US3777572A - Method and apparatus for automatic sample loading for chromatography columns - Google Patents

Method and apparatus for automatic sample loading for chromatography columns Download PDF

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Publication number
US3777572A
US3777572A US00747006A US74700668A US3777572A US 3777572 A US3777572 A US 3777572A US 00747006 A US00747006 A US 00747006A US 74700668 A US74700668 A US 74700668A US 3777572 A US3777572 A US 3777572A
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sample
fluid
column
chromatography column
inlet
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US00747006A
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J Hrdina
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Czech Academy of Sciences CAS
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1095—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices for supplying the samples to flow-through analysers
    • G01N35/1097—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices for supplying the samples to flow-through analysers characterised by the valves
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/24—Automatic injection systems

Definitions

  • ABSTRACT A device for metering successive samples in to the input of a chromatography column.
  • a device including plurality of sample receptacles, each of which can be loaded in advance, is connected by means of hydraulic switching means to a pump and the column.
  • the hydraulic switching means can either connect the pump directly to the column or direct eluent from the pump through receptacles and then-to the column.
  • FIG. 1 A first figure.
  • sample receptacles into which they have been brought manually within time periods convenient for the attending operator.
  • the sample receptacle should be placed as near as possible in front of the column, i.e., in the pressure branch of the pump forcing the eluent into the column.
  • the connecting tubing between the sample receptacle and the column should be capillary, as short as possible and its interior surface should retain the minimum possible amount of the sample. The same conditions apply to the sample receptacle itself.
  • the metering device should, within the shortest possible time, assure the transfer of the whole sample at the beginning of the analysis from the sample receptacle to the column. Finally, care must be taken, of course, to avoid any leakage in the apparatus under the pressure used enabling any escape of portions of the sample into parts outside the metering system.
  • the process and apparatus according to the present invention is designed to avoid the above mentioned difficulties.
  • the substance of the invention consists that the individual samples are preliminarily sucked into sample receptacles and remain at first outside the pressure input of the pump transporting the liquids through the apparatus and are placed into the pressure input not earlier than when the sample should be transferred into the chromatography column at the beginning of the analysis.
  • the transfer of the sample from the sample receptacle into the chromatography column is carried out by the action of the pump delivering the eluent into the column following a path which does not contain any of the said sample receptacles except in the metering period during which a single one of the said sample receptacles is connected into the flow circuit which is pressurized by the pump whilst the other sample receptacles remain unconnected with this pressurized circuit.
  • the sample receptacles are individually placed into the pressurized pump circuit via at least one multi-way value and at least one, by draulic distributor, such that in one: of the positions of at least one of the hydraulic distributors the pump is directly connected with the column and the individual sample receptacles are outside the pressure cycle of the pump.
  • FIG. 1 shows a front view partly in section of the metering device
  • FIG. 2 an embodiment of a multi-way value in'longitudinal section
  • FIG. 3 is a front view partly in section of the metering device with the sample receptacles
  • FIG. 4 illustrates an embodiment of the sealing
  • FIG. 5 shows a modification of the arrangement of the sample receptacle
  • FIG. 6 a modification of the sealing.
  • FIG. .1 shows one representative embodiment of design by which the principles outlined above can be realized.
  • a plurality of sample capillary tubes 1 are successively connected through capillary ducts 2 and 3, shown in dotted lines, to multi-way valves which have as many peripheral necks as there areperipheral capillary. tubes 1.
  • the peripheral necks are arranged to communicate with a central neck attachedtoduct 24 or 24', respectively.
  • the two multiway valves or. cocks 4 and 5, represented in FIG. 1 as six-way ones, may be arranged either on a single shaft or spindle, or may have the spindles separated from one another and mechanically interconnected by gears 6 and 7, or, alternately, by an additional connecting gear 8 in such a way that they perform identical motions.
  • This motion may be generated by a separate electric drive on the principle of an electric impulse transmitted from a centralized programming unit.
  • this motion may also be generated by a ratchet mechanism, a ratchet 9 of this type of mechanism being moved stepwise through a corresponding angular distance by a pawl 10,
  • This pawl is connected through an articulated joint to an oscillating arm 11, connected in its turn to a gear 12, engaging with gears 13, 14, which serve to transmit a motion to two additional hydraulic distributors 15 and 16.
  • These hydraulic distributors 15 and 16 may also be arranged on a single shaft and moved to and fro by one pitch distance by a corresponding electric drive of a known principle, not shown in FIG. 1, this drive being controlled from one centralized programming unit.
  • the hydraulic distributors 15 and 16 may occupy alternately one of two positions.
  • the pump 17 is connected with its delivery branch 18 through a port 19 provided in the rotatable core of a hydraulic changeover switch 15 to a connecting tubing 20 that communicates with one of the peripheralnecks of the hydraulic change-over switch 16.
  • the port 21 establishes a communication between a tubing 20 and a tubing 22 which leads straight to the column 23.
  • both the upper multiway cock 4 and the lower multiway cock 5 are dis connected from thepressure circuit and the pump 17 delivers the eluent to the column 23.
  • the communicating tubing 24 is, however, in the basic position inter connected, through an additional port 25 of the spindle of the hydraulic change-over switch 15, to the tubing 26 which leads to a mouthpiece, by means of which it is possible, using the mouth or another suitable suction device finely adjustable, to draw, by suction, the samples and covering buffers, together with the separating bubbles, into the respective reservoir or sample capillary tubes 1.
  • each reservoir that is filled in the above described way has to be connected to the communicating tubing 24, and thereby also to the tubing 26, by means of a suitable positioning of the multiway cock 4.
  • the bottom narrowed-down capillary mouth 27 has to be separated from the connecting member of the capillary tubing 3 in such a way as to make possible, through the narrow-down bottom capillary mouth 27, a drawing-in, by suction, of the sample and the covering buffers from the respective small vessels which are temporarily held under the reservoirs 1 in such a way as to situate the mouth 27 below the level of the corresponding fluid.
  • FIG. 1 there is further shown one of the numerous possible design alternatives of a suitable connector to the capillary tubing 3.
  • This tubing has one end flared in such a way that it cannot be pulled out of a bushing 28 made, for instance, of Teflon and fitted in its interior with a small sleeve 29 that can be made, for instance, of silicone rubber.
  • the bottom of the bushing 28 bears against a supporting member 30, whereby a tight joint is obtained between the reservoir capillary tube 1 and the capillary tubing 3.
  • the reservoir capillary tube 1 may lean here against an additional supporting member 31 which engages a expanded portion 32 of the bottom end of the full diameter of the reservoir capillary tube 1.
  • the entire equipment can remain in the aforesaid functional position for a longer time, and a thorough scouring of the entire equipment can be undertaken with a smaller amount of bubbles or even in their total absence.
  • a scouring may serve to remove residues of the samples from the walls of the equipment and is realized by a limited amount of the buffer, the latter being, however, divided by bubbles, this arrangement being an efficient means against any kind of sticking of the fluid to the walls of the tubing during the throughflow, particularly if the inside walls have been rendered water-repellent.
  • FIG. 2 shows an example of the design of multiway cocks, according to which the spindles 4 and 5 these cocks are merged into a single spindle 33, intermittently driven in the way described above.
  • the spindle 33 is mounted in a body 34 by means of three packing rings (made advantageously of Teflon). These three packing bushes 35, 36, 37 form, at the same time, the bearings that carry the spindle 33, and are spaced from one another by rigid metallic spacing tubes or rings 38 and 39; all these rings are, together with an additional metallic ring 40, forced together in the longitudinal direction by a packing set screw 41.
  • the spindle 33 tightly fills the hollow interior of the body 34, being forced to the left in the direction indicated by an arrow 42, whereby a bearing of the spindle 33 against the bottom 43 of the hole in the body 34 is ensured.
  • the spindle 33 In its left-hand extremity, the spindle 33 has a port drilled in an inclined position which enables the central capillary outlet tubing 45 to be connected to any one of the side tubings, one of the latter marked 46 being shown in FIG. 2.
  • FIG. 2 shows how each of these two tubings is separately packed by an elastic sleeve 47 which is clamped, together with a ring 48, by a clamping screw 49. Thereby not only a packing of the corres ponding capillary tubing is obtained, but it is additionally secured against any accidental pulling-out.
  • the tubing 46 corresponds to the capillary tubing 3 leading from the respective reservoir capillary tubes 1.
  • All tubings situated after the reservoir capillary tube have to be interconnected without any dead spaces, since otherwise a mixing-up of the zones of concentration would take place during the transmission of a sample and the surrounding protective buffers.
  • all tubings that supply the eluent to the reservoir capillary tube 1 need not necessarily be of a capillary type.
  • FIG. 2 further shows how the successive connection of the communicating tubing 24 to each of the capillary tubings 2 is arranged. Both these tubings are attached to the body 34 andpacked in a way similar to that of the tubings 45 and 46. Through a port 50 the tubing 24 is connected to a void that is formed in the annulus 51 between the ring 39 and the spindle 33, the ring 39 having a peripheral groove connected by one or more drilled holes to the interior surface so as to warrant a connection to the port 50 in any position whatever.
  • FIG. 3 is a diagrammatic view of the reservoir and dosing equipment, a characteristic of which is that the individual reservoir capillary tubes 1, 1' etc., are attached in a common body that may be either compact or formed by two strips 53, 54 with packing end faces ground to a smooth finish these faces being accurately contacted by the ends of sealed-on or otherwise attached individual reservoir capillary tubes.
  • the communicating tubing 24 or 25 bears tightly, in each case, against the two ends of one particular reservoir capillary tube, thus acting in subtitution of the multiway cocks 4 and 5 according to FIG. 1.
  • the design is, consequently simplified though, of course, the exacting requirements required of a perfect packing must now be satisfied in the case of the said tight joints between the tubing 24 or 25 and the two ends of the reservoir capillary tubes.
  • this entire equipment is arranged, as shown in FIG. 3, in such a way that the tubing 25 is switched over, at the timeof filling of the individual reservoir capillary tubes, by means of a communicating port 55 provided in the spindle of the hydraulic change-over switch 16 to an intake tubing 56 by which the sample is drawn to the reservoir. Otherwise the operation of the equipment is identical with that described in FIG. 1. Accordingly, as shown in FIG. 3, the reservoir capillary tubes 1, 1', etc., perform, together with the strips 53, 54, a step-by'step motion in the longitudinal direction.
  • the same effect is, however, obtained by not having the individual capillary tubes arranged on the same level, but, instead, in the form of a circle, spaced atregular angular intervals, whereby the strips 53, 54 will assume the shape of endwise discs.
  • the equipment shown in FIG. 3 can thus be regarded as a development of a cylindrical surface into a plane.
  • thecapillary tubes alternate in their connections to the respective tubings in such a way that with a ratchet mechanism and the corresponding pawl the entire system of reservoirs is arranged so as to be advanced by one pitch each time.
  • the arrangement of the packings which, with the exception of the functional position are not required to seal against any other pressure than the hydrostatic pressure of the sample column, is such that in the functional position the pressure circuit or its capillary tubing are secured also against high pressures.
  • the communicating tubing 24 is here made of a hollow capillary steel needle, similar to an injection needle, the end of which is situated close above the top surface of a slat 53, into which the respective capillary reservoirs I, 1', etc., are sealed.
  • a clamp 57 the needle 24 is tightly held in position, after a clamping screw 58 has been tightened, the clamp 57 being positively attached to the frame of the equipment.
  • One part of this frame is also formed by a non-movable plate 59 by which a seal 60 is pressed down so as to shut off tightly all orifices of the individual reservoir capillary tubes, with the exception of the one which is connected to the needle 24.
  • a sealing of the needle guides also against the pressures required by the column is accomplished by a small packing sleeve 61, made of silicone rubber, Teflon or a similar material, attached to the bottom end of the needle 24.
  • This little sleeve is held in position from without by a bushing 62, freely slipped upon the needle 24.
  • the bushing 62 is compressed by a threaded joint made up of a nut 63 and a screw 64. Whenthese two elements are screwed together, a proportional compression of the narrow sleeve 61 is produced.
  • the elastic seal 60 is also adequately compressed, this compression being produced in the vicinity of a tapered end of the bushing 62.
  • the individual reservoir capillary tubes 1, 1', etc., attached to slats 53 and 54 can be substituted by calibrated smooth holes drilled in one solid piece of perspex" transparent plastic or similar material.
  • FIG. 5 diagrammatically shows equipment that can be constructed in the form of a longitudinally adjustable bar 61 with transversally reversible calibrated ports 62 that can desirably alternate with other ports 63 which are provided only for rinsing of the capillary tubings 64 and 65.
  • the evacuation of individual ports 62 with samples at the time of transportation of the samples to the column is performed under a pressure generated by the main pump 17 by way of the tubings 18 and 24.
  • the seal can, at the same time, be arranged with a far lower resistance to pressure, for instance, through a mere light pressing-on of the sealing strips'66 7 made of silicone rubber.
  • FIG. 6 One of the numerous possibilities of a practical realization of this type of equipment is illustrated in FIG. 6 in a transverse sectional view at the outlet end of the intake tubing 65.
  • the sealing strip 66 is attached, for instance, by an adhesive to a stationary profiled slat 67, providing, in this position, with its narrow surface a seal to the upper end face of the body 61 with ports 62.
  • the entire equipment may be arranged not only for a linear displacement, but advantageously also for a rotary motion, in either case a motion of an intermittent character.
  • a method of sequentially chromatographically analyzing a plurality of liquid samples comprising:
  • each bore sequentially disposing each bore between an inlet fitting coupled to a source of eluting fluid and an outlet fitting coupled to the inlet of a chromatography column, and passing eluting fluid through said respective bore to convey said respective liquid sample into said column; and inhibiting the flow of eluent fluid to said inlet fitting while a bore is being disposed between said inlet and outlet fittings.
  • Automatic sample loading apparatus for a chromatography column having an inlet means, comprising in combination pump means injecting an eluting fluid into said column;
  • hydraulic switching means within said conduit means controlling the eluting fluid to flow from said pump means into the column either directly or through one of said receptacles transferring a respective sample.
  • Automatic sample loading apparatus for a chromatography column having an inlet means, comprising in combination a pressure pump for the eluent;
  • tubular capillary sample receptacles each having at its ends a first and a second opening, respectively;
  • each multiway valve having at least one outlet assigned to one sample receptacle
  • first hydraulic distributing valve governing the flow of eluent coming from the pressure pump and a second distributing valve governing the flow into the chromatographic column, said first and second distributing valve conductively associated with each other and each with one multiway valve;
  • said distributing valves in a first position connecting the pressure pump directly with the column bypassing any sample receptacle, and in a second position directing the flow of eluent from the pressure pump into and through one of the sample receptacles to transfer the sample therefrom into the column; suction means governed by the first distributing valve drawing a sample into one sample receptacle during the first position of the distributing valves; and
  • actuating means coordinating said multiway valves and said distributing valves for complementing action during said first and second position of the distributing valves.
  • Automatic sample loading apparatus for a chromatography column having an inlet means, comprising a pressure pump for the eluting fluid and a first conduit leading therefrom;
  • capillary sample receptacles each having at its ends a first and a second opening, respectively;
  • a first and a second hydraulic distributing valve operatively associated with each other and turnable into a first and a second position
  • the distributing valves in the first position the distributing valves establishing a direct communication of the pump with the column to deliver eluent thereinto, and in the second position connecting the pump with the column over and through one sample receptacle to directly carry. the sample therefrom into the column by the flow of eluting fluid originating in the pump.
  • first and second fluid coupling means disposed adjacent to said support means
  • said directing means further includes eluent fluid pumping means, and additional means for connecting said pumping means in fluid flow communication with said chromatography column along said first and second fluid coupling means and a bore positioned in flow communication therewith.
  • said directing means includes further means for connecting said pump means to said chromatography column in bypass of said first and second fluid coupling means while said introducing means is operative to introduce a sample into a bore disposed between said first and said second fluid coupling means.
  • Automatic sample loading apparatus wherein said introducing means is operative to introduce a sample and a covering buffer, separated by an air bubble, into a bore disposed between said first and second fluid coupling means.
  • each respective one of said plurality of bores is adapted to receive different unique liquid samples.
  • Automatic sample loading apparatus for a chromatography column comprising a chromatography column
  • first valving means having an inlet and first and second outlets
  • second valving means having a first and second inlet and an outlet
  • sample support means having a plurality of sample containing bores and first and second fluid coupling means, means for intermittently moving said sample support means to sequentially dispose each of said bores in flow communication with said first and second coupling means, fourth pressure line connecting said first coupling means to said first output of said first valving means, and a fifth pressure line connecting said second coupling means to said second input of said second valving means, and
  • said first and second valving means being operative to connect said pumping means to said chromatography column along said first and second coupling means when a bore is disposed therebetween whereby a sample contained in said disposed bore is discharged into said chromatography column by said pumping means.
  • Automatic sample loading apparatus as defined in claim 11 including further means for introducing sample into a bore disposed between said firstand second coupling means; said further means being operative while said pumping means is connected directly to said chromatography column along said first and second valving means and said first pressure line.
  • Apparatus for automatically loading samples for analysis on to a chromatography column comprising a transfer member movable along a path, a plurality of sample locating chambers located in a spaced relationship on said transfer member and extending through said member, a fluid inlet duct and a fluid outlet duct, said fluid outlet duct leading to said column, said ducts being located, relative to said transfer member to be connected with successive chambers, means for sealing both ducts to opposite sides of a fluid passageway through a given chamber with which the ducts are connected, such that fluid can flow from said inlet duct through the given chamber and through the outlet duct, and means for stepwise moving the transfer member such that each chamber is, in turn, simultaneously registerable with said inlet and outlet ducts.
  • Apparatus for automatically loading samples for analysis on to a chromatography column comprising a transfer member movable along a path, a plurality of apertures in a spaced relationship in said member and extending through said member, sample support means for locating and supporting samplein each aperture, means for stepwise moving the member such that each sample support means in each aperture is held in exclusive register with fluid inlet and outlet ducts and fluid sealing means between the member and each of the said ducts for sealing the ducts to opposite sides of a fluid passageway through the sample support means in exclusive register with the ducts, such that fluids flow from said inlet duct through the sample support means and through the outlet duct.
  • Apparatus for automatically loading samples for analysis onto a chromatography column comprising a transfer member having a plurality of sample locating chambers located in spaced relationship in said transfer member, each chamber being in the form of a bore extending through the transfer member, a fluid flow assembly including a fluid inlet duct and a fluid outlet duct, the two said ducts being located to be simultaneously registerable with a given chamber sealing the two ducts to the chamber such that fluid can flow from said inlet duct through the chamber to said outlet duct, said transfer member and said fluid flow assembly being relatively movable such that the two said ducts can register, in turn, with successive chambers of said transfer member.

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  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
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US00747006A 1964-06-01 1968-07-11 Method and apparatus for automatic sample loading for chromatography columns Expired - Lifetime US3777572A (en)

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US (1) US3777572A (de)
JP (1) JPS5233518B1 (de)
CH (1) CH447657A (de)
DE (1) DE1598223B2 (de)
FR (1) FR1435538A (de)
GB (1) GB1111664A (de)
SE (1) SE332903B (de)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3916692A (en) * 1973-11-14 1975-11-04 Waters Associates Inc Novel injector mechanism
US3999439A (en) * 1974-07-22 1976-12-28 Varian Associates High pressure sample injector and injection method
US4221568A (en) * 1979-01-17 1980-09-09 Jet Propulsion Laboratory Sample processor for chemical analysis
US5607581A (en) * 1995-10-31 1997-03-04 Systec, Inc. Debubbling and priming system for chromotography
EP1584922A2 (de) 2004-04-09 2005-10-12 Agilent Technologies Inc Bepacken einer Mikrovorrichtung mit Teilchen

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2656737A1 (de) * 1975-12-22 1977-07-14 Sandoz Ag Beschickungsvorrichtung fuer chromatographen

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2757541A (en) * 1956-02-27 1956-08-07 Perkin Elmer Corp Sampling apparatus
US2830738A (en) * 1956-06-28 1958-04-15 Standard Oil Co Fluid sample injection apparatus
US2973117A (en) * 1957-12-16 1961-02-28 American Oil Co Measuring and charging valve
GB947480A (en) * 1959-06-26 1964-01-22 Nat Smelting Co Ltd Improvements in or relating to the sampling of gases and liquids
US3160015A (en) * 1961-06-22 1964-12-08 Beckman Instruments Inc Gas chromatograph sampling valve
US3220246A (en) * 1961-07-13 1965-11-30 Technicon Chromatography Corp Apparatus and devices, especially for chromatography analysis
US3253468A (en) * 1964-09-24 1966-05-31 Dade Reagents Inc Ultramicro sampling device
US3487678A (en) * 1966-03-25 1970-01-06 Atomic Energy Authority Uk Sample loading apparatus

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2757541A (en) * 1956-02-27 1956-08-07 Perkin Elmer Corp Sampling apparatus
US2830738A (en) * 1956-06-28 1958-04-15 Standard Oil Co Fluid sample injection apparatus
US2973117A (en) * 1957-12-16 1961-02-28 American Oil Co Measuring and charging valve
GB947480A (en) * 1959-06-26 1964-01-22 Nat Smelting Co Ltd Improvements in or relating to the sampling of gases and liquids
US3160015A (en) * 1961-06-22 1964-12-08 Beckman Instruments Inc Gas chromatograph sampling valve
US3220246A (en) * 1961-07-13 1965-11-30 Technicon Chromatography Corp Apparatus and devices, especially for chromatography analysis
US3253468A (en) * 1964-09-24 1966-05-31 Dade Reagents Inc Ultramicro sampling device
US3487678A (en) * 1966-03-25 1970-01-06 Atomic Energy Authority Uk Sample loading apparatus

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3916692A (en) * 1973-11-14 1975-11-04 Waters Associates Inc Novel injector mechanism
US3999439A (en) * 1974-07-22 1976-12-28 Varian Associates High pressure sample injector and injection method
US4221568A (en) * 1979-01-17 1980-09-09 Jet Propulsion Laboratory Sample processor for chemical analysis
US5607581A (en) * 1995-10-31 1997-03-04 Systec, Inc. Debubbling and priming system for chromotography
EP1584922A2 (de) 2004-04-09 2005-10-12 Agilent Technologies Inc Bepacken einer Mikrovorrichtung mit Teilchen
EP1584922B1 (de) * 2004-04-09 2014-08-06 Agilent Technologies, Inc. Packen einer Mikrovorrichtung mit Teilchen

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DE1598223A1 (de) 1970-07-23
DE1598223B2 (de) 1973-02-08
CH447657A (de) 1967-11-30
JPS5233518B1 (de) 1977-08-29
FR1435538A (fr) 1966-04-15
SE332903B (de) 1971-02-22
GB1111664A (en) 1968-05-01

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