DE1598223A1 - Method and device for dosing samples in the pressure circuit, in particular for devices for column chromatography - Google Patents

Description

Dr. Expl.

• Ceskoslovenska akademie yed, Praha

Method and device for dosing samples in the pressure circuit, in particular for devices for column chromatography

The present invention relates to a method and a device for metering samples in the pressure circuit, in particular for column chroniatography.

In automatic apparatus for analytical purposes, such as the analysis of amino acid mixtures and similar substances by means of elution from ion exchangers, the samples must be automatically brewed into the column at the beginning of the analysis namely from storage containers, into which they have been put by hand in certain periods of time. At present Systems it is necessary to bring several samples into the storage container in order to utilize the full capacity of the apparatus. In modern apparatus this capacity is 5 to analyzes in 24 hours.

In very efficiently working modern apparatus, the undesirable effects of the separation of the Gemisohe on the column become subdued. The sample container must be used for this purpose

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possibly arranged close to the end of the column i.e. in the pressure branch of the pump which pushes the eluate into the column. In order to reduce blurring of the concentration zones, the connection line to the sample container and the column must be capillary and possibly be made short and the inner surface of the same may only hold back the minimum possible of the sample. The same Conditions apply with regard to the sample container itself. In addition, the dosing device must transfer the whole Sample can be ensured in the shortest possible time from the beginning of the analysis from the sample container to the column. In the end it must be ensured that there is no leakage in the apparatus comes about during the pressure curve and not the slightest Sample parts come out of the dispensing system.

Eliminate the method and device according to the invention these disadvantages. The essence of the invention is that the individual samples are first sucked into the sample container and temporarily remain outside the pressure circuit of the pump and only switched on in this circuit when the sample is to be applied to the column at the beginning of the analysis.

In the device according to this invention, the sample containers are individually in the pressure circuit of the pump via Mehrwe ^ - taps and hydraulic distributors, whereby in one

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the

80Λ η Tetteiler dl * Pump directly is unH the individual sample containers of the pimp are.

half dee

The further linearities of the misfinding can be found in the two Iiegen4 ^; :: l || i.iiji * i ^ «i ■ i ^ lij ^^^ 'ivi! T.' ^ί :? S '''^;'■",/' ^: ^ 'ii

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In Fig. 1, only one of these capillaries is common- · _.t |

sam shown with the associated capillary connections. -iff

The multi-way cocks 4 and 5, in Fig. 1 as six-way cocks .'t

shown, can either with a shaft or with a fj

Spindle, or the spindles can be separated from each other. S |

separated and mechanically connected to gears 6 »and 2 %

be, if necessary also by a further connection O

rad j3 in such a way that they perform corresponding movements · * · · ^: 'YJi

This movement can be carried out by an independent electrical - ■>,

“See drive on G-round caused by electrical impulses V

sent by a central programming device J '

will. The indicated movement can also be written, for example, by; V;

a ratchet mechanism must be produced, whereby the ■? ·

Ratchet wheels £ this mechanism gradually to the associated-. -ί

can be turned further by the pawl J _- O. This . fl

The ratchet wheel is hinged to the swing arm JJ _- the one with the tooth ί

wheel J _- O, which meshes with the gears JJ5 and 21,; i through which the movement of two further hydraulic

divide Jj ^ and J_6 is concerned. These hydraulic distributors' |

I _- ^ and JMS can also be executed with a single shaft jf - ¹ ^ i

to be and to be moved back and forth by a division by a ^Λ ί

associated, known per se and not shown in Fig. 1; r

_r? Neten electric drive, which is controlled by a central programming * · 'ff ·

■ ^>; · ■ M:

facility is controlled. The hydraulic distributors Jj? and · J ^ - 1 ^ 6 can take two positions. Is in one position
the pump 21 ni * its pressure line JJ3 with the channel Jjjl ^{in the} rotary.

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i | . core of the hydraulic switch 15 with the connecting line Si 20 connected, which to one of the circumferential stages of the hydraulic

\ -? -'- ν-; Switch _1j5 leads. In the basic position of the rotatable core / ^ of the holder _1j5, the channels 21 connect the line £ 0 to the r! _{: i;} . ■ "'■'" line-22, which leads directly into the column "22 In the Toggle:;: -;; so run basic position is both the upper Vielweghahn 4_, and the lower ^{Vielweghahn..!.} ji disconnected from the pressure circuit and the pump V7 pushes the Bluent into the column 2 ^. The connection line 2A is always connected in the basic position with the other channel 2 £ of the spindle of the hydraulic switch _1J5 with the line _26 leading to the mouthpiece, with soft by sucking in with the mouth or with an easily adjustable suction device, the samples and the protective buffers can be sucked into the individual storage capillaries J ^ together with the separating vesicles be be connected appropriately rotating the Vielweghahnes 4 ^. the narrowed down capillary orifice 2 £ needs of the connection piece of the capillary pipe 3 is separated so that the. possibility by the narrowed lower mouth 2J_ the ^: sample and the protective buffers from the associated small vessels, which are temporarily held under the containers ^ so that the mouth 2J. reaches below the surface of the associated liquid. j;

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Fig. 1 also shows one of the many possible Constructions of implementation of the connections of the capillaries Line 2 shown. This line is tapered at its end

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expanded so that it cannot be pulled out of the socket 28, which is made, for example, of leflon and can be designed inside with a small sleeve made of silicone rubber. On the socket 2j3 sits from below & 9T carrier ^ O, which ensures a sufficiently tight connection of the container capillary with the capillary line J5. The container capillary χ can be supported here on the further carrier jH, which with the extension £ 2 in the lower part of the full diameter supports the storage capillary 2.

In the basic position of the hydraulic distributors 1ft and JjS described, there is therefore the possibility of filling all the reservoirs of capillaries that are not connected to the pressure circuit, with the exception of one capillary. This is achieved by turning the hydraulic switches JJ and IjS to the second functional position. In this acceptance position, the pressure line JJ3 leading from the pump J_7 through the channel Jj) ₁ to the connecting line 2 £ switches over, the connecting line being connected to one of the connecting pieces of the multi-way valve 4 and thus also to the associated storage capillary J_. Through this connection to the pressure circuit, the content of the capillary J_ is printed through the capillary line J5 via the hydraulic switch 5, and the capillary line £ 4 to the hydraulic switch J_6 in the column 22. As can be seen, the connecting line 2 £ remains disconnected between the hydraulic switches and Jj5.

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In this functional position, the entire device can only remain in the most necessary time, which is necessary for the transport of the sample, which is particularly necessary in individual cases where several samples have been introduced _{into the container capillary J 1.} From this it is possible to further convey only one of the samples introduced together with the associated protective buffers, which are separated from the samples by bubbles and from one another. Under no circumstances should another sample be sucked in. If, however, only one sample was introduced into the storage capillary, the entire linear movement can remain in the stated position for a long time and it is possible to flush the entire device with a smaller number of bubbles or even without them. Such a flushing can do that Replace the expulsion of the sample residues from the walls of the equipment and is carried out with a limited amount of buffer, which is separated by bubbles, which is particularly effective, especially against the adherence of the liquid to the lines during flow and this especially when the inner walls are hydrophobized *

If several samples are entered in a fourth capillary container η can be one in Fig. 1 for its step-by-step emptying The device shown uses wexfen when moving the Hydraulic switch spindles JJ? and 1_6 and thus through Movement of the Sehwingansee JJ ^ the circuit of the wicked multi-way cocks £ and £ is carried out in a further position. at this process is therefore only made of each supply capillary J ^. the bottom sample passed | jr * Ugt »

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Of course, the reverse process is also possible, with all samples from each supply capillary being gradually conveyed to the column, whereby the multi-way valves 4_ and Jj must not be switched to a further position. Only after the end of emptying a capillary can the two multi-way cocks be switched to other ones. This process can also be implemented while maintaining the device according to Fig. 1, namely in that, for example, the flap 10 is lifted by the movement of the lifter JK) ¹ upwards against the force of the spring J_0 ″, so that the flap JO engages in the corresponding tooth of the ratchet wheel j) as long as the gradual removal of the samples from the storage capillary J ^ is not completed. A method for directly influencing the hydraulic distributors Jjj and JH5 by an independent electrical drive system is more suitable than this mechanical method; without using the mechanical connection of the flap JK) and the ratchet wheel j? the shifting of the spindle of the multi-way cocks £ and Jj into a further position is brought about at suitable moments in accordance with the instructions which the central programmer outputs in the form of pulses.

In Fig. 2 an example of the structural implementation of the multi-way cocks is shown in such a way that the spindle £ _ and Jj _- these cocks are combined in a single spindle, which is driven in pushes according to the method mentioned. The spindle is mounted in the body J34 ^{with the} aid of three sealing rings, which advantageously consist of Teflon. These three sealing bushings _3 £, _3J> and 37 simultaneously form the bearing for the spindle _3_3 and are of-

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• Metal spacer tubes or rings ^ 8 and J59 that are fixed to one another distanced, with all these rings duroh a further metal ring 40 together duroh the Stopfbuohsen-Reguliersohraube J52 are axially contracted. The spindle jj5_3 fills the space in Body J34 tight, in the direction to the left in the Direction of arrow £ 2 is pressed, ^ (gduroh ensured being that the spindle ^ 3_ on the bottom £ 5 of the hole in the body rests.

At its left end the spindle j $ 2 has an angled drilled channel H, which connects the central, capillary outlet of the line j4J? with any side lines, one of which is shown in Fig. 2 with the designation £ 6. In Fig. 2 it is shown how these two lines, each alone, are sealed with the help of the resilient sleeve 42, which together with the ring _4J3, by means of the pull-up tube are pulled together 4ft. In this way, not only is the associated capillary line sealed, but it is also adequately secured against accidental pulling out. The line 4 , 6 corresponds to the capillary line 2, ^d ^ ^{e of} the individual torrat scapillaries 1 leads. All lines after the storage capillary must be connected without any solder spaces, as otherwise the concentration zones would mix when the samples and the surrounding protection buffer are transferred. In contrast, not all of them have to

other lines that carry the eluent to the storage capillaries bring, be sure to be capillary.

Fig. 2 also shows how to connect one after the other the connecting line _2 £ to each of the capillary lines

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is carried out. Both lines are attached to body 34 and sealed in a manner similar to lines 45, 45 and 4j5. The line 2A is connected to the channel 5Ό with a cavity in the intermediate circuit 5J. between the ring 3Q and the spindle 33, arises, the ring 3j9 a. Has circumference srill e, which is connected with one or more through bores with the inner surface so that the connection with the channel J50 is secured in every position. In the intermediate circuit 5J., Which is permanently connected in this way to the line 2A , the eluent only passes through the obliquely drilled channel 52 to one of the, for example, six regularly distributed circumferential channels 53 which are connected to the individual capillary lines 2 and which lead to the individual capillary containers _1_.

The spindle of the hydraulic switches ^ 5_ and J_6 can also be carried out in a similar manner. In this case, the construction is simpler, since only the individual circumferential nozzles are connected, which are always in one plane for each of the two hydraulic switches.

In Fig. 3, the supply and metering device is shown schematically, which is characterized in that the individual supply capillaries ] _ _f V etc. are attached to a common body, which is compact or formed from two strips 53 and 54 can be, which have smooth end sealing surfaces on which exactly the ends of the cemented or otherwise fastened individual supply capillaries end.

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By '.de seal, as shown in Fig. 4, is achieves that the connecting lines 24 and 2 £ always close together sit on both ends of the specific supply capillaries and thus · take the puncture of the multi-way cocks £ and £ as shown in Fig. 1. So this construction is simplified around the multi-way cocks, even if of course the demands on the sealing of the cited ones Connections between lines j24 resp. 2y? and is transferred to both ends of the supply capillaries.

Instead of the samples or the protective buffer being sucked into the individual containers when they are filled by dipping their lower ends into the vessel containing the sample or the buffer, this device is carried out according to Fig. 3 in such a way that the line 2 £ is switched to the suction line during the filling of the individual supply capillaries with the aid of the connecting channel ££ in the spindle of the hydraulic switch Jj6, v / o through which the sample is sucked into the container. Otherwise, the punctures of the device correspond to the punctures, as they were explained with reference to Fig. 1. After the AbI. 3, the capillaries 1_ and V etc., together with the strips £ 3_ and ^ £, carry out a step-by-step longitudinal movement. The same effect is also achieved in that the individual capillaries are not arranged in the same plane, but in a circular line at the same angular intervals, the strips 3 and 54 being in the form of face plates. The device shown in Fig. 3 can therefore be viewed as a cylinder surface developed into the plane. In each case, the capillaries alternate in connection with the individual lines, via a ratchet mechanism and the associated flaps

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that the whole system of storage tanks is moved by one division.

The arrangement of the seal, except in the functional position against no pressure other than against the hydrostatic pressure of the column must seal the sample, is such that in the. Functional position of the pressure circle or its capillary line is also secured against high pressure.

The example of the structural implementation of the seal is shown in Fig. 4. The connecting line 2 £ is here made of a capillary steel hollow needle in the manner of an injection needle, which ends "just above the upper surface of the bar £ 2, into which the individual capillary storage containers J_ and V etc. are cemented. Through the ferrule ^ 2 is "fc the needle £ 4 firmly enclosed by tightening the drawcap jj8, whereby the clamp ££ is connected to the whole frame, the immovable plate £ £, by which the seal 6 £ is so pressed that it is connected, also suffered from this frame closes off all openings of the individual supply capillaries, with the exception of one which is connected to the needle 2 £.

The sealing of the needle line AUOH against pressure ,, requires wielohen column, is carried out so that the needle 2 at £

its lower end through the small sleeve 6j [made of silicone rubber, Teflon or similar. is sealed. This cuff is held from the outside by the bushing 62 which is loosely pulled onto the needle 2 . The bushing £ 2 'is through the screw connection formed from the nut 62 and the screw 6 £ , toeammeü-

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COPY

INSPECTED

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·. pressed. When tightening this screw device together an appropriately large compression of the tight man-

'sohette 6J _- evoked. In this case, the elastic seal J50 is expediently compressed in the vicinity of the conical end of the bushing 62 .

The individual supply capillaries _1_ and J_ 'etc., which are in the Slats 53, and 4 "pounds attached, can be calibrated through smooth openings, which are made in a piece of plexiglass or similar. drilled will be replaced.

In Fig.5 the device is shown schematically, the. in the form of a lengthwise displaceable rod §Λ_ with transversely drilled channels, calibrated channels 6 £, which can expediently be alternated by further channels _6J5, which only serve to flush the lines 64 and 65. The individual channels jS2 are emptied from the samples in the form of transferring the samples to the column by the pressure exerted by the main pump YJ_ via lines J_8 and _24.

In this way there is a substantial, simplification of the hydraulic! Distributor achieved compared to the previous constructions. Here, the seal can be achieved with a much lower resistance Resistance to pressure can be performed, for example: by lightly pressing on the silicone sealing strips j56 rubber. One of the numerous possibilities of practical implementation of this device is shown in Fig. 6 in cross section at the point where the suction pipe ends. Of the

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COPY -H-

ORlQiNAL INSPECTED

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Sealing strip 6j5 is fastened, for example, by gluing into the solid, shaped strip 67, with the narrow area at the upper end of the end face of the body 6j sealing this with the channels 62 . The entire device can be arranged not only for a linear movement but also for a rotational movement, in both cases with jerk movements.

- patent claims -

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Claims (8)

Sponsor claims 1. Method for dosing samples in the pressure circuit, in particular in devices for column ear chromatography, thereby marked that the individual samples are first sucked into the sample container and initially outside the Pressure circuit of the pump remain and are only switched into this circuit when the sample is at the beginning the analysis is to be applied to the column. 2. The method according to claim 1, characterized in that the transfer of the sample from the sample container into the column takes place by the action of a pump that carries the eluents outside of the time period during which a sample container in the pressure circuit of the pump is the only one that is switched on The sample container is located and the other sample containers remain outside of the print circle, on one of the none Ways containing sample containers are pressed into the column. 3. Device for executing the method according to the claims 1 and 2, characterized in that the sample containers (1) are fed individually into the pressure circuit of the pump (17) via multi-way taps (4, 5) and hydraulic distributors (15, 16) switch on are bar , the pump (17) being connected directly to the column (23) in one of the positions of the hydraulic distributors (15, 16). 009830 / U88 -U- is bound and the individual sample containers (1) aussertialb of the 'pressure circuit of the pump (17). 4. Device according to claim 3 »characterized in that the capillary line (3) attached to the sample container (1) is conically flared at its end and inserted into a socket (28) on which a two-dimensional carrier (30) is seated. 5. Device naoh claims 3 and 4, characterized by that the sample container (1) is provided on its circumference with an enlargement (32) which is supported against the carrier (31) is. ' 6. Device according to claims 3 to 51, characterized in that that the spindles of the multi-way cocks (4, 5) 'are designed as a single spindle (33), which is expediently carried out with the help of Spacers (38, 39) from one another held rings (35 »36, 37) are mounted in the body (34). 7. Device according to claims 3 to 6, characterized in that that the sample containers (1, 1 ') next to one another liquid- "..'" · f.- tightly in a common, optionally provided with two smooth '\\ ground end surfaces strips (53, 54) Sex / ■■ ;. \ ϊ formed bodies are stored, on which body sole- ' Fene surfaces of a Diohtung (60) rest that the openings. .. all sample containers (1, 1 ¹ ) with the exception of that container ' { (1 or 1 ¹ ) close the close to the supply line '1'ψ (24) and the discharge line (25) is connected, the h 009830 / U88 _ I. COPY - 17 - ) ORIGINAL SNSPECTED ι resting on the soled surfaces of the strips (53 »54) I Diohtung (61) along this surface together with the seal (60) is displaceable, the rest of which have not yet been emptied • Closing the sample container (1, 1 ¹ ). 8. Device according to claims 3 to 6, characterized marked that the sample containers (62) are formed by channels in a rod that is displaceable relative to the body (67), where- ¹ at in the body (67) and the seal attached to it ($ 6) the lines leading from the pump (17) and to the column (23) : ·· (24, 25) open. 009830/1488 ^ * · · ORIGINAL INSPECTED