviolet spectrogram of product is shown in Fig. 2, it can be seen that
Ultraviolet 215nm or so place has absorption maximum, illustrates that product is triterpenoid saponin.
Triterpenoid saponin concentration and dissolution rate is calculated in dissolution fluid in determination step (1):
Triterpenoid saponin dissolution rate (%)=triterpenoid saponin concentration × liquor capacity/gyro fruit seed quality × 100%.
Wherein triterpenoid saponin content is measured using vanillic aldehyde sulphate method, it is dense in maximum absorption wave strong point measurement different quality
The absorbance of degree, using triterpenoid saponin content as abscissa, absorbance is ordinate, and standard curve is as shown in Figure 1, can see
Out, the concentration of triterpenoid saponin shows good linear relationship within the scope of 0.7g/L, this explanation vanillic aldehyde-sulphate method can
Accurately quickly to measure triterpenoid saponin content.
In the present embodiment, triterpenoid saponin dissolution rate is 4.05% in triterpenoid saponin dissolution fluid.
Embodiment 2
In step (1), for the gyro fruit seed meal granularity used for 20-40 mesh, remaining is same as Example 1.Measurement dissolution
Triterpenoid saponin concentration and dissolution rate is calculated in liquid, dissolution rate 2.41%.
Embodiment 3
In step (1), for the gyro fruit seed meal granularity used for 40-60 mesh, remaining is same as Example 1.Measurement dissolution
Triterpenoid saponin concentration and dissolution rate is calculated in liquid, dissolution rate 3.1%.
Embodiment 4
In step (1), for the gyro fruit seed meal granularity used for 60-80 mesh, remaining is same as Example 1.Measurement dissolution
Triterpenoid saponin concentration and dissolution rate is calculated in liquid, dissolution rate 3.5%.
Embodiment 5
In step (1), for the gyro fruit seed meal granularity used for 100-120 mesh, remaining is same as Example 1.It measures molten
Triterpenoid saponin concentration and dissolution rate is calculated in liquid out, dissolution rate 4.07%.
Material size is also to influence gyro fruit it can be seen from the dissolution rate of triterpenoid saponin in the dissolution fluid of embodiment 1-5
One of the factor of seed meal hydrolysis result, with the increase of mesh number, i.e. material size is more and more tiny, the dissolution rate of triterpenoid saponin
Increase, 20-40 mesh dissolution rate only has 2.41%, and in 80-100 mesh, dissolution rate starts to tend towards stability, and is 3.8%.
Embodiment 6
In step (1), the solid-to-liquid ratio of gyro fruit seed meal and citric acid solution is 1:5, remaining and 1 phase of embodiment
Together.It measures triterpenoid saponin concentration in dissolution fluid and calculates dissolution rate, dissolution rate 3.25%.
Embodiment 7
In step (1), the solid-to-liquid ratio of gyro fruit seed meal and citric acid solution is 1:10, remaining and 1 phase of embodiment
Together.It measures triterpenoid saponin concentration in dissolution fluid and calculates dissolution rate, dissolution rate 3.38%.
Embodiment 8
In step (1), the solid-to-liquid ratio of gyro fruit seed meal and citric acid solution is 1:15, remaining and 1 phase of embodiment
Together.It measures triterpenoid saponin concentration in dissolution fluid and calculates dissolution rate, dissolution rate 3.3%.
Embodiment 9
In step (1), the solid-to-liquid ratio of gyro fruit seed meal and citric acid solution is 1:20, remaining and 1 phase of embodiment
Together.It measures triterpenoid saponin concentration in dissolution fluid and calculates dissolution rate, dissolution rate 3.26%.
Embodiment 10
In step (1), the solid-to-liquid ratio of gyro fruit seed meal and citric acid solution is 1:30, remaining and 1 phase of embodiment
Together.It measures triterpenoid saponin concentration in dissolution fluid and calculates dissolution rate, dissolution rate 3.3%.
Comparing embodiment 1,6-10 dissolution rate it is found that solid-to-liquid ratio under conditions of 1:5,1:10,1:15,1:20,1:30
The dissolution rate of triterpenoid saponin is all 3.3% or so, however triterpenoid saponin dissolution rate reaches highest under conditions of solid-to-liquid ratio 1:25.
Comparative example 1
In step (1), cellulase and beta-glucosidase are not added, remaining is same as Example 1.It measures in dissolution fluid
Triterpenoid saponin concentration simultaneously calculates dissolution rate, dissolution rate 2.2%.
The antioxidant activity of triterpenoid saponin is tested using DPPH radicals scavenging method:
DPPH free radical is a kind of stable free radical, is in organic solvent purple, has larger suction in 517nm wavelength
It receives, and a part of free radical can be made to be removed when antioxidant is added, so that the wavelength absorption weakens.The work of antioxidant
Property is higher, and the absorbance of the wavelength is then smaller, and the antioxidant activity of substance is evaluated with this.
Measuring method: the triterpenoid saponin that triterpenoid saponin that embodiment 1 is extracted is configured to 10mg/ml with dehydrated alcohol is molten
Liquid, when use, are diluted to a series of sample of concentration of 1mg/ml, 2mg/ml, 3mg/ml, 4mg/ml, 5mg/ml with dehydrated alcohol
Liquid.It takes 2mL testing sample solution that 2mLDPPH is added, 30min is kept in dark place in the 25 DEG C of water-baths of this solution, sample is measured at 517nm
The absorbance of liquid is A1；Taking 2mL ethanol solution that sample is replaced to measure absorbance is A0；It is anhydrous that 2mL is added into 2mL sample
Ethanol solution measures sample copy bottom absorbance A2；
Clearance rate K (%)=[A0-(A1 ﹣A2)]/A0× 100%
As a result see Fig. 3.As seen from Figure 3, triterpenoid saponin has the ability for eliminating free radical in gyro fruit seed meal, and
And its elimination ability also increases with the increase of concentration.Therefore it may be concluded that triterpenoid saponin has in gyro fruit seed meal
Certain inoxidizability, oxidation resistance increase with the increase of concentration.
Claims (8)
1. a kind of method for extracting triterpenoid saponin from gyro fruit seed meal, which comprises the steps of:
(1) it dissolves out: sequentially adding cellulase, β-Portugal in 2g gyro fruit seed meal after drying, granularity is 20-120 mesh
Polyglycoside enzyme and citric acid-sodium citrate buffer, are subsequently placed on shaking table, in 45-55 DEG C, the condition of 100-200r/min
Reaction solution is filtered the dissolution fluid to get triterpenoid saponin by lower reaction 0.5-2.5h；
(2) it adsorbs: 2g macroporous absorbent resin being added into the triterpenoid saponin dissolution fluid of step (1), in 45-55 DEG C, 100-200r/
1-5h is adsorbed under conditions of min, is filtered, is obtained the macroporous absorbent resin for being adsorbed with triterpenoid saponin；
(3) desorb: the addition ethanol solution in the macroporous absorbent resin of triterpenoid saponin that is adsorbed with toward step (2) is desorbed, so
Remove ethanol solution afterwards to get triterpenoid saponin sterling.
2. the method for triterpenoid saponin is extracted from gyro fruit seed meal as described in claim 1, which is characterized in that in step (1),
The granularity of gyro fruit seed meal is 80-100 mesh.
3. the method for triterpenoid saponin is extracted from gyro fruit seed meal as described in claim 1, which is characterized in that in step (1),
In step (1), the dosage of the cellulase and beta-glucosidase is the 1-2.5% of gyro fruit seed meal.
4. the method for triterpenoid saponin is extracted from gyro fruit seed meal as described in claim 1, which is characterized in that in step (1),
The citric acid-sodium citrate buffer is by the citric acid solution of 9.2mL 0.1mol/L and the lemon of 10.8mL 0.1mol/L
Lemon acid sodium solution mixes.
5. the method for triterpenoid saponin is extracted from gyro fruit seed meal as described in claim 1, which is characterized in that in step (1),
The solid-to-liquid ratio of gyro fruit seed meal and citric acid-sodium citrate buffer is 1:5-30.
6. the method for triterpenoid saponin is extracted from gyro fruit seed meal as claimed in claim 5, which is characterized in that gyro fruit seed
The solid-to-liquid ratio of the dregs of rice and citric acid-sodium citrate buffer is 1:25.
7. the method for triterpenoid saponin is extracted from gyro fruit seed meal as described in claim 1, which is characterized in that in step (1),
The reaction temperature is 50 DEG C, revolving speed 150r/min, reaction time 1h.
8. the method for triterpenoid saponin is extracted from gyro fruit seed meal as described in any one of claim 1 to 7, which is characterized in that
In step (3), the volumetric concentration of the ethanol solution is 80-100%.
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| CN201811457822.7A CN109316501B (en) | 2018-11-30 | 2018-11-30 | Method for extracting triterpenoid saponin from Gyroscope seed meal |
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| CN201811457822.7A CN109316501B (en) | 2018-11-30 | 2018-11-30 | Method for extracting triterpenoid saponin from Gyroscope seed meal |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102048885A (en) * | 2011-01-10 | 2011-05-11 | 广州市中医医院 | Extraction process for lychee seed saponin |
| CN103961388A (en) * | 2014-05-22 | 2014-08-06 | 江苏博益生物科技有限公司 | Method for preparing total saponins of radix cynanchi bungei |
| CN104971084A (en) * | 2015-07-21 | 2015-10-14 | 安徽师范大学 | Purifying method of gypenoside |
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2018
- 2018-11-30 CN CN201811457822.7A patent/CN109316501B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102048885A (en) * | 2011-01-10 | 2011-05-11 | 广州市中医医院 | Extraction process for lychee seed saponin |
| CN103961388A (en) * | 2014-05-22 | 2014-08-06 | 江苏博益生物科技有限公司 | Method for preparing total saponins of radix cynanchi bungei |
| CN104971084A (en) * | 2015-07-21 | 2015-10-14 | 安徽师范大学 | Purifying method of gypenoside |
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