CN105669877B - A kind of high-purity glucomannans preparation method - Google Patents
A kind of high-purity glucomannans preparation method Download PDFInfo
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- C08B37/0087—Glucomannans or galactomannans; Tara or tara gum, i.e. D-mannose and D-galactose units, e.g. from Cesalpinia spinosa; Tamarind gum, i.e. D-galactose, D-glucose and D-xylose units, e.g. from Tamarindus indica; Gum Arabic, i.e. L-arabinose, L-rhamnose, D-galactose and D-glucuronic acid units, e.g. from Acacia Senegal or Acacia Seyal; Derivatives thereof
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Abstract
The invention discloses a kind of preparation method of high-purity glucomannans, key step is as follows：From natural konjac polysaccharide or bletilla polysaccharide or its hydrolysate, choose the homogeneous polysaccharide raw material of 10~1000kD of molecular weight ranges, it is formulated as 0.5% 10.0% solution, filter aid processing is added, filtering uses the ultrafiltration membrane concentration of 100~1000nm micro-filtrate membrane filtrations, 2~100kD of molecular cut off successively, then concentrated, it is dry, obtain high-purity glucomannans or its liquid preparation that purity is more than 90%.Preparation process of the present invention is succinct, controllable, amplification production easy to implement.High, the related content of material of glucomannans purity obtained is low, can meet the quality standard of Medicinal crude drug, thus can be applied to prepare the related curable product such as drug and pharmaceutic adjuvant, medical instrument.
Description
Technical field
The present invention relates to a kind of glucomannans extracts by being extracted in natural plants, and present invention simultaneously relates to the Portugals
Application of the mannosan extract in preparing drug and curable product belongs to biomedicine field.
Technical background
Glucomannans is a kind of common natural stickiness polysaccharide, and main chain is that D-MANNOSE and the Portugals D- glucose form, one
As modified by 1-4 glucosides key connections of β, and with part branched structure and acetyl group.Such polysaccharide is mainly sent out in plant
It is existing, such as konjaku, aloe, bletilla (also known as bletilla striata, Bletilla striata) etc., in wholefood, cosmetics, medical supplies
Equal fields are widely used, and have notable economic value.
Bletilla polysaccharide is the main effective ingredient in traditional Chinese medicine bletilla, has the function of convergence, hemostasis, tonifying lung etc. and is used for
In related drug.Existing bletilla polysaccharide (also known as bletilla glue) generally uses the production technology of water extract-alcohol precipitation.Week is quick etc. to disclose " one
Kind bletilla polysaccharide and preparation method thereof and new application " (CN200810233680.6), which extracts through water, resin removes egg
Bai Houyong ethanol precipitations go out bletilla polysaccharide, and content reaches 70~74%.Song Lifei discloses a kind of " technique prepared by bletilla polysaccharide
Combination " (CN200910302937.3) prepares bletilla striata homogenate with 80~120 times at normal temperatures, through sedimentation centrifugation, media filtration
(filter aid talcum powder, activated carbon, on diatom) processing, then micro-filtration is carried out with the ceramic membrane of 2000~100nm, gained trapped fluid passes through
Alcohol precipitation, drying obtain bletilla polysaccharide.The present invention discloses a kind of " system of glucomannan acid hydrolyzed product in early-stage study
Preparation Method " (CN201010166718.X), the method uses molecular weight, and not less than 100kD, (1kD is 1000 dalton molecule
Amount) konjaku or bletilla glucomannans raw material, by being carried out in pH1.0-3.0 Acidity Ranges and 45-100 DEG C of temperature range
Sour water solution is controlled, through alcohol precipitation, the dry glucomannan acid hydrolyzed product for obtaining specified molecular weight (10~150kD).
Inventor is in the further development process of bletilla polysaccharide preparation process, although finding that aqueous extraction-alcohol precipitation technology can be effective
Bletilla polysaccharide is obtained, but since bletilla polysaccharide extracting solution viscosity is higher, the Polyphenols pigment that is coexisted in extracting solution during alcohol precipitation,
The impurity such as particulate matter can settle out together, to influence bletilla polysaccharide purity and color and luster；Oligosaccharides etc. in acid hydrolysis
Partial organic substances also can simultaneously be precipitated during alcohol precipitation.In the fields such as drug and other medical applications, to functional raw material
The quality such as purity, related substance have high requirements.For this purpose, the present invention finally found that through further investigation using membrane technology, and
In conjunction with pretreatment technologies such as filter aid aided filter, micro-filtrations, it is poly- that high-purity Portugal sweet dew of the purity more than 90% can be successfully obtained
Sugar product, so as to complete the present invention.
Invention content
The object of the present invention is to provide a kind of preparation methods of high-purity glucomannans.
In order to solve the above technical problems, present invention employs the following technical solutions.
A kind of preparation method of high-purity glucomannans, it is characterised in that use following steps：
(1) glucomannans raw material is chosen, the glucomannans solution GM-A of 0.5%-10.0% is prepared into；
(2) the glucomannans solution GM-A obtained by (1), is added filter aid processing, and filtering obtains glucomannans solution
GM-B；
(3) the glucomannans solution GM-B obtained by (2) obtains glucomannans using 100~1000nm micro-filtrate membrane filtrations
Solution GM-C；
(4) the glucomannans solution GM-C obtained by (3), at the ultrafiltration membrane concentration of 2~100kD of molecular cut off
Reason, obtains glucomannans concentrate GM-D；
(5) the glucomannans concentrate GM-D obtained by (4), it is dry through further concentrating, it is poly- to obtain high-purity Portugal sweet dew
Sugar；Or it through further preparing, dispenses, sterilizing obtains high-purity glucomannans liquid preparation.
High-purity glucomannans of the present invention is the homogeneous polysaccharide of 10~1000kD of molecular weight ranges, and purity is more than
90%, and molecular weight 2kD content of organics below is less than 5%.Glucomannans raw material is corresponding molecular weight in step (1)
The homogeneous polysaccharide of 10~1000kD of range, selected from natural konjac polysaccharide or bletilla polysaccharide or its hydrolysate；It is prepared
Method can refer to known references and be prepared using the methods of water extract-alcohol precipitation, sour water solution, also commercially obtain.Step
(2) filter aid described in is selected from the mixture of talcum powder, activated carbon, one or more of diatomite, wherein it is preferred that activated carbon,
The modes such as common plate-frame filtering, stud filtering can be used in its filter type, and can use centrifugation side before filtration as needed
Method removes activated carbon.Micro-filtration membrane aperture described in step (3) is selected from 100~1000nm ranges, more preferable 200~500nm ranges.
Ultrafiltration retaining molecular weight described in step (4) is in 2~100kD, and the 1/5 or less of glucomannans raw molecule amount.
The one kind of high-purity glucomannans drying means in freeze-drying, spray drying, vacuum drying described in step (5)；
The high-purity glucomannans liquid preparation is oral solution or injection.
The present invention further provides a kind of preparation methods of high-purity glucomannans, it is characterised in that uses following step
Suddenly：
(1) the glucomannans raw material for choosing 10~200kD of molecular weight ranges, is prepared into Portugal's sweet dew of 1.0%-10.0%
Glycan solution GM-A；
(2) the glucomannans solution GM-A obtained by (1), is added the processing of activated carbon filter aid, and filtering obtains glucomannans
Solution GM-B；
(3) the glucomannans solution GM-B obtained by (2) obtains glucomannans using 200~500nm micro-filtrate membrane filtrations
Solution GM-C；
(4) by glucomannans solution GM-C obtained by (3), using the ultrafiltration membrane concentration of 2~40kD of molecular cut off,
Obtain glucomannans concentrate GM-D；
(5) the glucomannans concentrate GM-D obtained by (4), it is dry through further concentrating, it is poly- to obtain high-purity Portugal sweet dew
Sugar；Or it through further preparing, dispenses, sterilizing obtains high-purity glucomannans liquid preparation.
The present invention is directed to low, the related impurities content height of bletilla polysaccharide product purity etc. prepared by traditional aqueous extraction-alcohol precipitation technology and lacks
It falls into, by testing repeatedly and numerous studies, using combination techniques techniques such as filter aid aided filter-micro-filtration-ultrafiltration-dryings, at
Work(develops the production technology of high-purity glucomannans, and the preparation process is succinct, controllable, amplification production easy to implement.Institute
The glucomannans product purity of acquisition is high, low in relation to content of material, stability is good, quality controllable, can meet Medicinal crude drug
Quality standard, thus the high-purity glucomannans can according to published glucomannans application document, be applied to system
The standby correlation such as drug and pharmaceutic adjuvant, medical instrument curable product.
The content of present invention is completed by system research and a large amount of creative experiments, is said with following specific embodiments
It is bright.
Specific implementation mode
High-purity glucomannans of the present invention is by such as by taking bletilla polysaccharide and konjac polysaccharide preparation method as an example
What the method represented by lower embodiment was manufactured or found, involved method is that those skilled in the art will appreciate that and transport
Technological means.But following embodiment must not be interpreted as the limiting to the claimed invention in all senses.
In embodiment of the present invention, the preparation of glucomannans raw material and associated sample analysis method have referred to inventor
Publication (CN201010166718.X), treatise [Zhang Yan, the sour water solution dynamics research of bletilla polysaccharide, University Of Suzhou master
Academic dissertation, 2010] etc. documents carry out.Glucomannans molecular weight of the present invention is average weight-average molecular weight (Mw), institute
The polydispersity coefficient (PI=Mw/Mn) of homogeneous polysaccharide is stated generally in 1.5~3.5 ranges.
Embodiment 1：The preparation of bletilla polysaccharide BT180
Bletilla striata medicinal materials are commercially available product, the homogeneous polysaccharide that bletilla polysaccharide contained therein is Mw186kD through HPGPC analyses.
(1) Bletilla striata medicinal materials medicine materical crude slice 4.0kg is weighed, 80kg, 70kg water is separately added into and extracts 2 times, each 1.5hr, extracting solution
It is concentrated into 40L, 1.5 times of 95% ethyl alcohol, which is added, makes precipitation, with 200 mesh filter-cloth filterings, obtains bletilla polysaccharide crude product GM-A (by Solid content
Count yield 15%)；Take GM-A60g (based on Solid content, similarly hereinafter), dissolved at 60 DEG C with 3L water, be configured to 2.0% GM-A it is molten
Liquid；
(2) 20g granular activated carbon filter aids are added in 2.0%GM-A, boil 30min, are stored at room temperature overnight, absorbent cotton
It is filtered with filter paper, obtains pale yellow solution GM-B 2.7L；
(3) it uses 450nm water system miillpore filters to filter GM-B, obtains clear, yellowish solution GM-C 2.6L；
(4) the polysulfone ultrafiltration membrane component of molecular cut off 40kD is used to carry out concentration to GM-C, while ultrafiltration recycles
Pure water is added in liquid amounts to 6L, it is final to obtain clear, colorless trapped fluid GM-D2.2L；
(5) GM-D obtained by (4), is further concentrated under reduced pressure into 0.6L, 60 DEG C or less baking ovens are dried under reduced pressure, obtain high through 80 DEG C
Purity glucomannans BT180 (42g).
The molecular weight and its purity of glucomannans are analyzed using HPGPC methods.Chromatographic condition：Chromatographic column TSK PW4000 columns
(7.8mm×300mm)；Mobile phase water, flow velocity 0.6mLmin-1；ELSD is detected；40 DEG C of column temperature；Test solution concentration 10mg/
ML, 20 μ L of sample size；Analysis time 20min.Bletilla polysaccharide (10~200kD) control series product are self-control, and molecular weight uses
HPGPC-RI/MALS (wyatt companies) is measured.The homogeneous polysaccharide that BT180 is Mw186kD is measured, 10kD is not detected in purity 92%
Related substances below.
Embodiment 2：The preparation of bletilla polysaccharide BT10
Bletilla polysaccharide 10 is prepared using acid-hydrolysis method, the homogeneous polysaccharide for being Mw16kD through HPGPC analyses.
(1) 10 crude product GM-A 20g of bletilla polysaccharide are weighed, is dissolved at 60 DEG C with 200mL waters for injection, is configured to 10.0%
GM-A solution；
(2) 0.2g injection activated carbon filter aids are added in GM-A, boils 30min, is stored at room temperature 4h, absorbent cotton and filter
Paper filters, and obtains nearly colourless solution GM-B 180mL；
(3) it uses 450nm, 200nm water system miillpore filter to filter GM-B successively, obtains transparent nearly colourless solution GM-C
170mL；
(4) the polysulfone ultrafiltration membrane component of molecular cut off 2kD is used to carry out concentration, while ultrafiltration circulation fluid to GM-C
In add water for injection and amount to 500mL, final clear, colorless trapped fluid GM-D 210mL；
(5) GM-D obtained by (4), it is freeze-dried, obtain injection high-purity glucomannans BT10 (16g).
It is analyzed using HPGPC methods, measures the homogeneous polysaccharide that BT10 is Mw16kD, 2kD phases below are not detected in purity 98%
Close substance.
Embodiment 3：The preparation of bletilla polysaccharide BT180 oral solutions
(1) 1 bletilla polysaccharide crude product GM-A 60g of embodiment are weighed, is dissolved at 60 DEG C with 3L water, is configured to 2.0% GM-A
Solution；
(2) 30g granular activated carbon filter aids are added in 2.0%GM-A, boils 30min, is stored at room temperature 6h, centrifuge, warp
Absorbent cotton and filter paper filtering, obtain pale yellow solution GM-B 2.8L；
(3) it uses 450nm, 220nm water system miillpore filter to filter GM-B successively, obtains clear, yellowish solution GM-C 2.7L；
(4) the polysulfone ultrafiltration membrane component of molecular cut off 40kD is used to carry out concentration to GM-C, while ultrafiltration recycles
Pure water is added in liquid amounts to 6L, it is final to obtain clear, colorless trapped fluid GM-D 2.7L；
(5) by GM-D obtained by (4), pure water is added to be formulated into final volume 5.4L, is packed as 100ml/ bottles of oral solutions, 120 DEG C
Sterilize 30min, obtains 1.0%BT180 oral solutions.
It is analyzed using HPGPC methods, measures the homogeneous polysaccharide that BT180 is Mw186 kD, 10kD or less is not detected in purity 94%
Related substances
Embodiment 4：The preparation of konjac polysaccharide KGM
Konjac polysaccharide crude product is commercially available product, the homogeneous polysaccharide that konjac polysaccharide contained therein is Mw926kD through HPGPC analyses.
(1) konjac polysaccharide crude product GM-A 15g (based on Solid content, similarly hereinafter) are weighed, is dissolved, is configured at 60 DEG C with 3L water
0.5% GM-A solution；
(2 are added 15g granular activated carbon filter aids in GM-A, boil 30min, are stored at room temperature overnight, absorbent cotton and filter paper
Filtering, obtains pale yellow solution GM-B 2.6L；
(3) 1000nm filtering with microporous membrane GM-B are used, clear, yellowish solution GM-C 2.5L are obtained；
(4) the polysulfone ultrafiltration membrane component of molecular cut off 100kD is used to carry out concentration to GM-C, while ultrafiltration recycles
Pure water is added in liquid amounts to 4L, it is final to obtain clear, colorless trapped fluid GM-D 2.8L；
(5) GM-D obtained by (4), it is spray-dried, obtain high-purity glucomannans KGM (11g).
It is analyzed using HPGPC-RI/MALS (wyatt companies), measures the homogeneous polysaccharide that KGM is Mw186 kD, purity
91%, 50kD related substances below are not detected.
Claims (8)
1. a kind of preparation method of high-purity glucomannans, it is characterised in that use following steps：
(1) glucomannans raw material is chosen, the glucomannans solution GM-A of 0.5%-10.0% is prepared into；
(2) the glucomannans solution GM-A obtained by (1), is added filter aid processing, and filtering obtains glucomannans solution GM-B；
(3) the glucomannans solution GM-B obtained by (2) obtains glucomannans solution using 100~1000nm micro-filtrate membrane filtrations
GM-C；
(4) the glucomannans solution GM-C obtained by (3) is obtained using the ultrafiltration membrane concentration of 2~100kD of molecular cut off
Glucomannans concentrate GM-D, wherein the ultrafiltration retaining molecular weight glucomannans raw molecule amount 1/5 with
Under；
(5) the glucomannans concentrate GM-D obtained by (4), it is dry through further concentrating, obtain high-purity glucomannans；Or
Through further preparing, dispense, sterilizing obtains high-purity glucomannans liquid preparation；Wherein, the glucomannans is molecular weight
The homogeneous polysaccharide of 10~1000kD of range, purity is more than 90%, and molecular weight 2kD content of organics below is less than 5%.
2. the preparation method of high-purity glucomannans according to claim 1, it is characterised in that Portugal is sweet described in step (1)
It is natural konjac polysaccharide or bletilla polysaccharide or its hydrolysate to reveal glycan raw material.
3. the preparation method of high-purity glucomannans according to claim 1, it is characterised in that drainage described in step (2)
Agent is selected from the mixture of one or more of talcum powder, activated carbon, diatomite.
4. the preparation method of high-purity glucomannans according to claim 3, it is characterised in that drainage described in step (2)
Agent is activated carbon.
5. the preparation method of high-purity glucomannans according to claim 1, it is characterised in that high-purity described in step (5)
Spend the one kind of glucomannans drying means in freeze-drying, spray drying, vacuum drying.
6. a kind of preparation method of high-purity glucomannans, it is characterised in that use following steps：
(1) the glucomannans raw material for choosing 10~200kD of molecular weight ranges, is prepared into the glucomannans of 1.0%-10.0%
Solution GM-A；
(2) the glucomannans solution GM-A obtained by (1), is added the processing of activated carbon filter aid, and filtering obtains glucomannans solution
GM-B；
(3) the glucomannans solution GM-B obtained by (2) obtains glucomannans solution using 200~500nm micro-filtrate membrane filtrations
GM-C；
(4) the glucomannans solution GM-C obtained by (3) obtains Portugal using the ultrafiltration membrane concentration of 2~40kD of molecular cut off
Mannosan concentrate GM-D, wherein the ultrafiltration retaining molecular weight in glucomannans raw molecule amount 1/5 or less；
(5) the glucomannans concentrate GM-D obtained by (4), it is dry through further concentrating, obtain high-purity glucomannans；Or
Through further preparing, dispense, sterilizing obtains high-purity glucomannans liquid preparation；Wherein, the glucomannans is molecular weight
The homogeneous polysaccharide of 10~200kD of range, purity is more than 90%, and molecular weight 2kD content of organics below is less than 5%.
7. according to the application of the high-purity glucomannans of any one of claim 1 to 6 the method preparation in medicine preparation.
8. the high-purity glucomannans prepared according to any one of claim 1 to 6 the method is in preparing curable product
Using.
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