CN107400150A - A kind of method that amarogentin is extracted from loquat seed - Google Patents
A kind of method that amarogentin is extracted from loquat seed Download PDFInfo
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- CN107400150A CN107400150A CN201710700305.7A CN201710700305A CN107400150A CN 107400150 A CN107400150 A CN 107400150A CN 201710700305 A CN201710700305 A CN 201710700305A CN 107400150 A CN107400150 A CN 107400150A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/18—Acyclic radicals, substituted by carbocyclic rings
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
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Abstract
The invention provides a kind of method that amarogentin is extracted from loquat seed, comprise the following steps：Loquat nut is chosen, is milled to powder, crosses 50 100 mesh；Fruit stone powder is sterilized, then mixed with ethanol solution, heating stirring；Ultrasonic wave extraction is carried out, centrifugate is distilled, reclaims ethanol, finally obtains extract solution；Maltose and dextrin are added in extract solution, is put into Trichoderma viride, tangerine ash mould, Aspergillus melleus and Rhizopus oryzae fermentation；Fermentate is freeze-dried；The fermentate that freeze-drying process is crossed crushes, grinding, obtains loquat yeast powder, adds n-butyl acetate extraction, freezing and crystallizing after extract filtering, crystal is vacuum dried, produces amarogentin.The present invention can improve extraction rate of amygdaloside and DNA purity.
Description
Technical field
The present invention relates to a kind of method that amarogentin is extracted from loquat.
Background technology
Amygdalin is primarily present in bitter apricot, almond, peach, nectarine (nectarines), loquat, plum, apple, black cherry
In the kernels and leaf such as peach, β-type glucosides that amygdalin is made up of gentiobiose and semen armeniacae amarae eyeball, molecular formula：
C20H27NO11, molecular weight：457.43.Its trihydrate crystallizes for rhombic prism shape, and 200 DEG C of fusing point, anhydride fusing point there are about 20
DEG C, [α]：20D-42℃.Amarogentin can specifically suppress blood glucose rise caused by alloxan, action intensity and semen armeniacae amarae in blood
Glycosides concentration is relevant.Amarogentin also has blood coagulation resisting function.
The content of the invention
The invention discloses a kind of method that amarogentin is extracted from loquat seed, recovery rate and DNA purity can be improved.
A kind of method that amarogentin is extracted from loquat seed, comprises the following steps：
（1）, choose loquat nut, be milled to powder, cross 50-100 mesh；
（2）, fruit stone powder sterilized, then mixed with the ethanol solution that mass concentration is 80%, it is quiet after heating stirring 10-15 minutes
Put half an hour；Ultrasonic wave extraction is carried out, extraction time is more than three times, each extraction time control is in 20-30 minutes, extraction temperature
Spend for 62-65 DEG C；Solution filtering, centrifugation after extraction, obtain centrifugate；Centrifugate is distilled under -0.1MPA pressure, returned
Ethanol is received, temperature is at 55-65 DEG C；Finally obtain extract solution；
（3）, maltose and dextrin, uniform stirring 5-8 minutes are added in extract solution；
（4）, be put into Trichoderma viride, tangerine ash mould, Aspergillus melleus and Rhizopus oryzae, ferment 2-3 days, must send out under the conditions of 42-44 DEG C
Ferment thing；
（5）, by fermentate at -35 DEG C precooling 1-2 hours, by the fermentate drying box after freezing, start to vacuumize, very
Reciprocal of duty cycle is in 15-20Pa, and temperature is between -20 to -15 DEG C, drying time 3-4h；Then, drying box temperature is brought up into 55-
60 DEG C, drying time 2h, dry vacuum degree is 10-15Pa；Finally, drying box temperature is reduced to 30-35 DEG C, drying time is
2h, dry vacuum degree are 15-20Pa, and freeze-drying terminates；
（6）, the fermentate of crossing freeze-drying process crush, grinding, obtain loquat yeast powder, add n-butyl acetate extraction, extraction
Take liquid filter after in -12 DEG C to -18 times freezing and crystallizing 16-18 hours, crystal is vacuum dried, produces amarogentin.
Further, described step（2）In, the weight ratio of ethanol solution and fruit stone powder is 6-8:1.
Further, described step（3）In, the weight ratio of maltose and dextrin is 0.2:1, and maltose and dextrin weight
Sum is the 8-9% of extract solution weight.
Further, the step of stating（3）In, Trichoderma viride, tangerine ash mould, the weight ratio of Aspergillus melleus and Rhizopus oryzae are 1:
0.6:0.5:0.1, and wheat Trichoderma viride, tangerine ash mould, Aspergillus melleus and Rhizopus oryzae weight sum are the 0.15- of extract solution weight
0.18%。
Further, described step（6）In, the weight ratio of n-butyl acetate and yeast powder is 8-12:1.
Amarogentin of the present invention mainly extracts from loquat nut, rather than the original that this cost of selection almond is higher
Material, from cost of material for, it is very cheap.
The present invention finally gives amarogentin by three extracting methods.Why select to extract three times, primarily to
Obtain more than 96% recovery rate and more than 99 purity.The selection of the present invention each parameter, ratio is also by true after many experiments
Fixed, its final purpose is to protect the maximization of main recovery rate and purity.
Embodiment
Embodiment 1
The method that amarogentin is extracted from loquat seed, step are as follows：
（1）, choose loquat nut, be milled to powder, cross 60 mesh；
（2）, fruit stone powder sterilized, then mix with the ethanol solution that mass concentration is 80%, the weight of ethanol solution and fruit stone powder
Than for 6.6:1, heating stirring stands half an hour after 12 minutes；Carry out ultrasonic wave extraction, extraction time is more than three times, every time
At 25 minutes, Extracting temperature was 64 DEG C for extraction time control；Solution filtering, centrifugation after extraction, obtain centrifugate；By centrifugate
Distilled under -0.1MPA pressure, reclaim ethanol, temperature is at 60 DEG C；Finally obtain extract solution；
（3）, to add weight ratio in extract solution be 0.2:1 maltose and dextrin, uniform stirring 8 minutes；Wherein, maltose and
Dextrin weight sum is the 8.3% of extract solution weight.
（4）, be put into weight ratio as 1:0.6:0.5:0.1 Trichoderma viride, tangerine ash mould, Aspergillus melleus and Rhizopus oryzae,
Fermented 3 days under the conditions of 43 DEG C, obtain fermentate；Wherein, wheat Trichoderma viride, tangerine ash mould, Aspergillus melleus and Rhizopus oryzae weight sum
For the 0.16% of extract solution weight.
（5）, by fermentate at -35 DEG C precooling 2 hours, by the fermentate drying box after freezing, start to vacuumize,
Vacuum is in 16Pa, and temperature is for -20 DEG C, drying time 4h；Then, drying box temperature is brought up to 60 DEG C, drying time
For 2h, dry vacuum degree is 15Pa；Finally, drying box temperature being reduced to 35 DEG C, drying time 2h, dry vacuum degree is 15Pa,
Freeze-drying terminates；
（6）, the fermentate of crossing freeze-drying process crush, grinding, obtain loquat yeast powder, add n-butyl acetate extraction, second
Sour N-butyl and the weight of yeast powder ratio are 9:1；Freezing and crystallizing 18 hours, crystal warp are true at -16 DEG C after extract filtering
Sky is dried, and produces amarogentin.
Extraction rate of amygdaloside is 96.2% in this implementation, and the purity that amarogentin white crystal is determined through HPLC is 99
.3%.
Embodiment 2
The method that amarogentin is extracted from loquat seed, step are as follows：
（1）, choose loquat nut, be milled to powder, cross 90 mesh；
（2）, fruit stone powder sterilized, then mix with the ethanol solution that mass concentration is 80%, the weight of ethanol solution and fruit stone powder
Than for 7:1, heating stirring stands half an hour after 13 minutes；Carry out ultrasonic wave extraction, extraction time is more than three times, carries every time
Taking time control, Extracting temperature was 64 DEG C at 26 minutes；Solution filtering, centrifugation after extraction, obtain centrifugate；By centrifugate-
Distilled under 0.1MPA pressure, reclaim ethanol, temperature is at 58 DEG C；Finally obtain extract solution；
（3）, to add weight ratio in extract solution be 0.2:1 maltose and dextrin, uniform stirring 7 minutes；Wherein, maltose and
Dextrin weight sum is the 8.6% of extract solution weight.
（4）, be put into weight ratio as 1:0.6:0.5:0.1 Trichoderma viride, tangerine ash mould, Aspergillus melleus and Rhizopus oryzae,
Fermented 2 days under the conditions of 42-44 DEG C, obtain fermentate；Wherein, wheat Trichoderma viride, tangerine ash mould, Aspergillus melleus and Rhizopus oryzae weight it
With 0.16% for extract solution weight.
（5）, by fermentate at -35 DEG C precooling 2 hours, by the fermentate drying box after freezing, start to vacuumize,
Vacuum is in 16Pa, and temperature is for -20 DEG C, drying time 4h；Then, drying box temperature is brought up to 60 DEG C, drying time
For 2h, dry vacuum degree is 15Pa；Finally, drying box temperature being reduced to 35 DEG C, drying time 2h, dry vacuum degree is 15Pa,
Freeze-drying terminates；
（6）, the fermentate of crossing freeze-drying process crush, grinding, obtain loquat yeast powder, add n-butyl acetate extraction, second
Sour N-butyl and the weight of yeast powder ratio are 10:1；Freezing and crystallizing 16 hours, crystal pass through at -13 DEG C after extract filtering
Vacuum drying, produces amarogentin.
Extraction rate of amygdaloside is 96.3% in this implementation, and the purity that amarogentin white crystal is determined through HPLC is 99
.4%.
Embodiment 3
The method that amarogentin is extracted from loquat seed, step are as follows：
（1）, choose loquat nut, be milled to powder, cross 80 mesh；
（2）, fruit stone powder sterilized, then mix with the ethanol solution that mass concentration is 80%, the weight of ethanol solution and fruit stone powder
Than for 7.3:1, heating stirring stands half an hour after 14 minutes；Carry out ultrasonic wave extraction, extraction time is more than three times, every time
At 22 minutes, Extracting temperature was 64 DEG C for extraction time control；Solution filtering, centrifugation after extraction, obtain centrifugate；By centrifugate
Distilled under -0.1MPA pressure, reclaim ethanol, temperature is at 62 DEG C；Finally obtain extract solution；
（3）, to add weight ratio in extract solution be 0.2:1 maltose and dextrin, uniform stirring 5-8 minutes；Wherein, maltose
With dextrin weight sum be extract solution weight 8.8%.
（4）, be put into weight ratio as 1:0.6:0.5:0.1 Trichoderma viride, tangerine ash mould, Aspergillus melleus and Rhizopus oryzae,
Fermented 2-3 days under the conditions of 42-44 DEG C, obtain fermentate；Wherein, wheat Trichoderma viride, tangerine ash mould, Aspergillus melleus and Rhizopus oryzae weight
Sum is the 0.15% of extract solution weight.
（5）, by fermentate at -35 DEG C precooling 2 hours, by the fermentate drying box after freezing, start to vacuumize,
Vacuum is in 16Pa, and temperature is for -20 DEG C, drying time 4h；Then, drying box temperature is brought up to 60 DEG C, drying time
For 2h, dry vacuum degree is 15Pa；Finally, drying box temperature being reduced to 35 DEG C, drying time 2h, dry vacuum degree is 15Pa,
Freeze-drying terminates；
（6）, the fermentate of crossing freeze-drying process crush, grinding, obtain loquat yeast powder, add n-butyl acetate extraction, second
Sour N-butyl and the weight of yeast powder ratio are 11:1；Freezing and crystallizing 16-18 hours, crystallization at -15 DEG C after extract filtering
Thing is vacuum dried, produces amarogentin.
Extraction rate of amygdaloside is 96.1% in this implementation, and the purity that amarogentin white crystal is determined through HPLC is 99
.3%.
Comparative example 1
It is in the difference of embodiment 1 in step（4）In, weight ratio is put into as 1:0.6 Trichoderma viride, tangerine ash mould, at 43 DEG C
Under the conditions of ferment 3 days, obtain fermentate；Wherein, wheat Trichoderma viride, 0.16% that tangerine ash mould weight sum is extract solution weight.
Extraction rate of amygdaloside is 93.8% in this implementation, and the purity that amarogentin white crystal is determined through HPLC is 99
.2%.
Comparative example 2
It is in the difference of embodiment 1 in step（4）In, weight ratio is put into as 1:0.1 Trichoderma viride, Rhizopus oryzae, at 43 DEG C
Under the conditions of ferment 3 days, obtain fermentate；Wherein, wheat Trichoderma viride, 0.16% that Rhizopus oryzae weight sum is extract solution weight.
Extraction rate of amygdaloside is 94.1% in this implementation, and the purity that amarogentin white crystal is determined through HPLC is 99
.2%.
Comparative example 3
It is in the difference of embodiment 1 in step（4）In, weight ratio is put into as 0.6:0.5:0.1 tangerine ash mould, Aspergillus melleus
And Rhizopus oryzae, fermented 3 days under the conditions of 43 DEG C, obtain fermentate；Wherein, tangerine ash mould, Aspergillus melleus and Rhizopus oryzae weight sum
For the 0.16% of extract solution weight.
Extraction rate of amygdaloside is 92.5% in this implementation, and the purity that amarogentin white crystal is determined through HPLC is 99
.1%.
The above described is only a preferred embodiment of the present invention, being not the limitation for making other forms to the present invention, appoint
What those skilled in the art changed or be modified as possibly also with the technology contents of the disclosure above equivalent variations etc.
Imitate embodiment.But it is every without departing from technical solution of the present invention content, the technical spirit according to the present invention is to above example institute
Any simple modification, equivalent variations and the remodeling made, still fall within the protection domain of technical solution of the present invention.
Claims (5)
- A kind of 1. method that amarogentin is extracted from loquat seed, it is characterised in that comprise the following steps：（1）, choose loquat nut, be milled to powder, cross 50-100 mesh；（2）, fruit stone powder sterilized, then mixed with the ethanol solution that mass concentration is 80%, it is quiet after heating stirring 10-15 minutes Put half an hour；Ultrasonic wave extraction is carried out, extraction time is more than three times, each extraction time control is in 20-30 minutes, extraction temperature Spend for 62-65 DEG C；Solution filtering, centrifugation after extraction, obtain centrifugate；Centrifugate is distilled under -0.1MPA pressure, returned Ethanol is received, temperature is at 55-65 DEG C；Finally obtain extract solution；（3）, maltose and dextrin, uniform stirring 5-8 minutes are added in extract solution；（4）, be put into Trichoderma viride, tangerine ash mould, Aspergillus melleus and Rhizopus oryzae, ferment 2-3 days, must send out under the conditions of 42-44 DEG C Ferment thing；（5）, by fermentate at -35 DEG C precooling 1-2 hours, by the fermentate drying box after freezing, start to vacuumize, very Reciprocal of duty cycle is in 15-20Pa, and temperature is between -20 to -15 DEG C, drying time 3-4h；Then, drying box temperature is brought up into 55- 60 DEG C, drying time 2h, dry vacuum degree is 10-15Pa；Finally, drying box temperature is reduced to 30-35 DEG C, drying time is 2h, dry vacuum degree are 15-20Pa, and freeze-drying terminates；（6）, the fermentate of crossing freeze-drying process crush, grinding, obtain loquat yeast powder, add n-butyl acetate extraction, extraction Take liquid filter after in -12 DEG C to -18 times freezing and crystallizing 16-18 hours, crystal is vacuum dried, produces amarogentin.
- A kind of 2. method that amarogentin is extracted from loquat according to claim 1, it is characterised in that described step （2）In, the weight ratio of ethanol solution and fruit stone powder is 6-8:1.
- A kind of 3. method that amarogentin is extracted from loquat according to claim 1, it is characterised in that described step （3）In, the weight ratio of maltose and dextrin is 0.2:1, and maltose and dextrin weight sum are the 8-9% of extract solution weight.
- A kind of 4. method that amarogentin is extracted from loquat according to claim 1, it is characterised in that the step of stating （3）In, Trichoderma viride, tangerine ash mould, the weight ratio of Aspergillus melleus and Rhizopus oryzae are 1:0.6:0.5:0.1, and wheat Trichoderma viride, Tangerine ash mould, Aspergillus melleus and Rhizopus oryzae weight sum are the 0.15-0.18% of extract solution weight.
- A kind of 5. method that amarogentin is extracted from loquat according to claim 1, it is characterised in that the step of stating （6）In, the weight ratio of n-butyl acetate and yeast powder is 8-12:1.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108178776A (en) * | 2017-12-22 | 2018-06-19 | 江南大学 | A kind of preparation of amarogentin enriched products |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1319431A (en) * | 2000-01-17 | 2001-10-31 | 丹羽耕三 | Method for improving antitumor activity of amarogentin substance and its composition and evaluation method |
| CN106509349A (en) * | 2016-10-31 | 2017-03-22 | 中国热带农业科学院热带作物品种资源研究所 | Decyanation method for cassava leaves |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108178776A (en) * | 2017-12-22 | 2018-06-19 | 江南大学 | A kind of preparation of amarogentin enriched products |
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