CN101332218B - Broken pollen extract and extracting method and use thereof - Google Patents
Broken pollen extract and extracting method and use thereof Download PDFInfo
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- CN101332218B CN101332218B CN 200710043270 CN200710043270A CN101332218B CN 101332218 B CN101332218 B CN 101332218B CN 200710043270 CN200710043270 CN 200710043270 CN 200710043270 A CN200710043270 A CN 200710043270A CN 101332218 B CN101332218 B CN 101332218B
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Abstract
The invention discloses an extract method of bee pollen powder, and the method includes the steps that the bee pollen powder is extracted by supercritical carbon dioxide and the part extracted supercritical is collected. The extract method is simple and easy to operate, and the prepared bee pollen powder extractive has better anti-inflammatory function as well as the efficacy of preventing and controlling the prostate diseases such as prostatitis and prostatic hyperplasia, etc.
Description
Technical field
The present invention relates to a kind of extract and method for distilling and application of pollen (processed by breaking wall).
Background technology
Pollen (pollen) is the male plant gametocyte.For example Pollen Helianthi is the male organ of multiplication of drying and ripening of feverfew Helianthi (Helianthus decapetalus Dari), and rape petal pollen is the male organ of multiplication of drying and ripening of crucifer Brassica campestris L (Bassica Campestros L).Existing research shows that pollen has estrogen-like effects, can be used for anti-prostatic hyperplasia, prostatitis etc. (Du Xu, Dou Qiulian, Li Chunyan etc. the pharmacodynamic study of Heilungkiang sunflower treated powder prostatic hyperplasia. Chinese Chinese medicine science and technology, 2002; 9 (2): 94～95; Wu Yanyin, Zhao Xiaoping, Lee pang. the progress of QIANLIEKANG treatment prostatic hyperplasia. the Chinese doctor of community, 2007,23 (2): 44).
Many discover pollen (processed by breaking wall) more not pollen (processed by breaking wall) have better curative effect, the wall-breaking method of existing pollen comprises: breaking cellular wall with enzyme method, machinery broken wall law, breaking cellular wall by temperature difference method etc. (Tang Wei, Zhang Xinghai. process for breaking wall of pollen progress, food and fermentation industries, 2003; 29 (2): 86～92).Yet better is the effective ingredient that will obtain in the pollen, to improve drug effect better.
Summary of the invention
The technical problem that the present invention will solve promptly provides a kind of extract and method for distilling thereof with pollen (processed by breaking wall) of better drug effect.
The inventor is surprised to find pollen (processed by breaking wall) through many tests, like the supercritical extract that pollen (processed by breaking wall) such as Helianthi, Brassica campestris L obtain behind supercritical extraction, has the effect of preferable antiinflammatory and anti-prostatic hyperplasia.
Therefore, the present invention solves the problems referred to above through following technical proposal.A kind of method for distilling of pollen (processed by breaking wall), it comprises pollen (processed by breaking wall) is adopted supercritical carbon dioxide extraction, collects the supercritical extraction part.
According to the present invention, the temperature of described supercritical carbon dioxide extraction is preferably at 35～55 ℃ of scopes, pressure 25～40MPa.
In order to improve the yield of extract, also adopt the extraction entrainer in the described supercritical carbon dioxide extraction.Described extraction entrainer can be polar organic solvent commonly used in the prior art, for example C such as methanol, various concentration ethanol, propanol, isopropyl alcohol, butanols
1～4Lower alcohol, acetone, ethyl acetate, chloroform etc.The preferred C of the present invention
1～4Lower alcohol, more preferably methanol and ethanol.
According to the present invention, the consumption of said entrainer is preferably pollen (processed by breaking wall) weight 5～10%.
With conventional, collection supercritical extraction part of the present invention can adopt isolating one or more levels separated and collected of decompression.For collecting fully also factors considerations such as composite technology time, cost, the temperature that preferred 25～45 ℃ of scopes of extract separation condition of the present invention are successively decreased, the decreasing pressure of 4～12Mpa as far as possible; More preferably secondary separates, and wherein the temperature of separation reactor I is 35～45 ℃, pressure 8～12MPa, and the temperature of separation reactor I I is 25～35 ℃, pressure 4～6Mpa.
Under the yield same case, along with CO in the supercritical carbon dioxide extraction
2The consumption that per hour circulates increases, and the extraction time can shorten, otherwise, CO in the supercritical carbon dioxide extraction
2The consumption that per hour circulates is more little, and the extraction time is long more.Factors such as the comprehensive yield of the present invention, process time, cost preferably adopt the CO of 25～45L/ hour circulation consumption
2Extracted about 1～3 hour.
The said pollen (processed by breaking wall) of the present invention is meant existing solid pollen (processed by breaking wall); It can commercially availablely obtain, also can adopt existing pollen wall breaking technology (as can reference: Zhang Xinghai etc. the microorganism wall breaking technology research of Pollen Brassicae campestris, Food Science; 2003,24 (3): 92～95; And publication number is the disclosed enzyme fermentation wall-breaking method of patent application document of CN1452888A) self-control obtain.By routine, pollen (processed by breaking wall) is carried out before the supercritical carbon dioxide extraction, as far as possible with pollen (processed by breaking wall), particularly moisture is removed in the commercially available prod, and is fully dry.
The preferred Helianthi pollen (processed by breaking wall) of the present invention, rape flower pollen (processed by breaking wall) are that example describes.
And a kind of pollen (processed by breaking wall) extract that the present invention will provide is made by the said extracted method, and the product merging that is about to two separating stills collections promptly gets pollen (processed by breaking wall) extract of the present invention.
The pollen (processed by breaking wall) extract that the present invention makes shows to have the effect of stronger antiinflammatory and anti-prostatic hyperplasia through animal experiment, can be prepared into antiinflammatory, and the medicine or the health food of prostatosis such as resistance prostatitis, prostatic hyperplasia.
The specific embodiment
Further specify the present invention with embodiment below, but the present invention is not limited.
The preparation of embodiment 1 Helianthi pollen (processed by breaking wall)
With reference to following document: Zhang Xinghai etc. the microorganism wall breaking technology research of Pollen Brassicae campestris, Food Science, 2003,24 (3): 92～95; And publication number is the patent application document of CN1452888A, and concrete steps are:
1. the preparation of culture medium: Semen Glycines is pulverized with pulverizer, add water by the part by weight of 1:10, steaming and decocting is 60～80 minutes in pressure cooker; Use filtered through gauze, or Semen Glycines is wrapped in steaming and decocting in the gauze, extract Semen Glycines juice; Press the 10g Semen Glycines, 3g sucrose, the ratio of 4g agar prepares culture medium;
2. sterilize, inoculate: the culture medium that will just prepare is divided in the culture dish while hot; Sterilization; Be cooled to room temperature; Inoculate test tube strains Aspergillus niger (Aspergillus3040sp.) and aspergillus oryzae (Aspergillus3514sp.) (available from institute of microbiology of Zhejiang University) respectively, put in 28～30 ℃ the constant incubator and cultivated 48 hours;
3. song preparation: adopt the Testa Tritici culture medium, according to the ratio of volume ratio 30%, the strain in the 2. middle culture dish of inoculation step fermented 2 days at 28～30 ℃ respectively, and after fermentation finished, 4000 leave the heart obtained aspergillus niger liquid and aspergillus oryzae liquid in 20 minutes respectively;
4. with above-mentioned aspergillus niger liquid and aspergillus oryzae liquid 2:1～3:1 constitutive enzyme liquid by volume; Exsiccant Pollen Helianthi and enzyme liquid is even according to the mixed of weight ratio 1:3; Put in the fermentation tank 37 ℃ of fermentations 24 hours; Treat pH value to 4.5, stop fermentation, the fermentation liquid convection drying is got Helianthi pollen (processed by breaking wall) 1.
5. fermentation liquid with 4000 rev/mins centrifugal 30 minutes, drying promptly gets Helianthi pollen (processed by breaking wall) 2.
The preparation of embodiment 2 Helianthi pollen (processed by breaking wall) SFE extract
The Helianthi pollen (processed by breaking wall) 1 that embodiment 1 is made is at 55 ℃; Extraction kettle pressure 30MPa; 45 ℃ of separation reactor I temperature, pressure 12MPa, 35 ℃ of separation reactor I I temperature, pressure 6MPa were with supercritical carbon dioxide (45L/ hour circulation consumption) extraction 1.5 hours; Obtain the supercritical extraction part, collect the supercritical extraction part.
The preparation of embodiment 3 Helianthi pollen (processed by breaking wall) SFE extract
The Helianthi pollen (processed by breaking wall) 2 that embodiment 1 is made is at 45 ℃; Extraction kettle pressure 25MPa, 35 ℃ of separation reactor I temperature, pressure 10MPa, 30 ℃ of separation reactor I I temperature, pressure 5MPa; With pollen weight 5% heavy methanol is entrainer; Carry out supercritical carbon dioxide (30L/ hour circulation consumption) extraction 2 hours, obtain the supercritical extraction part, collect the supercritical extraction part.
The preparation of embodiment 4 Helianthi pollen (processed by breaking wall) SFE extract
With 60 ℃ of drying under reduced pressure of commercially available Helianthi pollen (processed by breaking wall) 24 hours; Then with dried pollen (processed by breaking wall) at 40 ℃, extraction kettle pressure 40MPa, 35 ℃ of separation reactor I temperature, pressure 8MPa; 30 ℃ of separation reactor I I temperature, pressure 4MPa; With pollen weight 10% heavy 95% (v/v) ethanol is that entrainer carries out supercritical carbon dioxide (25L/ hour circulation consumption) extraction 2.5 hours, obtains the supercritical extraction part, collects the supercritical extraction part.
The preparation of embodiment 5 rape pollen with broken wall
With reference to following document: Zhang Xinghai etc. the microorganism wall breaking technology research of Pollen Brassicae campestris, Food Science, 2003,24 (3): 92～95; And publication number is the patent application document of CN1452888A, and concrete steps are:
1. the preparation of culture medium: Semen Glycines is pulverized with pulverizer, add water by the part by weight of 1:10, steaming and decocting is 60～80 minutes in pressure cooker; Use filtered through gauze, or Semen Glycines is wrapped in steaming and decocting in the gauze, extract Semen Glycines juice; Press the 10g Semen Glycines, 3g sucrose, the ratio of 4g agar prepares culture medium;
2. sterilize, inoculate: the culture medium that will just prepare is divided in the culture dish while hot; Sterilization; Be cooled to room temperature; Inoculate test tube strains Aspergillus niger (Aspergillus3040sp.) and aspergillus oryzae (Aspergillus3514sp.) (available from institute of microbiology of Zhejiang University) respectively, put in 28～30 ℃ the constant incubator and cultivated 48 hours;
3. song preparation: adopt the Testa Tritici culture medium, according to the ratio of volume ratio 30%, the strain in the 2. middle culture dish of inoculation step fermented 2 days at 28～30 ℃ respectively, and after fermentation finished, 4000 leave the heart obtained aspergillus niger liquid and aspergillus oryzae liquid in 20 minutes respectively;
4. with above-mentioned aspergillus niger liquid and aspergillus oryzae liquid 2:1～3:1 constitutive enzyme liquid by volume; Exsiccant Pollen Brassicae campestris and enzyme liquid is even according to the mixed of weight ratio 1:3; Put in the fermentation tank 37 ℃ of fermentations 24 hours; Treat pH value to 4.5, stop fermentation, the fermentation liquid convection drying is got rape pollen with broken wall 1.
5. fermentation liquid with 4000 rev/mins centrifugal 30 minutes, drying promptly gets rape pollen with broken wall 2.
The preparation of embodiment 6 extract of rape pollen with broken wall
The rape pollen with broken wall 1 that embodiment 5 is made is at 55 ℃; Extraction kettle pressure 30MPa; 45 ℃ of separation reactor I temperature, pressure 12MPa, 35 ℃ of separation reactor I I temperature, pressure 6MPa were with supercritical carbon dioxide (45L/ hour circulation consumption) extraction 1.5 hours; Obtain the supercritical extraction part, collect the supercritical extraction part.
The preparation of embodiment 7 extract of rape pollen with broken wall
The rape pollen with broken wall 2 that embodiment 5 is made is at 45 ℃; Extraction kettle pressure 25MPa, 40 ℃ of separation reactor I temperature, pressure 10MPa, 30 ℃ of separation reactor I I temperature, pressure 5MPa; With pollen weight 8% heavy acetone is entrainer; Carry out supercritical carbon dioxide (30L/ hour circulation consumption) extraction 2 hours, obtain the supercritical extraction part, collect the supercritical extraction part.
The preparation of embodiment 8 extract of rape pollen with broken wall
With 60 ℃ of drying under reduced pressure of commercially available rape pollen with broken wall 24 hours; Then with dried pollen (processed by breaking wall) at 35 ℃, extraction kettle pressure 40MPa, 35 ℃ of separation reactor I temperature, pressure 8MPa; 25 ℃ of separation reactor I I temperature, pressure 4MPa; With pollen weight 6% heavy isopropyl alcohol is that entrainer carries out supercritical carbon dioxide (25L/ hour circulation consumption) extraction 2.5 hours, obtains the supercritical extraction part, collects the supercritical extraction part.
Helianthi pollen (processed by breaking wall) in the foregoing description and not pollen (processed by breaking wall) be the product of strong rich first bio tech ltd in Beijing; Pollen Brassicae campestris and pollen (processed by breaking wall) are the product of Xuancheng City, Anhui Province hundred health apicultures.
The antiinflammatory action of test example 1 pollen (processed by breaking wall) extract of the present invention
The SD rat is by 10 every group of body weight random packet; Blank control group gives 0.4% tween 80; Test group gives corresponding test sample by the dosage of table 1; It is 0.12g/kg that positive group gives aspirin, and has the pollen medicine now: QIANLIEKANG (rape pollen with broken wall preparation) 0.5g/kg dosage and Prostat (naked barley pollen (processed by breaking wall) preparation) 0.015g/kg dosage is as contrast.Irritate the stomach volume and be the 10ml/kg body weight, successive administration 7 days, 1h measures the thickness of the left back sufficient sole of the foot of rat before the last administration with outside micrometer; The left back sufficient sole of the foot injection Ovum Gallus domesticus album 0.05ml/ pawl of 1h after the administration; Then behind the injection Ovum Gallus domesticus album 2,4,6 hours with outside micrometer each measures the once thickness of the left back sufficient sole of the foot, all at identical position, calculate the swelling degree of the left back sufficient sole of the foot of rat different time points after administration during measurement; Between its left back swelling degree of the paw organized relatively, the t check.
Swelling degree (mm)=left back sufficient sole of the foot thickness (behind the injection Ovum Gallus domesticus album)-left back sufficient sole of the foot thickness (before the injection Ovum Gallus domesticus album), the result is as shown in table 1 below.
Table 1 extract of the present invention causes the influence (X ± s) of rat paw edema degree to Ovum Gallus domesticus album
Compare with model group,
*P<0.05
*P<0.01
It is thus clear that, the antiinflammatory action of Helianthi pollen (processed by breaking wall) extract of the present invention and extract of rape pollen with broken wall, better like the existing antibiotic medicine aspirin commonly used of the curative effect of resistance prostatitis, also be superior to existing pollen (processed by breaking wall) preparation.
The anti-prostatic hyperplasia effect of test example 2 pollen (processed by breaking wall) extracts of the present invention
Get rat, get 9 at random and do pseudo-operation blank control group, other rats are under aseptic condition, and the pentobarbital sodium anesthesia with 2% is extractd bilateral testes, skin suture.Perform the operation after 1 week, will go the testis rat by the body weight random packet.Except that blank control group, all the other rats are pressed 0.1ml/200g body weight subcutaneous injection Testosterone Propionate every day, and dosage is 3mg/kg; Be total to 30d, test group and raw material matched group give corresponding embodiment or raw material test sample by the dosage of table 2, and positive component does not give QIANLIEKANG 0.5g/kg dosage and Prostat 0.015g/kg dosage; Blank control group and model group give isometric distilled water, all irritate stomach and give, and the administration volume is the 1.0ml/100g body weight; 1 time/d, successive administration 30d.2h puts to death animal after the last administration, cuts open and gets each leaf of prostate (abdomen, the back of the body+lateral lobe), absorbs surface liquid with filter paper, and electronic balance is claimed its weight in wet base, calculates each leaf ponderal index of prostate;
The result is as shown in table 2 below.
Table 2, extract of the present invention are induced the exponential influence of prostatic hyperplasia rat model prostate (X ± s) to Testosterone Propionate
Compare with model group,
*P<0.05
*P<0.01
It is thus clear that the anti-prostatic hyperplasia curative effect of pollen (processed by breaking wall) extract of the present invention is superior to existing pollen (processed by breaking wall) preparation, particularly common drug QIANLIEKANG and Prostat are better at present.
Claims (3)
1. the method for distilling of a pollen (processed by breaking wall) is characterized in that:
With Helianthi pollen (processed by breaking wall) 2 at 45 ℃; Extraction kettle pressure 25MPa, 35 ℃ of separation reactor I temperature, pressure 10MPa, 30 ℃ of separation reactor I I temperature, pressure 5MPa; With pollen weight 5% heavy methanol is entrainer; Carry out supercritical carbon dioxide extraction 2 hours with 30L/ hour circulation consumption, obtain the supercritical extraction part, collect the supercritical extraction part; Wherein, the concrete steps of described Helianthi pollen (processed by breaking wall) 2 preparations are:
1. the preparation of culture medium: Semen Glycines is pulverized with pulverizer, add water by 1: 10 part by weight, steaming and decocting is 60～80 minutes in pressure cooker; Use filtered through gauze, or Semen Glycines is wrapped in steaming and decocting in the gauze, extract Semen Glycines juice; Press the 10g Semen Glycines, 3g sucrose, the ratio of 4g agar prepares culture medium;
2. sterilize, inoculate: the culture medium that will just prepare is divided in the culture dish while hot; Sterilization; Be cooled to room temperature, inoculate test tube strains Aspergillus niger (Aspergillus 3040sp.) and aspergillus oryzae (Aspergillus 3514sp.) respectively, put in 28～30 ℃ the constant incubator and cultivated 48 hours;
3. song preparation: adopt the Testa Tritici culture medium, according to the ratio of volume ratio 30%, the strain in the 2. middle culture dish of inoculation step fermented 2 days at 28～30 ℃ respectively, and after fermentation finished, 4000 leave the heart obtained aspergillus niger liquid and aspergillus oryzae liquid in 20 minutes respectively;
4. with above-mentioned aspergillus niger liquid and 2: 1 by volume～3: 1 constitutive enzyme liquid of aspergillus oryzae liquid; Exsiccant Pollen Helianthi and enzyme liquid is even according to 1: 3 mixed of weight ratio; Put in the fermentation tank 37 ℃ of fermentations 24 hours; Treat pH value to 4.5, stop fermentation, the fermentation liquid convection drying is got Helianthi pollen (processed by breaking wall) 1;
5. fermentation liquid with 4000 rev/mins centrifugal 30 minutes, drying promptly gets Helianthi pollen (processed by breaking wall) 2;
Perhaps, with rape pollen with broken wall 2 at 45 ℃, extraction kettle pressure 25MPa; 40 ℃ of separation reactor I temperature, pressure 10MPa; 30 ℃ of separation reactor I I temperature, pressure 5MPa are entrainer with pollen weight 8% heavy acetone, carry out supercritical carbon dioxide extraction 2 hours with 30L/ hour circulation consumption; Obtain the supercritical extraction part, collect the supercritical extraction part; Wherein, the concrete steps of described rape pollen with broken wall 2 preparations are:
1. the preparation of culture medium: Semen Glycines is pulverized with pulverizer, add water by 1: 10 part by weight, steaming and decocting is 60～80 minutes in pressure cooker; Use filtered through gauze, or Semen Glycines is wrapped in steaming and decocting in the gauze, extract Semen Glycines juice; Press the 10g Semen Glycines, 3g sucrose, the ratio of 4g agar prepares culture medium;
2. sterilize, inoculate: the culture medium that will just prepare is divided in the culture dish while hot; Sterilization; Be cooled to room temperature, inoculate test tube strains Aspergillus niger (Aspergillus 3040sp.) and aspergillus oryzae (Aspergillus 3514sp.) respectively, put in 28～30 ℃ the constant incubator and cultivated 48 hours;
3. song preparation: adopt the Testa Tritici culture medium, according to the ratio of volume ratio 30%, the strain in the 2. middle culture dish of inoculation step fermented 2 days at 28～30 ℃ respectively, and after fermentation finished, 4000 leave the heart obtained aspergillus niger liquid and aspergillus oryzae liquid in 20 minutes respectively;
4. with above-mentioned aspergillus niger liquid and 2: 1 by volume～3: 1 constitutive enzyme liquid of aspergillus oryzae liquid; Exsiccant Pollen Brassicae campestris and enzyme liquid is even according to 1: 3 mixed of weight ratio; Put in the fermentation tank 37 ℃ of fermentations 24 hours; Treat pH value to 4.5, stop fermentation, the fermentation liquid convection drying is got rape pollen with broken wall 1;
5. fermentation liquid with 4000 rev/mins centrifugal 30 minutes, drying promptly gets rape pollen with broken wall 2.
2. the pollen (processed by breaking wall) extract that method for distilling as claimed in claim 1 makes.
3. pollen (processed by breaking wall) extract as claimed in claim 2 is in the medicine of preparation control prostatic hyperplasia or the application in the health food.
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| CN101856116B (en) * | 2009-04-10 | 2014-05-07 | 武汉小蜜蜂农产品加工工程技术研究有限公司 | Application of coumaroyl spermidine type compounds or plant extracts thereof |
| CN101856346B (en) * | 2009-04-10 | 2012-01-25 | 上海医药工业研究院 | Medical application of coumaroyl spermidine type compounds or plant extracts thereof |
| CN102178232B (en) * | 2011-04-21 | 2012-11-28 | 青海花宝蜂业股份合作公司 | Method for preparing rape pollen fatty acid soft capsules |
| CN102511786A (en) * | 2011-11-24 | 2012-06-27 | 郭强 | CO2 supercritical extraction method of sunflower disc nutrition ingredients |
| CN102671420A (en) * | 2012-05-17 | 2012-09-19 | 杭州天厨蜜源保健品有限公司 | Pollen extract production technology |
| CN102948496B (en) * | 2012-10-26 | 2014-06-11 | 北京知蜂堂蜂产品有限公司 | Method for pollen wall breaking by enzyme and extraction of pollen grease composition by supercritical CO2 |
| CN105520964A (en) * | 2016-01-30 | 2016-04-27 | 合肥华方医药科技有限公司 | Preparation method and application of royal jelly and bee pollen composition |
| CN108029918A (en) * | 2017-12-27 | 2018-05-15 | 浙江亚林生物科技股份有限公司 | One kind is rich in flavone compound solid beverage and preparation method thereof |
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