CN106432024A - Method for coupling extraction of astaxanthin from haematococcus pluvialis cells by rapid wall breaking - Google Patents
Method for coupling extraction of astaxanthin from haematococcus pluvialis cells by rapid wall breaking Download PDFInfo
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- CN106432024A CN106432024A CN201610854110.3A CN201610854110A CN106432024A CN 106432024 A CN106432024 A CN 106432024A CN 201610854110 A CN201610854110 A CN 201610854110A CN 106432024 A CN106432024 A CN 106432024A
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- haematococcus pluvialis
- astaxanthin
- broken wall
- cell according
- sand mill
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C403/00—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone
- C07C403/24—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone having side-chains substituted by six-membered non-aromatic rings, e.g. beta-carotene
Abstract
The invention discloses a method for coupling extraction of astaxanthin from haematococcus pluvialis cells by rapid wall breaking. The method comprises the following steps: (1) adding haematococcus pluvialis into an organic solvent, and stirring uniformly; (2) injecting the feed liquid obtained in the step (1) into a sand mill, and performing grinding and crushing to obtain a suspension; (3) performing solid-liquid separation on the suspension obtained in the step (2), and recovering a liquid phase; and (4) performing reduced pressure distillation on the liquid phase collected in the step (3), and recovering the organic solvent to obtain a residual wine ointment-shaped substance, namely an astaxanthin extract. According to the method disclosed by the invention, the wall breakage rate of the haematococcus pluvialis cells is extremely high, the solvent can directly extract the astaxanthin during wall breaking, and the whole breaking coupling extraction process greatly shortens the time consumption of an existing process, thereby effectively ensuring the biological activity of natural astaxanthin.
Description
Technical field
The invention belongs to technical field of bioseparation is and in particular to a kind of extract astaxanthin from haematococcus pluvialis cell
Method.
Background technology
Astaxanthin is 3,3 '-dihydroxy -4,4 '-diketo-β, β '-carrotene, is terpenes unsaturated compounds, is
One Carotenoids.Natural astaxanthin is one of the strongest antioxidant in nature, can oxygen effectively in scavenger-cell from
By base, strengthen cytothesis ability, the accumulation maintaining organism balance and reducing senile cell.The ability of its scavenging activated oxygen is
Ascorbic 6000 times, 700 times of anthocyanidin.Meanwhile, natural astaxanthin be uniquely a kind of can by the class of blood-brain barrier recklessly
Radish element.Since 2008, research discovery both at home and abroad has multiple physiological functions, such as non-oxidizability, antitumor, pre- anti-cancer,
Strengthen immunity, improve the aspects such as eyesight and have certain effect.
The main source of natural astaxanthin is marine products discarded object, phaffiafhodozyma and haematococcus pluvialis.The life of its moderate rain is red
Content astaxanthin in ball algae be far above in addition both, generally up to 1.5%-3%, be referred to as " concentrating of natural astaxanthin
Product ".Therefore, extracting natural astaxanthin from haematococcus becomes the focus of recent domestic research and produces natural astaxanthin
Main method.
Haematococcus pluvialis are subordinate to Chlorophyta, volvocales, haematococcus section, haematococcus, rain non-hibernating eggs.Its cell membrane is thicker, plus
The big difficulty therefrom extracting astaxanthin, by the way of organic solvent directly extracts, the recovery rate of astaxanthin is less than 50%
(CN103044304A), therefore the key that effective broken wall treatment is in Astaxanthin extraction technique is carried out to haematococcus pluvialis cell
Step.
The broken wall mode of existing report mainly includes：1. high-pressure homogeneous broken (CN104961668A).The method sporoderm-broken rate
For 90% about, sample needs through repeated multiple times broken wall treatment, take longer thus leading to effective ingredient oxidized；In addition red
Algae easily settles ball in the solution, leads to feed liquid uneven thus blocking instrument.2. flash extracter (CN105254551A).Should
Method crushing efficiency is not high, and treating capacity is less, and the recovery rate of final products is relatively low.3. high-pressure breaking (CN103787941A).
The method adopts 100-300mPa high pressure to crush, and, more than 95%, effect is preferably but higher to equipment requirement for sporoderm-broken rate, is difficult on rule
Modelling is used in producing.4. acidic treatment (CN103232375A).It is molten as broken wall that the method additionally with the addition of acid in system
Agent, increases the detached difficulty of product, and acid solution is not easy to recycle.5. at glacial acetic acid ice bath, refiner homogenate and liquid nitrogen
Manage the crush method (CN104529852A) that combines.The method sporoderm-broken rate is up to 97.7%, but operation sequence is complicated, relatively costly and
Recovery rate is relatively low, is not suitable for being applied to large-scale production.6. multigelation method (CN104557651A).The method is using addition sulphur
Sour aqueous ammonium multigelation broken wall, freezing and thawing cycle is up to 15-20h, and single broken wall inefficiency is not appropriate for actual production.7.
Ultramicro grinding method.The method belongs to dry grinding category, and broken wall efficiency is preferable, but due to the oily matter in broken rear haematococcus
It is released, lead to feed powder to stick in a large number in equipment kettle and be difficult to shift, have impact on the application of the method.
Content of the invention
The technical problem to be solved is the Astaxanthin extraction providing a kind of brand-new high-efficiency wall breaking to be coupled extraction
Technique.
For solving above-mentioned technical problem, the thinking of the present invention is to be coupled extraction process with wet grinding, using sand mill
Complete the extraction of astaxanthin while to haematococcus breaking-wall cell, feed liquid takes solution to carry out vacuum distillation after separation of solid and liquid and returns
Receive extractant, concentrate peony oil paste and be astaxanthin extract.This technological operation is easy, takes extremely short, and sporoderm-broken rate is high
Reach 99.9%, Astaxanthin extraction rate is up to more than 99%.
Specific technical scheme is as follows：
A kind of method of haematococcus pluvialis cell quick broken wall coupling extraction astaxanthin, it comprises the steps：
(1) haematococcus pluvialis are taken to add organic solvent to stir；
(2) feed liquid that step (1) obtains is injected in sand mill, after grinding, obtain suspension；
(3) suspension that step (2) is obtained carries out separation of solid and liquid, reclaim liquid phase；
(4) the liquid phase vacuum distillation that step (3) is collected, reclaims organic solvent, and residual dark reddish oil paste is shrimp
Blue or green extract.
In step (1), haematococcus pluvialis add organic solvent in the form of its dry haematococcus pluvialis algae powder, are dried
Haematococcus pluvialis algae powder, its water content is usually less than 2%.
In step (1), described organic solvent be in ethanol, acetone and n-hexane any one or two or more arbitrarily
The mixture of ratio.Organic solvent preferably these low viscosities, low boiling, the organic solvent of good fluidity.Higher boiling can not be adopted
Solvent or the higher solvent of viscosity or grease type solvent, this class solvent exists and is not easily recycled it is impossible to simple realization separation of solid and liquid
And the shortcoming affecting the content of effective ingredient in final astaxanthin extract.
In step (1), the ratio of haematococcus pluvialis and organic solvent is 1kg:1-10L.
In step (2), described sand mill is excellent pin-type sand mill or turbine type sand mill.
In step (2), described sand mill, in its grinding chamber, filler is ceramic fine bead, and in grinding chamber, microballon loading amount is to grind
The 75%-90% of mill chamber volume.The preferred material of described ceramic fine bead is zirconia ceramics etc., the size of described ceramic fine bead
Preferably diameter 0.5-5mm.
In step (2), described sand mill, grind cavity temperature and control between 10-35 DEG C, preferably 10 DEG C.
In step (2), the milled processed time is 10-600s/L feed liquid.
In step (3), the mode of separation of solid and liquid is centrifugation, plate compression, negative pressure leaching or natural subsidence.
In step (4), the temperature of vacuum distillation is 30-50 DEG C, preferably 40 DEG C.
In step (4), general vacuum distillation is till frozen-free solvent drippage.
In step (4), the organic solvent of recovery is circulated and is mixed with haematococcus pluvialis in step (1).
Beneficial effect：Inventive process have the advantage that：
1. haematococcus cell-wall breaking ratio is high, and while carrying out broken, solvent can directly extract astaxanthin, entirely crushes
It is coupled extraction process and substantially reduces the time-consuming of existing process, be effectively guaranteed the BA of natural astaxanthin.
2. obtained without any other solvent, recycling design in addition to the easy recycling design adding in whole technique
Astaxanthin extract does not have composition that other external sources add it is ensured that the quality of extract and be easy to the exploitation of subsequent product.
3. in whole technique, solvent reclaims the broken extraction process for a new round in addition to a small amount of volatilization completely, does not almost have
Consume.
4. in technique, solvent for use is low viscosity, low boiling, the organic solvent of good fluidity, on the one hand easily broken rear solid
The detached process of liquid, is on the other hand beneficial to the recycling of solvent.
5. grinding is minimum with microballon loss, and whole system operating cost is very low.
Brief description
Fig. 1 crushes front haematococcus pluvialis microscopy photo in a solvent；
Fig. 2 crushes rear haematococcus pluvialis microscopy photo in a solvent；
Fig. 3 raw material algae powder outward appearance photo；
Fig. 4 presses filter residue outward appearance photo after the extraction of embodiment 2 technique.
Specific embodiment
According to following embodiments, the present invention may be better understood.However, it is as it will be easily appreciated by one skilled in the art that real
Apply the content described by example and be merely to illustrate the present invention, and should not be also without limitation on basis described in detail in claims
Invention.
Sand mill model ESW-1.0 (excellent pin) used by following examples or EDW-1.0 (turbine type).
Haematococcus pluvialis algae powder used by following examples, its water content is below 2%.
Sporoderm-broken rate adopts microscope inspection technology, and the suspension before and after taking 100uL to crush makes sample, need not dye, directly exist
Observation of cell structure under 100 times of oil mirrors, cannot find intact cell configuration after crushing, therefore assert sporoderm-broken rate more than 99.9%.
The method that content astaxanthin assay method adopts GB GB/T 31520, recovery rate computational methods are extract Prawn
The ratio of astaxanthin total amount in blue or green element total amount and raw material.
Embodiment 1：
Take haematococcus pluvialis algae powder 1kg, add 1L absolute ethyl alcohol and stirring uniformly to inject afterwards stand-by in sand mill charging cylinder.Adopt
Use excellent pin-type sand mill, grinding chamber adds the 0.6mm zirconia ceramics microballon of 75% cavity volume, after 10 DEG C of precooling grinding chambers
Open charging pumping source, milling time 15s.Feed liquid 6000rpm refrigerated centrifuge 5min, takes 50 DEG C of vacuum distillations of supernatant extremely solvent-free
Drippage obtains astaxanthin extract, reclaims ethanol simultaneously and circulates the extraction for the astaxanthin in haematococcus pluvialis.Breaking-wall cell
Rate 99.9%, natural astaxanthin recovery rate 99.2%, content astaxanthin 4.2% in extract.Embodiment 2：
Take haematococcus pluvialis algae powder 0.5kg, stand-by in injection sand mill charging cylinder after adding 4L n-hexane to stir.Adopt
Use turbine type sand mill, grinding chamber adds the 1.0mm zirconia ceramics microballon of 85% cavity volume, after 10 DEG C of precooling grinding chambers
Open charging pumping source, milling time 60s.Feed liquid natural subsidence 1h, takes 40 DEG C of vacuum distillations of supernatant to come to solvent-free dripping
Astaxanthin extract, reclaims the extraction that n-hexane recycle is used for the astaxanthin in haematococcus pluvialis simultaneously.Cell-wall breaking ratio
99.9%, natural astaxanthin recovery rate 99.5%, content astaxanthin 5.1% in extract.(Fig. 1 crushes front microscopy photo, Fig. 2
Microscopy photo after broken, Fig. 3 raw material algae powder photo, filter residue photo after Fig. 4 extraction)
Embodiment 3：
Take haematococcus pluvialis algae powder 2kg, stand-by in injection sand mill charging cylinder after adding 20L acetone to stir.Using
Excellent pin-type sand mill, adds the 5.0mm zirconia ceramics microballon of 90% cavity volume in grinding chamber, beats after 10 DEG C of precooling grinding chambers
Drive material pumping source, milling time 600s into.Feed liquid negative pressure leaching, takes 30 DEG C of vacuum distillation to solvent-free drippings of filtrate to come to shrimp blue or green
Extract, reclaims the extraction that acetone recycle is used for the astaxanthin in haematococcus pluvialis simultaneously.Cell-wall breaking ratio 99.9%, natural
Astaxanthin extraction rate 99.8%, content astaxanthin 4.8% in extract.
Comparative example 1：
Operating procedure, with embodiment 1, is wherein added solvent and adopts sunflower oil to substitute ethanol, the suspension after grinding cannot
Directly separation of solid and liquid is realized using the method for centrifugation or negative pressure leaching or natural subsidence, after a large amount of water flocculation cell fragments need to be added
Again separation of solid and liquid is realized by centrifugation.Liquid phase after separating cannot reclaim the solvent of interpolation by way of vacuum distillation, entirely
Technique cell-wall breaking ratio 99.9%, natural astaxanthin recovery rate 85.8%, in extract, content astaxanthin is only 0.25%, unfavorable
Exploitation in subsequent product.
Claims (10)
1. a kind of method of haematococcus pluvialis cell quick broken wall coupling extraction astaxanthin is it is characterised in that it includes walking as follows
Suddenly：
(1) haematococcus pluvialis are taken to add organic solvent to stir；
(2) feed liquid that step (1) obtains is injected in sand mill, after grinding, obtain suspension；
(3) suspension that step (2) is obtained carries out separation of solid and liquid, reclaim liquid phase；
(4) the liquid phase vacuum distillation that step (3) is collected, reclaims organic solvent, and residual dark reddish oil paste is astaxanthin
Extract.
2. the method for haematococcus pluvialis cell according to claim 1 quick broken wall coupling extraction astaxanthin, its feature exists
In, in step (1), described organic solvent is any one or two or more arbitrary proportion in ethanol, acetone and n-hexane
Mixture.
3. the method for haematococcus pluvialis cell according to claim 1 and 2 quick broken wall coupling extraction astaxanthin, its feature
It is, in step (1), the ratio of haematococcus pluvialis and organic solvent is 1kg:1-10L.
4. the method for haematococcus pluvialis cell according to claim 1 quick broken wall coupling extraction astaxanthin, its feature exists
In, in step (2), described sand mill is excellent pin-type sand mill or turbine type sand mill.
5. the method that the quick broken wall of the haematococcus pluvialis cell according to claim 1 or 4 is coupled extraction astaxanthin, its feature
It is, in step (2), described sand mill, in its grinding chamber, filler is ceramic fine bead, and in grinding chamber, microballon loading amount is grinding chamber
The 75%-90% of volume.
6. the method that the quick broken wall of the haematococcus pluvialis cell according to claim 1 or 4 is coupled extraction astaxanthin, its feature
It is, in step (2), described sand mill, grinds cavity temperature and controls between 10-35 DEG C.
7. the method that the quick broken wall of the haematococcus pluvialis cell according to claim 1 or 4 is coupled extraction astaxanthin, its feature
It is, in step (2), the milled processed time is 10-600s/L feed liquid.
8. the method for haematococcus pluvialis cell according to claim 1 quick broken wall coupling extraction astaxanthin, its feature exists
In, in step (3), the mode of separation of solid and liquid is centrifugation, plate compression, negative pressure leaching or natural subsidence.
9. the method for haematococcus pluvialis cell according to claim 1 quick broken wall coupling extraction astaxanthin, its feature exists
In, in step (4), the temperature of vacuum distillation is 30-50 DEG C.
10. the method for haematococcus pluvialis cell according to claim 1 quick broken wall coupling extraction astaxanthin, its feature exists
In, in step (4), the organic solvent of recovery is circulated and mixed with haematococcus pluvialis in step (1).
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109369485A (en) * | 2018-12-06 | 2019-02-22 | 广州智特奇生物科技股份有限公司 | A kind of method of haematococcus pluvialis extraction astaxanthin ester |
| CN110301558A (en) * | 2019-08-06 | 2019-10-08 | 河南师范大学 | A kind of microcapsules fish feed additive and preparation method thereof rich in natural astaxanthin |
| CN110483358A (en) * | 2019-07-31 | 2019-11-22 | 杭州女舒生物科技有限公司 | A method of the rapidly extracting astaxanthin from haematococcus pluvialis |
| CN110511169A (en) * | 2019-07-31 | 2019-11-29 | 浙江九如堂生物科技有限公司 | The method for preparing Determination of Astaxanthin in Haematococcus Pluvialis based on Transdermal absorption |
| CN110732157A (en) * | 2019-10-18 | 2020-01-31 | 云南钰腾生物科技有限公司 | Extraction equipment and extraction method for astaxanthin with high stability |
| CN111217734A (en) * | 2018-11-27 | 2020-06-02 | 云南爱尔康生物技术有限公司 | Method for extracting astaxanthin from haematococcus pluvialis |
| CN111500461A (en) * | 2020-05-21 | 2020-08-07 | 南京理工大学 | Wall breaking method of haematococcus pluvialis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111217734A (en) * | 2018-11-27 | 2020-06-02 | 云南爱尔康生物技术有限公司 | Method for extracting astaxanthin from haematococcus pluvialis |
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| CN110732157B (en) * | 2019-10-18 | 2021-11-23 | 云南钰腾生物科技有限公司 | Extraction equipment and extraction method of high-stability astaxanthin |
| CN111500461A (en) * | 2020-05-21 | 2020-08-07 | 南京理工大学 | Wall breaking method of haematococcus pluvialis |
| CN111500461B (en) * | 2020-05-21 | 2022-08-09 | 南京理工大学 | Wall breaking method of haematococcus pluvialis |
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Application publication date: 20170222 |