CN107325177A - A kind of method for improving Bufrudin extract yield - Google Patents
A kind of method for improving Bufrudin extract yield Download PDFInfo
- Publication number
- CN107325177A CN107325177A CN201710780504.3A CN201710780504A CN107325177A CN 107325177 A CN107325177 A CN 107325177A CN 201710780504 A CN201710780504 A CN 201710780504A CN 107325177 A CN107325177 A CN 107325177A
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- China
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- hiruto
- bufrudin
- homogenate
- minutes
- extract yield
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/81—Protease inhibitors
- C07K14/815—Protease inhibitors from leeches, e.g. hirudin, eglin
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Biophysics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Tropical Medicine & Parasitology (AREA)
- Extraction Or Liquid Replacement (AREA)
Abstract
The present invention relates to a kind of method for improving Bufrudin extract yield, category hirudin extractive technique field.It is characterized in：Stirred by present invention homogenate, dilution and stirring, ultrasonic wave, mix chemical treatment, determination of activity 5 steps up to standard so that the extraction activity of Bufrudin brings up to 70~80% from original 40~55%.Pass through the exclusive physical characteristic of ultrasonic wave, and the processing of ultrasonic drilling machine intermittent running, hiruto cell tissue broken wall or deformation can be promoted, the cell of hiruto body is destroyed, chemical solvent rapid osmotic is allowed into hiruto body cell, the active ingredients of hiruto is extracted more abundant.Extracting method is simple and convenient to operate, and treating capacity is big, and extract yield is high, and separation, refining effect are good, using safety, is avoided wasting hiruto raw material to greatest extent, cost is reduced, so as to add economic well-being of workers and staff.Solve it is existing crush homogenate after, directly by trichloroacetic acid chemical method purification efficiency it is low, waste raw material the problem of.
Description
Technical field
The present invention relates to a kind of method for improving Bufrudin extract yield, category leech extractive technique field.
Background technology
A kind of anti-coagulate active components extracted out of hiruto body, with thrombolysis, suppress platelet aggregation, reduction blood viscosity
And the effect of blood fat, receive significant attention and apply on clinical medicine in recent years.The anti-coagulate active components of hiruto are mainly used
Waited indefinitely cardiovascular and cerebrovascular disease in treatment cerebral thrombus, heart infarction, coronary heart disease, cerebral arteriovenous malformation and cerebral infarction, it has wide medical science should
With prospect and market prospects.After Bufrudin active component is extracted at present typically using tissue homogenate polypide, trichloroacetic acid, acetone
Extract, but extracted by existing method after crude extract etc. chemical method, determine the active not ideal of Bufrudin, after measured luxuriant and rich with fragrance ox
The extraction of values of leech element is only the 40-55% of tested hiruto polypide total amount, and most of Bufrudin is still deposited in the luxuriant and rich with fragrance ox after homogenate
In leech body, a large amount of wastes of hiruto raw material are caused.Therefore, a kind of extracting method is researched and developed simple, easy to operate, extract yield
Height, the extraction activity up to 70~80% of Bufrudin, avoids wasting hiruto raw material to greatest extent, good in economic efficiency carries
The method of high Bufrudin extract yield is extremely necessary.
The content of the invention
It is an object of the present invention to, separation, purification high there is provided a kind of extract yield for above-mentioned the deficiencies in the prior art
Effect is good, and extracting method is simple and convenient to operate, and treating capacity is big, the extraction activity up to 70~80% of Bufrudin, uses peace
Entirely, avoid wasting hiruto raw material to greatest extent, reduce cost, the side of the raising Bufrudin extract yield of increase economic well-being of workers and staff
Method.
The present invention is to realize above-mentioned purpose by the following technical solutions：
A kind of method for improving Bufrudin extract yield, it is characterised in that：It is completed as follows：
1）By the hiruto body filter paper suck dry moisture cleaned up, crushed with pulverizer, place into refiner and be homogenized, homogenate
2~4 times, 2~3 minutes every time, often it is homogenized and is once spaced 1~2 minute afterwards, flowing water cools in refiner, keep homogenate temperature
Between 20 DEG C~25 DEG C；
2）With normal saline dilution hiruto homogenate, saline volume is 4~8 times of hiruto homogenate, passes through mixer
Stir, mixing time 2~3 minutes；
3）The hiruto fragment mixed liquor diluted after stirring is put into ultrasonic drilling machine and continues micronized pulverization processing, ultrasonic work(
Rate is 200~900W, 20~25KHz of supersonic frequency, 15~30min of total sonication time；Every ultrasound is spaced 1~3 second after 1~3 second；
4）10% trichloroacetic acid is added in hiruto solution after ultrasound adjusts pH value to 2.5,70 DEG C of water-baths 20 minutes, during which fits
Work as stirring, 4 DEG C of 10000rpm collect supernatant after centrifuging 10 minutes；The NaOH that supernatant is risen with 1mol/ adjusts pH value to 7.0,4 DEG C
12000rpm is centrifuged 10 minutes and is collected supernatant；
5）The Bufrudin purified to ultrasound and by chemical method carries out determination of activity, and the extraction activity for determining Bufrudin reaches
To the 70~80% of hiruto body total amount.
The sound wave of ultrasonic drilling machine can produce high speed, strong cavitation effect and stirring action, its exclusive physical characteristic energy
Promote hiruto cell tissue broken wall or deformation, destroy the cell of hiruto body, allow chemical solvent rapid osmotic to hiruto body
In cell, the active ingredients of hiruto are made to extract more abundant, extraction time is short, and treating capacity is big；Greatly improve recovery rate.
Meanwhile, Extracting temperature is low, saves energy consumption；Wide adaptability, it is adaptable to which most species Chinese medicines and different kind organism composition are carried
Take.In addition, ultrasonic wave extraction decoction impurity is few, active ingredient is easily isolated, purified；Extraction process operating cost is low, comprehensive warp
Ji remarkable benefit；Operation is simple for ultrasonic drilling machine, plant maintenance and easy maintenance.
The beneficial effect of the present invention compared with prior art is：
The method of the raising Bufrudin extract yield, high speed, strong cavitation effect and stirring action are produced by ultrasonic wave,
The cell of hiruto body is destroyed, chemical solvent is penetrated into the cell of hiruto body, shortens extraction time, separates, purify.
By 5 steps of the invention, the extraction activity of Bufrudin is brought up to 70~80% by original 40~55%, substantially increase
Recovery rate.Extracting method is simple, easy to operate, safe ready, and treating capacity is big, avoids wasting hiruto raw material, drop to greatest extent
Low cost, adds economic well-being of workers and staff.Solve it is existing crush homogenate after, directly by trichloroacetic acid chemical method purification efficiency it is low,
The problem of wasting raw material.
Embodiment
A kind of method for improving Bufrudin extract yield, it is characterised in that：It is completed as follows：
1）It is homogenized after the hiruto body pulverizer cleaned up is crushed with refiner, homogenate 2~4 times, 2~3 minutes every time, often
Homogenate is once spaced 1~2 minute afterwards, and flowing water cools in refiner, keeps homogenate temperature between 20 DEG C~25 DEG C；
2）Hiruto fragment after homogenate is put into physiological saline and diluted, physiological saline is 4~8 times of hiruto homogenate,
10min is stirred by mixer；
3）The hiruto fragment mixed liquor diluted after stirring is put into ultrasonic drilling machine and continues break process, ultrasonic power is
200~900W, 20~25KHz of supersonic frequency, 15~30min of total sonication time；Every ultrasound is spaced 1~3 second after 1~3 second；
4）10% trichloroacetic acid is added in hiruto mixed liquor after ultrasound and adjusts pH value to 2.5,70 DEG C of water-baths 20 minutes, during which
Appropriate stirring, 4 DEG C of 10000rpm collect supernatant after centrifuging 10 minutes；The NaOH that supernatant is risen with 1mol/ adjusts pH value to 7.0,4
DEG C 12000rpm is centrifuged 10 minutes and is collected supernatant；
5）The Bufrudin purified with the hiruto liquid of ultrasonic wave pre-treatment using trichloroacetic acid chemical method carries out active survey
Fixed contrast, the extraction activity for determining the Bufrudin extracted after ultrasound reaches the 70~80% of hiruto body total amount.
The method of the raising Bufrudin extract yield uses ultrasonic drilling machine assistant chemical solvent, passes through 5 steps of the present invention
Suddenly Bufrudin active matter is extracted from hiruto body, simple to operate, security is good, treating capacity is big, yield is high, shortens extraction
Time, substantially increase recovery rate, make the extraction activity of Bufrudin by original 40~55% improve till now 70~80%,
Avoid wasting hiruto raw material to greatest extent, effectively reduce cost, add the economic well-being of workers and staff of enterprise, while being Chinese medicine system
Medicine market provides the raw produce of high-quality.
Simply presently preferred embodiments of the present invention described above, the example above illustrates that the substantive content not to the present invention is made
Any formal limitation, technology of the person of an ordinary skill in the technical field after this specification has been read according to the present invention
Any simple modification or deformation that essence is made to above embodiment, and possibly also with the technology contents of the disclosure above
The equivalent embodiment of equivalent variations is changed or is modified to, in the range of still falling within technical solution of the present invention, without departing from
The spirit and scope of the invention.
Claims (1)
1. a kind of method for improving Bufrudin extract yield, it is characterised in that：It is completed as follows：
1）It is homogenized after the hiruto body pulverizer cleaned up is crushed with refiner, homogenate 2~4 times, 2~3 minutes every time, often
Homogenate is once spaced 1~2 minute afterwards, and flowing water cools in refiner, keeps homogenate temperature between 20 DEG C~25 DEG C；
2）Hiruto fragment after homogenate is put into physiological saline and diluted, physiological saline is 4~8 times of hiruto homogenate,
10min is stirred by mixer；
3）The hiruto fragment mixed liquor diluted after stirring is put into ultrasonic drilling machine and continues break process, ultrasonic power is
200~900W, 20~25KHz of supersonic frequency, 15~30min of total sonication time；Every ultrasound is spaced 1~3 second after 1~3 second；
4）10% trichloroacetic acid is added in hiruto mixed liquor after ultrasound and adjusts pH value to 2.5,70 DEG C of water-baths 20 minutes, during which
Appropriate stirring, 4 DEG C of 10000rpm collect supernatant after centrifuging 10 minutes；The NaOH that supernatant is risen with 1mol/ adjusts pH value to 7.0,4
DEG C 12000rpm is centrifuged 10 minutes and is collected supernatant；
5）The Bufrudin purified with the hiruto liquid of ultrasonic wave pre-treatment using trichloroacetic acid chemical method carries out active survey
Fixed contrast, the extraction activity for determining the Bufrudin extracted after ultrasound reaches the 70~80% of hiruto body total amount.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710780504.3A CN107325177A (en) | 2017-09-01 | 2017-09-01 | A kind of method for improving Bufrudin extract yield |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710780504.3A CN107325177A (en) | 2017-09-01 | 2017-09-01 | A kind of method for improving Bufrudin extract yield |
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| CN107325177A true CN107325177A (en) | 2017-11-07 |
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| CN201710780504.3A Pending CN107325177A (en) | 2017-09-01 | 2017-09-01 | A kind of method for improving Bufrudin extract yield |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107661357A (en) * | 2017-11-16 | 2018-02-06 | 广西远程水蛭养殖有限公司 | Leech medicinal liquor preparation method |
| CN107875175A (en) * | 2017-11-16 | 2018-04-06 | 广西远程水蛭养殖有限公司 | Leech capsule |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2191023A1 (en) * | 1995-11-24 | 1997-05-25 | Jorg Moller | Process for the utilization of biological matrices comprising peptides or proteins |
| CN102286098A (en) * | 2011-08-18 | 2011-12-21 | 周维官 | Production method of natural hirudin |
| CN104774259A (en) * | 2015-04-21 | 2015-07-15 | 虞龙 | Method for extracting hirudin from leeches based on membrane |
| CN104926937A (en) * | 2015-06-19 | 2015-09-23 | 广西复鑫益生物科技有限公司平南分公司 | Method for extracting hirudin from leech saliva |
-
2017
- 2017-09-01 CN CN201710780504.3A patent/CN107325177A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2191023A1 (en) * | 1995-11-24 | 1997-05-25 | Jorg Moller | Process for the utilization of biological matrices comprising peptides or proteins |
| CN102286098A (en) * | 2011-08-18 | 2011-12-21 | 周维官 | Production method of natural hirudin |
| CN104774259A (en) * | 2015-04-21 | 2015-07-15 | 虞龙 | Method for extracting hirudin from leeches based on membrane |
| CN104926937A (en) * | 2015-06-19 | 2015-09-23 | 广西复鑫益生物科技有限公司平南分公司 | Method for extracting hirudin from leech saliva |
Non-Patent Citations (2)
| Title |
|---|
| ABDUALKADER ET AL.: ""Leech Therapeutic Applications"", 《INDIAN JOURNAL OF PHARMACEUTICAL SCIENCES》 * |
| 汪波 等: "菲牛蛭抗凝血活性物质提取方法研究", 《中国农学通报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107661357A (en) * | 2017-11-16 | 2018-02-06 | 广西远程水蛭养殖有限公司 | Leech medicinal liquor preparation method |
| CN107875175A (en) * | 2017-11-16 | 2018-04-06 | 广西远程水蛭养殖有限公司 | Leech capsule |
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| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information |
Inventor after: Li Jun Inventor after: Yu Xiang Inventor after: Gong Yuan Inventor before: Gong Yuan |
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Application publication date: 20171107 |
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