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Nov. 10, 1970 K. LAUER ETAL 3,539,505

LARGE-SCALE COLUMN CHROMATOGRAPHY PROCESS AND APPARATUS THEREFOR Filed Oct. 22, 1968 2 Sheets-Sheet 1

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Nov. 10, 1970 K. LAUER ETAL 3,539,505

LARGE-SCALE COLUMN CHROMATOGRAPHY PROCESS AND APPARATUS THEREFOR Filed Oct. 22, 1968 2 Sheets-Sheet 2

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United States Patent Office

3,539,505

Patented Nov. 10, 1970

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This application is a continuation-in-part of application Ser. No. 552,934 filed May 25, 1966 and now abandoned.

The present invention is concerned with a process and apparatus for carrying out column chromatography on a large scale. More particularly this invention is concerned with a process and apparatus for carrying out liquid column chromatography on a large scale.

In recent years, column chromatography has become a conventional laboratory method for use in the purification and separation of materials. When attempts are made to use this method on a large scale, a number of difficulties arise which have heretofore rendered the advisability and practicability of large scale column chromatography questionable. Particularly serious difficulties arise with increasing length and diameter of the columns, by the "distorted running" of the fronts which lead to a poor utilization of the column capacity, and, especially in the case of low separation factors, to the impossibility of adaptation to large scale separation of the multi-component liquid.

The "distorted running" of the fronts is due to a great variety of different factors, such as non-uniform filling of the column, variations in temperature, viscosity and volume, canal formation, greater ease of flow between the column filling and the inner wall of the column, and the like. As has been established through experience, this "distorted running" cannot be avoided even when proceeding very carefully. Thus, whereas in the ideal case, the concentration time diagram of a liquid which has passed through a chromatographic column represents a more or less steep bell-shaped curve, in the case of columns used for large-scale chromatography, this diagram degenerates into completely irregular drawn-out shapes.

This invention has as an object a simple and practical method for carrying out liquid column chromatography on a large scale.

A further object of the invention is an apparatus for this purpose, i.e., applicable to carrying out liquid column chromatography on a large scale.

A still further object of the invention is a method and apparatus for accurately carrying out large-scale column chromatography, i.e., separating multi-component liquids into its components.

In accordance with the invention, it has now been found that these objects are accomplished and the disturbances arising in large-scale column chromatography due to the

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"distorted running" of the fronts avoided by radially mixing the axially flowing liquid in the column in small spaces arranged at predetermined intervals along the length of the column.

Concentration time diagrams obtained by analysis of multi-component liquids which have passed through a column constructed in accordance with the invention so as to provide for radially mixing of the axially flowing liquid at designated intervals along the column's length correspond to an almost ideal, steep, bell-shaped curve. Thus, the separation and purification effects which are observed in the considerably smaller dimensioned laboratory chromatography columns are also obtained in large scale operations and apparatus when processing according to the present invention.

This result is entirely unexpected and was not to have been foreseen as all radial mixing is automatically bound up with an axial mixing (return mixing). This return mixing invariably brings about a deterioration of the degree of separation achieved with the column and, in addition, leads to a flattening out of the concentration-time diagram. Thus, generally, an attempt is made to avoid return mixing and to keep it as low as possible. However, in the process according to the present invention, the radial mixing carried out at definite, i.e., predetermined distances leads, surprisingly, to a straightening out of the front and to such an increase in the performance of the column that the return mixing effect is no longer of importance.

The distances at which the radial mixing is to be carried out in the process according to the present invention are, in the main, independent of the column's diameter.

The mixing zones are advantageously located at distances of between 0.5 and 2 meters from one another and preferably are located at distances of 0.75 to 1.2 meters from one another. The height of the mixing zones is also independent of the column's diameter and advantageously amounts to from 1 to 5 cm. and preferably 2 to 3 cm.

In addition, the nature and particle size of the adsorption agent, the velocity of flow of the liquid phases and the specific separation factors are of importance. In practice, the optimum distances are determined empirically. If the distance between two mixing zones is too great, then disturbances due to "distorted running" of the fronts will take place. If, on the other hand, the distances are too small, then the elution curves are noticeably flattened, especially when the volumn of the mixing zones is too great in relation to the volume of the separation zones.

For a better understanding of the present invention, reference will be made to the accompanying drawings in which:

FIG. 1 shows one embodiment of a device for carrying out the process according to the present invention;

FIG. la shows in detail a part of a device for carrying out the process according to the present invention;

FIG. lb shows in detail a part of a device for carrying out the process according to the present invention;

FIG. lc shows in detail the deployment of a mixing means 7 within a space defined by upper and lower sieve, plates.

FIG. 2 shows in detail a preferred form of inserting a mixing zone into the column according to the present invention;

FIG. 2a shows another view of the construction of FIG. 2;

FIGS. 3a to c show concentration-time diagrams obtained according to the present invention and with conventional processes.

For carrying out the process according to the present invention, one or more pairs of sieve plates are incor

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porated into a column and a radial mixing of the eluting around the edges thereof. It is then possible for these agent takes place in the zone between the two sieve.plates combinations to be moved axially in the column correforming each pair. In the case of two or more pairs of sponding to the changes in volume of the column filling, sieve plates, the pairs are spaced apart. The adsorption Each sieve plate generally comprises a plate 1 conagent in the column is kept out of the mixing zone or taining holes and covered with a fine metallic or synzones by the sieve plates. The eluting agent, after having thetic resin gauze or mesh 2 which is not subject to attack been mixed in the mixing zone flows uniformly through by the elution agent, the mesh being arranged on that the lower sieve plate of the pair of sieve plates into the side of the sieve plate which is adjacent to the column next separation zone of the column. filling. The openings in the plate must be of such a size Generally, the mixing which automatically takes place ^ that the elution agent can pass through them without in the zone between the two sieve plates of a pair suf- encountering too much resistance but cannot be too large fices to produce satisfactory results. However, in the case in order that the mesh 2 is not subjected to very conof really large column diameters, this automatic radial siderable mechanical stresses. Opening diameters of 2-15 mixing alone is not sufficient and, therefore, it is advan- mm., preferably of 5-6 mm., have proved to be partageous in these instances to provide, between the sieve lg ticularly useful. The strength and mesh size of the meshes plates of each pair, a device which ensures a thorough to be used depend upon the particle size of the adsorpmixing of the liquid in the zone established between the tion agent used and, therefore, can be varied considertwo sieve plates of a pair. As mixing devices for this ably. Mesh sizes of 0.02-1 mm., preferably 0.1-0.2 mm., purpose, there are suitable, for example, mechanically- have been very successfully used. However, the dimendriven stirrers, magnetic stirrers, vibrators, ultrasonic 2o sions of the perforated plates, of the openings in these devices and the like, the choice of the possible and pref- plates and of the meshes must be so chosen that the erable mixing device depending in each instance, of adsorption agent cannot pass through and that the sieve course, on the dimensions and structural material of the plates do not become blocked up. In addition, of course, column. In actual practice, one embodiment of the inven- the sieve plates as a whole must be capable of withtion in which a centering plate 3 is arranged between 25 standing, without bending, the pressure load of the column a pair of sieve plates 1 and 1' has proved to be particu- filling, including the pressure due to the flow-through larly expedient. This embodiment has the particular of the elution liquid.

advantage, that, with structurally simple means which The dimensions of the mixing zones according to the

do not require supervision, a thorough radial mixing is present invention as noted above do not directly depend

ensured. Two instances of constructions of such center- gg upon the inner dimensions of the column and are advan

ing plates are shown in FIGS, la and lb of the accom- tageously 1 to 5 cm. in height and are located 0.5 to 2

panying drawings. In the case of a very large column meters apart. Clearly the distance between the two sieve

diameter, it may also prove advantageous to mount a plates should be as small as possible in order to avoid

distribution head below the centering plate. unnecessary remixing, but they must be large enough to

A radial mixing can also be achieved, with or without gg ensure complete axial mixing. When using sieve plates

the use of the centering plate by utilizing an arrange- wherein the openings in the sieve plates are not parallel

ment wherein the openings in the sieve plates are not to the axis thereof but rather are positioned at an angle

parallel to the axis thereof but rather are positioned at up to some 50°, axial mixing is established in a smaller

an angle to the axis so that fluid flowing through the area and the distance between the sieve plates of each

openings tends to rotate about the axis of the sieve plate. 40 pair can be reduced.

With a suitable thickness of the sieve plate and a suf- The material used in the construction of the mesh acficient angle of the openings, by the rotation of the cording to the present invention depends on the solvent column of liquid between the two sieve plates there is and the absorption agents as used within this process. Maensured a sufficient mixing and a uniform impinging of terials such as polyvinylidene chloride, polypropylene, the liquid over the whole of the lower sieve plate. 45 nylon, Perlon, Teflon, nickel-plated wire mesh and stain

The incorporation of the sieve plates into the column less steel mesh can be used with good success, can be achieved by flanging, that is, by constructing a The material of the gasket is used according to the column from a plurality of short columns joined together present invention can be chosen from the broad range of by pairs of sieve plates of the above-described type. How- compressible materials which are not affected by the elutever, it has proved to be especially advantageous to use 50 jng agent and include, for example, rubber synthetic rubone long column unit and to insert therein one or more ber, silicon, and Teflon.

combinations each comprising a pair of sieve plates and The optimum distances between any two mixing zones,

preferably also a centering plate. These combinations j.e.; the location of the mixing zones in the column is

(two sieve plates and possibly a centering plate) can comparatively independent of the column diameter and

be moved axially into any desired position and are held 55 advantageously is chosen so that the mixing zones are

in place by various means as, for example, by an annular located at distances one from the other at distances of

gasket 4. Furthermore, these combinations can be pro- o.5 to 2 meters and preferably at distances of 0.75 to 1.2

vided with one or two annular distance pieces or spacers meters. The dimensions, i.e., height of the mixing zones is

5, dimensioned so that their outer diameter is somewhat also aimost completely independent of the column diame

less than the inner diameter of the column, in order to 6Q ter and advantageously amounts to from 1 to 5 cm. and

combine these combinations to form a single unit. This preferably from 2 to 3 cm. in height. The installation,

preferred constructional form of the device according to ;e; the fastening of the individual mixing zones into

the present invention can be inserted at any desired posi- columns having fairly large diameters is preferably car

tion in the column. Consequently, a column which has ried out by screwing the mixing elements to the wall

already been set up, m this, manner, can be adapted 6_ througn narrow consoles fastened to the inner walls of

quickly and easily to the prevailing conditions resulting, a coiumn. In order t0 enable the installation and removal

for example, from the nature of the adsorption agent, of the m;xjng eiernents from the column, the elements

the flow-through rate and the like, so that it is always are formed with corresponding grooves which, if desired,

possible to work under optimum conditions. In the case can be sealed prior to operation of a column. The seal

of the use of column fillings which swell considerably 70 mg 0f the individual zones to the column wall is prefer

and, depending on the degree of charging, undergo con- ably carried out by constructing the mixing elements

siderable changes in volume (e.g., ion exchangers), it with conically tapering walls. Sealing rings are then in

is recommended only to draw on the compressible annular stalled and held through the conically tapering walls of

gasket to such an extent that adsorption agent and elution the mixing zone section in close abutting contact with the

liquid cannot flow past the sieve plate combinations 75 abutting wall.

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