DE2015818C3 - Process for the selective elution and removal of liquid substances and device for carrying out the process - Google Patents

Description

The invention relates to a method for the selective elution and withdrawal of liquid substances under Use of an adsorbent passed continuously through an elution zone, from which the eluted substance is continuously withdrawn. The invention also relates to a device for carrying out the method, consisting of a container filled with liquid, the an upper connection for giving up and a lower connection for discharging the loaded or unloaded adsorbent as well as connections for the supply and discharge of liquids having.

There are already various methods of eluting adsorbed on different carriers Known substances in which ion exchange resins are also used as adsorbents. One of these In professional circles, the process referred to as the "batch method" is generally used as the first elution stage or first cleaning stage used. Another known as the "chromatography method" Process is particularly suitable for obtaining eluted substances of higher purity. But when at the same time a very large amount of Adsorplionsmilteln used in the chromatography method there is a very low flow rate or a very low throughput of solvent liquid. Accordingly, the amount of adsorbent or carrier substances used in a »batch« limited. Because of the limited absorption capacity of the adsorbent itself, the elution can take place or the deposition cannot be carried out quickly and thoroughly on a large scale.

Processes for the continuous separation of aromatic and saturated hydrocarbons using silica gel or activated carbon have also already been described, in which the adsorbents in the form of a moving bed are passed through a column in countercurrent to the liquid mixing. The amount of aromatic hydrocarbons bound by the adsorbent is then removed in a desorption zone, ie elution zone, and the adsorbent is returned to the column. The equipment necessary to carry out this process, e.g. B for the transport of the adsorbent, are extensive and expensive (German Ausleseschrift 1 06 ^l > 602).

ίο Feiner is a device for treating Liquids known (German Auslegeschrift 1 294 933), which consists of a tubular loading container and a likewise tubular, downstream There is a regeneration tank and in which the circulated via a line system Ioncnaustauschermasse is cleaned in a further downstream washing tank. The usage two separate containers require a special one for the circulation of the adsorbent Line system with appropriate pumping or conveying devices.

The invention is based on the object of a method and a device for selective elution one or more substances from their Adas sorbents without the disadvantages mentioned above to create, which enable a steady work or a continuous throughput and with which the fluates are highly concentrated.

According to the invention, this object is achieved in that the adsorption material, together with the substance to be eluted, is moved through a narrowly delimited and stationary mixing zone N formed by a stationary buffer zone and a slightly more specifically dense stationary eluting liquid that can in principle be mixed with it, in which the substances attached to the adsorbent are selectively eluted and drawn off as a concentrate, while the other substances are moved with the adsorbent or are discharged with the overflow.

Due to the different specific weights of the two miscible liquids, d. H. due to the lighter buffer solution and the specifically heavier eluting liquid, the Mixing zone of both liquids, in which the specific gravity slowly increases to the larger value of the lower eluting liquid increases. The ones with the ones to be deposited Adsorbents loaded with substances sink through the upper buffer solution and are in this Mkch / one due to the increasing specific weight gradient and the associated associated reduction in weight difference gradually slowed down. Because of the greater affinity of the substances to be eluted to the lower eluting liquid takes place during the sinking through Mixing zone, a desorption takes place, with the cluted substances being concentrated in this mixing zone and can be withdrawn from it.

If in or on the carriers or the Adsorptionsniittcln two or more types of fabrics adhere, it is advantageous to have several mixing zones between to form each pair of adjoining fluids and these fluids to dig in such a way that each of the substances to be eluted as an eluate layer in a mixing zone due to the differences in affinities compared to ilen Liquids separated on both sides of the mixing zone and is focused. By aiifeiiumdeiT letiules

5 6

Sinking of the adsorbent through the ordered. Hin short inner cylinder 10 is at the

Mixing zones can all be defined by the inner wall of the carrier of the cylinder 1 with the aid of an O-ring 9

elute or extract adsorbed substances. attached, which has the task of, in addition to a sealing

With particular advantage, the device according to the invention can also have a suitable radial distance between

Method and device for cleaning a 5 of the inner wall surface of the cylinder I and the

Liquid by accumulation on the adsorption outer wall surface of the short inner cylinder 10

medium and downstream elution in continuicr- to be observed. The short inner cylinder lies above the

lichcm operation can be used. second outlet 8.

Further peculiarities and advantages as well as additional According to Fig. 1, two are in principle mixed

Features of the invention emerge from the "follow- io bare liquids of slightly different

the description of embodiments that are specific gravity, for example in the

on the embodiment shown in the drawing, differ in concentration, pH, etc., in

games relates. In the drawing shows the cylinder 1 filled separately, so that a

I- "i g. 1 a schematic axial section representation of the mixing zone between the first liquid and the

a device for carrying out the second liquid according to the invention. The level of the

according to the method, mixing zone is - as Fig. 1 shows - on the Ni

F i g. 2 is a schematic cross section taken along the level N , ie around the lower edge

Line H-II of Fig. 1, the short inner cylinder 10, through enlspre-

F i g. 3 shows a schematic cross section along the corresponding dimensioning of the volumes of the first and the

Line HI-III of Figs. 1, ²⁰ second liquid. It is assumed that the

F i g. 4 a schematic cross section along the first liquid, the liquid above the boundary

Line IV-IV of Fig. 1,. area N and the second liquid below the

F i g. 5 is a schematic cross-section along interface N.

Line VV of Fig. 1. Under these conditions, an adsorption Fig. 6 shows a schematic cross section along the means introduced into the first liquid, the line VI-VI in FIG. 1, the first liquid into the second liquid through FIG. 7 shows a schematic cross section along which the mixing zone N passes. A gravity line VII-VII of Fig. 1, sedimentation or a centrifugal force er-Fig. 8 shows a schematic cross section along the forced movement, which will be explained later, line VIII-VIIl of FIGS. 1, 30 can be used to generate the movement of the adsorbent. FIG. 9 shows a schematic vertical section through. If another embodiment of a device for several types of substances to be eluted is bound to the adsorbent and carrying out the method according to the invention, the different affinities towards Lö-Fi and 10 have a schematic cross section of a solvent liquid, and one of these substances is a cylinder section according to FIG . 9, 35 has a higher affinity compared to the second liquid F i s. 11 a plan view of the object of the kit, only this substance is in passage Fic. 10. Eluted through the mixing zone N. In this mixed zone Fi a. 12 shows a cross section along the line XII-XlI, an eluate layer with the concentrated derF "ig. 10. cluicrtcn substance is formed. You can now see this eluate layer through XHI of FIG. 10, "withdraw the first outlet6. Since the liquid wedge in FIG. 14 is a schematic vertical section through the annular space Π on the outer circumference of the inner another embodiment of a device for the pipe 10 against gravity flows upwards. The method according to the invention is carried out. no adsorbent from the cylinder 1 her-Fig. 15 shows a schematic vertical section through 45, unless the flow rate digkeii still another embodiment of a device when the eluate layer is drawn off by the method for carrying out the method according to the invention. Outlet 6 would be too big.

16 shows a schematic vertical section through. Of course, those substances remain which have a nui

yet another embodiment of a device with low affinity for the second liquid for carrying out the process according to the invention have 5 °, adsorbed by the carrier and sink mi

drive and "this adsorbent further down. You he

F i o. 17 finally a special modification reach the bottom part of the cylinder 1 and become

the device for the implementation of the invention from this together through the second outlet 8

appropriate forfeiture. ^{withdrawn with} their adsorbents. When the first embodiment of an eluate according to the invention is drawn off through the first outlet 6

Device for carrying out the method and the carrier or carriers through the second Aus

is determined according to the invention, is in Fig. 1 let 8 drain, the volume de is reduced

to 8 shown. The device consists of a second liquid. As a result, the Ni lowers

Cylinder 1 with an end open at the top and a height of the mixing zone TV. Such a level lockable lower end. The cylinder is with 60 change but can be eliminated by dal

an upper Scitcnauslaß 2. below it step by step through the refill inlet 7 second liquid

inlets 3, 4 and 5 arranged one below the other, additionally introduces in such an amount that si

a main outlet 6, a refill inlet ζ and compensates for the loss of the second fluid,

give away the lockable floor outlet 8. The material to be clued can already be sent to the odc These openings are the second outlet 8 at the bottom 65 to be adsorbed by the carrier before this adsorbent

of cylinder I and the other openings - mostly medium in the first liquid cingcnrav.lv «wci

to several distributed over the scope - to gccig- den. The material can also be used in the first FH'issk

η the level of the height in the wall of the cylinder I must be dispersed and on the basis of its affinity

8 3 δ

are adsorbed by the carriers before the carriers reach mixing zone N.

As stated above, those substances which have little or no affinity for the second liquid, together with their adsorbents, get down to the bottom of cylinder 1. However, if there are further liquids, a third, a fourth, up to a / 1-ICn -FlUsS ₁ SkCIt (where η denotes a positive integer of 3 or more) with correspondingly large affinities for certain substances from the bottom side of the second liquid downwards one above the other in the cylinder, so that mixing zones corresponding to the mixing zone N are present between pairs of these liquids are, the substances are eluted from the carriers according to their affinities in the mixing zones as soon as the carriers pass through the corresponding mixing zone. Eluate layers of the various substances then form in the respective mixing zones. These elual layers can be removed in the manner described above through additional outlets in the cylinder.

The present invention works mainly according to the principle outlined above. It should two exemplary embodiments will now be described, which are carried out using a device according to FIG the F i g. 1 to 8 were carried out:

example 1

Extraction of xanthine oxidase using calcium phosphate gels as a carrier:

A device according to FIGS. 1 to 8 was used. A phosphate buffer solution (0.1 M, pH 7.0). containing 10% by weight ammonium sulfate was placed in the lower part of cylinder 1 and a phosphate powder solution ((). 1 M, pH 7.0) was placed in the upper part of the cylinder, forming a mixing zone N between the two solutions . Calcium phosphate Gclc. dispersed in a phosphate buffer solution (0.1 M, pH 7.0), into the cylinder 1 through the inlets 3 and at the same time also a crude enzyme solution obtained by extracting cow's milk in a phosphate buffer solution (0.1 M, pH 7.0). introduced through the inlets 4. This crude enzyme solution was the material to be cleaned. It was adsorbed onto the supports. Furthermore, the phosphate buffer solution (0.1 M, pH 7.0) was introduced into the cylinder 1 through the inlets 5. When the carrier sank, the xanthine oxidase was eluted from the carrier in the mixing zone N and a brown, concentrated eluate liquid layer formed at the interface. Then the eluate liquid was withdrawn to the outside through the first outlets 6 and examined. The purity of the enzyme deposited in this way was about 95% and the yield about 90%. Provided that the solution or enzyme extracted from the cow's milk was 100%. The calcium phosphate gels that passed through the liquid layers the phosphate buffer solution and the buffer solution containing ammonium sulfate had passed through and had settled at the bottom of the cylinder, were withdrawn from the second outlet 8.

By adding phosphate buffer solution containing ammonium sulphate in the same additional amount as it approximately corresponded to the amounts taken from the cylinder via outlets 6 and 8 during withdrawal, ie by feeding this amount through inflow inlet 7, the mixing zone N kept at a constant level between the two liquids, so that destruction of the mixing zone by diffusion, adsorption of the liquid component from the adsorbent and by elution of the substance to be eluted into the liquid was prevented. A good result was achieved even with continuous operation over a long period of time. The impurities which did not remain adsorbed in the carriers were flushed out from the upper outlet 2, and the impurities which had not been eluted from a phosphate powder solution with ammonium sulfide flowed out through the second outlet 8 as they were attached the adsorbents remained bound.

Example 2

The elution was carried out in the same manner as in Example 1: however, they were different instead of calcium phosphate glycine, DEAE cellulose as a carrier and a phosphate sulfate solution (0.01 M, pH 7.0) in place of the phosphate solution (0.1 M, pH 7.0) was used. A pyrophosphate buffer solution (0.1 M, pH 8.3) formed the eluent solution to elute the adsorbed substance instead of the phosphate buffer solution containing ammonium sulphate. The purity of the enzyme thus obtained was about 95% and the yield was about about SO ". u.

In the F i g. 9 to 13 is another embodiment of an apparatus for practicing the invention Procedure illustrated. A cylinder 12 with an open-topped end and a The separable lower end consists of five cylindrical ones Unit pieces 13 to 17 and a bottom cylinder piece 18 with a basic outlet 24. The cylindrical Pieces 13 to 17 are of the same shape and size: they are more detailed in the Fig. K) to 13 shown. As from these figures, particularly from Fig. 10, each unit cylinder piece consists of a cylinder section with Inlets 19 and outlets 20 and an approximately funnel-shaped designed insert 22 which is supported on the cylindrical Einhcitsslück. The inlets 19 lie on the lower part of the cylindrical unit 21 and the outlets 20 on the upper part of this cylinder piece 21. The slightly funnel-shaped Inserts 22 are level with the outlets 20, with an annular space in front of these outlets 23 between its outer funnel-shaped surface and the inner wall surface of the cylindrical Leave unit 21 open. This free annular space 23 directs the flow against the Gravity flowing liquid withdrawn from outlet 20. As a result, the Carrier, which sinks downwards, not into this to be withdrawn flow, unless the flow velocity withdrawal from outlet 20 would be too great.

In this embodiment, five different liquids are used by each becomes greater specific gravity, and are formed in the lower opening of the inserts? .L four mixing zones N ₁ to N ₄ between pairs of these fünl liquids.

409609/133

■ 3 ο c

- > ο J υ

9 10

There will now be a use of this, without the mixing zones between the liquid

Device carried out example described: keitspaaren to disturb.

The advantages of the invention when using

Example 1 of the embodiments according to FIGS. 1 to 13 can be achieved

5 can be summarized as follows:

It was about the activation of DEAE cellulose by a) In the usual chromatography used up to now

Eluicrcn the substances bound to this cellulose phie method is a limitation of the throughput

with an acid, an alkali and sodium chloride in the amount of solid adsorbent or solid

to obtain. Support materials as the stationary phase, so that a

1. An aqueous solution of sodium ^1O separation and purification in larger scales chloride (2.0 M) can not be performed in the cylinder on the lower ™ ^l good efficiency, introduced most inlets 19e. In contrast, in the case of the measurement

2. Then an aqueous solution of NaOH method the stationary phase a liquid and the (1.0 M) and NaCl (0.5 M) via the ^inlets 19 d ^{bewff e} . ^Phase . ^Iie gf ^{as fes} ! ^es _u ^material ^ - Hence

b cht ^{l5 s} no difficulties with regard to the effective

3. Furthermore, an aqueous solution of NaCl was ^{samkeit u} . ^{nd the} throughput when applying the (1.0 M) present invention via the inlets 19r into the cylinder, i.e. the elu.e ^v . ,. /, ^J ren and cleaning can be quick and very effective in

4. Finally, a large scale aqueous solution of HCl was run.

(1.0N) via the inlets 19 for into the cylinder ^ao g Not only the separation or the elution of the

■ uu, substances, but also the cleaning of the carrier can

einoracni. ' be performed.

5. A suspension of DEAE cellulose on which. _. ^& _,, .... , ,,,
by an acid, an alkali and sodium J The separation or Elu.cren the StofTe or chloride to eluting ^ n St adsorbed ITe ₀ "f" ^ ZTZ ^{exchanger can KONTL "uierllc 'by'} and in. water was dispersed, was gerunri ^{in 5.}

introduced the cylinder via the inlets 19a. ^d > In the case of continuous separation or purification is

Because of the different specific needs, the muscle zones on a constant basis

mixed zones N ₁ , N. "N, and to maintain the level. This can be achieved by

/ V ₄ between the aqueous solutions: The in that one de volume throughputs of the liquids,

the adsorbent (DEAE cellulose) ad- ³ ° de removed in each unit of time and e.nge-

sorbed StofTe were eluted while the ^fuh _h ^r _t · ^are "" / Zander tunes or equal

Carrier slowly down through the four under- ™ ^aHL < ^d "'' ^{man kan} " P. ^um ^ ⁿ . use which

harmful fluid layers sank. The conditions contained ⁱⁿ the compilation (I)

Substance to be dissociated in an acid was then J »« * » ^e . ^rfullen · ^H ' ^{er2u can you} f **? £ ■ ««

in the mixing zone above the HCl solution ^{M de} / mixing zone by one of the following methods

and in the alkali solution going forward dissociating ^{ate 'to Danac!} l ^J "mpen control. the

Substance in the mixing zone in front of the NaOH solution altitude and the condition of the mixing zone can be determined

eluted. Eh in the respective mixing zones, for example by measuring an electromotive

ated substances were recorded continuously from the cy- ^ raft, by measuring an electrical wi-

tinder outwards by the corresponding ⁺ ° ^ ^ton * " ⁰ ^ '?"? ΐ ^^ ΤΓ '* "?

Outlets at the given level heights abee- a polarographic method by measuring the

leads. Then we the DEAE-cellulose activated f ^ M "Γ'ΐ h, ° _{fl ^H H} ^{Llc HTAL n]) TLON} 'fourth when ER- the bottom of the apparatus 2" ^ SS _11? "^"

had enough, from where the device “of the refraction of light, etc.

be withdrawn through the bottom outlet 24 " ^ln . ^F ' ^S _n J ⁴ _f f ^e _H ^lne * " ⁰ ^ * ^usfuh ™ ⁿ S sfonn emei.. E & device for carrying out the invention

^{KonnTe presented} according to the procedure. A container 25 loads

In this embodiment, the volume of a first chamber 27 and a second fluid flow rates (volume per time through a partition 26 from the first separation unit) were V 19a, V 19b, V 19c, F19d and F19e, the 5 <> th chamber 28. The two chambers 27 and 2 ″ via the corresponding inlets 19a, 19b, 19c. are at their upper and lower ends with one another 19 d and 19 e were introduced, and the volumes are in connection with one another. In the right-hand wall throughputs or amounts of liquid per unit time of the first chamber 27, ie the right-hand wall ^ 20a, V 20 b, V 20 c, V 20d and V 20e, which from the container 25, are an outlet 29, inlets 3f corresponding outlets 20 a, 20 b. 20c, 20d and 55 and 31, an outlet 33, an inlet 34, an outlet 3! 20e have been withdrawn, corresponding to the following and an inlet 36 at different levels

arranged with staggering downwards. In the left wall of the second chamber 28 are an upper one Inlet 37 and a lower outlet 38 provided 6o In the second chamber 28 is a carrier conveyor 39 arranged. It consists of rotating drives 40 to 43, such as gears, rollers, etc. unc an endless conveyor element 44 in the form of a chain of a belt or the like, which with these drives to-65 cooperates and to the appropriate sender

If the volume amounts are fixed according to the Obi vessels 45, n is the baskets, hooks. Gen like composition (I) have been observed, could have been. The whole thing is driven by an electric motor (not showing the DEAE cellulose activated continuously and evenly). The upper order

Conditions measured: V 20 a V 20b Compilation, 1 V 20c V 19a = V 20d V 19b = V 20e V 19c = V19d = K19e =

II 12

Return of the endless organ is to the side after the obe- solution through the inlet 31 with a volume At the end of the first chamber 27, through the salt ^ 31, add the phosphate buffer solution (0.1 M,

Embodiments will now be described below pH 7.0) through the inlet 34 with a volume use of this device, throughput ^ 34, 'the aqueous sodium solution through. 5 the inlet 36 with a volume ^throughput V 36 öeis P ' ^{el 4} and water through the inlet 37 with a volume

Purification of xanthine oxidase: a m throughput ^ 37; the solutions were provided by the

Device according to FIG. 14 is used, in which through the outlets 29, 33, 35 and 38 with volume flow rates

Inlet 36 an aqueous sodium chloride solution 1, OM from V 29, V 33, V 35 and V 38 withdrawn. The Vo-

was introduced. io lumen ratios corresponded to the following conditions

A phosphate buffer solution (0.1 M, pH 7.0) was used:

through inlet 34 and a crude xanthine oxidase assembly H

Solution introduced into the container via inlet 31- VlS - V 30 - ^L V 31

give. F33 = V 34

A phosphate buffer solution (0.01 M, pH 7.0) became 15 K 35 - K 36

through inlet 30 and through inlet 37 Was- ν "i% - V 37
filled with water.

In the first chamber 27, it was found that the impurities, the crude xanthine oxidase solution and the phosphate contained in the crude xanthine oxidase solution, phate buffer solution (0.1 M, pH 7.0) as well between the »0 were, but had no possibility of coupling with the phosphate buffer solution (0.1 M, pH 7.0) and the aqueous DEAE cellulose, continuously or with sodium chloride solution each a mixing zone / V ₁ non-continuous through the upper one Outlet 29 or N.,. In the second chamber 38 one could be guided. The solution of the purified mixed zone N ₃ between the water and the aqueous xanthine oxidase was removed through the outlet 33 of sodium chloride solution. Then a suitable one was drawn. The impurities, which were initially able to couple with nete amount of DEAE cellulose to the top of the DEAE cellulose and introduced from the end of the first chamber 27 as a carrier. aqueous sodium chloride solution (1.0M). The cellulose particles sank through the liquids, but could be withdrawn through the outlet 35; the one in the chamber 27 slowly downwards. The DEAE aqueous sodium chloride solution itself through the cellulose was first mixed with the phosphate buffer solution 30 38.

(0.01 M, pH 7.0) buffered, then with xanthine oxidase. When using this device, the and a portion of the impurities from the ro-DEAE cellulose as a carrier were not only coupled by the oxidase solution and then further after the second chamber 28 at the exit or task bottom. The DEAE cellulose was transported to the first chamber 27 at a point, where the phosphate buffer solution (0.1 M, pH 7.0), in which the xanthine oxidase no longer reacts to the DEAE cell within the second chamber 28. fourth. Furthermore, all work processes continued Iulose could remain bound. As a result, the xanthine oxidase was automatically eluted from the DEAE cellulose in the mixing zone N ₁ , so that the purification of the xanthine oxidase could be carried out quickly and effectively, while the DEAE cellulose could. In a practical application of this configuration together with impurities in the xanthine form, the chambers 27 and 28 oxidase can be lowered further down. In the mixed tube made from a bent U-tube, instead of a zone N., the impurities were eluted by dividing a single vessel with a separating aqueous sodium chloride solution (1.0M), and having to make up most of the wall activated DEAE cellulose settled at the bottom of the jar separating the first from the second compartment. Vessel from the device. 45 In Fig. 15 a further variant of a pre-

The endless conveyor 44 was illustrated with a method suitable for carrying out the inventive speed by means of the electric motor; driven in this device so that he has the deposed DEAE cellulose- are the same elements as in Fig. 14 with the same material with its buckets 45 in the second Kam- reference numerals or symbols. The Vormer 28 picked up and carried it upstairs. If the 5 ° direction according to FIG. 15 deviates from the design, the DEAE cellulose was passed through the mixing zone M, behind the device according to FIG. 14 in the following, took the place of the as dispersed points: The carrier flow mechanism 39 aqueous sodium solution to be viewed is available from a screw conveyor 47, which is now water from a chloride solution. In the upper section the motor 48 can be driven and a second pumping chamber 28 is obtained which forms a completely in 55 mechanism, such as DEAE cellulose, which is automatically known, which is washed with water by sludge pumps. The container 25 consists of a U-tube again in the upper section of the chamber 27. The first chamber 27 and the second chamber were brought and poured out there. The liquid withdrawn from the mer 28 at the upper free opening via an outlet 33 was a concentric connecting tube 48 which came to the first flow of xanthine oxidase, which is inclined in or on the mixer 27. The mode of operation of this pre-zone N _{1 had been} eluted and purified. If the direction according to FIG. 15 is essentially the same insertion and removal of the corresponding before as that of the device according to FIG took place, the.

the xanthine oxidase could be continuously purified 63 FIG. 16 shows yet another device

be, d. That is, the phosphate buffer solution (0.01 M, for carrying out the Veriah

pH 7.0) was through the inlet 30 with a Voren, in which the same elements

lumen flow rate I'30 introduced the crude oxidase as in Fig. 14 with the same reference numbers un <

13 14

ijmbolen are designated. The version according to The rotor 52 is allowed to rotate at 1500 rpm. 16 differs from the embodiment according to FIG. 14 and a phosphate buffer solution (0.1 M, pH 7.0), η from the following points: the carrier circulation, which contains 10 percent by weight of ammonium sulfate, to-Tieehanism or Umlaulmechanismus 39 only exists Introduced into the inlet compartment 54, from where it passes through the rotating drives 48 and 49 such as throws 5 through the connecting wheels, rollers and the like at the upper and lower end of the upper and lower end into the separating space 53. Then Jcr partition wall 26. Carrier 50 is through the two a solution of a small enzyme extract in the chambers 27 and 28 by means of endless elements of a phosphate buffer solution (0.1 M, pH 7.0) and there such a chain, belt, etc. or also passed through unmixed calcium phosphate gel into the inlet coupling with one another, entered io compartment 55, from which this mixture enters the separation space 53 via the channel group 65, is essentially the same as in the case of the A mixing zone N is formed between these processes according to FIGS. 14 and 15, so that the solutions in the separating space 53 also exist. that couples with xanthine oxidase. 15 or this oxidase adsorbed is called particles

In the Vot direction of FIG. 16, however, is thrown outward by the centrifugal force

compared to the devices according to FIGS. 14 and and passes through the mixing zone N. The GcI

15 the advantage that not only the orbital velocity particles lose their ability to

as a whole, but also to retain the sink oxidase as soon as it enters the phosphorus

speed of the carrier 50 freely selected and a 20 buffer solution with 10 percent by weight ammonium

can be asked. ■ sulfate, from which the xanthine oxidase: ni be

AIs material for the container that is rich with the liquid of the Mischzcnc N is duiert. Then will

wedges are filled, the gel particles w.-iterhin by centrifugal force

transparent material, such as transparent acrylic resin moved outside. Those components of the

or transparent glass is used so that the raw Εη / \ m solution, which is not affected by the calcium

conditions> 1cs device recognized immediately from the outside phosphate-Gcl have been adsorbed, d. H. the Vcr

can be. impurities, are removed from zone Ll of the separating

In FIG. 17, a further embodiment space 53 is conveyed to the outlet compartment 56 via the vanal mold for carrying out the compounding 61 according to the invention, and the solution of the xanthine process is illustrated. An oxidase on a drive shaft 30. The rotor 52 seated in the mixing zone N in the separation space elu-51 contains a double conical shape and is fed to the outlet compartment 57 via the channel-shaped rotor space or separation space 53, ring conveying group 62. The calcium phosphate gel is mige inlet compartments 54 and 55 and annular outlets not taken into the compartment 57, since the flow compartments 56 to 58. These compartments are concentric flow direction of the liquid in the channels 62 to the axis of the drive shaft 51. The inlet compartments 35 to Part opposite to the direction of centrifugal force 54 and 55 are at suitable intervals above the runs; the gel does not come into this channel and the separation space 53 is not arranged, and the outlet compartments 56 into it.

to 58 are at suitable intervals below the calcium phosphate gel or the Gclpartikel. Separation space 53 is provided. Furthermore, those that are dispersed in the phosphate buffer solution, the rotor52 groups of connecting channels 59 to 40 contain 10 percent by weight ammonium sulfate, whoever 63. The Kanalgruppc 59 goes from an opening 64 from the zone L 1 of the separation space 53 through the at the upper part of the outermost zone Ll of the separation connecting channels 63 in the outlet compartment 58 space 53 to the inlet compartment 54; the channel group directs. The impurities from the outlet compartment 60 lead from an opening 65 at the upper part of the 56, the xanthine oxidase solution from the outlet closer to the center of the separating space 53 and the 45 compartment 59 and the calcium phosphate gel particles from zone L1 to the inlet compartment 55 : the ducts to the outlet compartment 58 are from the rotor through group 63 leads from an opening 66 in the lower the outlet ducts 70, 71 and 72 using part of the zone IA to the outlet compartment 58: the duct of a suction device (not in the drawing group 61 from an opening 67 in the lower part of the illustrated) sucked. The mixing zone Λ 'can i; i middle zone L1 to the outlet compartment 56 and the channel group 62 of a constant certain position kept nalgruppe 62 leads from an opening 68 in the lower one by suitably placing the supply part approximately between the two zones A quantity of the solution reaches the inlet sections 54 and 55, in which - as will be shown later - one of the solutions is coordinated with the withdrawal quantity (in each case quantities in the time mixing zone N between the zones IA and 1.1. unit), which compartments 56 to 58 are withdrawn from the outlet to the outlet compartment 57, 55. Consequently

The device according to FIG. 17 is furthermore capable of separating and cleaning the to be obtained

Inlet lines 68 and 69 and outlet lines 70 .Xanthine oxidase without any interference.

give away his 72. Each end of the inlet conduits 68 can be routed through.

and 69 protrudes into an inlet compartment 54 and 55. Lim the mixed zone N in a certain position zi

without the rotation of the rotor 52 and the shaft 51 60 should hold the volume flow rates of the solutions

disturb. Likewise, each air duct 70 is connected to the inlet compartments 54 and 55. d. H. the Volumer

to 72 in the assigned outlet compartment 56. 57 or V 54 and I 55. as well as the volumetric flow rates. di (

58 led into it. can be withdrawn from outlet sections 56-58

The mode of operation of the device according to FIG. 17 i.e. the volumes V 56, V 57 and V 58. as well as there:

with reference to an implementation at 65 volume ratio of the calcium phosphate gels selbs

game of extracting and purifying xanthine to the dispersion solution containing the calcium phosphate

Oxidase illustrated, which contains calcium phosphate gels as gels and which are fed into the inlet compartment 55

Carriers used: will meet the following conditions:

Compilation III

V 54 f- KV 55 -
V 55 (lK)

approximately equal to V 58 \
approximately equal to V 56 ί

L ₁
L.

L ₁ (- L ₂ - roughly equal to K 57

The ones in the Auslali Abbey! The solution arriving in 57 comes partly from that layer of the liquid slightly outside the mixing zone N and partly from the layer of the liquid which is further inside than the mixing zone N ; these proportions of the volume per unit of time are given as L ₁ (from the outer layer) and L., (from the inner layer).

L ₁ and L, are usually extremely small compared to V 58 and V 56. The indication "approximately the same" proves to be necessary if a longer time unit is used to determine the comparison quantities.

In Fig. 17 is the drive shaft of the rotor vertical; but it can also be placed horizontally. If the wave is horizontal, the solutions will be taken within the inlet and outlet compartments 54 to 58 at approximately the same level, if the liquid flow rates are not exceptionally large.

When using the embodiment of the device according to FIG gel-like carrier adsorbed or coupled components at the separating gap between the liquid layers safely eluted, and the descent / speed of the carrier can be increased by increasing the centrifugal force compared to mil a settling that occurs solely due to gravity. Therefore, by applying a Centrifuge according to FIG. 17 significantly increases the total throughput and the total production time.

For this purpose 2 sheets of drawings

Claims (17)

Patent claims: 1. Method of selectively eluting and withdrawing liquid substances using an adsorbent passed continuously through an elution zone, from which the eluted substance is continuously withdrawn, characterized in that the adsorption material together with the eluting- io. the substance by one of a dormant Buffer zone and a slightly more specific dense resting zone that can in principle be mixed with it Eluting liquid formed, narrow and stationary mixing zone (Λ ') is moved, in which the substances attached to the adsorbent are selectively eluted and drawn off as a concentrate are, while the other substances are moved with the adsorbent or with can be discharged from the overflow. "O 2. The method according to claim 1, characterized in that the substances before their Treatment are adsorbed by the adsorbent. 3. The method according to claim 1, characterized in that unloaded adsorbent is introduced into a solution containing the substances and arranged above the buffer solution, that a selective adsorption takes place in this solution and that that loaded with the substance Adsorbent slowly moved through the buffer solution, the mixing zone and the eluting liquid The eluate concentrated here is withdrawn from the mixing zone. 4. The method according to any one of claims 1 to 3, characterized in that several eluting liquids of different density are arranged on top of one another with the formation of mutual, narrowly delimited mixing zones (N ₁ to / V ₄ ) and that an adsorbent to which several substances to be cluted are bound, moved through the liquids and their mixing zones and the eluate concentrates are withdrawn from these mixing zones. 5. The method according to claim 4, characterized in that that at least one of the elutable substances in the first liquid from the adsorbent material is adsorbed. 6. The method according to any one of claims 1 to 5, characterized in that the mixing zones in a Zcntrifugalkraftfcld are formed and the adsorption material by centrifugal forces is moved through the mixing zones. 7. The method according to any one of claims 1 to 6, characterized in that the eluate together is withdrawn from the mixing zone with parts of the eluting liquid and the buffer solution and that the resulting loss of liquid from the mixing zone by replenishing the same or corresponding amounts of the withdrawn liquid wedges is compensated. X. The method according to any one of claims 1 to 7, characterized in that the adsorbent is promoted by several misehings in communicating classes in circulation and that the liquid with dun to cluating substances continuously refilled and the concentrated liluate is continuously withdrawn. 9. Device for performing the method according to one of the preceding claims, consisting of a liquid-filled container which has an upper connection for feeding and a lower connection for discharging the loaded or unloaded adsorbent and connections for adding and removing treatment liquids, characterized in that in the container (1) several liquids of slightly different density are arranged one above the other to form narrowly delimited mixing zones (N) , at least one of which is an eluting liquid, that the outlets (6) for the egg are at the level of the upper mixing zone of the Eluting liquid are arranged on the container (1) and that the inflow connections (7) for refilling with eluting liquid open into the container (1) in the area of the eluting liquid column. 10. The device according to claim 1, characterized in that at the level of the mixing zone (N) in the container (1), a short inner cylinder (10) is arranged sealed against the container wall, which has a narrow ring channel (11) leading to the outlet (6) limited. 11. The device according to claim 1 or 2, characterized in that several eluent liquids of different densities are arranged one above the other in a container with the formation of mutual mixing zones and that an outlet at the level of each mixing zone (/ V ₁ , N.,) to withdraw the various eluates. is provided. 12. The device according to claim 3, characterized in that the container (25) is divided by a longitudinal wall (26) into two communicating via a lower and an upper flow chambers (27,28) that in one chamber (27) several Liquids of different densities are arranged to form narrowly delimited mixing zones (/ V ₁ , N.,) , of which at least one is a buffer solution and another is an eluting liquid, and that the draws (33, 35) for the eluate are at the level of these mixing zones and inflows (29, 30, 31) for replenishing buffer solution and adsorbent open into this container chamber (27) and that in the other chamber (28) a mechanical conveying device (39 to 45) for a continuous circulation of the adsorbent, two different densities, a mixing zone (Ν. _Λ ) forming liquids and at least one outlet (38) are provided. 13. The device according to claim 9, characterized in that that the conveyor (39 to 45) of the chamber (28) is an endless, driven Conveyor element (39), one part of which extends from the bottom of the chamber (28) to the upper one The end of the first chamber (27) extends and that with FördcrgcfäßL-n (45) for receiving and Unloading the adsorbent is provided. 14. The device according to claim ⁽ ), characterized in that the conveying device comprises a driven screw conveyor (46, 47) arranged in the second chamber (28). 15. The device according to claim 9, characterized in that that the conveyor has a drivable, endless row (39) of one another connected, by the first and the second Chamber (27 or 28) comprises circulating adsorbent carrier (SQ). 16. The device according to claim 9, characterized in that the first (27) and the second chamber (28) in the region of the Mischzohen (Λ / ,, N. " / Vj) each have a certain outlet for drawing off liquids, which in the are essentially opposite to the direction of movement of the adsorbent in the mixing zone. 17. Device for performing the method according to claim 6, characterized by a centrifuge rotor (52) with an interior (53) into which at least two liquids can be introduced via feed lines (68, 69) in order to create an approximately cylindrical mixing zone (/ V ₁ ) and to move an adsorbent through this mixing zone by means of centrifugal acceleration, and through discharge lines (71, 72) going out of the mixing zone and from the area of the largest interior diameter of the rotor interior to draw off the eluate and the adsorbent.