CN110302220A - A kind of preparation method of low phenolic acid ginkgo leaf neutrality extract - Google Patents
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Abstract
The invention discloses a kind of preparation methods of low phenolic acid ginkgo leaf neutrality extract, it is characterized in that, the following steps are included: (1) extracts waste powder as raw material using dry ginkgo leaf, organic solvent is added, is repeated refluxing extraction 3 times by soxhlet extraction；Combined extract recycles organic solvent, obtains extraction ointment；(2) aqueous slkali is added in the extraction ointment in step (1) and carries out saponification, obtain saponification liquor；(3) organic solvent is added in the saponification liquor in step (2) to extract 3 times, collects upper solution and merges, revolving removal organic solvent obtains low phenolic acid ginkgo leaf neutrality extract.The present invention prepares low phenolic acid ginkgo leaf neutrality extract using ginko leaves flavone, lactones extract production and processing waste as raw material, both reduced resource waste, and promoted the research and utilization to such compound, and play a significant role to the effective exploitation of gingko resource.
Description
Technical field
The present invention relates to a kind of preparation methods of low phenolic acid ginkgo leaf neutrality extract, belong to biomedicine technical field.
Background technique
Ginkgo is one of tree species most ancient in the world, and China is ginkgo plantation big country, and gingko resource accounts for about the world
70%, recent domestic a large amount of research carried out to ginkgo biloba p.e, ginkgo class drug also increasingly becomes the world
The natural active drug of upper popular treatment cardiovascular and cerebrovascular disease.
Flavones, lactones etc. were concentrated mainly on to the research of active skull cap components in ginkgo leaf in the past, and to ginkgo leaf
The research of the compounds such as neutral extract such as hydro carbons, polypenthylene alcohols, terpene alcohols, sterols is seldom, it was found that ginkgo
Contain a certain amount of ginkgo leaf neutrality extract and ginkgoic acid in leaf flavones, lactones extract production and processing waste, wherein silver
Apricot leaf neutrality extract also has obvious therapeutic action to diseases such as hypertension, hyperlipidemia, diabetes, and since ginkgoic acid may
It causes allergic reaction, the content in ginkgo biloba p.e is that a main control in international quality control index refers to
Mark.
Therefore, it is necessary to a kind of extracted from ginkgo leaf low ginkgo acid content neutral extract production method, thus will
Ginko leaves flavone, lactones extract production and processing waste are fully used, and are both reduced resource waste, and are promoted to such change
The research and utilization of object is closed, and is played a significant role to the effective exploitation of gingko resource.
Summary of the invention
It is raw using ginko leaves flavone, lactones extract that in view of the above existing problems in the prior art, the present invention provides a kind of
Produce the preparation method that processing waste produces low phenolic acid ginkgo leaf neutrality extract.
To achieve the above object, the present invention adopts the following technical scheme:
A kind of preparation method of low phenolic acid ginkgo leaf neutrality extract, comprising the following steps:
(1) waste powder is extracted as raw material using dry ginkgo leaf, organic solvent is added, is repeated by soxhlet extraction
Refluxing extraction 3 times；Combined extract recycles organic solvent, obtains extraction ointment；
(2) aqueous slkali is added in the extraction ointment in step (1) and carries out saponification, obtain saponification liquor；
(3) organic solvent is added in the saponification liquor in step (2) to extract 3 times, collects upper solution and merges, revolving is gone
Except organic solvent, low phenolic acid ginkgo leaf neutrality extract is obtained.
As the further improvement of invention, the partial size that the ginkgo leaf extracts waste powder is less than 5mm.
As the further improvement of invention, organic solvent used is n-hexane in step (1) or boiling range is 90~120 DEG C
Petroleum ether or ethyl acetate or acetone, it is 1kg:3 that the ginkgo leaf, which extracts waste powder and the w/v of organic solvent,
~8L, the Extracting temperature are as follows: 40~65 DEG C, each extraction time is 1~10h.
As the further improvement of invention, organic solvent used is n-hexane in step (1) or boiling range is 90~120 DEG C
Petroleum ether, it is 1kg:6L, the Extracting temperature that the ginkgo leaf, which extracts waste powder and the w/v of organic solvent, are as follows:
60 DEG C, each extraction time is 4h.
As the further improvement of invention, sodium hydroxide or potassium hydroxide that aqueous slkali described in step (2) is 5~20%
Methanol or ethanol solution, the w/v of ointment and aqueous slkali is 1kg:7.5~10L, the saponification condition are as follows: 55
Circumfluence distillation at~65 DEG C, mixing speed are 350~500r/min, and the time is 4~6h.
As the further improvement of invention, organic solvent used is n-hexane in step (3) or boiling range is 90~120 DEG C
The volume ratio of petroleum ether, saponification liquor and organic solvent is 1:2~4.
As the further improvement of invention, step rotates condition described in (3) are as follows: vacuum degree -0.1MPa, bath temperature 45
~50 DEG C.
The present invention has the advantage that compared with prior art
The present invention is first prepared using Soxhlet extraction using ginko leaves flavone, lactones extract production and processing waste as raw material
Ginkgo leaf extracts ointment, separates ginkgoic acid from extraction ointment further through saponification, will finally by multiple extraction
Ginkgo leaf neutrality extract extracts, and has not only effectively obtained the neutral extract in discarded ginkgo leaf, but also reduce neutrality and mention
The content of object middle acid substance is taken, to reduce resource waste, promotes the research and utilization to ginkgo leaf neutrality extract.
Specific embodiment
Below with reference to embodiment, the present invention will be described in detail, it is to be understood that protection scope of the present invention is not had
The limitation of body embodiment.
Screen influence of 1 solid-liquid ratio of embodiment to ointment is extracted
The ginkgo leaf for taking partial size 5mm or less dry extracts waste powder 5kg, and the petroleum that boiling range is 90~120 DEG C is added
Ether 15L, by soxhlet extraction, refluxing extraction 6h, repetition are extracted 3 times at 60 DEG C；Combined extract recycles organic solvent, obtains
Extract ointment.
To screen the weighing body for the petroleum ether that ginkgo leaf in embodiment 1 extracts waste powder and boiling range is 90~120 DEG C
Product ratio is screening embodiment 11, in the case where keeping other reaction conditions constant, is continued in fact for the w/v of feed liquid
Applied screening embodiment 12 to screening embodiment 15, and investigated it is each screening embodiment under gained extract ointment weight and extraction
The content of polypentenol in ointment, the results are shown in Table 1:
Influence of 1 solid-liquid ratio of table to ointment amount is extracted
As can be seen from Table 1, with the increase of solid-liquid ratio, gained ginkgo leaf extracts the weight of ointment and extracts in ointment
The content of polypentenol also gradually increases, and increases both when solid-liquid ratio is 1kg:6L and slows down and tend towards stability, therefore can select
Taking screening embodiment 14 is extraction stage best solid-liquid ratio.
Screen influence of 2 solvent of embodiment to ointment is extracted
The ginkgo leaf for taking partial size 5mm or less dry extracts waste powder 5kg, and the petroleum that boiling range is 90~120 DEG C is added
Ether 30L, by soxhlet extraction, refluxing extraction 6h, repetition are extracted 3 times at 60 DEG C；Combined extract recycles organic solvent, obtains
Extract ointment.
To screen organic solvent used in embodiment 2 as screening embodiment 21, keeping other reaction conditions constant
In the case of, continue to implement screening embodiment 22 to screening embodiment 24 for organic solvent, and investigated each screening embodiment
Lower gained extracts the weight of ointment and extracts the content of polypentenol in ointment, and the results are shown in Table 2:
Influence of 2 solvent of table to ointment amount is extracted
As can be seen from Table 2, the amount of ointment is extracted using ginkgo leaf obtained by different Extraction solvents and extract in ointment
The content of polypentenol is different, and when Extraction solvent is the petroleum ether or n-hexane that boiling range is 90~120 DEG C, ginkgo leaf extracts soft
The amount of cream and the content for extracting polypentenol in ointment are relatively high, therefore can choose screening embodiment 21 or screening implementation
Example 22 is extraction stage optimum solvent.
Screen influence of 3 temperature of embodiment to ointment is extracted
The ginkgo leaf for taking partial size 5mm or less dry extracts waste powder 5kg, and the petroleum that boiling range is 90~120 DEG C is added
Ether 30L, by soxhlet extraction, refluxing extraction 6h, repetition are extracted 3 times at 40 DEG C；Combined extract recycles organic solvent, obtains
Extract ointment.
To screen, reflux temperature is screening embodiment 31 in embodiment 3, in the case where keeping other reaction conditions constant,
Continue to implement screening embodiment 32 to screening for reflux temperature
Embodiment 36, and investigated gained under each screening embodiment and extracted the weight of ointment and the content of polypentenol, knot
Fruit is as shown in table 3:
Influence of 3 temperature of table to ointment is extracted
As can be seen from Table 3, ginkgo leaf extracts the amount of ointment and the content of polypentenol is gradually increased with temperature rising,
It when temperature is 60 DEG C, then improves temperature and both change less, therefore can choose and screen embodiment 35 as the extraction stage most
Good temperature.
Screen influence of 4 time of embodiment to ointment is extracted
The ginkgo leaf for taking partial size 5mm or less dry extracts waste powder 5kg, and the petroleum that boiling range is 90~120 DEG C is added
Ether 30L, by soxhlet extraction, refluxing extraction 2h, repetition are extracted 3 times at 60 DEG C；Combined extract recycles organic solvent, obtains
Extract ointment.
It is screening embodiment 41 to screen extraction time in embodiment 4, in the case where keeping other reaction conditions constant,
Continue to implement screening embodiment 42 to screening for extraction time
Embodiment 45, and investigated gained under each screening embodiment and extracted the weight of ointment and the content of polypentenol, knot
Fruit is as shown in table 4:
Influence of 4 time of table to ointment is extracted
As can be seen from Table 4, before 4h, ginkgo leaf extracts the amount of ointment and the content of polypentenol with extraction time
Increase gradually increase, and after 4h, ginkgo leaf extracts the amount of ointment and the content of polypentenol no longer becomes substantially
Change, therefore can choose screening embodiment 42 is extraction stage Best Times.
Embodiment 1
A kind of preparation method of low phenolic acid ginkgo leaf neutrality extract, comprising the following steps:
(1) ginkgo leaf for taking partial size 5mm or less dry extracts waste powder 5kg, and the stone that boiling range is 90~120 DEG C is added
Oily ether 30L, by soxhlet extraction, refluxing extraction 4h, repetition are extracted 3 times at 60 DEG C；Combined extract recycles organic solvent,
Ointment 260g must be extracted；
(2) sodium hydrate methanol solution that 2L10% is added in ointment is extracted in step (1), heat is returned at 60~65 DEG C
Stream saponification, mixing speed are 350~500r/min, and time 6h obtains saponification liquor；
(3) petroleum ether extraction 3 times that 5L boiling range is 90~120 DEG C are added in the saponification liquor in step (2), collect upper layer
Solution simultaneously merges, and boils off in 0.1MPa, 45~50 DEG C of water-bath backspins except organic solvent, obtains low phenolic acid ginkgo leaf neutrality extract.
Embodiment 2
A kind of preparation method of low phenolic acid ginkgo leaf neutrality extract, comprising the following steps:
(1) ginkgo leaf for taking partial size 5mm or less dry extracts waste powder 5kg, and the stone that boiling range is 90~120 DEG C is added
Oily ether 30L, by soxhlet extraction, refluxing extraction 4h, repetition are extracted 3 times at 60 DEG C；Combined extract recycles organic solvent,
Ointment 260g must be extracted；
(2) potassium hydroxide-ethanol solution that 2L 10% is added in ointment, heat at 60~65 DEG C are extracted in step (1)
Reflux saponification, mixing speed are 350~500r/min, and time 6h obtains saponification liquor；
(3) petroleum ether extraction 3 times that 5L boiling range is 90~120 DEG C are added in the saponification liquor in step (2), collect upper layer
Solution simultaneously merges, and boils off in 0.1MPa, 45~50 DEG C of water-bath backspins except organic solvent, obtains low phenolic acid ginkgo leaf neutrality extract.
Embodiment 3
A kind of preparation method of low phenolic acid ginkgo leaf neutrality extract, comprising the following steps:
(1) ginkgo leaf for taking partial size 5mm or less dry extracts waste powder 5kg, and the stone that boiling range is 90~120 DEG C is added
Oily ether 30L, by soxhlet extraction, refluxing extraction 4h, repetition are extracted 3 times at 60 DEG C；Combined extract recycles organic solvent,
Ointment 260g must be extracted；
(2) potassium hydroxide-ethanol solution that 2L 20% is added in ointment, heat at 60~65 DEG C are extracted in step (1)
Reflux saponification, mixing speed are 350~500r/min, and time 6h obtains saponification liquor；
(3) petroleum ether extraction 3 times that 5L boiling range is 90~120 DEG C are added in the saponification liquor in step (2), collect upper layer
Solution simultaneously merges, and boils off in 0.1MPa, 45~50 DEG C of water-bath backspins except organic solvent, obtains low phenolic acid ginkgo leaf neutrality extract.
Embodiment 4
A kind of preparation method of low phenolic acid ginkgo leaf neutrality extract, comprising the following steps:
(1) ginkgo leaf for taking partial size 5mm or less dry extracts waste powder 5kg, and the stone that boiling range is 90~120 DEG C is added
Oily ether 30L, by soxhlet extraction, refluxing extraction 4h, repetition are extracted 3 times at 60 DEG C；Combined extract recycles organic solvent,
Ointment 260g must be extracted；
(2) potassium hydroxide-ethanol solution that 2.5L 20% is added in ointment is extracted in step (1), at 60~65 DEG C
Heat reflux saponification, mixing speed is 350~500r/min, and time 6h obtains saponification liquor；
(3) petroleum ether extraction 3 times that 5L boiling range is 90~120 DEG C are added in the saponification liquor in step (2), collect upper layer
Solution simultaneously merges, and boils off in 0.1MPa, 45~50 DEG C of water-bath backspins except organic solvent, obtains low phenolic acid ginkgo leaf neutrality extract.
Embodiment 5
A kind of preparation method of low phenolic acid ginkgo leaf neutrality extract, comprising the following steps:
(1) ginkgo leaf for taking partial size 5mm or less dry extracts waste powder 5kg, and the stone that boiling range is 90~120 DEG C is added
Oily ether 30L, by soxhlet extraction, refluxing extraction 4h, repetition are extracted 3 times at 60 DEG C；Combined extract recycles organic solvent,
Ointment 260g must be extracted；
(2) potassium hydroxide-ethanol solution that 2L 20% is added in ointment, heat at 50~55 DEG C are extracted in step (1)
Reflux saponification, mixing speed are 350~500r/min, and time 6h obtains saponification liquor；
(3) petroleum ether extraction 3 times that 5L boiling range is 90~120 DEG C are added in the saponification liquor in step (2), collect upper layer
Solution simultaneously merges, and boils off in 0.1MPa, 45~50 DEG C of water-bath backspins except organic solvent, obtains low phenolic acid ginkgo leaf neutrality extract.
Embodiment 6
A kind of preparation method of low phenolic acid ginkgo leaf neutrality extract, comprising the following steps:
(1) ginkgo leaf for taking partial size 5mm or less dry extracts waste powder 5kg, and the stone that boiling range is 90~120 DEG C is added
Oily ether 30L, by soxhlet extraction, refluxing extraction 4h, repetition are extracted 3 times at 60 DEG C；Combined extract recycles organic solvent,
Ointment 260g must be extracted；
(2) potassium hydroxide-ethanol solution that 2L 20% is added in ointment, heat at 60~65 DEG C are extracted in step (1)
Reflux saponification, mixing speed are 350~500r/min, and time 4h obtains saponification liquor；
(3) petroleum ether extraction 3 times that 5L boiling range is 90~120 DEG C are added in the saponification liquor in step (2), collect upper layer
Solution simultaneously merges, and boils off in 0.1MPa, 45~50 DEG C of water-bath backspins except organic solvent, obtains low phenolic acid ginkgo leaf neutrality extract.
Embodiment 7
A kind of preparation method of low phenolic acid ginkgo leaf neutrality extract, comprising the following steps:
(1) ginkgo leaf for taking partial size 5mm or less dry extracts waste powder 5kg, and the stone that boiling range is 90~120 DEG C is added
Oily ether 30L, by soxhlet extraction, refluxing extraction 4h, repetition are extracted 3 times at 60 DEG C；Combined extract recycles organic solvent,
Ointment 260g must be extracted；
(2) potassium hydroxide-ethanol solution that 2L 20% is added in ointment, heat at 60~65 DEG C are extracted in step (1)
Reflux saponification, mixing speed are 350~500r/min, and time 6h obtains saponification liquor；
(3) it is added 5L n-hexane extraction 3 times, collects upper solution and merges in the saponification liquor in step (2), in
0.1MPa, 45~50 DEG C of water-bath backspins are boiled off except organic solvent, obtain low phenolic acid ginkgo leaf neutrality extract.
Embodiment 8
A kind of preparation method of low phenolic acid ginkgo leaf neutrality extract, comprising the following steps:
(1) ginkgo leaf for taking partial size 5mm or less dry extracts waste powder 5kg, and the stone that boiling range is 90~120 DEG C is added
Oily ether 30L, by soxhlet extraction, refluxing extraction 4h, repetition are extracted 3 times at 60 DEG C；Combined extract recycles organic solvent,
Ointment 260g must be extracted；
(2) potassium hydroxide-ethanol solution that 2L 20% is added in ointment, heat at 60~65 DEG C are extracted in step (1)
Reflux saponification, mixing speed are 350~500r/min, and time 6h obtains saponification liquor；
(3) petroleum ether extraction 3 times that 8L boiling range is 90~120 DEG C are added in the saponification liquor in step (2), collect upper layer
Solution simultaneously merges, and boils off in 0.1MPa, 45~50 DEG C of water-bath backspins except organic solvent, obtains low phenolic acid ginkgo leaf neutrality extract.
It weighs respectively to the ginkgo leaf neutrality extract that embodiment 1 is prepared into embodiment 8；Weigh 2.0mg neutrality
Extract is dissolved in 10mL methanol, using the content of total ginkgoic acid in high effective liquid chromatography for measuring ginkgo leaf neutrality extract,
Testing conditions are as follows: C8 chromatographic column (200mm × 4.6mm, 5 μm)；Mobile phase, which is that -0.4% phosphoric acid of methanol (55:45, v:v) is isocratic, washes
It is de-；Flow velocity 1.0mL/min；Column temperature is room temperature；Detect wave 360nm, 10 μ L of sample volume).The results are shown in Table 5:
Ginkgo leaf neutrality extract situation prepared by each embodiment of table 5
It is neutral to can be seen that the low phenolic acid ginkgo leaf prepared using the embodiment of the present invention 1 to embodiment 8 according to 5 result of table
Extract, for recovery rate 3.2% or so, the content of total ginkgoic acid is respectively less than 5ppm, meets the regulation in " Chinese Pharmacopoeia ".And
Comparative example 1 is to each parameter of embodiment 8 it is found that the concentration and dosage of aqueous slkali have an impact to the content of total ginkgoic acid, extraction
The dosage of agent then influences the recovery rate of neutral extract, but generally, and the ginkgo leaf using preparation of the embodiment of the present invention is neutral
The content of the recovery rate of extract and wherein total ginkgoic acid is floated little.
It is obvious to a person skilled in the art that invention is not limited to the details of the above exemplary embodiments, Er Qie
In the case where without departing substantially from spirit and essential characteristics of the invention, the present invention can be realized in other specific forms.Therefore, no matter
From the point of view of which point, the present embodiments are to be considered as illustrative and not restrictive, and the scope of the present invention is by appended power
Benefit requires rather than above description limits, it is intended that all by what is fallen within the meaning and scope of the equivalent elements of the claims
Variation is included within the present invention.In addition, it should be understood that although this specification is described in terms of embodiments, but it is not each
Embodiment only contains an independent technical solution, this narrating mode of specification only for clarity, this field skill
Art personnel should consider the specification as a whole, and the technical solutions in the various embodiments may also be suitably combined, form ability
The other embodiments that field technique personnel are understood that.
Claims (7)
1. a kind of preparation method of low phenolic acid ginkgo leaf neutrality extract, which comprises the following steps:
(1) waste powder is extracted as raw material using dry ginkgo leaf, organic solvent is added, repeat to flow back by soxhlet extraction
It extracts 3 times；Combined extract recycles organic solvent, obtains extraction ointment；
(2) aqueous slkali is added in the extraction ointment in step (1) and carries out saponification, obtain saponification liquor；
(3) organic solvent is added in the saponification liquor in step (2) to extract 3 times, collects upper solution and merges, revolving removal has
Solvent obtains low phenolic acid ginkgo leaf neutrality extract.
2. a kind of preparation method of low phenolic acid ginkgo leaf neutrality extract according to claim 1, which is characterized in that described
The partial size that ginkgo leaf extracts waste powder is less than 5mm.
3. a kind of preparation method of low phenolic acid ginkgo leaf neutrality extract according to claim 1, which is characterized in that step
(1) petroleum ether or ethyl acetate or acetone that organic solvent used is n-hexane in or boiling range is 90~120 DEG C, the ginkgo leaf
The w/v for extracting waste powder and organic solvent is 1kg:3~8L, the Extracting temperature are as follows: 45~65 DEG C, every time
Extraction time is 1~10h.
4. a kind of preparation method of low phenolic acid ginkgo leaf neutrality extract according to claim 3, which is characterized in that step
(1) petroleum ether that organic solvent used is n-hexane in or boiling range is 90~120 DEG C, the ginkgo leaf extract waste powder with
The w/v of organic solvent is 1kg:6L, the Extracting temperature are as follows: 60 DEG C, each extraction time is 4h.
5. a kind of preparation method of low phenolic acid ginkgo leaf neutrality extract according to claim 1, which is characterized in that step
(2) methanol or ethanol solution of sodium hydroxide or potassium hydroxide that aqueous slkali described in is 10~20%, ointment and aqueous slkali
W/v is 1kg:7.5~10L, the saponification condition are as follows: circumfluence distillation at 55~65 DEG C, mixing speed are
350~500r/min, time are 4~6h.
6. a kind of preparation method of low phenolic acid ginkgo leaf neutrality extract according to claim 1, which is characterized in that step
(3) volume ratio of the petroleum ether that organic solvent used is n-hexane in or boiling range is 90~120 DEG C, saponification liquor and organic solvent is
1:2~4.
7. a kind of preparation method of low phenolic acid ginkgo leaf neutrality extract according to claim 1, which is characterized in that step
(3) condition is rotated described in are as follows: vacuum degree -0.1MPa, 45~50 DEG C of bath temperature.
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| CN110973360A (en) * | 2019-11-27 | 2020-04-10 | 江苏三仪动物营养科技有限公司 | Livestock and poultry feed additive and preparation method thereof |
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Application publication date: 20191008 |