GB2301587A

GB2301587A - Astaxanthin carotenoid pigment recovery

Info

Publication number: GB2301587A
Application number: GB9610213A
Authority: GB
Inventors: Meshulam-Simon Galit; Gavrieli Jonah
Original assignee: IMI
Current assignee: IMI
Priority date: 1995-05-31
Filing date: 1996-05-16
Publication date: 1996-12-11
Anticipated expiration: 2016-05-16
Also published as: JPH09111139A; NO962225D0; AU5248396A; IL113935A0; CA2177752A1; NO962225L; IL113935A; DE19620471A1; GB9610213D0; GB2301587B

Description

2301587 PROCESS FOR THE RECOVERY OF CAROTENOID PIGMENT FROM ALGAE The

present invention relates broadly to the recovery of astaxanthin carotenoid pigment from algae. More particularly, the invention relates to the recovery of astaxanthin carotenoid from Haematococcus pluvialis.

BACKGROUND OF THE INVENTION.

As known, astaxanthin is a red-violet crystalline carotenoid pigment found in natural origin especially in the shells of crustaceans, yeasts, and accumulates in Haematococcus pluvialis algae.It is largely used as a pigment- ation supplement composition, generally for incorporation in feeds and particularly in animal feeds such as various types of fish. Since the cell walls of this type of algae are rigid and flexible, and therefore not easily disintegrated, the valuable pigment is available only after its recovery from the respective algae.

According to the PCT Publication No. 89/06910, the method involves a grinding of encysted Haematococcus after a prior drying, at cryogenic temperatures, such as liquid nitrogen usually followed by the addition of an antioxi- dant in order to inhibit the degradation of the pigment. The algae become impervious to many types of chemical and physical stresses and accordingly will withstand conventional grinding techniques.

Another method suggested for processing of Haematococcus pluvialis, applicable mainly on a laboratory scale, involves a vigorous vortexing with glass balls.

- 2 Another suggested method was to perform an enzymatic pretreatment which has the purpose of weakening the algae walls, preferably followed by glass balls grinding. The disa.,,antage of this method is that it requires a separate washing and an additional drying step, to remove the enzyme residue from the final product. However,a complete recovery of astaxanthin pigment was not achieved in any of the above mentioned methods.

A recent French Patent Application No. 2,703,692 claims a method for obtaining carotenoids from Haematococcus algae. The method involves freeze drying the encysted Haematococcus cells at a temperature below -500C followed by the addition of sodium chloride and comminnting the cells, in order to obtain particles with an average size below about 10 4. The carotenoids are further obtained from the comminuted Haematococcus algae by extraction using known solvents, such as:methylene chloride, hexane, ive berzene, acetone, etc. Although, this method might,I positive results on a small scale, it is doubtful whether it WOUld be applicable on a large scale, due to the requir,,:d stringent conditions. Furthermore, the use of organic solvents for this purpose seems to be quite una L I I-a,: t ilkle. it is an object of the present invention to provide a new process for tho recovery of astaxanthin carotenoid pigmerit Ut-oni Haematococcus pluvialis algae. it is another - 3 object of the present invention to provide a new process for an efficient rupture of algae cell walls without using stringent conditions. It is yet another object of the Present invention, to provide a simple process for an efficient rupture of algae cell walls, which can be easily implemented on an industrial scale.

BRIEF DESCRIPTION.OF THE INVENTION.

The invention relateh to a process for the recovery of astaxanthin carotenoid pigment from Haematococcus pluvi- alis algae, which comprises the steps of: comminution of the algae in a liquid medium by rupturing the cell walls algae containing said carotenoid and dispersing the resulted pignient from said algae, being characterized by the fact that said comminution is carried out i... a high pressure apparatus consisting of two consecutive units:

a pressure vessel and an interaction chamber, whereby a slurry, of said algae containing above 1% solids and gerierally between 5% to 10% by weight, enters into;-,aid pressure vessel maintained at a pressure of at least 2G 2,000 psi and preferably in the range of 3,000 to 6,000 psi and is conveyed into said interaction chamber mairtain.ed at a pressure of above 10,000 psi, and prefe rahly in the range of '1300) to 18000 psi said interaction (-..onta-nirig at least one channel, thi--,-,;ugit which lite luryy is passed ax: an increased velocity to effect a jet a rupture of the walls of the algae cells and a substantially decrease in the particles size of the algae, which impinge the disintegrated particles within a low pressure zone, whereby the astaxanthin car-otenoid pigment is released from the comminuted algae and is subsequently recovered. This recovery may be carried out either by drying the slurry containing the pigment, or by extraction of the pigment with an organic solvent. In the latter case one may also conceive to disintegrate the algae walls in the presence of an organic solvent which will subsequently extract the pigment. The particles size after the rupture of the algae cell walls decrease to about 20 to.15 g.

11 was found that by using this process, more than 95% of the aigae cell walls are disintegrated in a very short period of time. As a result,a high release of the pigment is achieved, as indicated by a very high extraction efficiency of the astaxanthin pigment, which is two to three times higher than the amount obtained by the known conventional methods.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1, repr-sents a flowsheet of the process for the recovery of astaxanthin carotenoid from Haematococcus pluvialis according to the present invention. iligure '.', iitistl,ates the percentage of the rupture as a function of the concent-ation of the algae in the slurry entering into the apparatus.

DETAILED DESCRIPTION OF THE INVENTION.

According to the present invention, the rupture of the algae cell walls is based upon performing a turbulent jet on the liquid mixture containing the algae in a high pressure Ppparatus. The turbulent jet is produced along a common boundary as formed by the mixture. In oider to achieve a better efficiency of breaking the algae walls, it is preferred to recycle at least a substantial portion of the mixture through said chamber at the pressure prevailing in the respective unit at the corresponding ve. I oc i t Y.

The mixture f lowing through the channels develops a high 15 velocity, being under a high rressure, linear shear cavitation and impact forcks. In this manner, the broken Palticies of the cell walls became smaller and are dislpersed through the liquid medium. The solid particles of the biotrass containing the pigment are separated out from the dispersion followed by extraction of the pigmenL. Alternatively, the entire biomass may be dried.

The cxtent of the algae. eel[ walls breakage and the r t:! 1 t: -l _,- C of ihe (-.arotenoids (;an be evaluated according to the colotur imparted to the liquid medium, or quantitativt_-1y determined by a microscopic or spectroscopic examina t o n - 6 :-cording to a most preferred embodiment, it is suggested to add an antioxidant reagent, such as butylated hydroxyanisole, in order to avoid any oxidation of the pigment.

The recovery of the pigment from the algae according to the present invention was found to be more efficient than other known methods. in the following Table 1 the results of the piEment recovery from algae are given, as compared with other known methods.

Table 1: Comparison of astaxanthin extraction.

The method used Free mg astaxanthin/per dry weight of aigae.

The present invention Grinding in the presence of liquid nitrogen water extraction + heat En7ymatic treatment Vortexing with glass balls 19 17 14 13 9 The astaxanthin released from the algae and quantitatively determined.

it may be concluded from the above Table that only the meitiod using liquid nitrogen reaches an extent of release which is of the same order of magnitude as the process according to the present invention. However, the method using, liquid nitrogen shows the following main disadvantages: it can not be casiy applied on an industrial scale., it---equires a prior drying of the algae, and it is much more expensive than the process according to the present invention.

DETAILED DESCRIPTION OF THE FTGURES

AS shovn in the Figure 1, the algae biomass enter into the pressure vessel (Cell 1). In this cell, the worPing pressure is maintained in the range of 2000 to 6000 psi and causes a separation of the algae aggregates.According to a prefeired embodiment, the algae biontass is recycled in this cell at least three times, in order to increase the yield -F pigment recovery. The re,4uiied slurry from Cell 1, j n7 rac. t ion chamber (Cell 2) is cor.,k.,eyej to the where a rup t u re of t,,, ase.te cell walls occurs, resulting in a dispersion (if the pignicent Into the.olution.

'ft;t, %,v(--srkint, pressure in Cell 2 is between 101OU to 230oO ps: and pi.e.ferably between 13000 to 180C10 psi. As in Cell it is al,,;o most preferable to recycle the resulted ax least four tintes.

- 8 As shown in Figure 1, the astaxanthin pigment may be extracted (step 3A) from the dispeision followed by formulation (step 4A) arJ drying (Step 'SA).

According to another embodiment, thr- blomass slurry con taining the pigment is directly conveyed to a formulation stel) (4), whereby an antioxidant is added in order to protect the oxidation and decay of the pigment.

According to another embodiment (not shown in said Figure 1), encapsulating agents, such as polysaccharides and gelatin are incorporated, thus recovering the desired pigment in a particular foi-m.

in ibe last step, the pignient is dried or crystallive.] Et c cord i ng t o t lie spec i f j c req u i remen t, c j- i t 8 t;n-,, i sagea Ilse.

Figure 2, illustrates in a graphic inariner, the results obtained in t fie Example presented be 1 ow. As can be observed, tbe pereentage of the rupture is a fune t; on c) f t he concen tra t i,,.n of t he a 1 cae i n t tie s 1 bi r ry.

at. a concentration of 8.5% w/w algae, the rupture was 6RT. after five passages compared with 9 CA' after a concentration of.1.5% w/w also after the five Of course, the paranieters of tbe ille t h od i 11 a kiepend and may varied according ro the tsrpe of the as well as on the pressure n!ziintained a t the inictk.,t;cjn chamber, as wi!l be shown in Fxample 2.

1) - 9

Claims

The invention will be hereafter illustrated by the following Examples, being understood that these Examples are not limiting the scope of the invention, and are gi,ven only fer a,etter understanding of the invention. A peison skilled in the ar-k, after reading the present specification, will be in a position to insert slight modifications, without being outside the scope of the invention as covered by the appended Claims. EXAMPLE 1. A slurry of red algae grown in an outside pond, having a cori,-entration of 10.9% (weight/voiime), was utilized in this experiment. The slurry was diluted to four different of solids concentrations: 2.5%, 5%, 7% and 8.5% (weight/volume). Each portion (2.50 cc) was introduccd into the pressure vessel (Cell 1) where a pressure of between 2500 to 3500 psi was maintained. in order to avoid oxidation, the process was cai-t-ied out below an ambient temperature. The flow rate was maintained between 400 to 500 mlAnin The resulted solution was conveyed into the interaction chamber (Cell 2), where a pressure of between 13000 to 170oO psi was The extent (if 8 C op 1 c c oli n t (het-nito(..ryte'l - maintained. algae breakage was determined by a microof the integer algae in the measuring cell - 10 The results obtained are summari zed in Table 2. the following I'ABI,F 2: Percentage of rupture as a function of the concentration of the algae. Concentration % ruptured algae 1"0 weight/,,olume) A B c 2. i 19 83 94 36 74 91 7 42 74 92 8.5 23.5 59 68 A: one cy-cle. B: two Cycles. C. five cycies.

1 to e (Ice I-e.i; se t 1 c) n o f F,. 5% in ihe extent of' rupture at the concentrais explained by the high density of the sftit-r-.-v, which caused clogging of some channels.

2, illustrates graphically the above results.

EXAMt-1.F 2.

n 0 A n of 250 ec. of a --luriy' of red algae, coiitaining.

llon basis) was introduced into the c e 11 1 ' Where a in the range of 2000 to 4500 psi was ma t r,, 1 i, ned and t he s 1 kir ry w as re c y c l t d four times. f he 1 1 temperature was maintained at about 250C, in order to avoid oxidation of the pigment.

The resulted solution was conveyed into the interaction chamber (cell 2), where a pressure of 11000 psi was maintained.

The extent of algae breakage was determined by a microscopic count of the integer algae of the solution in the measuring cell and was found to be 84%.

The same experiment was repeated, but the solution froij, was conveyed into the interaction chamber a pressure of between 13000 to 17000 psi the cel 1 1 ' (cell 2), where was maintained. The extent of algae breaking in this case, determined as mentioned above was above 9.5% C 1. A 1 M S 1. A,roce,-,s for the recovery of astaxanthin carotenoid pigment from Fi&ematocoizcxis pluvialis algae, which comprises the steps of: comminution of the algae in a liquid medium by rupturing the algae cell walls contain ing said carotenoid, and dispersing the resulted pigment from -iici algae,being characterized by the fact that -,Rid comminction is carried out in a high pressure apparatus consisting of' two consecutive units: a pressure vessel and an interaction chamber, whereby a slurry of said algae containing above 1% solids and generally between S?,7 to 10% by weight enterR into said pressure tained at a pressure of at least 2000 p into said interaction chamber maintained at vessel mainsi and conveyed a pressure of above 10000 psi. said interaction chamber containing at least one channel, through which the slurry is passed at an ine-reb2,etl! velocity to effect a turbulent jet caus 1 11 g a rtiptti,-t, of the walJs of the algae as a resil 1 t of t fie walls disintegriation, which impinges the disintegrated pit.lieles within a low pressure zone co i,ijrii 1 nu 1 e( astaxanthin car-tenoid pigment is released algae and subsequently recovered.

2. -.he process according to Claim 1, wherein the vesse i S m a i n t a i re c d a t a pressure in the range o

3.C;1-jo to h,000 pC;j.

whereby the from t fie 13 - 3. The process according to' Claims 1 or 2, wherein the pressure in the interaction chamber is maintained between 11000 to 23000 psi.

4. The process according to Claims 1 to 3, wherein the recovery of the pigment is carried out by drying the resulted disintegrated algae cells.

5. The process according to Claims 1 to 3, wherein the recovery of the pigment is carried out by its extraction from the resulted disintegrated algae cells by an organic solvent.

6. The process according to Claiins 1 to 3, wherein the breaking of the algae cell walls, is increased by recy- cling a predetermined portion of the mixture containing the algae biomass through said apparatus.

7. The process according to Claims 1 to 6, wherein an antioxidant reagent is added during the process.

8. The process according to Claim 7, wherein said ati;oxidant is butylated hydroxy anisole.

9. A process for the recovery of astaxanthin carotenoid pigment from Haematococcus pluvialis algae, substantially as described in the specification and in any one of Claims 1 to 8.

10. A process for the recovery of astaxanthin carotenoid pigment from Haematococcus pluvialis algae, substantially as hereinbefore described with reference to, and as shown in, Figure 1 of the accompanying drawings.

11. A process for the recovery of astaxanthin carotenoid pigment from Haematococcus pluvialis algae, substantially as described in Examples 1 and 2.