CN107812021A - A kind of method that saikoside is extracted from radix bupleuri - Google Patents
A kind of method that saikoside is extracted from radix bupleuri Download PDFInfo
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- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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Abstract
The present invention relates to a kind of method that saikoside is extracted from radix bupleuri.The present invention is that fresh radix bupleuri root is dried into radix bupleuri medicine materical crude slice, then a broken wall is carried out with wall-breaking machine, secondary broken wall is carried out with high pressure purified air cutting mill, radix bupleuri powder sodium acetate solution and absolute ethyl alcohol after broken wall extract saikoside and go the removal of impurity, obtained saikoside crude extract is refined by preparative high performance liquid chromatography instrument again, finally obtains saikoside powder by vacuum freeze drying.The present invention is using the technique that the cell progress broken wall of radix bupleuri is first extracted into saikoside again, cell-wall breaking ratio is up to 99.4%, saikoside recovery rate is up to more than 92%, low temperature broken wall and gentle extracting method, it can effectively avoid the chemical constitution of saikoside from changing, ensure the good pharmaceutical activity of saikoside.
Description
Technical field
Technical field of natural product extraction of the present invention, more particularly, to a kind of method that saikoside is extracted from radix bupleuri.
Background technology
Radix bupleuri, smoke, mountain dish, mushroom grass, faggot alias, be《Chinese Pharmacopoeia》The herbal medicine included, medicinal part are Umbelliferae
The dry root of plant radix bupleuri or radix bupleuri scorzoneraefolii.Radix bupleuri is bitter in taste, is slightly cold, Return liver, gallbladder channel, have with inducing diaphoresis, the work(of soothing the liver rising Yang
Effect, for cold, fever, fevers and chills alternate, malaria, stagnation of QI due to depression of the liver, sternal rib pain, prolapse of the anus, the prolapse of uterus, irregular menstruation.
Saikoside structure is pentacyclic triterpene oleanane type derivative, and its aglycon is divided into 7 kinds of different types：Epoxy
Ether (I), different ring diene (II), 12- alkene (III), with ring diene (IV), 12 alkene -28- carboxylic acids (V), different ring diene -30- carboxylic acids
(VI), 18- ene-type (VII).Typically containing only glucose, furan sugar, rhamnose and xylose in saikoside, in addition, removing any of the above sugar
There is pentitol (pentitol) outside.Up to now, have studied more than 20 kinds of radix bupleuri of the category altogether, from Bupleunim. L it is separated go out 90
A variety of saponin components, it was found that more than 30 kinds of noval chemical compound.It is 43 to separate within nearly 10 years the saponin(e identified, document report is more
It is saikosaponin a, b, c, d etc., saponin(e is mostly to get from radix bupleuri root, and the saponin(e that aerial part is got is few.
Various types of saikoside all has important pharmacological action：Saikosaponin a significantly inhibits Adenosine triphosphate
The platelet aggregation that glycosides induces, it is suitable with aspirin effect, and endogenous arachidonic acid generation blood is suppressed with dose-dependant
Bolt element.Saikosaponin a, d have the researchs such as class cyclopentanoperhydro-phenanthrene spline structure, Dobashi discovery:To Mouse oral saikoside 0.2~2.0
Find that the corticotropin (ACTH) in serum substantially increases after mg/kg, and ACTH precursors, inferior colliculus in adenohypophysis
The level of corticotropin generation hormone (CRF) mRNA in brain also substantially increases, and is in dose dependent, and radix bupleuri
Saponin(e a is but without this effect.Chiang LC etc. observe saikosaponin a, c, d work using the human hepatocarcinoma cells of HBV transfections
With discovery, saikoside c can substantially suppress the expression of HbeAg in culture medium, and HBV-DNA duplication has also obtained effective suppression,
Prompting saikoside can prevent conversion of the hepatitis B to liver fibrosis.In anti-GBM type ephritis and Heymann kidneys
In scorching rat model, saikoside d can mitigate Urine proteins, reduce blood fat in 2 disease early stage, late period stage administrations, discovery,
Mitigate glomerulus pathological change；IgG, C3 deposition substantially reduce compared with control group in glomerulus；Corticotrophins in serum
Glucocorticoid receptor (GR) has increased in element, cortisone, nephridial tissue.Prompt saikoside d may be by stimulating HPA
Axle, strengthen the function of glucocorticoid, suppress the generation of the metabolites such as arachidonic acid, so as to inhibit the generation of inflammation.
Saikosaponin a, d, f can increase mouse T, the activity of bone-marrow-derived lymphocyte and the secretion level of interleukins -2 (IL-2), radix bupleuri
Saponin(e a, d can also make the horizontal raising of IgA, IgG, IgM in blood plasma, most strong with saikoside d activity.Under saikoside d effect,
The autgmentability of macrophage, activity of acid phosphatase, chemiluminescence etc. are in dose dependent increase.Motoo Y etc. have studied bavin
Hu saponin(e different component is to liver, the effect of pancreatic tumor cell, it is found that when tumor cell of liver 50% suppresses, saikoside d concentration is
20 mg/mL, the concentration of saikosaponin a is 50 mg/mL, saikoside b2, c> 1 000 mg/mL.The mg/ of saikosaponin a 50
ML can suppress the growth of this 2 kinds of tumour cells and DNA synthesis.Chiang LC etc. research also found that saikoside d passes through work
Change Caspase3, Caspase7 induction HepG2 Human liver tumor's Apoptosis, so as to realize antineoplastic action.Tsai YJ etc.
Broken up with saikosaponin a, b1, b2, c, d inducing mouse C6 glioma cells.Saikosaponin a, d propagation capable of inhibiting cell, change
Cellular morphology, and saikosaponin a can significantly inhibit the activity of glutamine synthelase and cyclic nucleotide hydrolase.
The method for extracting saikoside from radix bupleuri at present is all without breaking-wall cell, directly using seepage pressure effects or is carried
The method for taking tank to extract.Chinese patent CN201210595558.X（Application number）Disclose one kind and radix bupleuri is prepared from bigleaf thorowax root
The method of saponin(e, this method comprise the following steps：(1) radix bupleuri raw material is ground into after coarse powder and adds continuous countercurrent extracting tank with third
Ketone aqueous solution refluxing extraction；(2) extract solution that will be handled through (1) filters, and obtains saikoside crude extract；(3) by gained radix bupleuri soap
Glycosides crude extract concentrates, and obtains the thick concentrate of saikoside；(4) the thick concentrate of gained saikoside is pumped into continuous extraction filling group material
Tank, extractant are ethyl acetate, and saikoside acetic acid ethyl acetate extract is obtained via extraction tower；(5) by saikoside acetic acid second
Ester extract concentrates, and obtains saikoside condensed cream；(6) saikoside condensed cream is mixed into silica gel, and by the mixture mixed as
Above silicagel column, eluted through ethyl acetate and petroleum ether mixed liquor, obtain eluent；(7) will be concentrated after eluent recycling design dry
It is dry to obtain saikoside semi-finished product；(8) saikoside semi-finished product are dissolved with aqueous acetone solution, filtered, reclaimed acetone, concentration, put
Precipitation crystal is put, filters to obtain saikoside finished product.The radix bupleuri of this method does not pass through breaking-wall cell, employs aqueous acetone solution
Refluxing extraction, it is also necessary to the step such as concentrated, extracted, during that acetone, ethyl acetate, petroleum ether etc. is used for multiple times is organic
Solvent, easily cause pollution and the residual of organic solvent.Chinese patent CN201610570811.4（Application number）Disclose one kind
The extracting method of saikoside, including：Step 1：Radix Bupleuri is crushed to 40~60 mesh；Step 2：It is with pH value by radix bupleuri powder
After 7~13 basic solvent soaks 10~50h, seepage pressure effects are carried out, obtain percolate, in temperature be 25~100 by percolate
Concentrated at DEG C；Step 3：By the mixture after concentration, ultrasound soaks 30~60min in ultrasonic extractor, then in microwave reactor
25~40min of middle heating；Step 4：Mixture after heating is cooled down into 30~50min；Step 5：Mixture after cooling is used
Ethanol water is dissolved, and macroporous resin adsorption post on lysate, is washed with water to neutrality, collection is washed buck elution removal of impurities
De- liquid；Step 6：Eluent is filtered, obtains saikoside powder after drying.This method does not pass through cell due to radix bupleuri
Broken wall, it is therefore desirable to seepage pressure effects, extraction effect are carried out after radix bupleuri powder is soaked into 10~50h with the basic solvent that pH value is 7~13
Rate is low, and the production cycle is long
Therefore, a kind of then method that radix bupleuri breaking-wall cell is first extracted to saikoside is developed, uses relatively mild extraction conditions
With nontoxic Extraction solvent, not only contribute to improve the production efficiency of saikoside, and saikoside can be ensured as natural
The security of extract.
The content of the invention
The present invention seeks to overcome to need high temperature distillation in the prior art and using more toxic organic solvents extraction bavin
A kind of deficiency of Hu saponin(e, there is provided method that saikoside is extracted from radix bupleuri.
To achieve the above object, the present invention uses following technical scheme：
A kind of method that saikoside is extracted from radix bupleuri, comprises the following steps：
（1）Fresh radix bupleuri root is cleaned, section, it is 8% ~ 12% to be dried to water content, obtains radix bupleuri medicine materical crude slice；This water content is advantageous to
Improve the broken wall efficiency of wall-breaking machine；
（2）Radix bupleuri medicine materical crude slice is put into -196 DEG C of liquid nitrogen 1 ~ 3 min of freezing, rapid take out is put into wall-breaking machine, with 20000 ~
40000 rpm rotating speed carries out the min of broken wall treatment 1 ~ 3, obtains the radix bupleuri powder of a broken wall；Wall-breaking machine is once under the conditions of this
Broken wall efficiency highest, up to 42 ~ 48%；
（3）The radix bupleuri powder of broken wall is adjusted into water content to 3% ~ 5%, -60 ~ -80 DEG C is refrigerated to, is cut with high pressure purified air
Pulverizer carries out secondary broken wall, and air pressure is 100 ~ 200 mPa, and air velocity is 600 ~ 800 m/s, and charging rate is 6 ~ 10
G/s, feed cycle are 2-5 s, and feed pressure is 0.2 ~ 0.3 mPa, obtains the radix bupleuri powder of secondary broken wall；It is secondary broken under the conditions of this
Wall efficiency highest, up to 97.1 ~ 99.4%；
（4）By the radix bupleuri powder of secondary broken wall, 2-5 mol/L sodium acetate solution and absolute ethyl alcohol according to 1:3~5:6 ~ 15 quality
Than mixing, 15 ~ 45 min are stirred at 40-60 DEG C, 30 ~ 60 min are then stood at 0 ~ 4 DEG C, then 10000 at 0 ~ 4 DEG C ~
15000 rpm centrifuge 10 ~ 30 min, take supernatant liquor, obtain saikoside crude extract；The extraction of saikoside under the conditions of this
The impurity-eliminating effect of efficiency highest, protein and nucleic acid is best；
（5）Saikoside crude extract is refined with preparative high performance liquid chromatography instrument, chromatographic column is C18 reverse phase silica gel colors
Post is composed, packing material size is 5 μm, and internal diameter is 7.8 mm, and length is 300 ~ 350 mm, and elution requirement is：0 ~ 10 min is 100% pair
Water is steamed, 10-20 min are volume ratio 10% ~ 20%：80% ~ 90% absolute ethyl alcohol and distilled water, 20-30 min be volume ratio 30% ~
40%：60% ~ 70% absolute ethyl alcohol and distilled water, 30-40 min are volume ratio 50% ~ 60%：40% ~ 50% absolute ethyl alcohol and double
Water is steamed, 40-50 min are volume ratio 70% ~ 80%：20% ~ 30% absolute ethyl alcohol and distilled water, 50-60 min be 100% it is anhydrous
Ethanol, eluent flow rate are 0.6 ~ 1 mL/min, and the elution composition containing saikoside is reclaimed, obtains refined saikoside
Eluent；Separation of saikoside and preparation effect are best under the conditions of this；
（6）Refined saikoside eluent is carried out into vacuum refrigeration under the conditions of -10 ~ -30 DEG C, -0.08 ~ -0.1 MPa to do
It is dry, obtain saikoside powder.
Preferably, step（1）In, the section is that radix bupleuri root is cut into the thin slice that thickness is 0.5 ~ 2 mm, this thickness
Radix bupleuri root thin slice is advantageous to wall-breaking machine broken wall.
Preferably, step（1）In, it is 9% to be dried to water content, this Water Content Conditions broken wall efficiency highest next time, is reached
48%。
Preferably, step（2）In, radix bupleuri medicine materical crude slice is put into -196 DEG C of liquid nitrogen and freezes 2 min, rapid take out is put into brokenly
Wall machine, the min of broken wall treatment 2 is carried out with 30000 rpm rotating speed, this contains condition broken wall efficiency highest next time, up to 48%.
Preferably, step（3）In, the radix bupleuri powder of a broken wall is adjusted into water content to 3%, -75 DEG C is refrigerated to, uses high pressure
Purified air cutting mill carries out secondary broken wall, and air pressure is 150 mPa, and air velocity is 700 m/s, and charging rate is
8 g/s, feed cycle are 3 s, and feed pressure is 0.2 mPa, secondary broken wall efficiency highest under the conditions of this, up to 99.4%.
Preferably, step（4）In, by the radix bupleuri powder of secondary broken wall, 3 mol/L sodium acetate solution and absolute ethyl alcohol according to
1:5:10 mass ratio mixing, 40 min are stirred at 55 DEG C, 40 min are then stood at 4 DEG C, then 12000 at 4 DEG C
Rpm centrifuges 20 min, the extraction efficiency highest of saikoside under the conditions of this, up to more than 92%, the impurity-eliminating effect of protein and nucleic acid
Preferably, more than 95% protein and more than 90% nucleic acid can be removed.
Preferably, step（5）In, described elution requirement is：0 ~ 10 min is 100% distilled water, and 10-20 min are body
Product ratio 15%：85% absolute ethyl alcohol and distilled water, 20-30 min are volume ratio 35%：65% absolute ethyl alcohol and distilled water, 30-
40 min are volume ratio 55%：45% absolute ethyl alcohol and distilled water, 40-50 min are volume ratio 75%：25% absolute ethyl alcohol and
Distilled water, 50-60 min are 100% absolute ethyl alcohol, and eluent flow rate is 1.2 mL/min, point of saikoside under the conditions of this
It is high and sharp from best, the eluting peak distinct of each saikoside, peak type with preparation effect.
Preferably, step（6）In, refined saikoside eluent is subjected to vacuum under the conditions of -20 DEG C, -0.1 MPa
Freeze-drying.
The present invention possesses the advantages of following and good effect：
（1）Using the cell progress broken wall of radix bupleuri is first extracted into the technique of saikoside again, cell-wall breaking ratio is up to the present invention
99.4%, saikoside recovery rate is up to more than 92%.
（2）The present invention uses low temperature broken wall and gentle extracting method, can effectively avoid the chemical constitution of saikoside from occurring
Change, ensure the good pharmaceutical activity of saikoside.
（3）The present invention prepares saikoside using gradient elution mode separation, good separating effect, it is high to prepare purity.
（4）Whole process of the invention does not add poisonous harmful chemical agents, and the saikoside of extraction is safe.
（5）Present invention process is simple, with short production cycle, workable, is adapted to a large amount of extractions and preparation of saikoside.
Embodiment
Below in conjunction with a kind of method that saikoside is extracted from radix bupleuri of the example description present invention, these descriptions are not pair
The restriction of present invention.
Embodiment 1
A kind of method that saikoside is extracted from radix bupleuri, comprises the following steps：
（1）Fresh radix bupleuri root is cleaned, is cut into the thin slice that thickness is 1 mm, it is 9% to be dried to water content, obtains radix bupleuri medicine materical crude slice；
（2）Radix bupleuri medicine materical crude slice is put into -196 DEG C of liquid nitrogen and freezes 2 min, rapid take out is put into wall-breaking machine, with 30000 rpm's
Rotating speed carries out the min of broken wall treatment 2, obtains the radix bupleuri powder of a broken wall, sporoderm-broken rate 48%；
（3）The radix bupleuri powder of broken wall is adjusted into water content to 3%, -75 DEG C are refrigerated to, with high pressure purified air cutting mill
Carry out secondary broken wall, air pressure is 150 mPa, and air velocity is 700 m/s, and charging rate is 8 g/s, feed cycle 3
S, feed pressure are 0.2 mPa, obtain the radix bupleuri powder of secondary broken wall, sporoderm-broken rate 99.4%；
（4）By the radix bupleuri powder of secondary broken wall, 3 mol/L sodium acetate solution and absolute ethyl alcohol according to 1:5:10 mass ratio mixes
Close, 40 min are stirred at 55 DEG C, 40 min are then stood at 4 DEG C, then 12000 rpm centrifuge 20 min at 4 DEG C, take
Layer clear liquid, obtains saikoside crude extract；
（5）Saikoside crude extract is refined with preparative high performance liquid chromatography instrument, chromatographic column is C18 reverse phase silica gel colors
Post is composed, packing material size is 5 μm, and internal diameter is 7.8 mm, and length is 350 mm, and elution requirement is：0 ~ 10 min is 100% pair of steaming
Water, 10-20 min are volume ratio 15%：85% absolute ethyl alcohol and distilled water, 20-30 min are volume ratio 35%：65% it is anhydrous
Ethanol and distilled water, 30-40 min are volume ratio 55%：45% absolute ethyl alcohol and distilled water, 40-50 min are volume ratio 75%：
25% absolute ethyl alcohol and distilled water, 50-60 min are 100% absolute ethyl alcohol, and eluent flow rate is 1.2 mL/min, will be contained
The elution composition recovery of saikoside, obtains refined saikoside eluent；
（6）Refined saikoside eluent is subjected to vacuum freeze drying under the conditions of -20 DEG C, -0.1 MPa, obtains radix bupleuri
Saponin(e powder.
Embodiment 2
A kind of method that saikoside is extracted from radix bupleuri, comprises the following steps：
（1）Fresh radix bupleuri root is cleaned, is cut into the thin slice that thickness is 0.5 mm, it is 12% to be dried to water content, obtains radix bupleuri drink
Piece；
（2）Radix bupleuri medicine materical crude slice is put into -196 DEG C of liquid nitrogen and freezes 1 min, rapid take out is put into wall-breaking machine, with 20000 rpm's
Rotating speed carries out the min of broken wall treatment 1, obtains the radix bupleuri powder of a broken wall, sporoderm-broken rate 45%；
（3）The radix bupleuri powder of broken wall is adjusted into water content to 5%, -60 DEG C are refrigerated to, with high pressure purified air cutting mill
Carry out secondary broken wall, air pressure is 100 mPa, and air velocity is 600 m/s, and charging rate is 10 g/s, feed cycle 2
S, feed pressure are 0.3 mPa, obtain the radix bupleuri powder of secondary broken wall, sporoderm-broken rate 98.6%；
（4）By the radix bupleuri powder of secondary broken wall, 3.5 mol/L sodium acetate solution and absolute ethyl alcohol according to 1:4:10 mass ratio mixes
Close, 45 min are stirred at 40 DEG C, 30 min are then stood at 4 DEG C, then 15000 rpm centrifuge 30 min at 4 DEG C, take
Layer clear liquid, obtains saikoside crude extract；
（5）Saikoside crude extract is refined with preparative high performance liquid chromatography instrument, chromatographic column is C18 reverse phase silica gel colors
Post is composed, packing material size is 5 μm, and internal diameter is 7.8 mm, and length is 300 mm, and elution requirement is：0 ~ 10 min is 100% pair of steaming
Water, 10-20 min are volume ratio 10%：90% absolute ethyl alcohol and distilled water, 20-30 min are volume ratio 30%：70% it is anhydrous
Ethanol and distilled water, 30-40 min are volume ratio 50%：50% absolute ethyl alcohol and distilled water, 40-50 min are volume ratio
70%%：30% absolute ethyl alcohol and distilled water, 50-60 min are 100% absolute ethyl alcohol, and eluent flow rate is 1 mL/min, will
Elution composition recovery containing saikoside, obtains refined saikoside eluent；
（6）Refined saikoside eluent is subjected to vacuum freeze drying under the conditions of -10 DEG C, -0.1 MPa, obtains radix bupleuri
Saponin(e powder.
Embodiment 3
A kind of method that saikoside is extracted from radix bupleuri, comprises the following steps：
（1）Fresh radix bupleuri root is cleaned, is cut into the thin slice that thickness is 2 mm, it is 8% to be dried to water content, obtains radix bupleuri medicine materical crude slice；
（2）Radix bupleuri medicine materical crude slice is put into -196 DEG C of liquid nitrogen and freezes 2 min, rapid take out is put into wall-breaking machine, with 40000rpm's
Rotating speed carries out the min of broken wall treatment 2, obtains the radix bupleuri powder of a broken wall, sporoderm-broken rate 44%；
（3）The radix bupleuri powder of broken wall is adjusted into water content to 4%, -80 DEG C are refrigerated to, with high pressure purified air cutting mill
Carry out secondary broken wall, air pressure is 180 mPa, and air velocity is 800 m/s, and charging rate is 6 g/s, feed cycle 2
S, feed pressure are 0.3 mPa, obtain the radix bupleuri powder of secondary broken wall, sporoderm-broken rate 99.0%；
（4）By the radix bupleuri powder of secondary broken wall, 4 mol/L sodium acetate solution and absolute ethyl alcohol according to 1: 3:10 mass ratio mixes
Close, 15 min are stirred at 60 DEG C, 30 min are then stood at 0 DEG C, then 15000 rpm centrifuge 10 min at 0 DEG C, take
Layer clear liquid, obtains saikoside crude extract；
（5）Saikoside crude extract is refined with preparative high performance liquid chromatography instrument, chromatographic column is C18 reverse phase silica gel colors
Post is composed, packing material size is 5 μm, and internal diameter is 7.8 mm, and length is 350 mm, and elution requirement is：0 ~ 10 min is 100% pair of steaming
Water, 10-20 min are volume ratio 15%：85% absolute ethyl alcohol and distilled water, 20-30 min are volume ratio 35%：65% it is anhydrous
Ethanol and distilled water, 30-40 min are volume ratio 55%：45% absolute ethyl alcohol and distilled water, 40-50 min are volume ratio 75%：
25% absolute ethyl alcohol and distilled water, 50-60 min are 100% absolute ethyl alcohol, eluent flow rate 0.8mL/min, will be contained
The elution composition recovery of saikoside, obtains refined saikoside eluent；
（6）Refined saikoside eluent is subjected to vacuum freeze drying under the conditions of -30 DEG C, -0.08 MPa, obtains radix bupleuri
Saponin(e powder.
Embodiment 4
A kind of method that saikoside is extracted from radix bupleuri, comprises the following steps：
（1）Fresh radix bupleuri root is cleaned, is cut into the thin slice that thickness is 1.5 mm, it is 12% to be dried to water content, obtains radix bupleuri drink
Piece；
（2）Radix bupleuri medicine materical crude slice is put into -196 DEG C of liquid nitrogen and freezes 3 min, rapid take out is put into wall-breaking machine, with 20000 rpm's
Rotating speed carries out the min of broken wall treatment 3, obtains the radix bupleuri powder of a broken wall, sporoderm-broken rate 42%；
（3）The radix bupleuri powder of broken wall is adjusted into water content to 5%, -75 DEG C are refrigerated to, with high pressure purified air cutting mill
Carry out secondary broken wall, air pressure is 150 mPa, and air velocity is 650 m/s, and charging rate is 9 g/s, feed cycle 4
S, feed pressure are 0.3 mPa, obtain the radix bupleuri powder of secondary broken wall, sporoderm-broken rate 97.1%；
（4）By the radix bupleuri powder of secondary broken wall, 2 mol/L sodium acetate solution and absolute ethyl alcohol according to 1:4:8 mass ratio mixing,
45 min are stirred at 55 DEG C, 60 min are then stood at 2 DEG C, then 12000 rpm centrifuge 10 min at 2 DEG C, take upper strata
Clear liquid, obtain saikoside crude extract；
（5）Saikoside crude extract is refined with preparative high performance liquid chromatography instrument, chromatographic column is C18 reverse phase silica gel colors
Post is composed, packing material size is 5 μm, and internal diameter is 7.8 mm, and length is 350 mm, and elution requirement is：0 ~ 10 min is 100% pair of steaming
Water, 10-20 min are volume ratio 20%：80% absolute ethyl alcohol and distilled water, 20-30 min are volume ratio 40%：60% it is anhydrous
Ethanol and distilled water, 30-40 min are volume ratio 60%：40% absolute ethyl alcohol and distilled water, 40-50 min are volume ratio 80%：
20% absolute ethyl alcohol and distilled water, 50-60 min are 100% absolute ethyl alcohol, and eluent flow rate is 0.6 mL/min, will be contained
The elution composition recovery of saikoside, obtains refined saikoside eluent；
（6）Refined saikoside eluent is subjected to vacuum freeze drying under the conditions of -25 DEG C, -0.09 MPa, obtains radix bupleuri
Saponin(e powder.
Claims (8)
- A kind of 1. method that saikoside is extracted from radix bupleuri, it is characterised in that comprise the following steps：（1）Fresh radix bupleuri root is cleaned, section, it is 8% ~ 12% to be dried to water content, obtains radix bupleuri medicine materical crude slice；（2）Radix bupleuri medicine materical crude slice is put into -196 DEG C of liquid nitrogen 1 ~ 3 min of freezing, rapid take out is put into wall-breaking machine, with 20000 ~ 40000 rpm rotating speed carries out the min of broken wall treatment 1 ~ 3, obtains the radix bupleuri powder of a broken wall；（3）The radix bupleuri powder of broken wall is adjusted into water content to 3% ~ 5%, -60 ~ -80 DEG C is refrigerated to, is cut with high pressure purified air Pulverizer carries out secondary broken wall, and air pressure is 100 ~ 200 mPa, and air velocity is 600 ~ 800 m/s, and charging rate is 6 ~ 10 G/s, feed cycle are 2-5 s, and feed pressure is 0.2 ~ 0.3 mPa, obtains the radix bupleuri powder of secondary broken wall；（4）By the radix bupleuri powder of secondary broken wall, 2-5 mol/L sodium acetate solution and absolute ethyl alcohol according to 1:3~5:6 ~ 15 quality Than mixing, 15 ~ 45 min are stirred at 40-60 DEG C, 30 ~ 60 min are then stood at 0 ~ 4 DEG C, then 10000 at 0 ~ 4 DEG C ~ 15000 rpm centrifuge 10 ~ 30 min, take supernatant liquor, obtain saikoside crude extract；（5）Saikoside crude extract is refined with preparative high performance liquid chromatography instrument, chromatographic column is C18 reverse phase silica gel colors Post is composed, packing material size is 5 μm, and internal diameter is 7.8 mm, and length is 300 ~ 350 mm, and elution requirement is：0 ~ 10 min is 100% pair Water is steamed, 10-20 min are volume ratio 10% ~ 20%：80% ~ 90% absolute ethyl alcohol and distilled water, 20-30 min be volume ratio 30% ~ 40%：60% ~ 70% absolute ethyl alcohol and distilled water, 30-40 min are volume ratio 50% ~ 60%：40% ~ 50% absolute ethyl alcohol and double Water is steamed, 40-50 min are volume ratio 70% ~ 80%：20% ~ 30% absolute ethyl alcohol and distilled water, 50-60 min be 100% it is anhydrous Ethanol, eluent flow rate are 1 ~ 2 mL/min, and the elution composition containing saikoside is reclaimed, refined saikoside is obtained and washes De- liquid；（6）Refined saikoside eluent is carried out into vacuum refrigeration under the conditions of -10 ~ -30 DEG C, -0.08 ~ -0.1 MPa to do It is dry, obtain saikoside powder.
- 2. the method according to claim 1 that saikoside is extracted from radix bupleuri, it is characterised in that step（1）In, it is described Cut into slices as radix bupleuri root is cut into the thin slice that thickness is 0.5 ~ 2 mm.
- 3. the method according to claim 2 that saikoside is extracted from radix bupleuri, it is characterised in that step（1）In, dry It is 9% to water content.
- 4. the method according to claim 1 that saikoside is extracted from radix bupleuri, it is characterised in that step（2）In, by bavin Hu medicine materical crude slice, which is put into -196 DEG C of liquid nitrogen, freezes 2 min, and rapid take out is put into wall-breaking machine, is broken with 30000 rpm rotating speed Wall handles 2 min.
- 5. the method according to claim 1 that saikoside is extracted from radix bupleuri, it is characterised in that step（3）In, by one The radix bupleuri powder of secondary broken wall adjusts water content to 3%, is refrigerated to -75 DEG C, is carried out with high pressure purified air cutting mill secondary broken Wall, air pressure are 150 mPa, and air velocity is 700 m/s, and charging rate is 8 g/s, and feed cycle is 3 s, feed pressure For 0.2 mPa.
- 6. the method according to claim 1 that saikoside is extracted from radix bupleuri, it is characterised in that step（4）In, by two The radix bupleuri powder of secondary broken wall, 3 mol/L sodium acetate solution and absolute ethyl alcohol are according to 1:5:10 mass ratio mixing, is stirred at 55 DEG C 40 min are mixed, 40 min are then stood at 4 DEG C, then 12000 rpm centrifuge 20 min at 4 DEG C.
- 7. the method according to claim 1 that saikoside is extracted from radix bupleuri, it is characterised in that step（5）In, it is described Elution requirement be：0 ~ 10 min is 100% distilled water, and 10-20 min are volume ratio 15%：85% absolute ethyl alcohol and distilled water, 20-30 min are volume ratio 35%：65% absolute ethyl alcohol and distilled water, 30-40 min are volume ratio 55%：45% absolute ethyl alcohol And distilled water, 40-50 min are volume ratio 75%：25% absolute ethyl alcohol and distilled water, 50-60 min are 100% anhydrous second Alcohol, eluent flow rate are 1.2 mL/min.
- 8. the method according to claim 1 that saikoside is extracted from radix bupleuri, it is characterised in that step（6）In, by essence The saikoside eluent of system carries out vacuum freeze drying under the conditions of -20 DEG C, -0.1 MPa.
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|---|---|---|---|---|
| CN109363027A (en) * | 2018-10-22 | 2019-02-22 | 蓝妙田 | The preparation method of mango beverage |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101757553A (en) * | 2008-12-04 | 2010-06-30 | 郑州豫密药业股份有限公司 | Method for preparing depression alleviating pills |
| CN102659903A (en) * | 2012-05-10 | 2012-09-12 | 青川德康源药业有限公司 | Method for extracting and purifying Chinese thorowax root saponin a or Chinese thorowax root saponin d |
| CN104127491A (en) * | 2014-07-29 | 2014-11-05 | 成都乾坤动物药业有限公司 | Pharmaceutical composition with fever relieving effect and preparation method and application thereof |
-
2017
- 2017-11-17 CN CN201711146228.1A patent/CN107812021A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101757553A (en) * | 2008-12-04 | 2010-06-30 | 郑州豫密药业股份有限公司 | Method for preparing depression alleviating pills |
| CN102659903A (en) * | 2012-05-10 | 2012-09-12 | 青川德康源药业有限公司 | Method for extracting and purifying Chinese thorowax root saponin a or Chinese thorowax root saponin d |
| CN104127491A (en) * | 2014-07-29 | 2014-11-05 | 成都乾坤动物药业有限公司 | Pharmaceutical composition with fever relieving effect and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 高立超等: "《仲景源中医食方食品丛书 药食同源本草素食方库》", 30 June 2016, 上海科学技术文献出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109363027A (en) * | 2018-10-22 | 2019-02-22 | 蓝妙田 | The preparation method of mango beverage |
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